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Turfgrass and EnvironmentalResearch Online

Volume 7, Number 23December 1, 2008

The 2008 USGA Turfgrass and EnvironmentalResearch Summary is an annual compilation of proj-ects currently funded by USGA’s Turfgrass andEnvironmental Research Program. The summary con-tains research summaries of 75 projects funded in2008.

PURPOSE

The purpose of USGA Turfgrass and Environmental Research Online is to effectively communicate the results ofresearch projects funded under USGA’s Turfgrass and Environmental Research Program to all who can benefitfrom such knowledge. Since 1983, the USGA has funded more than 350 projects at a cost of $29 million. The pri-vate, non-profit research program provides funding opportunities to university faculty interested in working on envi-ronmental and turf management problems affecting golf courses. The outstanding playing conditions of today’sgolf courses are a direct result of using science to benefit golf.

Editor

Jeff Nus, Ph.D.1032 Rogers PlaceLawrence, KS [email protected](785) 832-2300(785) 832-9265 (fax)

Research Director

Michael P. Kenna, Ph.D.P.O. Box 2227Stillwater, OK [email protected](405) 743-3900(405) 743-3910 (fax)

Permission to reproduce articles or material in the USGA Turfgrass and Environmental Research Online (ISSN 1541-0277) isgranted to newspapers, periodicals, and educational institutions (unless specifically noted otherwise). Credit must be given tothe author(s), the article title, and USGA Turfgrass and Environmental Research Online including issue and number. Copyrightprotection must be afforded. To reprint material in other media, written permission must be obtained from the USGA. In anycase, neither articles nor other material may be copied or used for any advertising, promotion, or commercial purposes.

USGA Turfgrass and Environmental Research Committee

Steve Smyers, ChairmanJulie Dionne, Ph.D.

Ron DodsonKimberly Erusha, Ph.D.

Ali Harivandi, Ph.D.Michael P. Kenna, Ph.D.

Jeff Krans, Ph.D.Brigid Shamley Lamb

James MooreJeff Nus, Ph.D.

Paul Rieke, Ph.D.James T. Snow

Clark Throssell, Ph.D.Ned Tisserat, Ph.D.Scott Warnke, Ph.D.

James Watson, Ph.D.Chris Williamson, Ph.D.

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Since 1983, the United States Golf Association has funded more than 350 university researchprojects at a cost of $29 million. The Turfgrass and Environmental Research Program provides directionto these intuitions and employs science as the foundation to benefit golf in the areas of turfgrass andresource management, sustainable development, and environmental protection. At the end of each year,the USGA provides a summary of the research conducted under this important national program and thisreport summarizes the results from 2008.

There are two primary goals of the research program. The first is to develop turfgrasses and cul-tural systems with better stress tolerance and reduced water requirements and pesticide use. To addressthe USGA's first research goal, 49 research projects were funded in integrated turfgrass management,physiology, breeding, genetics, and course construction practices. The second goal is to investigate envi-ronmental issues and sustainable resource management for golf courses. Five research projects thatinvestigate the environmental impact of golf courses are reported.

The research program actively coordinates and supports research, associated educational pro-grams, and other partnerships to benefit golf, the environment, and people. For example, the USGA,GCSAA, and National Turfgrass Evaluation Program (NTEP) together have developed turfgrass varietytesting programs conducted on golf courses throughout the United States. In addition, the USGA workswith state research foundations and superintendent chapters to fund applied research through the Grant-in-Aid Research Program. Ten research projects are included in this report under the Grant-in-AidResearch Program. USGA’s Turfgrass and Environmental Research Program also provides funding tothe National Fish and Wildlife Foundation to support the Wildlife Links Research Program. Seven proj-ects are currently being funded and summaries of their progress are included in this report.

2008 USGA Turfgrass and Environmental Research Summary

Jeff L. Nus

USGA Turfgrass and Environmental Research Online 7(23):1-84.TGIF Record Number: 145333

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http://www.lib.msu.edu/cgi-bin/flink.pl?recno=145333

Table of Contents

Course Construction Practices ..............................................................................................................................................................1

A Comparison of Water Drainage and Storage in Putting Greens Built Using Airfield Systems and USGAMethods

of Construction (McInnes and Thomas)..................................................................................................................................................2

Integrated Turfgrass Management ......................................................................................................................................................3

Biological and Biorational Management Options for the Annual Bluegrass Weevil on Golf Courses (McGraw and Koppenhöfer) ......................4Developing Best Management Practices for Anthracnose Disease on Annual Bluegrass Putting Green Turf (Murphy, Clarke, and Roberts)........5Relative Pathogenicity and Fungicide Sensitivity of Isolates of Rhizoctonia and Other Fungal Pathogens and the

Disease Responses of Seashore Paspalum and Zoysiagrass Cultivars (Stiles, Harmon, and Kenworthy) ............................................6Natural Enemies and Site Characteristics Affecting Distribution and Abundance of Native and Invasive White Grubs

on Golf Courses (Potter and Redmond) ..................................................................................................................................................7Interpreting and Forecasting Phenology of Annual Bluegrass Weevil in Golf Course Landscapes (Peck, Seto, and Diaz) ....................................8Seasonal Life History and Suitability of Horticultural Plants as Nectar Sources for Larra bicolor, a Parasitoid

of Mole Crickets in the Northern Gulf Coast (Held) ..............................................................................................................................9Rooting and Carbohydrate Metabolism in Creeping Bentgrass Putting Greens in Response to

Summer Irrigation and Aeration (Fu and Dernoeden) ..........................................................................................................................10Infection and Colonization of Bermudagrass by Ophiosphaerella herpotricha, the Causal Agent of Spring Dead Spot (Walker) ........................11Silicon Amendment: A Component of an Integrated Gray Leaf Spot Management Strategy (Uddin) ....................................................................12Optimization of Foliar Nitrogen Nutrition to Improve Turf Performance Under Energy Stress (Henning, Branham, and Mulvaney) ..................13Use of a Baculovirus for Season-long Control of Black Cutworms on Golf Courses and

Compatability with Soil Insecticides and Insect-resistant Turfgrasses (Bixby and Potter) ..................................................................14Accurate Identification and Gene Expression in Relation to Virulence of Rhizoctonia Isolates Infecting Turfgrasses (Lakshman) ......................15Mole Cricket Sensory Perception of Insecticides (Kostromytska and Buss) ............................................................................................................16The Efficacy of Spring Fungicide Applications Plus Organic Fertilizer for Controlling Spring

Dead Spot of Bermudagrass (Tomaso-Petersen) ..................................................................................................................................17Developing Best Management Practices for Bermudagrass Control in Zoysiagrass Fairways (McElroy and Doroh)............................................18Salinity Management in Effluent Water Irrigated Turfgrass Systems (Qian) ..........................................................................................................19Cultural Practices, Environment, and Pathogen Biology: Studies for Improved Management of

Large Patch of Zoysiagrass (Kennelly, Fry, St. John, and Bremer) ......................................................................................................20Optimizing Oriental Beetle Mating Disruption Through a Better Understanding of Dispersal Behavior (Koppenhöfer and Fournier) ................21Improvement of Water Management Strategies and Practices (Morris, Johnson, Bushman, and Johnson) ............................................................22Enhancement of Soil and Soil Management Practices (Morris and Zobel)..............................................................................................................23

Breeding, Genetics, and Physiology ..................................................................................................................................................24

Identification of the Colonial Bentgrass Contribution to Dollar Spot Resistance in Colonial x CreepingBentgrass Interspecific Hybrids (Belanger) ..........................................................................................................................................25

A Bentgrass Breeding Consortium to Support the Golf Industry (Casler, Jung, Warnke, Bonos, Belanger, and Bughrara) ..................................26Development of Seeded Turf-type Saltgrass Varieties (Christensen and Qian) ......................................................................................................27Perennial Ryegrass Anti-freeze Protein Genes Enhances Freezing Tolerance in Plants (Fei)..................................................................................28Breeding and Evaluation of Kentucky Bluegrass, Tall Fescue, Perennial Ryegrass, and Bentgrass for Turf (Funk, Meyer, and Bonos) ..............29Production, Maintenance, and Evaluation of Triplolid Interspecific Bermudagrass Hybrids for QTLAnalysis (Hanna and Paterson)................30Resistant Turfgrasses for Improved Chinch Bug Management on Golf Courses (Heng-Moss, Baxendale, Shearman, Sarath, and Twigg)..........31Development and Application of Molecular Markers Linked to Heat Tolerance

in Agrostis Species (Huang, Belanger, Bonos, and Meyer) ..................................................................................................................32Quantitative Trait Loci (QTL) Mapping of Resistance to Gray Leaf Spot in Lolium (Jung) ..................................................................................33Breeding and Evaluation of Turf Bermudagrass Varieties (Wu, Martin, and Taliaferro) ........................................................................................34Accelerated Discovery of Cynodon Genes and DNAMarkers by cDNA Sequencing (Paterson) ..........................................................................35Multiple Stress Tolerance, Seed Dormancy Breaking, and Establishment of Seeded Saltgrass (Qian and Christensen) ......................................36Breeding Seashore Paspalum for Recreational Turf Use (Raymer) ..........................................................................................................................37

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Germplasm Development and Management of Buffalograss Varieties (Shearman..................................................................................................38Genetic Improvement of Prairie Junegrass (Watkins) ..............................................................................................................................................39Evaluating Poverty Grass (Danthonia spicata L.) for Use in Tees, Fairways, or Rough Areas on

Golf Courses in the Midwest (Navarrete-Tindall, Fresenburg, and Van Sambeek) ............................................................................40Evaluation of Perennial Ryegrass, Creeping Bentgrass, and Kentucky Bluegrass Cultivars

for Salt Tolerance (Bonos, Honig, Gianfagna, and Huang)..................................................................................................................41Confirmation of QTL Markers for Dollar Spot Resistance in Creeping Bentgrass (Bonos, Honig, and Kubik) ....................................................42Linking Drought Tolerance Traits and Candidate Genes in Perennial Ryegrass through

Association Mapping (Jiang) ................................................................................................................................................................43Selection of Bermudagrass Germplasm that Exhibits Potential Shade Tolerance and Identification of

Techniques for Rapid Selection of Potential Shade Tolerant Cultivars (Bell and Wu) ........................................................................44Identification of Quantitative Trait Loci (QTL) Associated with Drought and Heat

Tolerance in Creeping Bentgrass (Huang, Bonos, and Belanger) ........................................................................................................45Evaluation of the New England Velvet Bentgrass Collection (Brown, Percivalle, Jung, and Dowgiewicz) ..........................................................46Evaluating Methods for Vegetative Propagation and Enhancement of Seed Production of Greens-type Poa annua Cultivars (Huff) ..................47Evaluation and Development of Salt Tolerant Poa Germplasm (Johnson, Robins, and Bushman) ........................................................................48Genetic Enhancement of Turfgrass Germplasm for Reduced-input Sustainability (Morris and Warnke) ..............................................................49Breeding Turf-type Annual Ryegrass for Salinity Tolerance (Nelson) ....................................................................................................................50Bermudagrass (Cynodon spp.) and Seashore Paspalum (Paspalum vaginatum) Cultivar Response

to the Sting Nematode (Belonolaimus longicaudatus) (Pang, Crow, and Kenworth) ..........................................................................51Collection and Evaluation of Native Grasses from Grazed Arid Environments for

Turfgrass Development (Kopec, Smith, and Pessarakli) ......................................................................................................................52

Environmental Impact ..............................................................................................................................................................................53

Pesticide Matrix Project: Developing a Data Base Tool to Guide EnvironmentallyResponsible Pesticide Selection (Branham, Fermanian, Cohen, and Grant) ......................................................................................54

Utilizing Reduced-risk Pesticides and IPM Strategies to Mitigate Golfer Exposure andHazard (Clark, Putnam, and Doherty) ................................................................................................................................................55

Optimization of Vegetative Filter Strips for Mitigation of Runoff from Golf Courses Turf (Clark, Doherty, Lanza, and Parkash)......................56Nitrogen and Phosphorus Fate in a 10+ Year Old Kentucky Bluegrass Turf (Frank)..............................................................................................57Attenuation of Pharmaceuticals, Personal Care Products, and Endocrine-disrupting

Compounds by Golf Courses Using Reuse Water (McCullough) ........................................................................................................58

Audubon International ..............................................................................................................................................................................59

The Audubon Cooperative Sanctuary System Program for Golf Courses (Dodson)................................................................................................60

Grant-in-Aid Research Program ........................................................................................................................................................61

Assessing the Usefulness of Physical Water Conditioning Products to Improve Turfgrass Quality and ReduceIrrigation Water Use (Leinaeuer, Serena, and Schiavon) ......................................................................................................................62

Preventive and Curative Control of Algae on Putting Greens Using Products Other than Daconil (Kaminski)......................................................63Bermudagrass Control Programs in Kentucky Bluegrass (Willis and Askew) ..........................................................................................................64Organic Matter Dilution Programs for Sand-based Putting Greens (Ervin, Horvath, and Henderson) ..................................................................65Impact of Sand Type and Application Rate of Fairway Topdressing on Soil Physical Properties, Turfgrass

Quality, Disease Severity, and Earthworm Castings (Henderson) ........................................................................................................66Best Management Practices for the Conversion of Established Bermudagrass to Buffalograss (Henry) ................................................................67Comparison of Chlorothalonil, Propiconazole, and Iprodione Products for Control

of Dollar Spot and Brown Patch Diseases (Landschoot and Fidanza) ................................................................................................68Identification, Pathogenicity, and Control of Leaf and Sheath Blight of Bermudagrass Putting Greens (Martin and Park)..................................69

Nitrogen Nutrition of Distichlis (Saltgrass) under Normal and Salinity Stress Conditions Using 15N (Pessarakli) ..............................................70Leaf Cuticle Characteristics and Foliar Nutrient Uptake by a Cool-season

and Warm-season Turfgrass (Richardson, Stiegler, Karcher, and Patton) ............................................................................................71

(Continued on next page)

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Wildlife Links Program............................................................................................................................................................72

Reproductive Success and Habitat Use of Painted Buntings on Golf Courses in Coastal SouthCarolina (Jodice, Freeman, Gorzo, and Marsh) ..................................................................................................................................73

Abundance and Diversity of Stream Salamanders on Golf Courses (Semlitsch) ....................................................................................................74Brown-headed Nuthatch Enhancement Study (Stanback) ........................................................................................................................................75Avian Pesticide Exposure on Golf Courses (Cristol and Burdge) ............................................................................................................................76Puget Sound Salmon-Safe Golf Course Program (Burger) ......................................................................................................................................77Using Buffer Zones to Promote Amphibian Populations (Boone) ............................................................................................................................78Population and Community Responses of Reptiles to Golf Courses (Goode) ........................................................................................................79

Product Testing ..............................................................................................................................................................................................80

Evaluation of Plant Growth Regulators and Biostimulants for Use in Managing Summer Bentgrass Decline (Huang and Xu) ..........................81Evaluation of Cytokinian Plant Extract Biostimulants, Iron, and Nitrogen Products

for Their Effect on Creeping Bentgrass Summer Quality (Dernoeden and Settle) ..............................................................................822007 Bufalograss Expermental Line and Cultivar Evaluation (Shearman, Abeyo, Kopec, Johnson

Goss, Koski, Goatley, St. John, and Stahnke) ......................................................................................................................................84

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http://usgatero.msu.edu

The United States Golf AssociationTurfgrass and Environmental

Research Program

VisionUse science as the foundation to benefit golf in the areas ofturfgrass and resource management, sustainable develop-ment and environmental protection.

MissionCoordinate and support research, associated educationalprograms, and partnerships to benefit golf, the environment,and people.

GoalsDevelop turfgrasses and cultural systems with enhancedstress tolerance and reduced supplemental water require-ments, pesticide use and costs.

- Course Construction Practices

- Integrated Turfgrass Management

- Breeding, Genetics, and Physiology

Investigate environmental issues and sustainable resourcemanagement for golf courses.

- Environmental Impact of Golf Courses

- Wildlife and Habitat Management·

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USGAGreen Section Turfgrass and Environmental ResearchProject Grants in 2008

Project Area Number Grant $ % of Total

Course Construction Practices 2 60,000 3.8 %Integrated Turfgrass Management 22 444,438 28.1 %

Sustainable Management 7 121,605 7.7 %Pathology 8 181,667 11.5 %Entomology 7 141,166 8.9 %

Breeding, Genetics, and Physiology 30 640,599 40.4 %Cool-season Grasses 17 379,921 24.0 %Warm-season Grasses 6 141,049 8.9 %New or Native Grasses 7 119,629 7.5 %

Environmental Impact 12 311,668 19.6 %Fate and Transport 5 111,668 7.0 %Wildlife Links Program 7 200,000 12.6 %

Outreach Programs 27 127,000 8.0 %Grant-in-Aid Research Program 17 58,000 3.7 %Product Testing 7 34,000 2.1 %Cooperative Research-Allied Assoc.s 3 35,000 2.2 %

Total 93 1,584,205 100.0%

Location of 2008 projects funded by the USGATurfgrass and Environmental Research Progam

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Julie Dionne, Ph.D.Royal Canadian Golf Association272 Ile de FranceSaint Basile le GrandQuebec, CANADAv: 514-705-3889 f: [email protected]

Ronald DodsonAudubon International, Inc.46 Rarick RoadSelkirk, NY 12158v: 518-767-9051 f: [email protected]

Kimberly Erusha, Ph.D.USGA Green SectionP.O. Box 708Far Hills, NJ 07931-0708v: 908-234-2300 f: [email protected]

Patrick R. Finlen, CGCSThe Olympic Club524 Post StreetSan Francisco, CA 94102-1229v: [email protected]

Ali Harivandi, Ph.D.University of CaliforniaCooperative Extension1131 Harbor Bay Parkway, #131Alameda, CA 94502v: 510-639-1271 f: [email protected]

Michael P. Kenna, Ph.D.USGA Green Section ResearchP.O. Box 2227Stillwater, OK 74076v: 405-743-3900 f: [email protected]

Jeff Krans, Ph.D.1258 LaFave RoadManitowish Waters, WI 54545v: 715-543-2114 f: [email protected]

Brigid Shanley Lamb290 Pleasant Valley RoadMendham, NJ 07945v: 973-543-6221 f: [email protected]

James MooreUSGA Green Section Construction Educ.770 Sam Bass RoadMcGregor, TX 76657v: 254-848-2202 f: [email protected]

Jeff Nus, Ph.D.USGA Green Section Research1032 Rogers PlaceLawrence, KS 66049v: 785-832-2300 f: [email protected]

Paul Rieke, Ph.D.2086 Belding CourtOkemos, MI 48824v: 517-349-2784 f: [email protected]

James T. SnowUSGA Green SectionLiberty Corner RoadP.O. Box 708Far Hills, NJ 07931-0708v: 908-234-2300 f: [email protected]

Clark Throssell, Ph.D.Golf Course Supts. Assoc. of America3280 Banff AvenueBillings, MT 59102v: 406-656-1986 f: [email protected]

Ned Tisserat, Ph.D.Biological Sciences and PestManagementColorado State UniversityFort Collins, Colorado 80523-1177v: (970) [email protected]

Scott Warnke, Ph.D.USDA-ARSRoom 110, Bldg 010A BARC-West10300 Baltimore AvenueBeltsville, MD 20705-2350v: 301-504-8260 f: [email protected]

James Watson, Ph.D.3 Larkdale DriveLittleton, CO 80123v: 303-794-5346 f: [email protected]

Chris Williamson, Ph.D.University of Wisconsin246 Russell Labs1630 Linden DriveMadison, WI 53706v: 608-262-4608 f: [email protected]

2008 USGA Turfgrass andEnvironmental Research Committee

Chairman

Steve SmyersSteve Smyers Golf Course Architects, Inc.

2622 W. Memorial Blvd.Lakeland, FL 33815

v: 863-683-6100 f: [email protected]

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Course Construction Practices

To take advantage of new ideas and technologies and to address the environmen-tal and economic challenges of the coming decades, USGA sponsors research studies onthe construction and establishment of golf courses. Preference is given to research studiesthat address issues related to:

1. Materials testing procedures for putting green rootzones and bunker sands2. Alternative construction methods and materials3. Effects of irrigation water quality on the selection of construction materials4. Moisture-related problems in putting green rootzones5. Calcareous sands6. Various organic matter sources7. Greens/surrounds interface construction and management8. Alternative irrigation methods9. How to amend various-textured native soils for use as tees (and possibly low-cost greens).

Interdisciplinary approaches are expected including the disciplines of soilphysics, soil chemistry, soil microbiology, physiology, management, and pathology.Studies should give due consideration to environmental issues and to the use of alterna-tive water sources for golf course irrigation.

Locations of the following projects funded in 2008 by the USGA Turfgrass andEnvironmental Research Program under the category of Course Construction Practices

1

This research investigates the dynam-

ics of water movement through, and stor-age within, the rootzone of putting greensconstructed using a geotextile atop a plas-tic support acting as the drainage structure(Airfield System's design) compared to agreen constructed with a gravel-baseddrainage structure (USGA recommended).

Constructed to USGA specifica-tions with a 100-mm layer of gravel, sand-based rootzone mixture placed over gravelholds water at the rootzone-gravel inter-face at tensions between 0 and 100 mmwater when watered to such a degree thatdrainage occurs. The Airfield Systems'design replaces the gravel with a geotextileatop a 25-mm deep porous plastic supportfor drainage. Our laboratory observationshave shown that the bottom of the rootzonein the Airfield Systems design is typicallyat about 50 mm less tension after drainage.

One hundred eighty permeame-ters were constructed from 150-mm innerdiameter by 350-mm tall PVC pipe andend caps. Each permeameter was filledwith 300-mm deep rootzone mixture over

a geotextile supported on a 25-mm deepplastic geogrid (Airfield Systems'AirDrain). A tensiometer was placed ontop of the geotextile in each test cell beforeit was filled with the rootzone mixture.

Ten geotextiles were evaluated:two woven, three spunbond, five needlepunch. The woven materials wereWM104F (Propex) and FW404 (Tencate).The spunbond fabrics were Typar 3301L(Fiberweb), Typar 3341G (Fiberweb), andLutradur 130g (Freudenberg). The needlepunch materials were NW351 (Propex),NW401 (Propex), NW1001 (Propex),NW10 (Gundle/SLT Environmental), andNW16 (Gundle/SLT Environmental).Apparent opening sizes (AOS) of the tengeotextiles were 212, 425, 300, 250, 122,300, 212, 150, 150, 150 micrometers,respectively.

Six rootzone mixtures with differ-ing distributions of fines were created byblending three parent materials with differ-ing particle size distributions, two sandsand a sandy clay loam (SCL). One parent-material sand's particle size distributionfell in the middle of the USGA specifica-tions (Sand A). The other parent-materialsand exceeded the specified limits for fineand very fine sand (Sand B). Total fines<150 micrometer ranged from 6% to 39%for the six rootzone mixtures. Each geo-textile was tested in combination with allsix rootzone mixtures.

Test cells were irrigated at ratesrepresentative of cultural practices forestablishment and maintenance of a put-ting green. Greater amounts of water wereapplied periodically to simulate stormevents and to observe changes in whole-system permeability and drainage.Changes in whole-system permeability anddrainage were assessed by measuring thetemporal distributions of drainage ratefrom an instantaneous application of 25mm (1 inch) of water to the top of the testcell.

We did not observe positive pres-sure atop the geotextiles for any apprecia-ble time. When drainage rates from thetest runs were averaged across rootzonemixtures for each of the geotextiles, no sig-nificant differences in drainage rates wereobserved. We conclude that geotextiles inthe test cells have not clogged. Whendrainage rates from the test runs were aver-aged across geotextiles for each of therootzone mixtures, significant differencesin drainage rates were observed. We con-clude that the particle size distribution ofthe rootzone mixtures affects drainage rate,as expected, but not through clogging ofthe geotextiles.

A Comparison of Water Drainage and Storage in Putting GreensBuilt Using Airfield Systems and USGA Methods of Construction

Start Date: 2007Project Duration: three yearsTotal Funding: $90,000

Kevin McInnes and James ThomasTexas A&M University

Objectives:1. Investigate potential for clogging of geotextile pores from fines migrating out of the rootzone mixture.2. Investigate the changes in the temporal distribution of drainage and spatial distribution of water holding capacity ofa green constructed with a geotextile compared to a green constructed with gravel.

3. Investigate construction methods designed to reduce down-slope movement of water in sloping greens.4. Develop criteria for selecting appropriate rootzone mixtures for putting greens constructed with geotextiles.

Permeability and drainage of tests cells were affectedby rootzone mixtures (above), but not through cloggingof geotextiles.


Summary PointsClogging of geotextiles was not

observed

Permeability and drainage of tests cellscontaining rootzone mixtures with totalfines <150 micrometers ranging between 6and 39% were not affected by geotextileswhose apparent opening size ranged from150 to 400 micrometers.

Permeability and drainage of tests cellswere affected by rootzone mixtures, butnot through clogging of geotextiles.

2

Ten geotextiles were evaluated: two woven, three spun-bond, five needle punch. Permeameter tests showedthat none of the geotextiles clogged during the test peri-od using 1 inch (25 mm) of irrigation water.

Integrated Turfgrass Management

Improved turfgrasses developed for use on golf coursesrequire management practices that provide quality playing surfaceswhile conserving natural resources and protecting the environment. Aseries of research projects are being funded with the aim of conserv-ing natural resources by reducing the use of water, pesticides, and fer-tilizers. The objectives of these studies will focus on the following:

1.

2.

3.

4.

The results of these studies should lead to the developmentof turfgrass management programs that conserve our natural resourcesand reduce costs, with minimal impairment of playing quality condi-tions or aesthetic appeal. We encourage regional cooperation amongresearchers where similar climactic and soil conditions exist.

Location of the following projects funded in 2008 by the USGA Turfgrass andEnvironmental Research Program under the category of Integrated Turfgrass Management

3

Develop cultural practices that allow efficient turfgrassmanagement under unique conditions, such as poor quali-ty soils, shade, and marginal quality water.

Determine the range of adaptability and stress tolerance ofturfgrasses.

Evaluate direct and interacting effects of two or three cul-tural practices like mowing, irrigation, fertilization, cultivation, compost utilization, and develop programs to con-trol pests and organic matter accumulation (thatch).

Investigate pest management practices such as biological,cultural, and mechanical controls, application of turf man-agement practices utilizing IPM and reduced inputs, andpest modeling and forecasting.

The annual bluegrass weevil (ABW),

Listronotus maculicollis, formerly'Hyperodes weevil', is a serious andexpanding pest of close-cut annual blue-grass on golf courses through much of theNortheast. Adult ABW emerge from over-wintering sites in leaf litter and tall roughin early April and migrate to short mowedturfgrasses (greens, tees, fairways) to feedand mate. Females lay eggs directly intothe stem of the turfgrass plant from lateApril through May in New Jersey.

The young larvae are initiallystem borers, feeding internally on theplant, ultimately tunneling through thecrown and destroying the turfgrass plant.Later instars feed externally on crowns androots which leads to the most extensiveturf loss typically around early to mid June(Figure 1). Damage caused by the 2nd and3rd generations is usually less severe andmore localized as peak larval densitiestend to be lower than in the 1st generation.In previous project years we found that

entomopathogenic nematodes (EPNs) nat-urally occurring in golf course fairwaysnot treated for ABW control can cause upto 50% generational ABW mortality.Additionally, EPNs have provided highlevels of larval control (65-100%) whenapplied to field-infested turf cores in thelaboratory.

We examined the efficacy ofcommercial EPN products against 1st gen-eration ABW immature stages in field tri-als in 2008. Products based onSteinernema feltiae, S. carpocapsae, andHeterorhabditis bacteriophora were cho-sen for closer examination in field trialssince they have provided at least 70% con-trol in past field trials (Figure 2). Sincelower control levels in 2007 than in 2006were believed to be linked to high ABWlarval populations in the treated areas, weincluded higher EPN rates [0.5, 1.0, and 2billion infective juveniles (IJs) per acre] inthe 2008 trials. However, despite lowerlarval densities, the higher EPN rates didnot provide greater or even equivalent con-trol than in 2007. Only the high rate of S.carpocaposae provided statistically signif-icant control (86%).

In the 2007 trials, treatments con-sisting of combinations of species andapplications split into half doses applied inconsecutive weeks provided higher levelsof control than the 1 billion IJs per acrerate alone. In 2008, the three combinationsof two species treatments failed to improveupon the levels of control observed at thefull rate (1 billion IJs/acre) of either of thespecies applied alone. Similarly, splittingapplications into two consecutive half-ratetreatments (0.5 billion IJs/acre) did notprovide better control than the full or halfrate applied once. The inconsistent controlof ABW by EPNs could be due to numer-ous abiotic (e.g., weather, soil type) andbiotic (e.g., EPN formulation, persistence,

ABW density) factors. Another year offield trials will be conducted to solve someof the inconsistencies in control of ABWwith EPNs.

Biological and Biorational Management Options forthe Annual Bluegrass Weevil on Golf Courses


Figure 1. First generation larval feeding damage to anedge of a fairway (June 23, 2008).

Benjamin A. McGraw and Albrecht M. KoppenhöferRutgers University

Objectives:1. Conduct surveys for entomopathogenic nematodes in annual bluegrass weevil (ABW) infested areas and adultABW hibernation sites on golf courses.

2. Determine the virulence to annual bluegrass weevil of entomopathogenic nematodes, Bacillus thuringiensis (Bt)strains, and several biorational compounds.

3. Determine the field efficacy of promising entomopathogenic nematodes, Bacillus thuringiensis, and several biora-tional compounds.

Figure 2. Density of ABW stages (average ± SE) in afield experiment 2 weeks after application of the nema-todes S. carpocapsae (Sc), H. bacteriophora (Hb), H.megidis (Hm), S. feltiae (Sf), and S. kraussei (Sk) forfield trials conducted in 2006, 2007, and 2008.Nematodes were commercial strains (BU = BeckerUnderwood) except for one field isolated (Hb PB).Figures above bars are percent reduction compared tountreated control (UTC). Asterisces (*) indicate signifi-cant mean separation from the UTC (P < 0.05).


Summary PointsEntomopathogenic nematodes can pro-

vide significant control of ABW larvaeunder laboratory conditions. However, infield trials, control was inconsistentbetween years.

The level of suppression achieved inthe field is likely affected by factors suchEPN concentration, ABW larval densities,and timing of application.

An additional field season should helpclarify the effects of application timingand concentration, and ABW density onthe ability of EPNs to suppress ABWbelow damaging thresholds.

4

Anthracnose is a destructive disease of

annual bluegrass and bentgrass puttinggreen turf throughout the United States.The disease, caused by the fungusColletotrichum cereale, begins as smallareas of yellowed turf (1 to 2 inches indiameter) with individual leaf blades even-tually senescing. The frequency and sever-ity of anthracnose outbreaks on golf courseputting greens has increased over the pastdecade perhaps due to practices employedto improve playability and increase ballroll distances on putting greens.

An evaluation of how irrigationmanagement practices influence anthrac-nose was initiated in 2006. Three consec-utive years indicate that deficit irrigation(40% daily ETo replacement) generally

had the greatest amount of disease com-pared to plots with higher soil water con-tent. Disease tended to decrease as irriga-tion increased; that is, 60% ETo replace-

ment had less disease than 40% ETo, and

80% ETo had less disease than 60% ETo.

Replacing 100% ETo daily (excessive soil

water) had levels of disease similar to 40%ETo treatments in August 2006 and 2008.

While this relationship was not observed in2007, turf quality of 100% ETo plots was

greatly reduced by August in all threeyears of study. Irrigation at 80% ETo often

resulted in the least disease and best turfquality. An additional study was initiatedin 2008 to evaluate the effect of mowingtiming relative to daily irrigation at 100%ETo. Disease severity was the same

regardless of whether irrigation wasapplied before or after mowing.

A study to follow up on previouswork was initiated in 2006 and comparedthe effect of lightweight roller type onanthracnose. Both roller types (i.e.,sidewinder and triplex-mounted vibratory)reduced disease compared to non-rolledturf under moderate disease pressure in2007 and 2008. The heavier sidewinderroller had less disease than the triplexmounted vibratory roller on 4 of 13 ratingdates over two years. Turf areas whererollers changed direction of travel andreceived a "clean-up" mowing at the edgeof a putting green had less disease com-pared to the area where rollers traversedwith no clean-up mowing on 6 of 13 ratingdates.

A study was initiated in 2007 todetermine the effect of variable rate andfrequency of summer soluble N fertility onanthracnose severity. Nitrogen applied at0.1 lb per 1000 ft2 every 7 days or 0.2 lbper 1000 ft2 every 14 days had the greatestreduction in anthracnose severity through-out both years. Nitrogen applied at 0.1 lb

per 1000 ft2 every 14days was the lowest Ntreatment to reduce dis-ease severity. Summersoluble N fertilization at0.1 lb per 1000 ft2 every28 or 56 days had thegreatest anthracnoseseverity.

A study was initi-ated in 2007 to evaluatethe effects of sand top-dressing and foot traffic

(golf shoes) on anthracnose severity. Sandtopdressing initially increased anthracnosein 2007; however, continued weekly appli-cations of sand reduced anthracnose sever-ity by August 2007 and throughout 2008.Sand topdressing reduced anthracnoseseverity regardless of foot traffic, and foottraffic decreased anthracnose in the pres-ence or absence of sand topdressing. Thecombination of daily foot traffic withweekly sand topdressing resulted in thelowest disease severity and best turf quali-ty in both 2007 and 2008.

Developing Best Management Practices for AnthracnoseDisease on Annual Bluegrass Putting Green Turf

Start Date: 2006Project Duration: two yearsTotal Funding: $60,000

Light frequent sand topdressing buries and protects crowns and leaf sheaths.Note depth of crowns in the middle (1 ft3 per 1000 ft2 per week) and right (2 ft3

per 1000 ft2 per week) profile samples are deeper than the profile sample onthe left (no topdressing).

James A. Murphy, Bruce B. Clarke, and Joseph A. RobertsRutgers University

Objectives:1. The multiple objectives of this research were organized into four field studies on annual bluegrass putting green turfthat were designed to evaluate the main effect and interactions of : 1) irrigation quantity; 2) lightweight rollers andmowing equipment; 3) topdressing and foot traffic, and 4) nitrogen fertilization on anthracnose disease.


Summary PointsDeficit irrigation (40% ETo) induced

wilt stress and intensified anthracnoseseverity. Irrigation at 80% ETo often

resulted in the lowest anthracnose severityand best turf quality.

Lightweight rolling every other daywith either roller type (i.e., sidewinder ortriplex-mounted vibratory) effectivelyincreased ball-roll distance and decreasedanthracnose severity under moderate dis-ease pressure.

Adequate nitrogen fertility to sustainmoderate growth was needed to reduceanthracnose severity. Nitrogen appliedevery 7 (0.1 lb per 1000 ft2) or 14 days(0.2 lb per 1000 ft2) provided the greatestreduction in anthracnose severity.Applications every 14 days at 0.1 lb per1000 ft2 also reduced disease, but to a less-er extent.

Sand topdressing initially increasedanthracnose in 2007. Continued weeklyapplications of sand reduced anthracnoseseverity by August 2007 and throughout2008. Sand topdressing reduced anthrac-nose severity regardless of foot traffic.Moreover, daily foot traffic decreasedanthracnose severity. The combination ofdaily foot traffic with weekly sand top-dressing resulted in the lowest diseaseseverity and best turf quality in both 2007and 2008.

5

A diverse group of Rhizoctonia

pathogens cause disease on warm-seasonturfgrasses in Florida. Diagnosing theserelated diseases can be difficult becausethey produce very similar symptoms onclosely-mown turf during overlappingperiods of disease-favorable conditions.

Seashore paspalum isolates col-lected during sampling efforts have provento be very diverse. Isolates that fall withinthe Waitea circinata species complex thatincludes R. zeae, R. oryzae, W. circinatavar. circinata and others are especiallyinteresting. Additional research needs tobe done to further characterize this groupof pathogens.

Rhizoctonia zeae isolates havebeen used to inoculate seashore paspalumcultivars and symptoms similar to thoseobserved in the field (and similar to leafand sheath spot on bermudagrass) resultedThe pathogens were re-isolated from thesymptomatic turf confirmingpathogenicity.

Some broad-spectrum fungicideproducts may be management tools for allof the Rhizoctonia diseases, but our fungi-cide-amended media assays indicate thatspecies of the pathogens differ in sensitiv-ities to most products. These differencesmay help to explain reports by superin-tendents of varying management resultsand efficacy windows for the same productused to treat similar disease symptoms.

Isolates of R. zeae were comparedto isolates of R. solani that cause largepatch, those that cause brown patch, R.

cerealis, and some related species. Isolatesof R. zeae were less sensitive to all fungi-cides tested (azoxystrobin, flutolanil, ipro-dione, propiconazole, pyraclostrobin, tebu-conazole, thiophanate methyl, and tri-adimefon) than R. solani isolates. Ourresults agreed with previously publishedfindings in that R. zeae was completelyinsensitive to thiophanate methyl, but wealso discovered than even with fungicidesthat are effective for both large patch andleaf and sheath spot, leaf and sheath spotisolates are less sensitive as a group thanlarge patch isolates.

Additional differences betweenisolates were identified that may havemanagement implications. Yellow patchisolates were sensitive as a group to ipro-dione, but some R. zeae and Marasmiusspp. isolates were moderately to complete-ly insensitive according to our test.

Fungicide evaluation trials onseashore paspalum for control of largepatch, dollar spot, and fairy ring haveshown that DMI fungicides such as tri-adimefon, propiconazole, triticonazole,

metconazole, and combinations of strobil-urins with these fungicides are safe andeffective as disease management tools onseashore paspalum. The products Headwayand Tartan have been among the best treat-ments evaluated and offer broad-spectrummanagement options from single products.

Progress has been made towardidentifying Fusarium isolates that affectseashore paspalum. Some of the isolateshave been identified as previouslydescribed Fusarium species and othersmay be previously undescribed.Considerable work and additional collec-tions will need to go into investigatingthese apparently new species prior to pub-lishing descriptions and names; however,pathogenicity trials have been completedand management trials are beginning toprovide insight. Research is ongoing.

An inoculation of Dr.Kenworthy's zoysiagrass evaluation trialresulted in little disease. Additional iso-lates are also being tested on a small num-ber of cultivars to find the most pathogen-ic and consistent isolate to use in furtherscreening and selection research.

Relative Pathogenicity and Fungicide Sensitivity of Isolates ofRhizoctonia and Other Fungal Pathogens and the Disease

Responses of Seashore Paspalum and Zoysiagrass Cultivars


Carol M. Stiles, Philip F. Harmon, and Kevin E. KenworthyUniversity of Florida

Objectives:1. To confirm pathogenicity of Rhizoctonia zeae on seashore paspalum and screen isolates of R. solani and R. zeae

to different fungicides.2. To confirm whether isolates of Fusarium or Microdochium spp. obtained from diseased seashore paspalum sam-ples are pathogenic. If so, identify and characterize these isolates and develop diagnostic responses and man-agement recommendations.

3. Screen existing seashore paspalum and zoysiagrass cultivars for resistance to various pathogens.4. Screen zoysiagrass germplasm accessions to select for resistance to Rhizoctonia spp.


Summary PointsIsolates of several Rhizoctonia spp.

have been characterized from warm-sea-son turf in Florida and initial results sug-gest a more diverse group than expected.

Amended-media studies indicate thatRhizoctonia spp. vary in sensitivity tomany different fungicides and isolatesoften vary within species, as well.

A diverse group of pathogens havebeen isolated from seashore paspalum turfin Florida. We suspect at least two of thespecies we have confirmed pathogenicitywith are previously undescribed.

6

Isolates of several Rhizoctonia spp. have been charac-terized from warm-season turf in Florida and initialresults suggest a more diverse group than expected.

Biological insecticides and natural

enemy conservation can reduce the needfor chemical inputs on golf courses. Thiswork is the first survey of white grubs andtheir natural enemies on golf courses in thetransitional climactic zone, and the firstanywhere for masked chafers(Cyclocephala spp.), the most importantnative grub pests. We seek new pathogenshaving promise as bio-insecticides and toclarify how site characteristics might bealtered to enhance natural suppression ofgrub populations.

Grub survey kits were sent to 34golf superintendents throughout Kentuckyin late summer asking them to collect 30grubs and a soil sample from their worstnon-treated grub site. Samples werereturned via overnight mail. Six addition-al Lexington-area golf courses were inten-sively sampled in late August, mid-September, and early October to track nat-ural enemy incidence over time.

Grubs were identified, incubatedfor 30 days, and dissected to assess mortal-ity from bacterial, fungal, or protozoanpathogens. Masked chafers (MC) andJapanese beetles (JB) accounted for 64 and30%, respectively, of grubs sent in bysuperintendents. MC also predominatedon Lexington courses. Grub populationsdeclined from about 18 per 0.1 m2 in lateAugust to about 5 per 0.1 m2 in Octoberand 2 per 0.1 m2 the following springowing to natural mortality agents. Tiphiawasps, Metarhizium fungus, Serratia(amber disease) and Paenibacillus (milkydisease) bacteria, and entomopathogenic

nematodes (EPN) infected 2, 5, 8, 20, and18% of the MC grubs, and 0, 3, 5, 12, and19% of the JB grubs sent in by superin-tendents.

Ovavesicula, a protozoan thatcauses debilitating disease in adult JB, wasuncommon in KY, but gregarines(Stichtospora) infected 26% of JB grubs inthe spring. The latter two pathogens wereabsent or uncommon in MC grubs.Additional Stichospora-infected grubswere reared to determine their fate whichhas not previously been studied. Suchgrubs emerged as adults although theirmaturation was slightly delayed. EightyEPN strains, 40 each from MC and JBgrubs, are being processed for PCRidentification.

Replicated stands of irrigatedturfs used in fairways or roughs of transi-tion zone golf courses were sampled forgrub species preference and incidence ofparasitoids and pathogens. Plots main-tained at fairway height (5/8") consisted ofcreeping bentgrass (CB), perennial rye-grass (PR), zoysiagrass (Z), and bermuda-grass (B). Plots at rough height (2.5 in)

consisted of turf-type tall fescue (TF),Kentucky bluegrass (KB), PR, or a TF/KBmix. Of the fairway-height grasses, Z andB had the highest incidence of MC grubs.MC predominated in CB, whereas JBfavored PR. JB populations were highestin rough-height grasses, outnumbering MC2-4 fold. Skunk damage was greatest inCB and PR at fairway height. There waslittle skunk digging in fairway-height Z orB, or in rough-height grasses. Parasitismby Tiphia occurred in all grasses but wasgreatest in zoysiagrass.

Grub collection kits were sent tocooperators in 22 states to survey naturalenemy load in JB populations throughoutthe species range in the USA. Collectedgrubs are being analyzed for pathogens totest whether impact of particular agents isassociated with how long JB has beenestablished in a given area.

Natural Enemies and Site Characteristics Affecting Distribution andAbundance of Native and Invasive White Grubs on Golf Courses


Grubs were identified, incubated for 30 days, and dis-sected to assess mortality from bacterial, fungal, or pro-tozoan pathogens.

Daniel A. Potter and Carl T. RedmondUniversity of Kentucky

Objectives:1. Determine identity and incidence of microbial pathogens and parasitoids of Japanese beetle and masked chafergrubs on golf courses across Kentucky, the first such study in the transitional turfgrass zone. Quantify site char-acteristics associated with particular grub species and natural enemy incidence.

2. Evaluate how grass species, mowing height, and golf course pesticides affect incidence of pathogen infection andparasitism of naturally occurring white grubs in the field and if impact of naturally-occurring pathogens can beenhanced.

3. Test the hypothesis that natural enemy load on Japanese beetle grub populations on golf courses is greater in geo-graphical regions where populations have stabilized than in regions into which the pest has more recently spread.


Summary PointsGrub species and associated pathogens

and parasitoids were surveyed on 27Kentucky golf courses, the first such studyin the transitional climactic zone. Maskedchafers and Japanese beetle grubs account-ed for about 66 and 30% of the grub infes-tations, respectively. Insect-pathogenicnematodes, Tiphia wasps, milky disease,and other pathogens account for moderateto high natural mortality at some sites.

Nematodes isolated from MC and JBgrubs are currently being identified.

Turfgrass species and mowing heightinfluence incidence of grubs and naturalenemies.

Pathogens of JB grubs are being sur-veyed across the eastern and centralUnited States to determine if naturalenemy load can predict area-wide cyclesof decline of grub populations.

7

The annual bluegrass weevil (ABW)

is a burgeoning pest throughout theNortheast and Mid-Atlantic regions. Thestem-boring and crown-feeding larvaecause highly visible damage to short-cutPoa annua, a major component of golfcourse playing surfaces. Managementoptions are largely limited to pyrethroids,and applications may be made 2-5 times aseason. Poor targeting often leads to con-trol failures.

Detailed population surveys wereconducted at two sites in Upstate NY todescribe patterns of variation in populationecology across year, site, and habitat.Population data were collected from 2004-2006. Based on extractions of larvae fromsoil cores and collections of adults flushedby a disclosing solution, 3,838 larvae and8,576 adults were sampled. Five larvalinstars were confirmed based on head cap-sule width. Because there is no overlap,instar can be determined with a simplemeasurement. Adults were also identifiedas male and female, callow and mature.Because there is no divergence in male andfemale population curves, assessing gen-der does not help to interpret adultfluctuations.

Most population parameters (e.g.fluctuation curves, abundance, synchrony,number and timing of generations) variedmore between years than between sites.

Sex ratio and abundance varied betweenmanagement habitats. For instance, maleswere more abundant in the rough wherethe mean sex ratio (male:female) was1.7:1, versus 1:1 in the fairway. In termsof insect load, larvae and adults were 8-9times more abundant on the fairway thanthe rough. Across the fairway itself, abun-dance was consistently greater near theedge at one site, but insects were evenlydistributed across the fairway at the othersite.

An analysis of three years of pop-ulation data shows that degree-day may bea better fit than Julian date at predictingoccurrence of the first generation. Givenlow variation in R2 values, using the mostconvenient base-temperature model maybe feasible. In 2008, we partnered withcollaborators across NY to validate thismodel through focused populationsurveys.

Field surveys showed that over-wintering adults tend to settle along thetree line adjacent to the fairway, establish-ing up to 60 meters from the fairway and10 meters into the woods. Little or nooverwintering occurs on the fairway oradjacent rough. In a choice experiment,we showed that white pine litter is not apreferred overwintering substrate. Adultspreferred to settle in rough-mown grassand a combination of pine and deciduouslitter over fairway-mown grass and pinelitter alone. Captures in linear pitfall trapswere greatest in spring. At one site, direc-tional movement toward the fairway inspring was confirmed, but there was noevidence for reverse movement in the fall.

Results lead us to propose a newconceptual model of flux between habitatsand overwintering site selection. Our the-ory is that spring immigration occurs most-ly by walking, with orientation to low-mown turf. Fall emigration occurs throughflight with orientation to defined tree lines.Through a "snow-fence" effect, adults stopflying at the tree line, drop to the ground,

and settle into overwintering substratesaccording to preferences. In 2008, labora-tory and field observations confirmed thatABW is capable of vigorous flights. Bettertargeting of ABW will depend on futurestudies that emphasize dispersal behavior,host plant associations, and habitatpreferences.

Interpreting and Forecasting Phenology of theAnnual Bluegrass Weevil in Golf Course Landscapes


Annual bluegrass weevil launching into flight from thetop of a stick

Daniel C. Peck, Masanori Seto, and Maria DiazCornell University

Objectives:1. Describe patterns of variation in population fluctuations and phenology.2. Describe the overwintering strategy by establishing the factors that affect site selection and success.3. Document the relationship between overwintering sites and developmental sites.4. Develop and validate a degree-day model to forecast phenology.


Summary PointsMost population parameters (e.g. shape

of the fluctuation curve, number of gener-ations, and generation time) vary morebetween years than between sites.

Sex ratio and abundance vary dramati-cally between rough and fairway habitats.Across the fairway, insect distribution doesnot explain the prevalence of damagealong the edge.

Degree-day accumulation may be bet-ter than calendar date at predicting phenol-ogy, and a preliminary model has high pre-dictive power for timing of the first gener-ation.

Overwintering adults tend to settlealong tree lines adjacent to the fairway, asfar as 10 m into the woods and 60 m fromthe edge of the fairway. Little to no over-wintering occurs on the fairway and bor-dering rough. Pine litter is not preferredover other overwintering substrates.

It is hypothesized that adults immigrateto fairways in spring largely by walkingwith orientation to low cut turf, but theyemigrate in fall largely by flying with ori-entation to defined tree lines where theysettle into overwintering substrates.

8

Annual bluegrass weevil eggs laid in a stem of annualbluegrass

Larra bicolor, an introduced ectopar-

asitoid of mole crickets, has spread into thenorthern Gulf region (Mississippi andAlabama) from where it was originallyintroduced (Florida). This creates anopportunity for golf course superintend-ents in this area to utilize this biologicalcontrol agent.

We are monitoring wasp activitymonthly on four sites on the MississippiGulf Coast where where L. bicolor isknown to occur. Across all three years,wasps were active beginning in June andgenerally ending in November.Comparison of activity of foraging waspson turf is comparable to foraging wasps onnectar sources. This suggests that the pres-ence of wasps on nectar sources will indi-cate wasps foraging in turf. Wasps werenot active at St. Andrews in 2008. This sitewas one of the first along the Mississippicoast where L. bicolor was detected.

In 2007 and 2008, we establisheda replicated garden of flowering herba-ceous plants at the Coastal REC. The gar-den consists of four replicates of the 15plant taxa arranged in 0.5 × 0.5 m plots.Between 1100-1300 h during peak flight,numbers of L. bicolor on flowering plantswere recorded. Of all the species presentand flowering, only two, Pentas and

Spermacoce, were visited by L. bicolor.White pentas was readily foraged upon byL. bicolor and numbers of wasps werecomparable to Spermacoce. Among pen-tas, white-flowered pentas was significant-ly different from either pink or red-flow-ered pentas.

In 2007, both red and pink flow-ered pentas were comparable, however,they were significantly different in 2008.There may be volatiles or ultraviolet visu-al cues recruiting wasps to these flowers.In 2008, attempts to observe recruitment toflowers or host mole crickets under con-trolled conditions in a wind tunnel wereunsuccessful.

In 2007 and 2008, diel activity ofL. bicolor was studied by monitoring waspactivity on flowers of blooming plots ofSpermacoce verticillata and white-flow-ered Pentas lanceolata. These plots werepart of the garden experiment previouslydescribed. This study was conducted fromAugust 7-10, 2007 and October 9-11, 2008when wasp populations were abundantenough to collect data. The number andsex of wasps on flowering plants in 8 plotsin the garden were taken hourly from0500-2000 h each day.

In both years, wasps begin active-ly foraging about 2-3 hours after sunrise tonear dusk. Sunrise, and not temperature,likely triggers activity. Air temperatures inAugust when wasps became active were27-28 C, whereas wasps were active whenair temperatures were 18.3 C in Oct.Males and females are present throughoutthe day with females being more transient.Populations were more clearly male-biasedin October than in August samples. It isgenerally thought that L. bicolor flowersare male-biased, but these data suggest thatmay depend on time of year. At certaintimes during the August 2007 observa-tions, numbers of male and females werealmost equal.

In May and June, plots were

established on a local golf course (GreatSouthern Golf Course) and at the CoastalREC. The objective was to determine ifproximity to pentas could increase para-sitism of mole crickets. Plots of floweringplants were established in the center of 11-m transects. Once wasps were active onthe flowers, containers of mole cricketswould be buried in the ground at 1, 3, and5 meters from the center. Unfortunately,wasps were not present in the floweringplots during July, August, or September.

Seasonal Life History and Suitability of Horticultural Plantsas Nectar Sources for Larra bicolor, a Parasitoid of

Mole Crickets in the Northern Gulf Coast


David W. HeldMississippi State University

Objectives:1. Determine the seasonal life history of Larra bicolor in the northern Gulf Coast.2. Determine the suitability of flowering plants as nectar sources for Larra bicolor.3. Determe if incorporating wildflowers on golf courses will facilitate the establishment of Larra wasps on that site.


Summary PointsLarra bicolor have a 4-5 month activi-

ty in the northern Gulf region to parasitizemole crickets. Wasps are active mostlyduring daylight hours. Although theimpact of insecticides on L. bicolor havenot been evaluated, turf managers wantingto conserve these biological controlsshould consider making insecticide appli-cations later in the day as numbers of for-aging wasps wane.

In two trials, pentas, particularly whitepentas, recruit equivalent numbers of L.bicolor as flowering Spermacoce verticil-lata. Attempts to assess the impact of nec-tar sources on parasitism were not success-ful in 2008 and will be repeated in 2009.

9

Numbers of wasps recorded on plants in the commongarden experiment. Data from 6 days in 2007 and 10days in 2008 during the peak of activity were selectedand analyzed. Only four taxa, two species, wereobserved with wasps. There were significantly morewasps on Spermacoce and the white-flowered pentasthan either of the other two flowering pentas in bothyears.

Common garden experiment conducted in 2007 and2008 to assess preference of L. bicolor for floweringornamental plants

There has been little study on the

impact of irrigation and aeration manage-ment on rooting in creeping bentgrassgrown in a sand-based rootzone under fieldconditions.

'Providence' creeping bentgrasswas grown on a sand-based rootzone meet-ing USGA recommendations. Plots weresubjected to two irrigation programs: lightand frequent versus deep and infrequent.Light, frequent plots were irrigated dailyon rain-free days to maintain a moist con-dition in the upper 4-6 cm (1.6-2.4");whereas, deep, infrequent irrigated plotswere irrigated at leaf wilt to a depth > 24cm (>9.5").

A majority of roots (55%) werefound in the upper 2.4" (6 cm) of soil at theend of the summer, regardless of irrigationregime. Deep, infrequent irrigated bent-grass produced a greater number of roots,longer roots, and a larger root surface areaand a smaller root diameter (2007) vs.light, frequent irrigated bentgrass.

Soil temperatures were on aver-age < 0.7 C (< 1.4 F) greater in light, fre-quent irrigated bentgrass. Deep, infrequentirrigated bentgrass had lower canopy pho-tosynthetic rates, but respiration was simi-lar to light, frequent irrigated bentgrass.Canopy temperatures were 2.2 C (4.0 F)higher in deep, infrequent vs. light, fre-quent irrigated bentgrass.

Deep, infrequent irrigated bent-grass had lower color and quality andlower chlorophyll levels in 2006 and mostof 2007. By late summer, however, colorand quality and higher chlorophyll levelswere detected in deep, infrequent vs. light,frequent irrigated bentgrass. Deep, infre-quent irrigated bentgrass developed a less

thick thatch-mat layer, which containedless organic matter versus light, frequentirrigated bentgrass.

Deep, infrequent irrigated bent-grass leaves had higher water soluble car-bohydrate and total non-structural carbo-hydrate levels in 2006, but higher storagecarbohydrate levels in both years. Deep,infrequent irrigated bentgrass had higherstorage carbohydrate and non-structuralroot carbohydrate levels than light, fre-quent irrigated bentgrass in both years.Deep, infrequent irrigated bentgrass accu-mulated proportionately more non-struc-tural carbohydrate in roots versus leaves.Nearly twice as much water was applied tolight, frequent versus deep, infrequentplots in both years.

Regarding aeration, three regimeswere assessed: spring only, spring plusthree summer corings, and a non-coredcheck. Spring core aeration holes werefilled to the surface with topdressing, butin summer, aerated plots cores werebrushed to re-incorporate soil and no addi-tional topdressing was applied.

In 2005, total root counts andtotal root length were increased by sum-mer coring vs. spring coring. Total rootcounts and total root length generally weregreater in the entire profile in spring plussummer-cored versus spring or non-cored

bentgrass in 2007.Data indicated that summer core

aeration should be avoided the first sum-mer of establishment. If necessary, onlycore aerate to the depth of the thatch-matlayer. The % total root counts in the 0-6cm (0-2.4") of soil ranged 61to 74%, 58 to59%, and 62 to 77% among all three cor-ing treatments in late summer of 2005,2006, and 2007, respectively.

Spring and spring plus summer-cored plots developed a thicker thatchlayer than non-cored bentgrass. Theamount of organic matter (loss on ignition)in the thatch-mat layer increased in allthree regimes, but the levels remained thesame among regimes. However, the organ-ic matter concentration (gravimetric organ-ic: dry wt. of cores) was lower in coredplots. Organic matter concentration lessthat 110 g kg-1 was associated with betterturf performance.

Spring and spring plus summercoring reduced quality for about twoweeks, but generally had higher color rat-ings than non-cored bentgrass. Late sum-mer quality was better in cored plots.Chlorophyll a and a+b levels were higherfor spring and spring plus summer coredbentgrass in both years.

Rooting and Carbohydrate Metabolism in Creeping BentgrassPutting Greens in Response to Summer Irrigation and Aeration


Jinmin Fu and Peter H. DernoedenUniversity of Maryland

Objectives:1. Evaluate physiological processes and rooting of putting green-height creeping bentgrass in response to two irriga-tion management and three aeration regimes.

2. Determine the influence of aeration and irrigation frequency on creeping bentgrass summer performance androot longevity during periods of supraoptimal temperature stress.

3. Provide information on the effects of soil temperature and soil water content on carbohydrate metabolism and itsrelationship to summer bentgrass decline.

Soil temperature was measured by installing tempera-ture sensors about 0.8 inches below the soil surface.


Summary PointsDeep, infrequent irrigation produced a

greater number of roots, longer roots, alarger root surface, lower soil temperature,less thatch, higher water soluble and totalnon-structural carbohydrates than light,frequent irrigation.

Data indicated that summer core aera-tion should be avoided the first summer ofestablishment.

Spring and spring plus summer-coredplots exhibited reduced quality for abouttwo weeks, but generally had higher colorratings and chlorophyll a and b levels thannon-cored plots.

10

Spring dead spot is the most devastat-

ing and important disease of bermudagrassthat undergoes winter dormancy. The dis-ease is caused by one or more of three fun-gal species in the genus Ophiosphaerella(O. herpotricha, O. korrae, or O. nar-mari). The disease results in unsightlydead patches on fairways, tees, andbermudagrass greens, giving way to theencroachment of weeds and costly man-agement efforts to eliminate weeds andreestablish grass in the affected area.

A critical limitation to the studyof turfgrass root diseases is the inability ofresearchers to rapidly and easily study theplant-fungus disease interaction because ithappens below ground and often inside ofroots. The overall goal of this study is toenhance our understanding of the interac-tion between O. herpotricha and itsbermudagrass host and how environmentaland host factors influence this interaction.

We have inserted two differentfluorescent reporter genes (red and green)into the fungus and examined root and

stolon infection of various bermudagrass-es. For the interspecific hybrid (Cynodondactylon × C. transvaalensis) cultivars‘Tifway’ and ‘Midlawn’, root cortical cellswere rapidly colonized. However, theroot's stele remained uninfected 10 daysafter inoculation. For a C. transvaalensisaccession, both the root cortex and stelewere colonized by eight days after inocula-tion. In general, ‘Tifway’ roots exhibitedgreater colonization and necrosis than themore tolerant cultivar ‘Midlawn’ and C.transvaalensis, which, though its rootswere heavily colonized, exhibited very lit-tle necrosis.

Additional studies have been con-ducted on the stolons of these bermuda-grasses. When stolons were inoculated onunwounded internodes, very limited epi-dermal and cortical infections wereobserved after 28 days for all threebermudagrasses. However, stolons thatwere root-inoculated six weeks earlierbecame infected through cut end of thestolon.

For ‘Tifway’ stolons, infectionresulted in internal necrosis and cavities,similar, but less severe response wasobserved for ‘Midlawn’, however in C.

transvaalensis, stolon tissues were colo-nized without any apparent necrosis.

Ongoing studies using the confo-cal scanning laser microscope will optical-ly 'section' infected roots and produce 3-dimentional images of the fungus as itmoves in and on bermudagrass roots. Thisbasic information on how the cultivarsreact to the fungus will improve our abili-ty to enhance and deploy host-plant resist-ance through traditional breeding efforts atOklahoma State University.

Infection and Colonization of Bermudagrass by Ophiosphaerellaherpotricha, the Causal Agent of Spring Dead Spot


Spring dead spot is the most devastating and importantdisease of bermudagrass.

Nathan R WalkerOklahoma State University

Objectives:1. To transform O. herpotricha to express fluorescent protein genes.2. Evaluate infection and colonization of bermudagrass cultivars at different temperatures.3. Evaluate differences in infection and colonization between bermudagrass cultivars that vary in disease

susceptibility.

For a C. transvaalensis accession, both the root cortexand stele were colonized by eight days after inoculation.


Summary PointsFungal colonization of root tissues var-

ied among the bermudagrass cultivarsexamined.

Colonization and necrosis of roots wasextensive for ‘Tifway’, less for ‘Midlawn’,however, limited necrosis occurred for theC. transvaalensis accession even though itwas extensively colonized.

Internal colonization of stolon resultedin severe necrosis for ‘Tifway’ but not forthe C. transvaalensis accession.

This information will be used toenhance host-plant resistance through tra-ditional breeding efforts at OklahomaState University.

11

Root cortical cells were rapidly colonized in the inter-specific hybrids ‘Tifway’ and ‘Midlawn’.

Gray leaf spot, caused by

Magnaporthe oryzae, is a serious diseaseof perennial ryegrass turf that can causeextensive damage to turf in golf coursefairways. The disease is effectively man-aged by fungicide applications. However,turf managers often explore the possibili-ties of various cultural practices that can berelatively easily integrated along with fun-gicide into a broader disease managementstrategy.

Soil silicon amendment havebeen proven effective in controlling bothsoilborne and foliar fungal diseases of sev-eral plants including some turfgrassspecies. However, the effects of silicon ongray leaf spot development of perennialryegrass turf were not known. This studywas undertaken to investigate the effects ofsilicon on gray leaf spot incidence andseverity in perennial ryegrass.

The experiments were set up inWillow Hollow Golf Course and LuluTemple Country Club in Pennsylvania in asplit-plot design with rate of silicon as themain-plot factor and rate of fungicides asthe split-plot factor. Silicon (calcium sili-cate) was applied at six different rates: 0,

0.5, 1, 2, 5, and 10 metric ton/ha. Thefungicides selected for the study were: asystemic fungicide, azoxystrobin, and acontact fungicide, chlorothalonil. Eachfungicide was sprayed at low, medium, andhigh label rates - 0.31, 0.46 and 0.61 a.i.kg/ha azoxystrobin, and 4.53, 6.29 and8.18 a.i. kg/ha chlorothalonil, respectively.Calcium silicate was broadcast as a topdressing in each treatment plot (76.2 x 45.7cm) and incorporated by core aerationusing a Toro procore 660 open-tine. Thecores were 1.9 cm in diameter and 7.6 cmin depth at 7.6 cm x 7.6 cm spacing.

The grass in the plots was main-tained at a 3.5 cm height and mowed twicea week. The grass was fertilized with (5%N, 5% P2O5 and 20% K2O) at 1.22 x 10-3

kg/m2 with Team for crabgrass controlbefore the experiments was initiated.Fungicides were applied six weeks afterapplication of silicon and repeated after 14days to deliver two applications prior toinoculation with Magnaporthe oryzae (60x103 conidia/ml H2O with 0.1% Tween

20).Fungicides were applied in water

equivalent of 814 L/ha (2 gallons/1000 sqft). Disease severity (index 0-10; 0=turfarea asymptomatic, 10=91-100% of turfarea necrotic) was assessed weekly for sixweeks. Disease progress over time wasevaluated using the parameter estimates,rate r, andAUDPC, where r=rate of diseaseprogress and AUDPC = area under the dis-ease progress curve. Data were analyzedusing regression and ANOVA procedures.

The results showed that effectssilicon and fungicide on gray leaf spotseverity were significant (P=0.05).Assessment of the disease showed thatgray leaf spot severity (weekly), rateparameter, r, and the area under diseaseprogress curve (AUDPC; severity-weeks)were significantly reduced by applicationof silicon and fungicides.

Although all three rates of each of

the systemic or contact fungicide reducedgray leaf spot severity similarly, combina-tion of silicon and the fungicide providedfurther reduction of the disease. Increasedrate of silicon application in combinationwith fungicide generally provided greaterreduction of gray leaf spot.

This study demonstrates that con-trol of gray leaf spot in perennial ryegrassby fungicide application may be augment-ed by amendment of soil with silicon. Thisstudy suggests that silicon may be used asan important component of an integratedgray leaf spot management strategy thatincludes cultural and chemical approaches.

Future studies are warranted todetermine the residual effects of silicon ongray leaf spot development in silicon-amended soil over time and interactionsbetween silicon and various commonlyused fungicides for gray leaf spot control.

Silicon Amendment: A Component of an IntegratedGray Leaf Spot Management Strategy


Wakar UddinPennsylvania State University

Objectives:1. To evaluate accumulation of silicon in perennial ryegrass plants.2. To determine the effects of soil type, source of silicon, and rate of silicon amendment on gray leaf spot severity

and incidence.3. To devise a management strategy for gray leaf spot through integration of silicon into a fungicide program.

Figure 2. Comparison of AUDPC for the non-fungicide-treated and non-amended control, low, medium andhigh rates of chlorothalonil, and all treatments ofchlorothalonil combined with calcium silicate treatmentsin Site 1.


Summary PointsGray leaf spot severity was significant-

ly reduced by amendment of soil with sili-con.

Effects of fungicides on gray leaf spotwere augmented by application of silicon.

Increased rate of silicon applicationgenerally provided greater suppression ofthe disease within the fungicidetreatments.

12

Figure 1. Comparison of AUDPC for the non-fungicide-treated and non-amended control, low, medium andhigh rates of azoxystrobin, and all treatments of azoxy-strobin combined with calcium silicate treatments atSite 1.

Plants primarily assimilate nitrogen

through root uptake of nitrate from the soilsolution. Plants must convert nitrate toammonia before it can be incorporated intonucleic acids, proteins, and other nitrogencontaining compounds. It is estimated thatplants can use almost 25% of their photo-synthetically derived energy in this con-version process.

One possible way to reduce thisenergy-intensive process is to providenitrogen in the ammonium form, typicallyas urea or ammonium sulfate. Our researchseeks to understand the practice of foliarfertilization and determine the positive ornegative impacts of this process. By apply-ing fertilizers enriched with a stable, non-radioactive isotope of nitrogen (15N) as atracer, nitrogen use can be directly trackedthroughout the plant.

Beginning in early 2008, we ana-lyzed the tissue from several experimentsconducted in the summer of 2007 thatcompared urea and ammonium sulfateapplied to creeping bentgrass maintained

at a 0.5" height of cut. The application rateof each fertilizer was 0.2 lbs N/1000 ft2

with a spray volume of 40 GPA. Studieson the time course of 15N absorption sug-gest that nitrogen applied to the foliage isabsorbed by the plant within 4-6 hours. Wehave measured uptake efficiencies from10-35% under field conditions, suggestingthat uptake efficiency in the field can bequite variable.

Our studies to determine theeffect of spray gallonage on uptake effi-ciency indicate that lower spray gallonagesincrease uptake of foliar applied N. Resultssuggest that applying fertilizer at gallon-ages above 20 GPA does not result in sig-nificant foliar feeding.

Studies to determine if tank-mix-ing adjuvants with fertilizer wouldincrease uptake efficiency indicated thatthe addition of an adjuvant to foliar-applied fertilizer significantly increaseduptake efficiency although results did notindicate that a particular class of adjuvantis most effective. Results of other tank-mixing studies suggest that tank-mixing offertilizer with other leaf-applied chemicalsmay affect uptake efficiency.

In the summer of 2008, we initiat-ed a field study to determine whether foliarapplications of NH4-N have a positive

effect on plant metabolism. Plots of creep-ing bentgrass (Agrostis stolonifera L. 'PennA-4') were maintained at three heights ofcut, 0.125, 0.1 and 0.085", on a native soilputting green topdressed (10-day intervals)with sand and treated with foliar nitrogen.Foliar urea (as an NH3-N source), and cal-

cium nitrate (as a NO3- source), are being

applied weekly at annual rates of applica-tion of 2.4 lbs N/1000 ft2/year.

Nitrogen applications are divided

evenly into weekly applications applied inApril through October. Clippings are col-lected once per week during the study, andthe plot area is mowed six days per week.For evaluation and comparison of tradi-tional soil applied N-fertilization, bothforms of N fertilizer are applied weekly asa soil drench at the same annual N rates asthe foliar applications.

Data collected include leaf tissueN concentration, clipping production, per-cent coverage, turf quality, turf color,chlorophyll concentration, root mass, anddisease incidence.

Optimization of Foliar Nitrogen Nutrition toImprove Turf Performance Under Energy Stress


Mean fertilizer uptake over a 24-hour period as affectedby spray gallonage (20, 40, 60, 80, and 100 GPA). Errorbars indicate one standard deviation.

S. Henning, B. E. Branham, and R. L. MulvaneyUniversity of Illinois

Objectives:1. To determine the efficiency of ammoniacal-N uptake in the field from foliar feeding, particularly when done incombination with other turf care products.

2. To determine the positive or negative benefits of foliar ammoniacal-N applications on highly managed turfgrass3. To determine fertilizer use efficiency through foliar feeding versus conventional fertilization.4. To determine the effect of foliar ammoniacal-N applications on root and shoot growth, development, and perform-ance in the field

5. To determine if the practice of foliar application of nitrogen fertilizer to a crop that has a portion of its foliageremoved frequently is useful.

Mean fertilizer uptake over a 24-hour period as affect-ed by adjuvants. Error bars indicate one standard devi-ation.


Summary PointsFoliar applied N enters the leaf within

4-6 hours of application.

Lower spray gallonages are more effec-tive at promoting N-uptake through theleaf.

Addition of an adjuvant for foliar fertil-ization is recommended for optimizingfoliar N-uptake of urea.

Tank-mixing fertilizer with commonlyleaf-applied chemicals has limited impacton N-uptake through the leaf.

13

Biological controls, once established,

have the potential to provide prolongedsuppression of insect pests on golf courses.In 2003, a former graduate student discov-ered a caterpillar-specific virus decimatingpopulations of black cutworms (BCW) onKentucky golf courses. The virus, identi-fied as Agrotis ipsilon multiple nucle-opolyhedrovirus (AgipMNPV), wasamplified by feeding it to healthy BCW,harvesting their cadavers, and mixing theconcentrated virus particles with water.

This crude biological insecticidewas applied to turf to see if it would infectresident larvae. Initial testing showed thisAgipMNPV suspension to quickly killyoung larvae, but that larger ones requirehigher dosages and continue to feed forseveral days before being killed. Virus-infected BCW rupture in death and spreadmillions of infective virus particles ontofoliage and thatch. Virus sprays gave goodcontrol of BCW in small-plot field trials infairway- and collar-height creeping bent-grass.

Thousands of laboratory-rearedBCW were infected with AgipNMPV dur-ing the winter, harvested, and stockpiled toprepare enough virus suspension for a real-istic field trial. Six whole tees at each oftwo central Kentucky golf courses, plus 2meters into the surrounds, were sprayedwith virus suspension in spring 2008, withsix untreated tees on each course used forcomparison. Efficacy was determined atmonthly intervals by sampling natural den-

sities of BCW and by implanting sentinellarvae and eggs, using irritant (soap)drenches to recover short-term survivors,and rearing them to determine how manyultimately died from virus.

The virus provided 78% controlof young larvae hatched from eggs and 45-56% control of third instars placed in theturf one week after application. We arecurrently examining hundreds of BCWblood smears on microscope slides todetermine how many weeks of suppressionthe virus application provided.

Studies were initiated in 2008 toinvestigate the virus' compatibility withhost-plant resistance. BCW that developon endophytic grasses or Kentucky blue-grass show delayed growth and develop-ment. Such stress may increase their sus-ceptibility to virus residues encountered inthe turf.

BCW were allowed to feed in thegreenhouse for 3 days on 'Rosalin' perenni-al ryegrass with (E+) or without (E-) endo-phyte. Four rates of virus were thenapplied to the grass blades. Larvae were

left to feed on grass for 10 days and thensurvival, growth, and incidence of viruswere assessed. Main effects of endophyteand virus were highly significant. Larvaefeeding on E+ grass were stunted, yet thevirus caused similarly high (85-97%) mor-tality of BCW irrespective of grass type.This shows that AgipMNPV is not deacti-vated by alkaloids in the E+ grasses.

Inactivation of microbes by solarradiation can be a major hurdle to biologi-cal insecticides under field conditions.Optical brighteners have been used asadjuvants for insect pathogens in other sys-tems because they absorb UV light and canprotect original virus activity for weeksafter application.

In summer 2008 we evaluatedtwo optical brighteners for enhancing effi-cacy and persistence of AgipNMPVagainst BCW in fairway-height creepingbentgrass field plots. The virus providedand average of 89, 44, and 18% mortalityafter 1, 3 and 5 weeks, respectively, butthe brighteners did not significantly syner-gize or prolong infectivity.

Use of a Baculovirus for Season-long Control of BlackCutworms on Golf Courses and Compatability with Soil

Insecticides and Insect-resistant Turfgrasses


Daniel A. Potter and Andrea BixbyUniversity of Kentucky

Objectives:1. Evaluate AgipMNPV, a naturally occurring baculovirus, as a bio-insecticide for season-long and multi-year pre-ventive control of black cutworms (BCW) on golf courses.

2. Compare infectivity and persistence of AgipMNPV to BCW in sand-based and soil-based putting green and fair-way height creeping bentgrass habitats.

3. Investigate compatibility and possible synergism of AgipMNPV with soil insecticides used for grub control ongolf courses.

4. Investigate compatibility of endophytic and other insect-resistant turfgrasses with biological control of black cut-worms by AgipMNPV.

Once infected by the virus, black cutworm larvae can beused to prepare virus suspensions and applied to thefield to further infect resident larvae.


Summary PointsAgipMNPV gave good short-term con-

trol of BCW on golf course tees underactual play conditions. Extent to which itgave season-long control is still beingevaluated.

AgipMNPV is compatible with insect-resistant endophytic turfgrasses.

AgipMNPV also gave good control infairway-cut creeping bentgrass, but addi-tion of optical brighteners to the virus fromUV degradation did not detectably prolonginfectivity or synergize its efficacy atlower rates.

14

Rhizoctonia blight (brown patch),

caused by R. solani is a disease of cool-season grasses, including bentgrasses, fes-cues, and ryegrasses. Anastomosis groups(AG) 1, 2, 4 and 5 have been previouslyisolated from blighted grasses. Rhizoctonialeaf and sheath spot of both warm and coolseason grasses are caused by both R. zeaeand R. oryzae.

Rhizoctonia species and AGs arereported to differ in sensitivity to commonfungicides. Also, the prevalence and sever-ity of Rhizoctonia diseases on turfgrassesdepends, among other factors, on infectionby a particular species and AG ofRhizoctonia. Thus, minimization of chem-ical use as well as consistent and reliablemanagement of Rhizoctonia diseases withgenetic and biological methods will large-ly depend on identification of Rhizoctoniaisolates to species and subspecies level andknowledge of its virulence-regulatinggenes.

During the summer of 2008, wecollected more Rhizoctonia samples fromNorthern Virginia and Beltsville area ofMaryland. We were able to collect 136 iso-lates from Leesburg and Woodbridge ofVA, and 83 isolates from Beltsville, MD.Dr. Brandon Horvath and his research teamalso helped in collecting additional 22samples from Richmond and VirginiaBeach. The total Rhizoctonia isolates wehave at Beltsville USDA-ARS, so far,amounts to 448. Out of that, around 7% ofisolates belong to either R. zeae or R.oryzae. The nuclear staining of suspectedR. cerealis isolates from Blacksburg (col-lected in 2007) proved to be R. solani sinceall of them were multi-nucleate.

The molecular identification ofRhizoctonia isolates will be carried out by

employing AFLP, UP-PCR, and sequenc-ing of rDNA-ITS regions. Sequencing ITSregions will be utilized to group sampleisolates together with tester strains in orderto come up with their phylogenetic rela-tionships. The banding patterns of UP-PCR and AFLP will be used to constructphenograms of the tested isolates. Similarto the analysis of ITS regions, the bandsgenerated will be analyzed with the resultsof tester trains to find differences at molec-ular level.

Out of the total isolate collection, weselected a random sample of approximate-ly 10% encompassing different geographiclocations. The colony morphology of theseisolates was recorded after 10 days incuba-tion on PDA. The color of mycelial matand color and size of sclerotia of the iso-lates differed according to their speciesand anastomosis group (AG). The averagehyphal diameter of all the isolates werewithin 5-11µm which agrees with the pre-vious published data for R. solani, R. zeae,and R. oryzae.

Presently, we are extracting DNAfrom the selected isolates for carrying outmolecular detection techniques.Simultaneously, we are planning to per-form hyphal fusion experiments with testerstrains on water agar plates in order to

determine AGs of the isolates. Theresearch work carried out previously hasrecorded the AGs of 1, 2, 3, 4, 5 and 6 fromdiseased turfgrasses. However AG1, AG2-2 and AG4 are the most common. The datagenerated from molecular techniques willbe considered for accurate identification ofbrown patch causal agents into theirspecies and intraspecific level.

We are also investigating thepathogenicity of R. solani throughExpressed Sequenced Tag (EST) analyses.Approximately 1,000 clones each fromtwo virulence-differentiated EST librarieshave been sequenced and analyzed. Thenumber of unigenes identified from viru-lent and avirulent libraries are 214 and590, respectively. We are conducting arudimentary analysis of genes under thetwo virulence differentiable conditions.However, as in any gene expression proj-ect, we need to sequence more ESTs tomake a comprehensive bioinformaticsanalysis of the two EST libraries to identi-fy up- or down-regulated genes related tovirulence.

Accurate Identification and Gene Expression in Relation toVirulence of Rhizoctonia Isolates Infecting Turfgrasses


Dilip K. LakshmanUSDA-ARS

Objectives:1. Molecular identification of Rhizoctonia solani isolates pathogenic to turfgrasses using Universally Primed-

Polymerase Chain Reaction (UP-PCR) and nucleic acid hybridization analysis.2. Expressed Sequence Tag (EST) analysis for surveying genes and creating a gene database for R. solani with

emphasis on genes affecting virulence and pathogenicity.

Differences in colony morphology of a few isolates after10 days incubation on PDA. Isolates A (R. solani AG2-1), B (R. oryzea), C (R. solani AG1-1A), D (R. zeae) andE (R. solani AG4) were collected either from Richmond,Beltsville, or Leesburg area.


Summary PointsWe have collected a total of 448

Rhizoctonia isolates from golf courses andlawns of Virginia and Maryland. Out ofthem, around 7% of isolates belong toeither R. zeae or R. oryzae. The colonymorphology, color of mycelium, color andsize of sclerotia of a random sample of10% of those isolates have been character-ized so far.

We have conducted initial experimentswith UP-PCR technique and amplificationof ITS regions and selected the primers forfuture full-scale experiments.

From the initial analysis of the two ESTlibraries thus far, about 214 and 590 uni-genes have been identified from the viru-lent and avirulent EST libraries, respec-tively.

15

A B C

D E

Mole crickets (Scapteriscus spp.) are

mobile subterranean insects that damagegrass by their subsurface tunneling androot feeding. Mole crickets avoid areastreated with insect pathogens, but theirresponse to insecticide-treated soil is poor-ly understood.

The sensitivity of insects tochemical stimuli is affected by the pres-ence and abundance of chemosensorystructures (sensilla) on their bodies. Wecollected 10 adult males and females eachof tawny and southern mole crickets fromsound and linear pitfall traps in spring2007, examined their antennae and mouth-parts with a scanning electron microscope(SEM), and identified and counted thesensilla present. Southern mole cricketadults had ~80 flagellomeres (antennalsegments), and tawny mole crickets had~70. Females had more flagellomeres thanmales for both species.

Two-dimensional laboratory as-says were conducted to demonstrate iftawny and southern adult mole cricketscould detect acephate, bifenthrin, fipronil,and indoxacarb. Behavioral observationsoccurred for the first 90 minutes in thedark under red light, and the amount andlength of tunneling within 24, 48, and 72hours were recorded.

Tunneling initiation points werechosen randomly by either mole cricketspecies in the acephate experiment. Theamount of tunneling in the acephate treat-ed arenas did not significantly differ fromthe control for tawny mole crickets.However, southern mole cricket tunnelingafter 72 hours was significantly reduced bythe high rate of acephate. At the lowestrates, southern mole crickets tunneled lessin the treated areas.

Tunneling initiation was random

in arenas with all rates of bifenthrin andthe control. During the first 90 minutes,mole crickets in bifenthrin-treated sandmade more tunnel branches, closed moretunnels, and had rapid, erratic movementscompared to the controls. However, by 72hours, tunneling activity was greater inuntreated compared to treated sand.Avoidance of treated areas was notobserved, and the overall length of tunnel-ing in treated areas did not differ from thelength of tunnels in untreated areas.

In arenas with the full rate offipronil, 80% of mole crickets started tun-neling into the untreated area, which indi-cated a potential repellent effect.Tunneling initiation was random at the twolower rates. The total amount of tunnelingby 72 hours in the control was greater thanin arenas treated with any of the three ratesof fipronil. Most mole crickets (about70%) that tunneled through fipronil-treatedsand and then entered untreated sand didnot return to the treated sand. After expo-sure to the full fipronil rate, adults madetunneling leg movements, but remained inone place, making existing tunnels wider.Exposure to fipronil did not result inincreased tunneling.

Responses of tawny and southernmole crickets to indoxacarb were similar.

Tawny and southern mole crickets closed~30% and ~40% of their tunnels during thefirst 90 minutes, respectively. After 90minutes of exposure, mole cricketstremored and had erratic leg and wingmovements. After ~48 hrs of exposure,mole crickets were motionless, butresponded with kicks and tremors if dis-turbed. Total amount of tunneling by 72hours was significantly reduced in arenaswith the highest rate of indoxacarb for bothspecies. Mole crickets started tunnelingpredominantly in the untreated area andmost (~90%) egg laying occurred inuntreated sand. However, mole cricketstunneled equally on both treated anduntreated halves of the arenas.

All mole crickets survived in con-trol arenas. Mortality was 90% after 48hours for the full rates of acephate, bifen-thrin, and fipronil. Mortality of bothspecies was 60% in the arenas treated withthe highest rate of indoxacarb. In general,southern mole crickets tend to move fasterin tunnels than tawny mole crickets andmake more tunnels during the first hour ofa test.

Mole Cricket Sensory Perception of Insecticides


Tawny mole cricket nymphs tunneled less in areastreated with bifenthrin and fipronil (¼ of labeled rate),than in untreated sand.

Olga Kostromytska and Eileen A. BussUniversity of Florida

Objectives:1. Determine the type, location, and abundance of different sensilla on the antennae and mouthparts of S. vicinus and

S. borellii.2. Demonstrate the physiological effect of insecticides on the mole cricket nervous system and/or ability of mole

crickets to detect chemical stimuli.3. Demonstrate the behavioral response of mole crickets to sub-lethal insecticide doses.


Summary PointsSouthern and tawny mole cricket

antennae and palps have similar numbersand types of sensilla, but southern molecrickets have more flagellomeres, and thusmay be more sensitive to chemical stimuli.

Females of both species have more fla-gellomeres than males, possibly becausemales are more aggressive and clip offantennae.

All of the insecticides tested reducedoverall tunneling activity if evaluated after3 days. However, females preferred to layeggs on untreated areas within arenas, notin treated sand.

Tawny mole cricket nymphs tunneledless in areas treated with bifenthrin andfipronil (¼ of labeled rate), than in untreat-ed sand. Avoidance to other insecticideswas not observed.

16

Spring dead spot is a serious root-rot

disease of bermudagrass and is the mostimportant disease of hybrid bermudagrass-es managed as putting green and fairwayturf. Aesthetically undesirable necroticpatches ranging from a few inches to sev-eral feet in diameter are evident in thespring and early summer in bermudagrassswards that experience a dormant period.

Three fungal species in the genusOphiosphaerella (O. korrae, O. her-potricha, and O. narmari) are identified asthe causal organisms throughout theUnited States andAustralia. In Mississippi,O. korrae has been identified as the causalorganism of spring dead spot.

Results of a previous study con-ducted at Mississippi State University sug-gests the frequency of O. korrae inbermudagrass roots was greatest duringspring transition compared to summer andfall transition growth periods. As a resultof the observed fungal activity in bermuda-grass roots during spring transition, thisstudy was initiated in the spring of 2007 ina ‘Tifway’ bermudagrass fairway with ahistory of spring dead spot. Symptoms ofspring dead spot were observed throughoutthe study area in the spring of 2007. Thetreatment plots (15 ft × 10 ft) werearranged in a split-plot randomized com-plete block design replicated four times.

Fungicide treatments were thewhole-plot factor and nitrogen (N) sourceis the sub-plot factor (7.5 × 10 ft sub-plots). Fungicide treatments were appliedduring the spring and fall transitions. TheN sources include 12-2-12 organic fertiliz-er and 12-2-12 blend of inorganic fertilizerincluding ammonium sulfate (21-0-0),triple super phosphate (0-46-0) and muri-ate of potash (0-0-60) applied at 1.0 lb of

N per 1000 ft2 per month (May-October).Spring dead spot severity was

visually rated (1 to 9; 9 = no disease) in thespring of each year and quantified usingdigital image analysis. Recovery of symp-tomatic patches was monitored throughoutspring transition. Turfgrass quality wasrecorded each month of the growingseason.

Fungicide applications had a sig-nificant effect on spring dead spot ratingsin the spring of 2008. Neither the N source(organic vs inorganic) nor the interactionof fungicide and N source had an effect onspring dead spot severity. Rubigan treat-ments applied in April, September,October, or September and October;September application of Rubigan tank-mixed with thiophanate-methyl; Octoberand November applications of propacona-zole and myclobutanil, respectively, wereall similar with spring dead spot ratings of> 8.5. Rubigan applied in March, April,May or March, April, September, October,and the untreated control were similar withan average spring dead spot rating of 7.3.

Spring green-up was similar forall treatments. However, some plotsreceiving organic N initially appearedgreener than the inorganic N plots.Turfgrass quality was similar for all treat-

ments each month and averaged 6.0 (scale1 to 9; 9=best) for the growing season. Inthe spring and summer of 2008, the soil pHin plots receiving organic N (pH=5.63)was significantly higher than the soil pH ofplots receiving inorganic N (pH=5.21).

We anticipate the results of thisstudy will identify a fungicide/fertility dis-ease management program efficacious forcontrolling spring dead spot of bermuda-grass managed as fairway turf. Theseresults will also allow us to determinewhether there is an added benefit of usingan organic nitrogen source that includesbiostimulants and microbes as compared toan inorganic, acidifying fertilizer forreducing spring dead spot incidence andseverity.

The Efficacy of Spring Fungicide Applications Plus OrganicFertilizer for Controlling Spring Dead Spot of Bermudagrass


Untreated control plot, March 2008. Half the plot was treated with an organic N source (foreground) and the otherhalf with an inorganic N source (background) thorught the 2007 growing season.

Maria Tomaso-PetersonMississippi State University

Objectives:1. Determine the efficacy of spring and fall fungicide applications for reduction of spring dead spot incidence and

severity.2. Determine the effect of organic fertilizer for the reduction of spring dead spot incidence and severity and overall

improvement of turf quality.


Summary PointsFungicide treatments, including fenari-

mol (4.0 fl oz/1000 ft2) applied April,September, and October (2007), signifi-cantly reduced spring dead spot in 2008

N source had a significant effect on soilpH.

Spring green-up and turfgrass qualitywere similar for all treatments.

Turf recovery from spring dead spotsymptoms was evident in late June 2008.

17

Zoysiagrasses (Zoysia spp.) are

becoming a popular choice of turf for golfcourses in the transition zone. Their rela-tively slow growth rate, high density, andtolerance to shaded areas make them agood choice for a golf course setting.However, one of the most difficult weedsto control in zoysiagrass is bermudagrass(Cynodon dactylon). Using herbicides tocontrol bermudagrass in zoysiagrass hastraditionally been ineffective due to theirphysiological similiarites as C4, warm-sea-

son grasses. Therfore, zoysiagrass is typi-cally susceptible to the same herbicidesused to control bermudagrass.

Recent research has been con-ducted to evaluate new aryloxyphe-noxypropionate (AOPP) herbicides fortheir efficacy on bermudagrass control.Favorable results have been observed forbermudagrass control and zoysiagrasssafety when AOPP herbicides are tank-mixed with triclopyr. Additional researchis now being conducted to evaluate otherAOPP herbicides for bermudagrasscontrol.

The fairway conversion studywas initiated in May 2008. Plots meas-ured 5' X 10' and were arranged in a ran-domized complete block design. 'Zorro'zoysiagrass was sprigged at a rate of 10bushels/1000 ft2. Siduron applied atestablishment provided the poorestbermudagrass control. Treatments withEPTC and dazomet applied before zoysi-agrass establishment yielded the lowestbermudagrass groundcover. Treatmentswith good bermudagrass control yieldedhigher zoysiagrass cover than those withpoor control.

To evaluate the influence of cul-

tural practices on zoysiagrass competitive-ness with bermudagrass, a standard cupcutter was used to transplant 'Common'and 'Tifway' bermudagrass plugs into'Zorro' zoysiagrass plots. Treatmentsincluded increasing rates of nitrogen withand without trinexpac-ethyl. Plugs wererated monthly for maximum diameterspread.

At one and two months after ini-tial treatment, there was no significant dif-ference among treatments. This is designedas a long-term study and it is reasonable toassume that greater separation in treat-ments will be observed in time. We will beinitiating this study in a japonica andmatrella type next year to evaluate thiseffect.

A greenhouse study was initiatedin October 2008 to evaluate the competi-tive effects of various weed species onseeded 'Zenith' zoysiagrass (Zoysia japon-ica). It has been our observation thatDigitaria spp. are the most detrimental tozoysiagrass development, while goose-grass and sedge species are not as compet-itive. The study involves an additivedesign in which zoysiagrass seeding rate isheld constant while weed seeding rate isincreased. Ratings of percent weed and turf

cover will be measured over an 8-weekperiod.

The evaluation of newAOPP her-bicides for bermudagrass control studywas initiated in June 2008. Plots werevisually rated for bermudagrass controland zoysiagrass injury. Digital images uti-lizing a light box were also taken and ana-lyzed for percent green cover using SigmaScan Pro.

When applied at 3-week inter-vals, clodinafop and fenoxaprop are notsafe to apply to zoysiagrass turf and do noteffectively control bermudagrass.Metamifop tank-mixed with triclopyr issafe to apply to zoysiagrass and effectivelycontrols bermudagrass.

Developing Best Management Practices forBermudagrass Control in Zoysiagrass Fairways


Scott McElroy and Mark DorohAuburn University

Objectives:1. Evaluate the best integrated practices for fairway conversion of bermudagrass (Cynodon dactylon) to 'Zorro'zoysiagrass (Zoysia matrella) turf through various cultural and chemical methods.

2. Evaluate the influence of various cultural practices on zoysiagrass competiveness with bermudagrass.3. Evaluate the competitive effects of various weed species on seeded 'Zenith' zoysiagrass (Z. japonica).4. Evaluate new aryloxyphenoxypropionate (AOPP) herbicides for control of bermudagrass in zoysiagrass turf.

Research has been conducted at Auburn University to evaluate new aryloxyphenoxypropionate (AOPP) herbicidesfor their efficacy on bermudagrass control.


Summary PointsEPTC and dazomet in mixtures provid-

ed the best management strategy for fair-way conversion of bermudagrass to zoysia-grass.

Studies have been initiated to evaluatethe competitive effects of weed species on'Zenith' zoysiagrass.

Metamifop tank-mixed with triclopyris safe to apply to zoysiagrass and effec-tively controls bermudagrass.

18

In many large urban areas in the aridand semiarid regions of the U.S., the pop-ulation increase has not only increased thefresh water demand but also increased thevolume of wastewater generated. Effluentwater appears to be the only water resourcethat is increasing as other sources aredepleted. Effluent water has emerged as areliable and consistently available sourceof water for golf course irrigation. Thevery nature of effluent water warrants spe-cial management steps to be taken to min-imize the possible drawbacks associatedwith its constituents. The constituents aredependent on the source of effluent water.Some of the main constituents include:salts of different types, nutrient elements,and organic compounds. Effluent waterhas relatively high levels of sodium con-centration relative to calcium and magne-sium. Golf course managers are often con-cerned about salinity and sodicity issuesassociated with effluent water irrigation.Long-term and continued use of effluentwater may lead to increased soil sodicity,with a resulting reduction of soil infiltra-tion, permeability, and aeration, especiallyin clayey soils, exacerbating salinityproblems.

We sampled and tested soil fromfairways of 10 golf courses that were nearmetropolitan Denver and Fort Collins, CO.Among these courses, five had been irri-gated exclusively with effluent water for 5-33 years. The other five with similar turfspecies, age ranges, and soil textures hadused surface water for irrigation. Ourresults indicated that soils from fairwayswhere effluent water was used for at least5 years exhibited slightly higher soil pHand higher concentrations of extractableNa, B, and P. Compared to sites irrigatedwith surface water, sites irrigated witheffluent water exhibited higher EC and ahigher sodium adsorption ratio (SAR) ofsaturated paste extracts.

In 2008, we installed a series ofin-ground salinity and soil water contentsensors on two fairways at the Heritage atWestmoor Golf Course in Westminster,Colorado. At each fairway, 6 plots wereestablished. On each plot, two 5TE sen-sors were installed at 15 cm and 30 cmbelow soil surface. A total of 12 sensorswere installed on each fairway. Wire leadsfrom each sensor were buried and connect-ed to a data logger located at the edge ofthe fairway. These sensors measure threeparameters: temperature, volumetric watercontent, and the electric conductivity of thesoil.

The installation process was per-formed over the course of several weeks

followed by technical testingand calibration. Prior to auto-matic data collection, sensor-measured soil electrical con-ductivity (EC) was compared toconventional saturated pasteextracted soil EC to assess dataaccuracy. Significant linear cor-relation was observed betweensensor-measured soil salinityvs. saturated paste extracted soilsalinity (r = 0.77).

After calibration, dataloggers have been programmedto collect data once a day to

provide information on spatial and tempo-ral salt accumulation patterns.

Data collected thus far showedthat in early August when the weather wasrelatively dry, a well-drained fairway hadan average soil salinity of 1.8 and 1.3 dS/mat 15 and 30 cm below soil surface, respec-tively, whereas the poorly drained fairwayhad 2.95 and 2.43 soil EC at 15 and 30 cmbelow soil surface, respectively. A signifi-cant thunderstorm in mid-August effec-tively leached soil salts. Our preliminarydata indicated that the accumulation ofsalts appears to relate to precipitation pat-terns, soil texture, and drainageeffectiveness.

Salinity Management in Effluent Water Irrigated Turfgrass Systems


Yaling QianColorado State University

Objectives:1. To determine spatial and temporal salinity accumulation patterns in soil profiles on a golf course fairway site

with effluent water irrigation.2. To evaluate different management practices for reducing sodium and salt accumulation in the soil.

In 2008, a series of in-ground salinity and soil water content sensors ontwo fairways at the Heritage at Westmoor Golf Course in Westminster,Colorado were installed.


Summary PointsEffluent water is a reliable and consis-

tently available source of water for golfcourse irrigation.

Salinity and sodicity are major con-cerns associated with effluent water irriga-tion.

Compared to sites irrigated with sur-face water, sites irrigated with effluentwater exhibited higher EC and higher sodi-um adsorption ratio (SAR) of saturatedpaste extract.

Significant linear correlation wasobserved between 5TE salinity sensor-measured soil salinity vs. saturated pasteextracted soil salinity

Accumulation of salts appears to relateto precipitation patterns, soil texture, anddrainage effectiveness.

19

A total of 12 sensors were installed on each fairway tomeasure temperature, volumetric water content, andelectrical conductivity of the soil.

Large patch, caused by Rhizoctonia

solani AG 2-2, is the most common andsevere disease of zoysiagrass in the transi-tion zone. Knowledge is lacking about theinteraction of cultural techniques, weather,and disease development. In this project,we are conducting field experiments atseveral sites to investigate these interac-tions.

In preliminary experiments,spring aeration, verticutting, and sand top-dressing surprisingly led to higher levels oflarge patch. As work progresses, we willbetter elucidate the influence of cultivationpractices on large patch, and we will mon-itor the effects of weather on disease devel-opment. We will also investigate fungicideapplication timing and correlate it withenvironmental data to develop a model foroptimal fungicide deployment if fungi-cides are used.

In spring and fall 2008, we start-ed to build our collection of large patchisolates to use in further studies. We cur-rently have 60 isolates, stored on oat seeds,for long-term stability.

We carried out the first set of cul-tural and fertility practices for Objectives 1and 2. Plots were established at 3 sites(Manhattan, Olathe, and Haysville, KS).At all 3 sites, plots are set up as a split-plotwith 4 replications. The main treatmentplots are 12 x 20 feet. Treatments are culti-vation (aeration + verticutting + topdress-ing) versus non-cultivated. The subplot isfertility, either spring + fall or summer fer-tilization. For the spring + fall treatment,plots were treated with 1 pound N/1,000 ft2

as urea (46-0-0), in both spring and fall.

The summer treatment was 2 lbs N/1,000ft2 as polymer coated urea.

To induce disease development,all plot areas were inoculated in September2008 by taking out small cores, insertinglarge-patch-infested oats, and replacing thecores. Disease will be assessed in spring2009.

We deployed temperature andwetness sensors in several experimentalplots in fall 2008. The data will be ana-lyzed this winter, and the same type ofmeasurements will be taken in the comingseasons.

We have started to propagate the34 zoysiagrass lines in the greenhouse.The zoysiagrass is propagated by taking 2-3 node sections of stolons and placing in asoilless potting mix under mist for severalweeks, then transferring to the greenhouse.

In addition, field plantings of the zoysia-grass progeny were inoculated in bothManhattan and Olathe. Disease symptomswere apparent in some plots this fall.Disease will be assessed in the spring,along with time-to-recovery.

We established fungicide timingtrials at two sites, an inoculated site at ourresearch facility and a naturally-infestedarea at a local golf course.

Cultural Practices, Environment, and Pathogen Biology: Studiesfor Improved Management of Large Patch of Zoysiagrass


Megan Kennelly, Jack Fry, Rodney St. John, and Dale BremerKansas State University

Objectives:1. Determine the effects of aeration, verticutting, and sand topdressing on large patch and investigate the biology

of the interaction of cultural practices and disease.2. Determine the effects of nitrogen source and time of application on disease development.3. Study the environmental conditions associated with disease development in the field.4. Compare large patch susceptibility of 34 new freeze-tolerant zoysiagrass genotypes.5. Study the effects of different preventative fungicide application timing and correlate with weather conditions to

develop better guidelines for fungicide deployment.

Large patch, caused by Rhizoctonia solani AG 2-2, is the most common and severe disease of zoysiagrass in thetransition zone.


Summary PointsFirst set of cultural practices was car-

ried out and plots inoculated for diseaseratings in spring 2010.

Zoysiagrass progeny were inoculatedin the field, and plant material is beingmultiplied in the greenhouse.

Fungicides were applied for first tim-ing experiments, and disease will be ratednext spring.

20

The oriental beetle (OB) is the most

important turfgrass insect pest in NewJersey, Connecticut, Rhode Island, andsoutheastern New York. Our overallobjective is to investigate the dispersalbiology of oriental beetle adults in order toimprove the efficacy of mating disruptionto control oriental beetle grubs in turfgrass.

Before effective applicationparameters for oriental beetle mating dis-ruption using sex pheromone can be rec-ommended, we need to understand how faroriental beetle females move. Sincefemales should not be affected by their sexpheromone, they may mate outsidepheromone-treated areas and migrate intotreated areas to deposit eggs.

Female dispersal studies are ham-pered by the lack of female attractants andthe fact that females are active arounddusk. Females were placed into the turf-

grass between 5 and 7 p.m. in early July.Most females either dug into the soil oronly crawled short distances. Few femalesalighted but only flew 3-12 feet, afterwhich they either did not alight again orcould not be found. We will test fluores-cent markers in future observations andlook for other improved tracing methods.

Oriental beetle collected frommultiple black light traps placed at multi-ple locations during early to mid-July con-tained on average 52.0 ± 2.4% females.These trap captures suggest that femalemay fly significant distances. Femalestrapped from selected traps were immedi-ately frozen and will be dissected to deter-mine their mating status.

Oriental beetle male attraction todifferent pheromone sources was investi-gated in release and recapture field studies.Trécé Japanese beetle traps were placed inthe ground with only the funnel part aboveground in areas that had been treated withMerit in the previous year. The traps werelured with a virgin female placed in a metalmesh cage, red rubber septa with 10 or 30µg sex pheromone, or one pellet (approx.weight 28 mg) of a dispersible pheromoneformulation containing 5% pheromone.This pellet formulation effectively disrupt-ed oriental beetle mating in previous fieldstudies.

Color-marked males werereleased 12.5 to 200 feet downwind fromthe traps (100 males per distance).Recapture rates were determined after 24hours. For each treatment, one replicatewas tested on each of three consecutivedays with trap locations switched daily.Analysis of variance showed that recapturewas higher for the pellet and the septa thanthe virgin females. Recapture ratesdeclined logarithmically with distance forall pheromone sources. The low recapturerates with virgin females are in part due tothe fact that females produce less

pheromone and call only for a few hoursaround dusk. Pheromone production mayalso have been affected by the unnaturalfemale placement in cages.

Males were attracted to femalesfrom as far as 100 feet distance. Recaptureof unmarked males from the backgroundpopulation was the highest for the pellet(1,388 ± 295 per day), intermediate for thesepta (30 µg; 682 ± 261; 10 µg: 231 ±172), and the lowest for the virgin female(96 ± 41). Future studies will examinehow pheromone treatment of an areaaffects oriental beetle dispersal.

Optimizing Oriental Beetle Mating DisruptionThrough a Better Understanding of Dispersal Behavior


Albrecht M. Koppenhöfer & Valerie FournierRutgers University

Objectives:1. Examine the dispersal pattern of non-mated and mated oriental beetle females.2. Explore the dispersal behaviors of male oriental beetle in pheromone-treated and non-treated areas.3. Determine the distance male oriental beetles travel in response to different pheromone rates and/or sources.

Males were attracted to females from as far as 100 feetdistance in controlled studies.


Summary PointsOB males are attracted to pheromone

lures and formulation pellets from at least200 feet distance, but attraction to femalesis weaker and over shorter distances.

A 1:1 sex ratio in black light trap cap-tures suggest that female OB may dispersesimilarly as males. Limited direct observa-tion suggest more limited dispersal.

21

The oriental beetle (OB) is the most important turfgrassinsect pest in New Jersey, Connecticut, Rhode Island,and southeastern New York.

This research involves the collection

of seeds of promising reduced-input turf-grasses in Eurasia and their subsequentevaluation in representative field environ-ments in Utah. The objective of thisresearch is to identify germplasm that maybe used for reduced-input turf in the west-ern U.S.

Dr. Douglas Johnson from theFRRL joined staff from the N.I. VavilovInstitute of Plant Industry (VIR) in St.Petersburg, Russia to collect seeds ofreduced-input turfgrass species inKyrgyzstan (2006), northwestern Russia(2007), and the southern Ural Mountainsof Russia (2008). Seeds were collected inthe field, cleaned, and cataloged, and thenexported to the U.S.

During 2006, 92 collections ofAgrostis, Festuca, Trisetum, Poa,

Puccinellia, and other species used in theU.S. turf industry were made inKyrgyzstan. These were planted in repli-cated field trials at Evans Farm at Logan inJune 2007. During June 2008, Dr. RobSoreng from the Smithsonian Institutionobtained vouchers and taxonomically veri-fied these collections. These collectionswill be evaluated for their drought toler-ance and turf quality characteristics under50% ETo irrigation in 2009 and 2010.

During 2007, 49 collections ofAgropyron, Agrostis, Festuca, Koeleria,and Poa were made in northwestern Russiathat may have potential for reduced-inputapplications in the western U.S. In 2008,42 collections in the genera Agrostis,Festuca, Koeleria, and Poa were alsomade in the South Ural Mountains ofRussia.

These 91 collections from Russiawill be evaluated in replicated field trials atthe Greenville Farm at Logan and theWasatch Front at the Utah BotanicalCenter near Kaysville. Transplants of these

Russian collections will be established inJune 2009 and maintained under 50% EToirrigation. During 2010 and 2011, the mostdrought-tolerant materials with favorableturf quality characteristics will be identi-fied under water-limited conditions.

A greenhouse salt screeningexperiment was completed, evaluating pri-marily accessions of Poa pratensis andsome other Poa species. The experimentwas conducted twice with sufficient repli-cates, and results are nearing publication.

Stimulated by a grant from theUnited States Golf Association, webrought on a Ph.D. student (ShyamShridhar) to identify genes that respond tosalt stress in Poa. Shyam completed theRNA-based differential gene expressiontechnique and is currently cloning andsequencing the candidate genes. Poa col-lections, intended for water deficit stressresponse evaluations, are established in areplicated trial in the field.

A field study of fine fescue col-lections from central Asia was initiated toevaluate turf potential under low mainte-nance conditions characteristic of theIntermountain West.

Improvement of Water Management Strategies and Practices


Kevin Morris Douglas Johnson Shaun Bushman Paul JohnsonNational Turfgrass Federation, Inc. Utah State University

Objectives:1. Collect, evaluate, and improve grass germplasm to reduce water use and other inputs on turfgrass stands.2. Develop an understanding of the ecology and physiology of promising plant materials for the western U.S.3. Evaluate promising North American and Asian plant materials for their use on public and private lands.4. Facilitate germplasm exchange with Asian scientists.

Dr. Douglas Johnson joined staff from the N.I. Vavilov Institute of Plant Industry (VIR) in St. Petersburg, Russia tocollect seeds of reduced-input turfgrass species in Kyrgyzstan (2006), northwestern Russia (2007), and the south-ern Ural Mountains of Russia (2008).


Summary PointsMany grasses of various species,have

been collected in natural settings inKyrgyzstan (2006), northwestern Russia(2007), and the southern Ural Mountainsof Russia (2008). Many of these grasseshave been planted in field trials to evaluatetheir turf potential and low water use effi-ciency.

USDA germplasm and other Poagermplasm was screened in the green-house for salt tolerance.

A Ph.D. student is using biotechnologyto identify genes that confer salt tolerancein Poa.

A collection of fine fescues were estab-lished in a field trial to evaluate their per-formance under low maintenanceconditions.

22

Topsoil is often unavailable insufficient amounts during urban soil con-structions or restoration of degraded soils.Therefore, there is an increasing use ofmineral and organic by-products fromagricultural and manufacturing industriesas topsoil replacements, or as amendmentsin constructed soils.

A selected suite of municipal andagricultural organic by-products (anaerobi-cally digested poultry litter, poultry littercompost, yard waste compost, and turkeylitter compost), and quarry industry prod-ucts were evaluated as topsoil replace-ments in a growth chamber experiment.The objective was to determine which by-product mix(es) provided the most desir-able soil properties for turfgrass growthcompared to a reference commercial top-soil mix.

Plant biomass was similar in mosttreatments, however it was consistentlyand statistically significantly lower intreatments where yard waste compost wasused. Microbial activity varied amongtreatments. We found that the substratespresent in the constructed soils promotedbacterial dominance compared to the fun-gal dominance in the reference commercialmix. It is also likely that the higher pH ofthe quarry mixes promoted the observedbacterial dominance in those treatments.

Similarly, soil enzyme activitiesalso varied among treatments. Organicamendments increased soil ß-glucosidaseactivity in all treatments over their non-organic amended controls. A strong corre-lation was found between ß-glucosidaseand available phosporus. Results of thisstudy suggest that selected waste streamscan be effectively used in constructedtopsoil.

The growing residential and com-mercial development of urban and ruralareas results in an increased demand forborrowed topsoil for landscaping purpos-es. Yet, topsoil definition itself varies

among authoritativesources and include theA horizon surface layer,the A master horizon(Am), or a mixture of A

and E master horizons.This study was aimed atevaluating how differenttopsoil definitions affectthese properties of theborrowed topsoil. Thehypothesis was that amixture of A and E hori-zons will result in larger salvaged soil vol-ume while having minor, if any, adverseeffects on the borrowed topsoilcharacteristics.

Of the over 100,000 entries of theUSDA-NRCS National Soil Survey Center(NSSC) database, 59,300 entries of differ-ent soil orders (excluding histosols,oxisols, and andisols) from the 48 contigu-ous states were used. AE-mix topsoilresulted in an average reduction of 38% inorganic carbon and negligible changes inaverage sand, silt, and clay content com-pare to its respective Am topsoil (i.e.

among Am and AE-mix constructed from

the same pedon). Yet, average thickness ofAE-mix topsoil was over 2.5 times that ofits respective Am topsoil.

The study provides average top-soil characteristics on a soil order basisthat can serve as a reference in developingand/or refining guidelines for topsoil char-acteristics and specifications for borrowedtopsoil.

Recent implementation of phaseII of USEPA National Pollutant DischargeElimination System (NPDES) requires theparticipation of urban municipalities ofless than 100,000 people in the program.This means that most municipalities inurbanized areas will be required to captureand treat storm water. Soil-based infiltra-tion systems, such as rain-gardens, aresome of the recommended practices forreduction and for on-site treatment ofstorm water. Hence, at minimum, rain gar-den soil needs high infiltration rates andthe ability to promote and sustain cover

vegetation.Research was conducted to iden-

tify local materials, and mixes thereof, indeveloping recipes for rain garden plantingsoil from locally available materials. Localtopsoil, three quarry sands (<1/4" sand-stone, washed <1/4" limestone, and nonwashed <1/4" limestone), USGA sand, andtwo commercially available composts(coarse and fine texture) were evaluated.

The mixture of sandstone:topsoil:yard waste (55:25:20) with saturatedhydraulic conductivity of 2.5 inches/hourand pH of 7.5 (6.8 in 10mM CaCl2) was

selected for further use. The above mixrecipe was used in a large-scale operationto create a planting soil for the construc-tion of a rain garden at a nearby munici-pality (Beckley, WV).

The vegetation development andsoil chemical and physical properties aremonitored and evaluated at different timesfrom inception. In addition, a water quali-ty monitoring system is expected to beinstalled in the near future for further per-formances evaluations.

Enhancement of Soil and Soil Management Practices


Kevin MorrisNational Turfgrass Federation, Inc.

Objectives:1. The purpose of this research is to develop improved soils and or soil management practices for turfgrass.

Soil-based infiltration systems, such as rain-gardens, are some of the recom-mended practices for reduction and on-site treatment of storm water.


Summary PointsGood turfgrass growth can result from

soil constructed from organic wastes andquarry industry by-products

Mixing A and E horizon soil profilesfrom a construction site can create a “top-soil” that performs much like a natural top-soil

Several soil mixes are being evaluatedfor use in rain gardens (ie., on-site treat-ment of storm water).

23

Richard ZobelAppalachian Farming Systems Research Center

7-1-086-30-08

Breeding, Genetics, and PhysiologyThe quality and stress tolerance of turf is a product of the environment, man-

agement practices, and genetic potential of the grass plant. In many cases, major limi-tations to turf quality are stress effects, many of which can be modified or controlledthrough plant improvement. Projects are directed toward the development of turf culti-vars that conserve natural resources by requiring less water, pesticides and fertilizers.Research projects that apply new biotechnological methods toward turfgrass improve-ment are considered. Among the characteristics most desirable in the new turfgrassesare:

1. Reduced need for pesticides by increasing resistance to disease, insects,nematodes, and weed encroachment

2. Increased shade tolerance3. Reduced requirements for mowing, irrigation, and fertilization4. Tolerance of non-potable water5. Ability to survive high- and low-temperature extremes6. Increased drought tolerance7. Tolerance of intensive traffic8. Tolerance of poor quality soils.

Research in the fields of biotechnology, genetics, cytogenetics, cytology, ento-mology, genetics, microbiology, nematology, pathology, physiology, and other sciencesthat support the project objectives and provide improved techniques for improving golfturf species will be considered.

Locations of the following projects funded in 2008 by the USGA Turfgrass andEnvironmental Research Program under the category of Breeding, Genetics, and Physiology

24

Dollar spot disease is one of the major

management problems encountered withcreeping bentgrass. The related species,colonial bentgrass, has good resistance todollar spot and may be a source of novelgenes or alleles that could be used in theimprovement of creeping bentgrass.Despite the dollar spot resistance of colo-nial bentgrass, creeping bentgrass is gener-ally preferred because colonial bentgrassdoes not have the desirable aggressivestoloniferous growth habit of creepingbentgrass, which aids in repair of the turffrom the damage incurred during play.

Interspecific hybridization hasbeen used by breeders of many crops tointroduce beneficial traits from relatedspecies into crop species. However, it hasnot yet been used in bentgrass breedingand so presents a great opportunity forcreeping bentgrass improvement. The ulti-mate goal of such an approach would be todevelop bentgrass cultivars with thestoloniferous growth habit of creepingbentgrass combined with the dollar spotresistance of colonial bentgrass.For the past few years we have been

investigating the possibility of using inter-specific hybridization between colonialbentgrass and creeping bentgrass to intro-duce the dollar spot resistance of colonialbentgrass into creeping bentgrass. Wehave generated dollar spot resistant colo-nial bentgrass x creeping bentgrass inter-specific hybrids. One of the hybrids wascrossed with a creeping bentgrass plantand progeny were evaluated over twoyears for dollar spot resistance to assessthe feasibility of introgression of dollarspot resistance from colonial bentgrassinto creeping bentgrass. Of the 271 back-cross genotypes in the test, 31 (11%)exhibited 20% or less disease, similar tothe parental interspecific hybrid, in both

years of testing. From this we can estimatethat three colonial bentgrass genes may berequired for the observed resistance.

One approach to eventual identi-fication of genes conferring important phe-notypic traits is genetic linkage mapping ofa segregating population. We used 93backcross individuals as the mapping pop-ulation for genetic linkage mapping of thecolonial bentgrass genome. This is thefirst genetic linkage map of colonial bent-grass. As part of the project, we generatedcolonial bentgrass expressed sequence tag(EST) resources that were used for map-ping genes, and we developed a newmethod of marker identification calleddideoxy polymorphism scanning. We used

the established rice-wheat chromosomalrelationships to make the colonial bent-grass linkage group assignments. Most(81%) of the mapped ESTs conformed tothe expectation of chromosomal locationbased on the location of the most similargenes in rice. The linkage map covers1156 cM and it consists of 212 amplifiedfragment length polymorphic markers and110 gene-based markers. Colonial bent-grass is an allotetraploid species (2n = 4X= 28, A1 and A2 subgenomes). The mapconsisted of the expected 14 linkagegroups, which could be assigned to eitherthe A1 or A2 homoeologous subgenomes.

To identify colonial bentgrassgenomic regions conferring the dollar spotresistant phenotype, we searched for colo-nial bentgrass markers found in all of theresistant plants. A cluster of 14 markers onlinkage group 2A1 and a single marker ongroup 3A1 were found in the resistantplants suggesting that these regions may bethe chromosomal locations of the colonialbentgrass genes required for dollar spotresistance.

In the future it may be possible todevelop molecular markers linked to thecolonial bentgrass dollar spot resistancegenes that could be used in marker-assist-ed selection.

Identification of the Colonial Bentgrass Contribution to Dollar SpotResistance in Colonial x Creeping Bentgrass Interspecific Hybrids


Faith C. BelangerRutgers University

Objectives:1. To construct a genetic linkage map for colonial bentgrass.2. To identify the regions of the colonial bentgrass genome that confer dollar spot resistance to the colonial xcreeping bentgrass interspecific hybrids.

Colonial bentgrass linkage group 4A1. Most of thegenes mapped to this linkage group are similar togenes on rice chromosomes 3 and 11.


Summary PointsColonial bentgrass is resistant to dollar

spot disease. Hybridization with creepingbentgrass may be a way to improve thedollar spot resistance of creeping bent-grass.

Field evaluations of the backcrossprogeny from a cross of an interspecifichybrid with creeping bentgrass suggestthat three colonial bentgrass genes may berequired for the observed dollar spot resist-ance.

We developed the first genetic linkagemap of colonial bentgrass. Our data sug-gests the colonial bentgrass dollar spotresistance genes may reside on linkagegroups 2A1 and 3A1.

25

Crosses have been made between

creeping bentgrass and creeping x colonialbentgrass hybrid plants selected duringPhase I of this project, creating new prog-eny populations. Progeny populations willbe screened for resistance to dollar spotand snow mold pathogens and will also beevaluated for DNA markers previouslyassociated with resistance to these two dis-eases. Plants will be selected based onboth DNAmarkers and disease evaluationsto test the effectiveness of DNAmarkers asa selection tool. Selected plants will becrossed with additional elite plants to cre-ate additional disease-resistant popula-tions.

This research will build upon pre-vious results of Phase I of this project andwill tie into previous results that havemapped putative genes for disease resist-ance in creeping bentgrass. Elite clones ofcreeping bentgrass and creeping x colonialhybrids will be released to private breeders

for use in developing new varieties for thegolf industry.

Activities for 2008 focused on theestablishment of replicated clonal evalua-tion plots of progeny from elite crosses,extraction of DNA from leaves, and theinitial assay of DNA markers. Replicatedfield evaluations were established in

Massachusetts, New Jersey, and Marylandin 2007 and 2008.

In 2009, data will be collected ondisease resistances of field-grown plantsand DNAmarkers. Selected plants will beused to create new hybrid plants, progenypopulations, and releases of individualclones or genotypes to bentgrass breederswho are developing new varieties.

A Bentgrass Breeding Consortium to Support the Golf Industry


(three institutions)

Michael Casler Geungwha Jung Scott Warnke Stacy Bonos and Faith Belanger Suleiman BughraraUniversity of Wisconsin University of Massachussets USDA-ARS Rutgers University Michigan State University

Objectives:1. To develop elite clones of creeping bentgrass with multiple pest resistances and stress tolerances that can be deliv-

ered to the seed industry for use in synthesizing new creeping bentgrass cultivars broadly adpated to a range ofecological and environmental conditions including reduced pesticide application.

Creeping bentgrass clones originating from Illinois, Michigan, or Wisconsin tended to have better resistance ratingsto both dollar spot and snow mold when evaluated in their state of origin compared to clones from outside that state.Four creeping bentgrass clones demonstrating various levels of resistance to the dollar spot fungus, Sclerotiniahomeocarpa, at Gateway Golf Club, Land 'O Lakes, WI., are shown above.


Summary PointsCreeping bentgrass plants with

improved resistance to both dollar spot andsnow mold fungi have been identified andpropagated.

These plants will be released to privateturf breeders for use in developing newand more disease-resistant creeping bent-grass varieties.

This research has contributed to theidentification of specific genes for resist-ance to snow mold and dollar spot dis-eases, which can be used to design moreefficient and effective breeding methodsbased using DNA marker technologies.These methods will be tested in the currentphase of this project.

26

Selection of the most disease resistant clones at the O.J. Noer Turfgrass Research and Education Center, Verona,WI.

The native grass, desert saltgrass, has

potential in western US golf coursesbecause of its traffic resistance and toler-ance of salty soils and water. Under theseconditions it outperforms other droughtresistant species such as buffalograss,bermudagrass, and blue grama.

The 3000 Cycle 2 plugs, plantedAugust 2007, grew into their 3 by 3 footplots this year. Each plug represents a dif-ferent phenotype. We took measurementson percent grow-in, flowering dates, spikenumbers, height, verdure, and leaf shredfrom mowing.

Resistance to leaf rust (Pucciniasubnitens) has been an important trait forselection, as the disease can render salt-grass leaves brown the first week ofAugust. This past year, natural infectionalong the Front Range of Colorado hasbeen spotty and we have resorted to mak-ing collections of uredia (spores) through-out northeast Colorado. About 50 poundsof infected leaves are washed in 100 gal-lons of water with a commercial spray

adjuvant/surfactant. The material isscreened through cheese cloth, the cloth iswashed, and all supernatant is put into asprayer and sprayed onto nursery plantsimmediately.

Natural disease pressure can usu-ally be quantified in June, so that collect-ing and artificial inoculation can beplanned, if necessary, for the first week ofAugust. We speculate the daily morningdews in August aid infection, since this isthe time of year of natural re-infection byurediospores. Susceptible check pheno-types showed telia development inSeptember. Ratings on telia on all plantscan be made in October, using a modifiedwheat leaf rust rating. Inheritance of resist-ance is complex, and progeny testing isused to track resistance genes.

It is established that the size of an organis proportional to the size of the meristemfrom which it develops. Based on this,large shoot meristems should give rise tolarge shoots, and small shoot meristemsshould give rise to small shoots.Correlations also exist between smallshoots and high shoot density and largeshoots and low shoot density. Between1960 and 1980, there was a flurry of activ-

ity in measuring correlations and correlat-ed responses in many species. Some of thesuccess in breeding for turf traits can beattributed to positive correlations amongturf traits.

Much of the effort in a breeding pro-gram is planting and maintaining the nurs-eries. If the material going into the outdoornursery were higher caliber, it would pres-ent increased efficiency in selection. Weset up an experiment in which we meas-ured the diameter of 2-cm long shoots ofrandom new progeny seedlings in thegreenhouse. We transplanted this experi-ment into the field and, after a year, meas-ured height and shoot density. Seedlingswith small diameter shoots matured intoturf with shorter height and higher shootdensity.

Development of Seeded Turf-type Saltgrass Varieties


Wild population behind chairs, Cycle 2 turf in front ofchairs

Dana Christensen and Yaling QianColorado State University

Objectives:1. Evaluate new collections and Cycle 2 population. Select parents from these and intercross for the Cycle 3

population.2. Screen the collection for rhizome depth.3. Evaluate selection’s potential for commercial seed harvest.4. Correlate meristems to plant size.


Summary PointsPhenotypes planted in August 2007

were evaluated for percent fill-in, flower-ing date, height, and mowing quality.

Artificially inoculate the nursery withleaf rust and rate response.

Seedlings with small diameter shootsmature into turf-type plants.

27

Mature turf from seedling with smaller shoot diameter

Harvesting 2 cm long shoots to measure diameter

Cold acclimation or cold hardening is

an effective mechanism to increase freez-ing tolerance in temperate plants.Seedlings of two annual ryegrasses culti-vars, ‘Gulf’ (early flowering) and ‘Tur’(late flowering), and two perennial rye-grass cultivars, ‘Caddyshack’ (cold toler-ant) and ‘Lafayette’ (cold sensitive) wereused for measuring leakage induced byfreezing. Five-week-old seedlings wereplaced into growth chambers at 23 ºC fortwo weeks and then acclimated at 4 °C forvarious periods before they were used forion leakage test, a measurement of cellmembrane damage.

Freezing tolerance tests showedthat after 14 days of cold acclimation at 4°C, freezing tolerance greatly increased inboth cultivars of perennial ryegrass, but itdid not change in the two annual ryegrasscultivars. This suggests that perennial rye-grass can successfully cold acclimate,whereas annual ryegrass cannot.Increasing the length of cold acclimationbeyond 14 days, however, did not increasefreezing tolerance in perennial ryegrass.

Genes that are highly responsiveto cold are detected in higher copy num-bers in cold-acclimated perennial ryegrassthan in non-cold-acclimated controls.Many genes are highly up-regulated bycold, while some genes are down-regulat-

ed. Among the highly upregulated genes,two ice recrystallization inhibition protein(IRIP) genes, LpIRIP1 and LpIRIP2 wereidentified from a cDNA library constructedfrom cold-acclimated perennial ryegrass‘Caddyshack’. Reverse transcriptase PCR(RT-PCR) results showed that both geneswere highly up-regulated after one day orseven days of acclimation at 4 ºC.

The DNA sequences of bothgenes suggest that they probably sharesimilar functions with known antifreezeproteins from other plant species. BothLpIRIP1 and LpIRIP2 were transformedinto Arabidopsis ecotype ‘Columbia’through Agrobacterium-mediated transfor-mation for overexpression. Transgenicplants are allowed to advance to the T2generation. We randomly selected six T2

transgenic Arabidopsis lines carryingLpIRIP1 and another six lines carryingLpIRIP2 gene for freezing and survivaltest. One-month-old T2 seedlings were

subjected to 70 minutes of freezing tem-peratures in a programmed freezer.Survival rates were recorded after theplants were recovered in a 22 ºC growthchamber for a week. Without cold-accli-mation, two LpIRIP1 transgenic lines(LpIRIP1-7 and LpIRIP1-9) and twoLpIRIP2 transgenic lines (LpIRIP2-4 andLpIRIP2-8) showed improved survivalrates after being subjected to -4, -6, and -8ºC for 70 minutes compared to a wild type(Col-0) or seedlings carrying the emptyvector alone without the antifreeze genes

(EV-6).The survival rates of the same

transgenic lines were also evaluated aftercold acclimation. One-month-oldseedlings from the selected T2 lines were

acclimated at 4 ºC for eight days beforebeing subjected to freezing temperatures.All seedlings, including wild type (Col-0)and seedlings carrying an emptor vector(EV-6) showed improved survival ratesafter being subjected to -10, -12, and -14 ºC for 70 minutes. The selected LpIRIP1and LpIRIP2 transgenic lines showedmuch higher survival rates than exhibitedby either the wild type control or the plantscarrying the empty vector alone after coldacclimation.

Ion leakage test of these trans-genic lines indicated that under non-cold-acclimated condition, only seedlings fromLpIRIP1-7 and LpIRIP2-4 showedreduced ion leakage than the control plantsCol-0 or EV-6 after freezing at -4 or -5 ºC.However, after cold acclimation, all select-ed transgenic lines showed reduced ionleakage under either -8, -9 or -10 ºC offreezing comparing to the control plants,although the ion leakage for the controlplants also decreased.

Perennial Ryegrass Anti-freeze Protein GenesEnhances Freezing Tolerance in Plants


Transgenic Arabidopsis lines ines showed improved survival rates after being subjected to -4, -6, and -8 ºC for 70minutes compared to a wild type or seedlings carrying the empty vector alone without the antifreeze genes.

Shui-zhang FeiIowa State University

Objectives:1. Measure and compare freezing tolerance of perennial ryegrass cultivars with annual ryegrass cultivars before andafter cold acclimation.

2. Characterize functions of the antifreeze protein genes IRIP1 and IRIP2 by overexpressing them in the modelspecies Arabidopsis and assess membrane damages and overall survival rate of the transgenic plants under bothcold-acclimated and non-acclimated conditions.


Summary PointsIon leakage test indicated that perennial

ryegrass can successfully cold acclimate,while annual ryegrass is unable to coldacclimate. Two weeks of cold acclimationis sufficient for perennial ryegrass toachieve freezing tolerance, and lengtheningthe cold acclimation beyond two weeks didnot increase freezing tolerance further.

Overexpressing the two perennial rye-grass antifreeze protein genes, LpIRIP1and LpIRI P2, in the model speciesArabidopsis enhanced freezing toleranceas evidenced by the increased survival rateand reduced ion leakage after the trans-genic lines were subjected to freezing tem-peratures.

28

Wild Type Transgenic TransgenicWithoutantifreeze genes

As of October 30, 2008, over 1,150

promising turfgrasses and associated endo-phytes were collected in France, Italy, andIslands of Finland.. Many of these associ-ated endophytes should be new and uniqueand should have properties to enhance turf-grass performance. Over 9,434 new turfevaluation plots, 92,000 plants in spaced-plant nurseries, and 30,000 mowed single-clone selections were established in 2008.

Over 200,000 seedlings fromintra- and inter-specific crosses ofKentucky bluegrass were screened forpromising hybrids under winter green-house conditions of short daylengths andcool temperatures. Over 19,500 tall fes-cues, 34,000 perennial ryegrasses, 9,000bentgrasses and 8,200 fine fescues werealso screened during the winter in green-houses, and the superior plants put intospaced-plant nurseries. The progenies of150 new hybrid Kentucky bluegrasseswere screened in spaced-plant nurseries todetermine apomixis levels and otherimportant turf and seed production charac-teristics.

The following crossing blockswere moved in the spring of 2008: 5 hardfescues, 2 strong creeping red fescues, 2Chewings fescues, 16 perennial ryegrass-es, 20 tall fescues, 4 velvet bentgrasses, 4colonial bentgrass, and 3 creeping bent-grass crossing blocks.

The 30 perennial ryegrasses iden-tified in two different locations of the 2004National Turfgrass Evaluation Trial inNew Jersey have continued to displayresistance to gray leaf spot (Pyriculariagrisea) through 2008. These were devel-oped in collaboration with other organiza-tions since the fall of 2000 when the firstsevere epidemic occurred at Adelphia,New Jersey.

We are making continuousprogress with annual cycles of recurrentselection in perennial ryegrass for gray leafspot, dollar spot (Sclerotinia homoco-carpa), red thread (Latisaria fusiformis),and crown rust (Puccinia coronata). Someof the newly released perennial ryegrassesreleased this year are 'Defender', '1GSquared', 'Derby Extreme', 'Amazing GS','Manhattan 5 GLR', 'Gray Star', 'KokomoII', 'Primary', 'Uno', 'Transformer', 'GrayGoose', 'Gray Fox', and 'Palmer GLS'.

New promising Kentucky blue-

grasses and Texas x Kentucky bluegrasshybrids are 'Ridgeline', 'Aries', 'Aura','Delight', 'Futurity', 'Katie', 'Rhapsody','Hampton', 'Juliet', and 'Gaelic'.

Continued developments of turf-type tall fescue are being released withimproved brown patch resistance. Theyinclude 'Falcon NG, 'Jamboree', 'Terrier','Rocket', 'Essential', 'Fat Cat', 'Bullseye','Virtuosa', 'Mustang 4', 'Firecracker LS','Titanium LS', and 'Cochise IV'.

'Foxfire' strong creeping red fes-cue and 'Intrigue II' Chewings fescue werereleased in 2008.

Breeding and Evaluation of Kentucky Bluegrass, TallFescue, Perennial Ryegrass, and Bentgrass for Turf

Start Date: 1982Project Duration: ContinuousTotal Funding: $10,000 per year

C. Reed Funk, William A. Meyer, and Stacy A. BonosRutgers University

Objectives:1. Collect and evaluate potentially useful turfgrass germplasm and associated endophytes.2. Continue population improvement programs to develop improved cool-season turfgrass cultivars and breedingsynthetics.

3. Develop and utilize advanced technologies to make current breeding programs more effective.

Summary PointsContinued progress was made in

obtaining new sources of turfgrassgermplasm from old turf areas in Europe.These sources are being used to enhancethe Rutgers breeding program.Modified population backcrossing and

continued cycles of phenotypic and geno-typic selection combined with increasingsources of genetic diversity in turfgrassgermplasm and beneficial endophytesenables significant improvements in theperformance of new cultivars. Thirteennew perennial ryegrasses were releasedduring 2008 with improved gray leaf spotresistance.Twelve new improved tall fescues were

released in 2008.Substantial progress was made in

developing intra- and inter-specifichybrids of Kentucky bluegrass. Ten newpromising Kentucky bluegrass cultivarswere released in 2008.

Two new fine fescue for low mainte-nance turf were released.

One creeping bentgrass was releasedcalled 13M with dollar spot resistance.Colonial bentgrass clones with improvedbrow patch resistance were used to devel-op 4 populations. Clones of velvet bent-grass found with resistance to copper spot,dollar spot, brown patch, and pythiumwere used in velvet population increases.

2008 USGA Turfgrass and Environmental Research Summary29

Thirteen new perennial ryegrasses were released dur-ing 2008 with improved gray leaf spot resistance.

Continued developments of turf-type tall fescue arebeing released with improved brown patch resistance.

Hand pollinations of the Cynodon

transvaalensis-T574 x C. dactylon-T89cross were made in the spring of 2007.Crosses were harvested and the products ofthis cross were planted in the greenhousein the spring of 2008.

Progeny were transplanted to 5-cm pots, and single plants were establishedin 2m x 2m plots (methyl bromide fumi-gated soil) in the field in 2008. We wereable to produce 20 new hybrids (con-firmed with flow cytometry). Plant sam-ples have been made available to A.Paterson for the molecular study.

The maintenance nursery for thetriploid hybrids was re-established in 2008in newly fumigated soil to maintain thepurity of the stocks.

After analyzing the data on seed

head formation, longest stolon, plantradius, texture, and vigor which we col-lected on the hybrids during the past threeyears and provided to Dr. Paterson, hefound 105 (however, he indicated that thismay be a little over-estimated) statisticallysignificant QTL marker trait associations.

Production, Maintenance, and Evaluation of TriploidInterspecific Bermudagrass Hybrids for QTL Analysis

Start Date: 1999Project Duration: ongoingTotal Funding: $90,000

The interspecifice triploid hybrids ranged in color from 1 to 3 (LSD0.05 =1.5), in turf quality from 4 to 8.5 (LSD0.05=1.8)

Wayne Hanna and Andrew PatersonUniversity of Georgia

Objectives:1. Increase the size of the T574 x T89 mapping population by 100 or more triploid interspecific hybrids.2. Evaluate the hybrids for characteristics important in turf improvement and provide the information to Dr.

Paterson for association to the molecular map.

The products of the cross between C. tranvaalensis-T574 and C. dactylon-T89 are being propagated andestablished in field plots on 2- meter by 2-meter centersat the University of Georgia research facilities.


Summary PointsQTL molecular marker associations

(105) were found for seed head formation,longest stolon, plant radius, plant texture,and vigor in the bermudagrass hybrids.

New hybrids have been developed toadd to the population for use in the genet-ic map.

30

The overall goal of this research is to

identify chinch bug-resistant turfgrasses,investigate the mechanisms of this resist-ance, and identify specific genes contribut-ing to the resistance. Knowledge of spe-cific resistance mechanisms is valuable foridentifying biochemical and physiologicalmarkers for use in germplasm enhance-ment programs, and for characterizingplant defense strategies to insect feeding.

We continue to evaluate selectedwarm-season turfgrasses for resistance tochinch bugs. Of the 100 buffalograssgenotypes evaluated in greenhouse andfield studies, four have been categorized ashighly resistant (‘Prestige’, NE 184, NE196, and NE PX 3-5-1) and 24 as moder-ately resistant. Of the resistant buffalo-grasses studied, ‘Prestige’ exhibited thehighest level of resistance even though itoften became heavily infested with chinchbugs. Subsequent choice and no-choicestudies characterized ‘Prestige’ as tolerant.

A second component of thisresearch investigated the underlying bio-chemical and physiological mechanismsresponsible for chinch bug resistance. Theimpact of chinch bug feeding on the phys-iological responses of resistant and suscep-tible buffalograss has been evaluatedthrough gas exchange and chlorophyll flu-orescence measurements at specific timeintervals using established procedures.These studies have demonstrated thatresistant plants can generate energy forrecovery from chinch bug feeding.Susceptible plants appear unable to main-tain compensatory photosynthesis and, as aconsequence, suffer substantially more tis-sue damage from chinch bug feeding.

Our research also focused oncharacterizing protein changes, specifical-ly oxidative enzymes, in resistant and sus-

ceptible turfgrasses challenged by chinchbugs and explored the value of thesechanges as protein-mediated markers toscreen for insect-resistant turfgrasses.Reactive oxygen species (ROS), such ashydrogen peroxide, are known to beimportant early signals for altering geneexpression patterns in plant cells inresponse to abiotic and biotic stressors.

Despite the benefits gained fromreactive oxygen species molecules likehydrogen peroxide as defense signals,accumulation of these molecules can betoxic to cells. To protect themselves fromthe effects of ROS accumulation, plantshave developed oxidative enzymes thatbreak down ROS. Peroxidase, catalase,and superoxide dismutase have all beendocumented as ROS scavengers in plantsstressed by insects and pathogens.

Research by our group has docu-mented increased levels of peroxidases fol-lowing chinch bug feeding in the resistantbuffalograss, ‘Prestige’, and a loss of cata-lase activity in the susceptible buffalo-grass, ‘378’. These findings support ourworking hypothesis that an initial plantdefense response to chinch bug feeding isto elevate the levels of specific oxidativeenzymes, such as peroxidase, to helpdetoxify peroxides that accumulate as aresult of plant stress (see figure).

Native gels stained for peroxidasehave identified differences in the isozymeprofiles of resistant and susceptible buf-falograsses. Studies are currently under-

way to identify these specific per-oxidases and measure theirexpression over time in buffalo-grasses challenged by chinchbugs using qRT-PCR.

Our group has success-fully constructed and character-ized two subtracted cDNAlibraries. These libraries wereprepared from the resistant buf-falograss ‘Prestige’ and the sus-ceptible ‘378’ buffalograss 5 days

after initiation of chinch bug feeding.Several transcripts showed significant lev-els of change in ‘Prestige’ (resistant) whencompared to the chinch bug susceptiblebuffalograss ‘378’.

These transcripts were cloned,sequenced, and categorized according toputative function. These assays indicatedup-regulation of three defense-related tran-scripts in the resistant ‘Prestige’ buffalo-grass, but not in the susceptible buffalo-grass ‘378’.

Resistant Turfgrasses for ImprovedChinch Bug Management on Golf Courses

Start Date: 2003Project Duration: six yearsTotal Funding: $60,000

Tiffany Heng-Moss, Fred Baxendale, Robert Shearman, Gautam Sarath, and Paul TwiggUniversity of Nebraska

Objectives:1. Evaluate selected cool- and warm-season turfgrasses for resistance to chinch bugs in the Blissus complex.2. Investigate the biochemical and physiological responses of buffalograss to chinch bug feeding.3. Identify genes conferring resistance to chinch bugs.

To protect themselves from the effects of ROS accumulation, plantshave developed oxidative enzymes that break down ROS.


Summary PointsWarm-season grasses with resistance to

chinch bugs in the Blissus complex havebeen identified.

Commercial production of ‘Prestige’provides consumers with a high qualitybuffalograss with improved chinch bugresistance.

We hypothesize that peroxidases andother oxidative stress enzymes are playingmultiple roles in the resistant plant'sdefense response, including the down-stream signaling of plant defense reactionsto chinch bug injury and/or efficientremoval of reactive oxygen species.

This research identified genes differen-tially expressed in response to chinch bugfeeding and will serve to increase thegenomic resources available for buffalo-grass and facilitate development ofimproved turfgrasses with resistance tochinch bugs.

31

Two independent studies, suppression

subtractive hybridization (SSH) and pro-teomic profiling, were conducted toexplore the molecular mechanisms ofhigher heat tolerance of A. scabra vs.creeping bentgrass at both RNA and pro-tein levels. A total of 143 unique geneswith higher expression level in heat-stressed thermal A. scabra were identifiedusing differential display (DD-PCR) orsuppression subtractive hybridization(SSH) approaches.

An ongoing study on proteomicprofiling of heat-stressed vs. un-stressed A.scabra and creeping bentgrass hasrevealed 70 differentially expressed pro-teins. A large portion of identified geneshave functions involved in protein and car-bon metabolism, signaling / transcription,and stress defense. Therefore, these genesand their protein products are highly likelyto be associated with the increased heattolerance observed in thermal A. scabracompared to creeping bentgrass.

Several common genes withknown functions in heat tolerance wereidentified from both gene expression andproteomic profiling, including thoseencoding for proteins involved in carbonmetabolism (fructose 1,6-bisphosphatealdolase), stress and defense responses(phenylalanine ammonia-lyase, disulfideisomerase, and glutathione S-transferase).All of these genes code for products thatare suggested to function in heat stress tol-erance by either increasing glucose utiliza-tion or amplify signaling molecule biosyn-thesis. In addition, four heat-responsivegenes encoding heat shock protein 70(HSP70), heat shock protein 16 (HSP16),cysteine protease (AsCP1), and expansion(AsEXP1) were selected based on theirheat-inducible patterns in both geneexpression (northern blot) and protein

(western blot) analyses. These genes mayplay the critical roles in the heat toleranceand could be used as potential candidategenes for marker development.

Eight heat-responsive genes wereselected based on results from previousstudies of differential gene expressionanalysis and proteomics. The apparentconservation of gene function acrossspecies provides a unique opportunity totranslate discoveries in model species tocreeping bentgrass. Availability ofgenome sequences from closely relatedspecies (rice, maize, wheat, and barley) inthe public domain will dramatically short-en the effort to obtain the full length cDNAfragments of gene orthologs and increasethe efficiency of creeping bentgrass mark-er development. Orthologous regions forthe chosen candidate genes were identifiedfrom public databases.

SSR primer pairs were designedfrom the A. scabra EST sequences or theidentified orthologous genes of relatedspecies. SSRs within each of the selectedgenes were found using the SSR identifi-cation tool Primer3 based on the A. scabrafull-length cDNA. The same software wasused to design primers amplifying frag-ments that contained the SSR region fol-lowing main parameters and all otherparameters in default settings.

Screening of two candidate geneprimers (cysteine protease and expansin)indicated that one of the expansin primersshowed polymorphisms. This primer pairamplifies a 166 bp, and a 188 bp fragmentin 7418-3 and L93-10 genomes, respec-

tively, which harbor the tetra-nucleotiderepeat motif "AGCT".

Development and Application of Molecular MarkersLinked to Heat Tolerance in Agrostis Species


Bingru Huang, Faith Belanger, Stacy Bonos, and William MeyerRutgers University

Objectives:1. To identify molecular markers linked to heat tolerance for heat tolerance genes from a thermal Agrostis species.2. To develop a marker-assisted selection system for improving heat tolerance in creeping bentgrass.


Summary PointsSuppression subtraction hybridization

(SSH) cDNA libraries from the heat-toler-ant A. scabra identified several important'heat tolerance' genes or molecular markersfrom thermal A. scabra. Heat-responsivegenes in thermal bentgrass were catego-rized into five functional groups: proteinmetabolism, signaling/transcription, car-bon metabolism, stress defense, and othermetabolism. Some genes have unknownfunctions. The largest group of heat-responsive genes is involved instress/defense, followed by the group ofgenes related to protein metabolism.

Proteomic profiling revealed that theup-regulation of sucrose synthase, glu-tathione S-transferase, superoxide dismu-tase, and heat shock protein Sti (stressinducible protein) may contribute to thesuperior root thermotolerance of A. scabra.Phospho-proteomic analysis indicated thattwo isoforms of fructose-biphosphatealdolase were highly phosphorylated underheat stress, and thermal A. scabra hadgreater phosphorylation than A.stolonifera, suggesting that the aldolasephosphorylation might be involved in rootthermotolerance.

Several PCR-based SSR markers fromheat-responsive genes were developed,which could be used in marker-assistedselection of heat-tolerant bentgrass andother cool-season turfgrass species.

32

Agrostis scabra (NTAS) and creeping bentgrass (Cv. Penncross) exposed to 35oC for 12 days in a growthchamber.

NTAS Penncross

Despite the fact that perennial ryegrass

(Lolium perenne) is a valuable cool-seasonturfgrass, gray leaf spot caused byMagnaporthe grisea has become a seriousproblem on perennial ryegrass fairways ongolf courses. Under favorable environ-mental conditions, gray leaf spot (GLS)can completely destroy ryegrass stands in ashort period of time.

The use of host resistance is anenvironmentally sound method to controlgray leaf spot which has been well studiedand utilized in other hosts (mainly rice) ofM. grisea. With the use of recentlyimproved perennial ryegrass cultivars(‘Paragon GLR’, ‘Palmer GLS’, ‘PantherGLS’, ‘SR 4600’, ‘Protégé’, ‘Gray Star’,‘Gray Fox’, ‘Gray Goose’, and’Manhattan5’) with resistance to gray leaf spot, therehas been some concern that those resistantcultivars might break down due to selec-tion of pathogen isolates that can over-come the resistance genes. Research oninteraction between pathogen variabilityand host resistance in perennial ryegrassneeds immediate attention.

Previously detected GLS-resist-

ant QTLs in MFA and MFB plants arerelated with partial resistance. The combi-nation of partial resistance has proven tobe more durable to attacks by differentGLS pathogen races and so is likely to lastlonger than race-specific resistance. Toevaluate interaction between pathogenvariability and host resistance in GLS dis-ease, two ryegrass mapping parent clones(MFA, MFB), five commercial resistantcultivars, ‘Paragon GLR’ (two selectedGLS-resistant, Paragon-1R, Paragon-2Rand susceptible plants, Paragon-3S andParagon-4S based on the previous inocula-tion experiment), ‘Gray Star’, ‘Gray Fox’,‘Grey Goose’, ‘Manhattan 5’ (obtainedfrom Dr. Stacy Bonos, Rutgers Universityand commercial turfgrass seed compa-nies), and two breeding lines, 2COL-07and 2NKM-1 (from seed company) wereinoculated using twelve perennial ryegrassand one rice isolates. The isolates were:GG9, GG11, GG12 (Dr. M. Farman, Univ.of Kentucky), BL00, LP97, Lin00 (Dr. A.Hamblin, Univ. of Illinois), 6082 (Dr. S.Leong, Univ. of Wisconsin), 05T-04, 02V-23.1, 04S-01, 06T-02, 11W-03, and 11W-07 (Dr. W. Uddin, Penn. State Univ.).

The ryegrass plants were grownin French Hall greenhouse at University ofMassachusetts-Amherst and then inoculat-ed with the isolates under growth chamberconditions. As previously evaluated, MFAand MFB were resistant to all those iso-lates and Paragon-3S and Paragon-4S werevery susceptible as expected. All commer-cial cultivars and experimental lines testedwere highly resistant to the isolates but2NKM-1 and ‘Gray Star’ were moderatelyresistant. A marginal significant interac-tion between isolates and genotypes wasobserved, but more inoculation experi-ments are required to confirm the result.

The results encourage furtherinvestigation. Some individual plants (5-10) will be randomly selected from those

resistant cultivars and breeding lines inorder to test whether there is any signifi-cant interaction between pathogen vari-ability and host resistance of differentsources. Further inoculations using clonal-ly replicated plants and 13 geographicallydiverse isolates will be performed to checkwhether the resistance in commercial culti-vars differs from ones in MFA and MFB.

If pathogen variability does havea significant interaction with host resist-ance, crosses will combine differentsources of resistance genes so that thegenetic basis of gray leaf spot resistancecan be improved in perennial ryegrass.Further, DNAmarkers significantly associ-ated with QTLs for GLS resistances of var-ious sources will be developed for marker-assisted selection. Multiple disease resist-ance genes will be incorporated into anelite perennial ryegrass cultivar which willincrease the use of host resistance as anintegrated pest management strategy forturfgrass managers.

Quantitative Trait Loci (QTL) Mapping ofResistance to Gray Leaf Spot in Lolium


Geunhwa JungUniversity of Massachusetts

Objectives:1. To generate populations by crosses of resistant clones (from the MFA x MFB and L4B-5 x MF-8 populations) withresistant plants to be selected from commercial perennial ryegrass cultivars and breeding lines including ‘ParagonGLR’, ‘Gray Star’, ‘Gray Fox’, ‘Grey Goose’, ‘Manhattan-5’, 2COL-07, and 2NKM-1.

2. To develop perennial ryegrass plants having a broad spectrum of GLS resistance by pyramiding multiple resistantgenes originated from various sources of Lolium species and cultivars.

QTL mapping for gray leaf spot resistance is essentialin developing resistant (left) versus susceptible (right)perennial ryegrass cultivars.


Summary PointsSignificant difference in pathogenicity

among 13 gray leaf spot (GG9, GG11,GG12, BL00, LP97, Lin00, 05T-04, 02V-23.1, 04S-01, 06T-02, 11W-03, and 11W-07) and one rice (6082) isolates under thecontrolled greenhouse conditions.

Seven perennial ryegrass cultivars andbreeding lines with improvement of GLSresistance showed a high level of resist-ance to the 13 geographically diverse iso-lates, which might indicate race non-spe-cific resistance in perennial ryegrass.

Preliminary results indicated a margin-ally significant interaction between grayleaf spot isolates and ryegrass germplasmsunder controlled greenhouse conditions.However, more inoculation experiments atsingle individual plant rather than a culti-var of mixed genotypes are needed to con-firm the finding.

33

Bermudagrass is the most widely used

turfgrass in the southern USA and through-out tropical and warmer temperate regionsof the world. With USGA financial sup-port, the OSU turf bermudagrass geneticimprovement program made new progressin the enhancement of turf bermudagrassgermplasm and the development of exper-imental cultivars in 2008.

Initial screening of 1,080 putativeF1 progeny plants (C. dactylon x C. trans-vaalensis), field established in 2006, wascontinued in 2008 by evaluating wintercolor retention, spring green-up, win-terkill, foliage color, texture, sod density,seedhead abundance, and overall turf qual-ity. Large variations were observed for turfperformance and adaptation trait descrip-tors. We will select the best 1-2% of theplants for advancement to a replicated per-formance trial under fairway managementto begin in 2009.

A turf bermudagrass germplasmnursery, field planted in summer 2007, wasfully established on the OSU AgronomyFarm in 2008. The germplasm nurserycontained 298 genotypes consisting oforiginal accessions from geographicalregions in the world, selected promising

breeding lines, and commercial standardcultivars representing C. dactylon and C.transvaalensis and inter-specific hybridsof the two taxa. A progeny selection nurs-ery of a tetraploid bermudagrass popula-tion in initial cycle (C0) was planted in thesummer 2008.

The broad-based breeding popu-lation was formed by polycrossingtetraploid and desirable Chinese Cynodongermplasm accessions. A second selectionnursery consisting of approximately 1,500clonal progeny plants was planted in thesummer, as well. The putative F1 hybrids

plants were derived from inter-specifichybridizations of Chinese C. dactylonaccessions (4x=36 and 6x=54) with eliteOSU C. transvaalensis (2x=18) breedinglines. Plants in each of these two popula-tions will be evaluated over the next 2 to 3years for turf performance traits and forseed yield and seed quality traits for theChinese tetraploid population.

A field trial was continued tocomprehensively evaluate eight OSUexperimental synthetics for turf perform-ance traits against clonal and seeded stan-dard cultivars at the Turfgrass ResearchCenter in 2008. Standard field perform-ance parameters for fairway-type ber-mudagrass were assessed in this trial. Inaddition to the trial, a 2007-2012 NTEP

ancillary bermudagrass trial was inoculat-ed with Ophiosphaerella herpotricha, oneof the casual agents of spring dead spot infall 2008. A second 2007 NTEP ancillarybermudagrass trial was also inoculatedwith O. korrae, an additional causal agentof the disease, in 2008.

Tolerance to the disease will beassessed over the next four years in thesetrials as well as in a 2004-2012 compre-hensive evaluation of 32 clonal selections.Preliminary testing of an in-field mini-lysimeter technique for water use rate wasinitiated on 20 bermudagrass varieties infall 2008. These trials are managed undergolf course fairway conditions.

Breeding and Evaluation of Turf Bermudagrass Varieties


A turf bermudagrass germplasm nursery, planted in summer 2007, was fully established on the OSU AgronomyFarm in 2008.

Yanqi Wu, Dennis L. Martin, and Charles M. TaliaferroOklahoma State University

Objectives:1. Assemble, evaluate, and maintain Cynodon germplasm with potential for contributing to the breeding of improved

turf cultivars.2. Improve bermudagrass germplasm for seed production potential, cold tolerance, leaf firing resistance, and other

traits that influence turf performance.3. Develop, evaluate, and release seed- and vegetatively-propagated turf bermudagrass varieties.

Summary PointsA clonal bermudagrass selection nurs-

ery was evaluated and screened for turfperformance and adaptation traits.

Two new selection nurseries were fieldplanted in 2008.

A new turf bermudagrass germplasmnursery was fully established in 2008.A field trial to comprehensively evalu-

ate eight new experimental synthetics wascontinued.

Spring dead spot disease toleranceevaluation has been expanded in 2008.

Water use rate testing of selectionbegan in fall 2008.

2008 USGA Turfgrass and Environmental Research Summary 34

A field trial was continued to comprehensively evaluateeight OSU experimental synthetics for turf performancetraits against clonal and seeded standard cultivars atthe Turfgrass Research Center in 2008.

As a fringe benefit of the Human

Genome Project, it has become possible todiscover most of the genes that comprisethe genetic blueprint of a plant or animal atmanageable cost and in a short timeframe.Such 'gene encyclopedias' empowerresearchers to determine the role(s) of eachgene in the life-cycle of an organism, iden-tify hereditary differences among individu-als, and engineer genotypes that better suithuman needs. This provides an alternativeto controversial biotechnology approachesusing genetically modified organisms(GMOs) and may permit society to reapthe potential benefits of many researchgoals in a publicly acceptable and environ-mentally safe manner.

While most genes have beenidentified for some crops such as maize,rice, and sorghum, Cynodon (bermuda-grass) lags far behind. Our recently com-pleted USGAproject yielded sequences forportions of nearly 5,000 Cynodon genes,and a partner project (A. Guenzi) hasyielded nearly 4,000 more for a total ofabout 9,000. While seemingly large, thesenumbers are tiny in comparison to 868,456known for wheat, 472,163 for barley, andsimilarly high numbers for many other

crops (per GenBank, as of this writing).Studies in other organisms suggest thatEST sequencing is the most cost-effectivegene discovery method up to at least100,000 sequences. In other words, thereis much more to learn about Cynodongenes from this efficient approach.

We have more than doubledknowledge of the Cynodon transcriptome(set of genes that encode an mRNA) toabout 20,000 ESTs (sequences for portionsof genes). While still far smaller than thenumbers available for many other crops,this will considerably improve our knowl-edge of the Cynodon gene set, identifyingmany additional genes for which functionscan be deduced based on analogy to geneswith similar sequence ('spelling') in otherplants.

The resulting increased numberof genes will be sufficient to begin pursu-ing important applications with a reason-able expectation of success, some of whichwe elaborate on below. Further, the pro-posed tools are central to establishing suf-ficient 'critical mass' to warrant a 'turfgenomics initiative' of national scope, andwill immediately leverage two NSFawards in making the results web-accessi-ble and user-friendly.

A Cynodon 'gene encyclopedia'will provide the community with the com-plete set of tools needed to locate and iso-late important and unique Cynodon genes,setting the stage for a new era in breedingand genetics research in Cynodon andother turfgrasses.

In addition to enriching knowl-edge of the Cynodon gene set, we havealso provided on-line resources to fosterdevelopment of large numbers of DNAmarkers suitable for a wide range of stud-ies. Conserved intron scanning primers(CISP) provide large numbers of PCR-based 'pan-grass' tools suitable for linkinggenomics research in many crops of criti-

cal economic importance but which lackappreciable sequence information (such asCynodon), to burgeoning knowledge inbotanical models and better-studied crops.Because CISPs are based on PCR, theyrequire little DNA. This makes them suit-able to varying levels of technology andcost associated with practice in a widerange of scenarios ranging from high-throughput application in breeding pro-grams to targeted study in molecular biol-ogy labs.

Because CISPs are designedwithin genes, this approach permits us totarget variation in genes directly, ratherthan indirectly via a proxy DNA marker.This increases the likelihood of finding thespecific mutation responsible for a trait,rather than merely a diagnostic tool.

Accelerated Discovery of Cynodon Genesand DNA Markers by cDNA Sequencing


Examples of eight conserved-intron-scanning primerPCR products using the same primers for (from left ineach set) rice, maize, sorghum, pearl millet, tef,bermudagrass, wheat, and barley.

Andrew H. PatersonUniversity of Georgia

Objectives:1. We will more than double the Internet-accessible 'gene encyclopedia' for Cynodon by sequencing 12,000

Cynodon expressed sequence tags (ESTs).2. To nurture application of EST resources to many questions in Cynodon biology and improvement, we will devel-op a freely-available online resource of ‘conserved intron scanning primers’.


Summary PointsThis project increased the number of

Cynodon gene sequences to about 20,000,more than doubling our knowledge of itsgenes and their functions.

Identification of correspondingrice/sorghum genes will permit scientiststo deduce the probable functions of manyCynodon genes and also reveal featuresthat are present in the genomic DNA thatsurrounds Cynodon genes but are absentfrom the ESTs, such as the 'promoters'(on/off switches) that regulate theirexpression.

Large numbers of 'conserved intronscanning primers,' (PCR-based markers)are ideal for many applications in Cynodonresearch and improvement.

While a complete sequence of theCynodon genome remains far in the future,our work will build the framework for effi-cient progress by linking this project, themost extensive Cynodon sequencingeffort, to the first detailed Cynodon genet-ic map.

35

Saltgrass seeds have a low germination

rate due to their seed dormancy. We evalu-ated different seed treatments and foundthat stratification and machine scarifica-tion improve germination and establish-ment of seeded saltgrass. However, infor-mation is lacking concerning the effects ofseeding date and seeding rate on establish-ment of saltgrass.

Growing degree days (GDD) orheat accumulation units are useful in pre-dicting suitable seeding time for warm-season grasses. We have completed astudy to 1) determine the effect of seedingrate, seeding date, and two different seedtreatments on saltgrass germination andestablishment, and 2) determine therequired accumulative GDD for saltgrassto establish adequate coverage (80%) afterseeding. Seeding dates tested were May15, June 15, and July 15 at two locations(Horticulture Research Center and a golfcourse in Denver).

This field study showed thatmachine scarification and stratificationbroke saltgrass seed dormancy equallywell. Saltgrass seeded in May establishedadequate coverage (80%) in Septembereven using the lowest seeding rate (74 kgha-1). For plots seeded in June, only thehigher seeding rates (123 and 170 kg ha-1)established adequate coverage by the endof growing season. For plots seeded inJuly, however, even the highest seedingrate failed to establish adequate coveragein September.

The accumulated GDD to achieveadequate coverage was 1,748, 1,663, and1,435 for 74, 123, and 172 kg ha-1 seedingrates, respectively. When seeded on July15, total accumulated GDD toward the first

frost was 889 and 1,220 at our two experi-mental sites. Therefore, it was unlikely toachieve adequate coverage when saltgrasswas seeded in July in Northern Colorado.However, a higher seeding rate may beused to compensate for the June seedingdate to achieve adequate coverage by theend of the growing season.

The accumulated GDD assess-ment provided in our study suggests thatsaltgrass has higher establishment GDDrequirements than bermudagras and buf-falograss, but lower or similar GDDrequirements as zoysiagrass. Our resultswill aid in selecting appropriate seedingdates for different climates for successfulestablishment of saltgrass.

Saltgrass has been classified as ahalophyte, however, saltgrass is less salin-ity tolerant during germination than estab-lished turf stands. Our lab and growthchamber tests showed that Proxy solution(at 5-10 mM a.i.) and thiourea (30 mM)increased saltgrass germination under arange of salinity treatments (0-30 dS/m).Experiments have been conducted in 2008to scale up growth chamber studies to fieldconditions in order to confirm the effec-tiveness of seed pretreatments with Proxyand thiourea in enhancing saltgrass seedgermination under saline conditions.Water (control), Proxy (5-10 mM a.i.), andthiourea (30 mM) were used as pretreat-ment agents to soak scarified saltgrass

seeds for 24 to 48 hours.After pre-treatment, saltgrass was

seeded under four different salinity condi-tions in the field. Data were collectedweekly on soil salinity, the number ofseeds germinated, and plot coverage.Saltgrass germination percentage was 51%under non-saline condition, which wasreduced to 14% when soil salinityincreased to 18 dS m-1. Soaking saltgrassseeds in Proxy solution (5-10 mM a.i.)increased saltgrass germination percentageunder all salinity levels. However,thiourea pre-treatment did not increasesaltgrass germination in the fieldconditions.

Multiple Stress Tolerance, Seed DormancyBreaking, and Establishment of Seeded Saltgrass


Yaling Qian and Dana ChristensenColorado State University

Objectives:1. Quantify cold hardiness of potential varietal releases and advanced seeded lines.2. Continue to determine the level of salinity tolerance during germination (seeded type only) and as mature turf

for potential new cultivars.3. Evaluate different seed treatments to break seed dormancy.4. Evaluate saltgrass seeding establishment in the field with high and moderate levels of soil salinity.5. Determine saltgrass rooting characteristics and soil moisture extraction patterns under two irrigation regimes.

The May seeding date is best for saltgrass establishment in Colorado, with seeding rates as low as 74 kg ha-1achieving optimal establishment of saltgrass in one season.


Summary PointsThe May seeding date is best for salt-

grass establishment in Colorado, withseeding rates as low as 74 kg ha-1 achiev-ing optimal establishment of saltgrass inone season.

Saltgrass has higher establishmentGDD requirements than bermudagras andbuffalograss but lower or similar GDDrequirements as zoysiagrass.

Soaking scarified saltgrass seeds with5-10 mM Proxy solution increased thesubsequent germination of saltgrass at 5 -20 dS m-1soil salinity.

Thiourea pre-treatment did not increasesaltgrass germination in the fieldconditions.

36

The University of Georgia has an

established seashore paspalum breedingeffort for the development of improvedcultivars suitable for use by the golf courseindustry. Thus far, our cultivars have beenwell accepted by the turf industry bothdomestically and internationally. TheUniversity of Georgia turfgrass breedingprogram has the largest and most diversecollection of seashore paspalum ecotypesin the world. We are now utilizing thisgrowing germplasm collection to generatenew genetic variation throughrecombination.

This approach allows us to gener-ate thousands of unique individuals eachyear. Individual plants are hand-trimmedin the greenhouse and undesirable plantseliminated. In 2008, more than 6,000 indi-viduals were also screened for salt toler-ance in the greenhouse and approximately2,000 salt-tolerant individuals were latertransplanted to field plots for further eval-

uation of turf quality and resistance to dol-lar spot. This approach allows our breed-ing program to efficiently evaluate largenumbers of individuals for important traitsand should insure continued improvementin turf quality, disease resistance, and salttolerance in our future cultivar releases.

Over the past two years, a numberof turf trials were established including aVegetative Preliminary Trial, a FairwayVariety Trial, and a Green Variety Trial atGriffin, as well as a Fairway Variety Trialat Tifton. In addition, in 2007, threeadvanced breeding lines were entered intothe first National Turfgrass EvaluationProgram (NTEP) multi-state trials forseashore paspalum. All of these trials arenow generating valuable data on the per-formance of several advanced lines in ourprogram.

Based on the data that we haveaccumulated thus far, it appears that sever-al of these lines represent significantimprovements over any existing cultivarsand therefore are being positioned forrelease as our next cultivar. Vegetativeincrease of these lines is now underwayand plans are to begin more broad-scaleevaluation during 2009.

In 2007, Dr. Bob Carrow and hisstaff initiated a replicated field study with24 entries under our automated rain outshelter. These plots were subjected to three

dry-down cycles during the 2008 growingseason. During these dry down events,individual plots were rated twice weeklyfor turf quality, leaf firing, and normalizeddifference vegetative index (NDVI).

When the data were averagedover all dry-down periods, one of ouradvanced lines topped the lists for NDVI,turf quality, and color under drought stress.The top group for turf quality also includ-ed ‘SeaIsle 2000’, ‘Sea Dwarf’, ‘SeaIsle1’, and several other advanced lines. Thebottom group for turf quality included‘Aloha’, ‘Salam’, ‘Tifway’ bermudagrass,and a number of our advanced lines.

During the summer of 2009, weplan to continue this evaluation for droughtresistance as well to as establish a newdrought tolerance experiment with ournewest advanced lines. Data generatedfrom such studies will be essential indeveloping the next generation of droughttolerant seashore pasaplum cultivars.

Breeding Seashore Paspalum for Recreational Turf Use


Paul RaymerUniversity of Georgia

Objectives:1. Develop superior quality turf cultivars suitable for use on golf courses and recreational venues.2. Document and improve disease resistance.3. Further improve salt tolerance.4. Develop improved weed management strategies.5. Develop molecular tools to support breeding.

Automated rain-out-shelter covers field plots during natural rainfall events allowing us to impose drought conditions.


Summary PointsExtensive greenhouse and field testing

for salt tolerance, turf quality, and diseaseresistance is used to identify new cultivarswith a combination of improved traits.

Drought studies conducted under arain-out shelter help us to identify poten-tial cultivars that can maintain turf qualityeven during periods of prolonged waterstress.

37

Dr. Paul Raymer develops superior seashore paspalumcultivars for use on golf courses.

During 2008, several trials were con-

ducted to address these objectives.Twenty-one single crosses were construct-ed to assess seed yield potential and con-tinue to create variation in our germplasm.Significant yield differences were observedamong the crosses, and high yielding com-patible parents were identified.Approximately 1,300 individual plantswere selected from these progeny. In addi-tion, 25 collections were made from fivestates. These selections and collectionswere established in the greenhouse andtransplanted to the field in June for turf-grass performance evaluations. Hybrid-ization and collection have enriched thegermplasm as a major source of new geno-types for cultivar development.

A new set of buffalograssadvanced lines (Series IV), consisting of104 genotypes was established in replicat-ed plots at the John Seaton AndersonTurfgrass Research Facility located nearMead, NE. The genotypes in theAdvanced Lines IV trial were from previ-ous advanced line trials, selection trials,and recent collected materials. These geno-types will be evaluated for turfgrass per-

formance characteristics and seed yieldpotential for three or more years.Outstanding genotypes will be identifiedand further evaluated for possible release.

An experimental line and cultivarevaluation trial, consisting of 20 genotypeswas initiated in 2007 at five locations toassess the performance of these elite geno-types over a wide range of environments.Some genotypes did not establish well insome locations in 2007, so those sites werere-established in 2008 along with fouradditional locations, providing nine loca-tions in eight states. Data from each loca-tion were analyzed separately for seededand vegetative genotypes.

Buffalograss performance is bestmet through genetic improvement anddevelopment for optimum managementpractices. Even though buffalograss is anideal low-input turfgrass, it is still impor-tant to know its response to different mow-ing heights and fertilizer application rates.Management studies involving nitrogenrates, mowing heights, and cultivars wereconducted. Results from these studiesindicated significant differences among Nrates, mowing heights, and genotypes.Interactions were mostly non-significant.Buffalograss genotypes responded differ-ently to the management practices.

The effects of growing degreedays (GDD) of harvested sprigs, environ-ment, and their interaction on buffalograssvegetative establishment from sprigs werestudied. Though genotype and environ-ment do play a role, GDD proved to be themajor contributor in successful establish-ment suggesting that sprigging after 1,050GDD will not be successful. Further studyis needed to better understand the GDDinfluence on pre- and post-harvest physiol-ogy of sprigs.

Developing a buffalograssgenome map will be helpful to betterunderstand the function, location, andinheritance of genes involved in pestresistance, stress tolerance, and improved

turfgrass quality. Crosses were madebetween two diploid (NE 3297 x NE 2871)parents. An F1 mapping population was

generated. Sequence Related AmplifiedPolymorphism (SRAP) markers were usedto evaluate the pattern of polymorphism inthe parents and their progenies. Based onmarker segregation, F2 and backcross

types of segregations were observedamong the progenies.

Germplasm Development and Management of Buffalograss Varieties


Robert ShearmanUniversity of Nebraska

Objectives:1. Breed, select, and evaluate seeded and vegetative genotypes with improved turfgrass quality, pest resistance, andstress tolerance.

2. Improve our basic knowledge of the genetics of buffalograss through modern molecular marker technologies.3. Expand understanding and use of efficient management practices for best genotypic performance.4. Develop protocols for best turfgrass establishment.

Summary PointsGermplasm was enriched through col-

lection and hybridization. New selectionsand advanced lines were established.

Elite genotypes were evaluated under awide range of environments. Significantdifferences were observed among geno-types tested for most traits. The trial willcontinue and seeds of some elite genotypeswill be increased.

Results from management studiesshowed significant differences among Nrates, mowing heights, and genotypes, butnot for their interactions. The trials willcontinue for conclusive results.

Growing degree-day accumulation is amajor contributor to successful establish-ment. Further study is needed to furtherunderstand the nature of the influence.

Framework mapping of diploid popula-tion of buffalograss will be developed tobetter understand this grass.


USGA’s research committee members view improvedbuffalograss selections at the University of Nebraska.

Buffalograss performance is best met through geneticimprovement and development for optimum manage-ment practices.

Grass species that are native to North

America should be better able to cope withour environment and could lead to overallreductions in inputs such as fertilizers, pes-ticides, and water. Prairie junegrass(Koeleria macrantha), which is native tothe Great Plains of the United States, hasshown the potential to be successfully usedas a turfgrass in lower-input environments.The species is widely distributed through-out much of the western United States andit can also be found throughout much ofEurope and Asia. Based on datathat has been collected in recentyears, this species appears to per-form well in Minnesota underlow-input conditions (no irriga-tion, limited nitrogen application,and no fungicide or insecticideapplications).

Prairie junegrass hasseveral attributes that make it auseful low-input turfgrass inMinnesota including tolerance ofdroughty and alkaline soils, toler-ance of sandy areas, survival of low andhigh temperature extremes, and reducedgrowth rate. 'Barkoel' was the first cultivarof this species specifically developed foruse as a turfgrass. However, this cultivarwas developed with ecotypes from Europe.We are proposing the development of acultivar using germplasm native to North

America.Developing a high quality turf-

grass is not, by itself, adequate. In order tobe used by consumers, an economicallyviable turfgrass cultivar must be able toproduce sufficient quantities of seed. Non-selected populations of the species canproduce seed for 4-5 years. Collections ofnatural ecotypes made in 2005 suggest thatindividual genotypes may possess the abil-ity to be highly productive. However, it isunknown if it can produce economicallyadequate amounts of seed.

In order for a cultivar of thisspecies to be used on a wide scale, two cri-teria must be met: 1) the cultivar must pos-

sess adequate turfgrass quality in a medi-um to low maintenance management situa-tion, and 2) the cultivar must possess ade-quate seed production traits so that a suffi-cient supply of seed can be produced at areasonable cost.

We have collected native prairiejunegrass germplasm from Minnesota,South Dakota, North Dakota, Colorado,and Nebraska. These germplasm collec-tions have been established in breedingnurseries, and in some cases, experiencedone cycle of selection. We have estab-lished several spaced-plant evaluationsthat will be used to determine the geneticvariation present in our populations forvarious turfgrass and seed productioncharacteristics.

In 2007, two experiments wereestablished in both St. Paul and Becker,MN. The first experiment evaluated the

USDA collection of Koeleria macranthafor seed production potential in Minnesota.The second experiment evaluated the samecollections for turf potential as mowed

spaced plants.The seed production

study is now complete and wefound significant variation amongaccessions. Collections thatshowed high levels of seed pro-duction potential includedgermplasm collected in Iowa.Collections with low levels ofseed production were generallyfrom areas of southwestern Asia.Based on these results, we willplace additional focus on theinclusion of local collections in

our breeding program in order to improveseed production potential of the species.

Preliminary results from themowing study indicate that sufficient vari-ation for many important turf traits existsin the USDA collection. Of particularinterest to our program are differences inmowing quality and the ability to maintaingreen color through summer stress periods.The mowing study will continue through2009 and top-performing accessions willbe integrated into our breeding program.

Genetic Improvement of Prairie Junegrass


Eric WatkinsUniversity of Minnesota

Objectives:1. Determine the genetic potential of native prairie junegrass (Koeleria macrantha) germplasm for use as low-inputturfgrass.


Summary PointsGreat diversity exists in public collec-

tions of Koeleria macrantha.

Local collections will result inimproved seed production characteristics.

Integration of traits from diversegermplasm should be effective in thedevelopment of a low-input cultivar.

39

Poor mowing quality of a Koeleria macrantha genotype

Collecting prairie junegrass in southeastern Minnesoata

Variation in seed production characteristics of threeKoeleria macrantha accessions.

Poverty grass (Danthonia spicata L.),

a native-cool season perennial grass withwide distribution in the United States, isbeing evaluated for its suitability for useon golf courses. Potential benefits of usingpoverty grass include reduced dependencyon fertilizers and irrigation and less fre-quent mowing. The goal is to identify prac-tices to improve seed germination and suc-cessfully establish field plots as monocul-tures or with other native species.

This grass is found in mostprairies, savannas, and open woodsthroughout Missouri. However, seed com-mercially available is very limited. Seedhas been collected every year since 2005from different locales in Missouri in coop-eration with seed producer MervinWallaceof Missouri Wildflowers nursery. This seedis being used for trials and is also availablefor private seed producers in Missouri.

Seed averages 400,000PLS/pound and is highly dormant.Untreated seed collected in Missouri in2006 was 89% dormant with less than 5%

germination under 25oC. Seed collected in2008 was also mostly dormant.

Different combinations of stratifi-cation (moist storage), scarification(mechanical disruption of seed coat), light,and temperature were evaluated to break

seed dormancy andimprove germina-tion. Lightimproved germina-tion of scarifiedseed (63 % vs.46% in dark) andnon-scarified seed(30 % vs. 4 % indark). Germinationafter 40 days at

25oC for scarifiedseed exposed tocold, moist stratifi-

cation (5oC) for10, 20, 30, and 40days averaged 64,66, 62, and 59%,respectively, incontrast to 35, 36, 53, and 59%, respec-tively, for non-scarified seed. The best seedtreatment combinations will be evaluatedon existing seed collections to determine ifdormancy varies among seed sources andlength of storage.

Seedlings were started in June2006 from scarified seed sown in traysfilled with a commercial soil mix or a siltloam soil for seed production plots.Seedlings from Minnesota, but notMissouri seed sources, were infected withDreschslera leaf spot. Seedlings estab-lished 3-4 inches apart on Mexico silt loamsoil at the University of Missouri-BradfordResearch and Extension Center in June2007 produced an average of 146 lb/acreof seed collected manually in June 2008.Plots established with scarified seed atthree seeding rates (1000, 2000, and 3000PLS/sq. ft.) in fall 2007 had 24, 25, and30% poverty grass ground cover respec-tively in May 2008. Bare ground was 60%average for all rates.

Because these plots were infectedwith white clover and a few other broad-leaves (5-15% cover), 2,4 D and Remedyherbicides were sprayed at a rate of 0.5 or1 pint/acre. Remedy reduced clover and

2,4 D did not have much effect. Povertygrass did not seem to be affected by eitherherbicide. Additional weed control wasdone by hand. Ground cover will again beevaluated in 2009.

Shade tolerance studies were ini-tiated in summer 2008 in pots filled with asoil mix at Bradford Farm and will bereplicated at Carver Farm at LincolnUniversity in 2009. Four shade levels (0,30, 45, and 80%) are being evaluated onthe growth rate and seed production ofpoverty grass using structures coveredwith shade cloth.

Evaluating Poverty Grass (Danthonia spicata L.) for Use in Tees,Fairways, or Rough Areas in Golf Courses in the Midwest


Nadia E. Navarrete-Tindall, Brad Fresenburg, and J.W. Van SambeekUniversity of Missouri

Objectives:1. Identify best practices to optimize seed germination, seed production, and field seeding rates.2. Determine tolerance to shade and herbicides.

Plots established with scarified seed at three seedingrates (1000, 2000, and 3000 PLS/sq ft) in fall 2007 had24, 25, and 30% poverty grass ground cover respec-tively in May 2008.


Summary PointsNatural prairies or savannas in

Missouri are a good source of seed ofpoverty grass.

Mechanical scarification and lightincreased seed germination of povertygrass.

Poverty grass grown with completeweed control produced 146 lb. seed peracre. However, more work is needed toreduce weed competition.

Broadleaf herbicides 2,4-D andRemedy do not seem to affect povertygrass growth when applied during thegrowing season in May-June.

40

Four shade levels (0, 30, 45, and 80%) are being evaluated on the growth rate and seedproduction of poverty grass using structures covered with shade cloth.

Water conservation is a necessary and

responsible practice, especially in highwater using urban landscapes and golfcourses. As water conservation effortsincrease, the need for identifying turfgrass-es with salt tolerance is necessary.

The goal of this project is to eval-uate and screen commercial cultivars ofperennial ryegrass, creeping bentgrass, andKentucky bluegrass cultivars for salt toler-ance and initiate inheritance and physio-logical studies. A greenhouse and a fieldscreening technique for salt tolerancedeveloped at Rutgers will be compared fortheir ability to evaluate salt tolerance incool-season turfgrasses.

Cultivars of perennial ryegrass,creeping bentgrass, and Kentucky blue-grass are being tested under both green-house and field conditions. Two green-house screening runs were conducted onclones from five perennial ryegrass culti-vars at four salinity levels (0, 5, 10, and 15dS/m). Significant differences wereobserved between salinity treatments withthe highest salinity treatments causing themost injury to perennial ryegrass plants.Significant differences were also observedbetween clones. Clones of ‘Palmer III’exhibited the highest percent green ratingscompared to other cultivars.

Two greenhouse screening runswere conducted on 21 Kentucky bluegrasscultivars at four salinity levels (0, 3, 6, and9 dS/m). The cultivars exhibiting the high-est percent green ratings were ‘Eagleton’,‘Liberator’, and ‘Cabernet’. The cultivarsand selections with the lowest percentgreen were a Texas x Kentucky bluegrassselection, A03TB-246, ‘Baron’, and theKentucky bluegrass selection A03-84.

One greenhouse run was conduct-

ed on clones from eight bentgrass cultivarsat four salinity levels (0, 3, 6, and 9 dS/m).The second run is scheduled for the springof 2009.

A field study to evaluate cultivarsof perennial ryegrass, Kentucky bluegrass,and creeping bentgrass was established inthe fall of 2006. Twenty-one Kentuckybluegrass cultivars, 22 perennial ryegrasscultivars, and 15 bentgrass cultivars wereestablished. They were evaluated for salttolerance in the summer of 2007 and 2008by treating with a salt solution (EC=10dS/m) three times per week throughout thesummer.

By the end of the season in 2007,the soil EC reached levels above 3 dS/mwhich caused significant stress on the turf-grass plants. Significant differences wereobserved among cultivars and selectionsunder field conditions. ‘Bewitched’, theexperimental selection, A03-84, ‘Langara’,‘Bedazzled’, ‘Jefferson’, ‘Diva’, P105,‘Rhythm’, and ‘Liberator’ had the highestpercent green leaf tissue under these con-ditions while ‘Julia’ had the least.

The field results were not strong-ly correlated to greenhouse salt chamberresults. We hope to identify the critical

factors influencing salinity tolerance underfield conditions in order to develop effi-cient selection techniques for improvingsalinity tolerance in cool-season turfgrass-es. Clones of perennial ryegrass wereestablished in the fall of 2007 and treatedwith a salt solution with an EC of 10 dS/m.Initial broad-sense heritability was esti-mated to be 0.78, indicating a large pro-portion of the variation could be con-tributed to genetic effects. Narrow-senseheritability studies have also been estab-lished and will be calculated after the datahas been analyzed.

Evaluation of Perennial Ryegrass, Creeping Bentgrass, andKentucky Bluegrass Cultivars for Salt Tolerance


Stacy A. Bonos, Josh A. Honig, Thomas Gianfagna, and Bingru HuangRutgers University

Objectives:1. Evaluate salinity stress tolerance of cultivars of several turfgrass species, including perennial ryegrass,

creeping bentgrass, and Kentucky bluegrass using a combination of greenhouse and field screening techniques.2. Begin studies to understand physiological basis for salt tolerance among these cool-season turfgrass cultivars.3. Develop cultivar recommendations of salt-tolerant cultivars for turfgrass managers.4. Initiate inheritance studies of salt tolerance.

Salt injury on cool season turfgrasses evaluated underfield conditions.


Summary PointsSignificant differences were observed

in five perennial ryegrass cultivars treatedwith 4 different salinity levels (0, 5, 10,and 15 dS/m) under greenhouse condi-tions.

Clones of ‘Palmer III’ exhibited thepercent green ratings compared to othercultivars.

Significant differences were observedin 21 Kentucky bluegrass cultivars treatedwith 4 different salinity levels (0, 3, 6, and9 dS/m) under greenhouse conditions. Thecultivars exhibiting the highest percentgreen ratings were ‘Eagleton’, ‘Liberator’,and ‘Cabernet’. The cultivars and selec-tions with the lowest percent green were aTexas x Kentucky bluegrass selection,A03TB-246, ‘Baron’ and the Kentuckybluegrass selection A03-84.

Twenty-one Kentucky bluegrass culti-vars, 22 perennial ryegrass cultivars, and15 bentgrass cultivars were established ina field trial in the fall of 2006 to evaluatefor salinity tolerance under field condi-tions. Data was collected in 2007, 2008,and will continue through 2009.

Initial broad-sense heritability of salin-ity tolerance in perennial ryegrass wasestimated to be 0.78 from replicatedclones.

41

Previous research projects have iden-tified QTL (Quantitative Trait Loci) forimportant traits in mapping populations ofcreeping bentgrass. However, all initialQTL experiments have several limitations.They may detect ghost QTLs, fail to detecta real QTL, and may over- or underesti-mate the true effects of QTLs.Futhermore, QTLs can vary in differentenvironments and different genetic back-grounds, and QTLs may not be linked tothe quantitative trait after further rounds ofrecombination.

The confirmation of initial QTLsis necessary to successfully utilize QTLmarkers in marker-assisted selectionbreeding programs. Almost every QTLconfirmation study finds some discrepancywith initial QTL experiments. The goal ofthis project is to confirm putative QTL fordollar spot resistance identified in a previ-ous study (funded by the USGA, OJ NoerFoundation, and USDA) by evaluating theQTL markers in three subsequent mappingpopulations (two backcross populationsand a second generation [pseudo F3] popu-

lation) developed from the initial mappingpopulation.

This confirmation step is neces-sary because it is important to determine ifthe QTLs remain linked to dollar spotresistance after further rounds of recombi-nation. This project will result in easilyuseable, reproducible, SSR QTL markersfor dollar spot resistance that will be pub-licly available to turfgrass breeding pro-grams interested in using them for marker-assisted selection.

We have identified approximately200 unique SSR markers. Approximatelytwo-thirds of the markers have been geno-typed in the backcross and pseudo F3 gen-

erations. Initial genetic linkage maps ofeach original parents based on approxi-mately 200 SSR markers were developed.Three putative QTL markers have beenidentified for the ‘Crenshaw’ isolate andone different putative QTL marker wasidentified for the perennial ryegrassisolate.

We analyzed the backcross andsecond generation populations for theseputative QTLs and are in the process ofscoring the markers and genotyping theprogeny for presence or absence of theQTLs. This data will be compared to thephenotype data to determine whether theQTL remains linked to dollar spot resist-ance in these populations.

A replicated, mowed spaced-plant

trial of the pseudo F2, pseudo F3 and back-

cross populations was planted in the springof 2008 to reevaluate the population to adifferent virulent isolate of S. homoeo-carpa than was previously evaluated. Wewill inoculate the plants next spring andevaluate the populations for two years fordollar spot resistance.

Confirmation of QTL Markers for DollarSpot Resistance in Creeping Bentgrass


Stacy A. Bonos, Joshua Honig, and Christine KubikRutgers University

Objectives:1. Genotype backcross and pseudo F3 generations for the 180 SSR markers identified to date.2. Evaluate the populations for dollar spot resistance to a different isolate of Sclerotinia homoeocarpa than previ-ously evaluated.

3. Confirm QTLs in the backcross and pseudo F3 generations to provide definitive SSR QTL markers that can bedirectly utilized for marker-assisted selection in turfgrass breeding programs.

A replicated, mowed spaced-plant trial of the pseudoF2, pseudo F3 and backcross populations was plantedin the spring of 2008 to reevaluate the population to dif-ferent virulent isolate of S. homoeocarpa.


Summary PointsThree putative SSR QTL markers

were identified in the initial population forthe ‘Crenshaw’ isolate. One was identifiedin the susceptible parent and three wereidentified in the resistant parent. One QTLhas been identified in the resistant parentfor the perennial ryegrass isolate.

Two of the three QTLs for the‘Crenshaw’ isolate show evidence of dom-inance for susceptibility and only oneexhibits dominance for resistance. This isconsistent with gene number estimations(2-5 genes).

Selection for a combination of markersidentified genotypes with good dollar spotresistance. These markers may be usefulin marker-assisted selection breeding pro-grams to develop new cultivars withimproved dollar spot resistance.

Classical genetics and molecular mark-ers support evidence for dominant genesfor susceptibility.

42

Host resistance for plant diseases is an important part of improving turfgrass cultivars. Researchers at RutgersUniversity are using molecular techiniques to identify genetic components that are responsible for resistance todolar spot caused by Sclerotinia homoecarpa, prevalent on golf course turf.

Production and utilization of drought

tolerant turfgrass materials is an importantapproach for water conservation and forimproving turfgrass performance in water-limited environments. However, theprogress of breeding drought tolerant turf-grass can be restricted due to complexnature of drought stress, the large genotypeby environment interactions, and the limit-ed knowledge of the genes or traits linkedto drought tolerance.

In recent years, association map-ping has been developed as a novel andmore powerful mapping technique. It usesa natural plant population to providemolecular markers associated with a phe-notypic trait of interest and serves as analternative method for mapping quantita-tive traits loci (QTL). Compared to linkagemapping in traditional bioparental popula-tions, association mapping offers threemain advantages: increased mapping reso-lution, reduced research time, and greaterallele numbers.

A mapping population of 192diploid perennial ryegrass accessions(Lolium perenne L.) was established.These grasses were selected from 43 coun-tries and regions to maximize geneticdiversity. The populations were planted inthree different locations in Indiana: WestLafayette, Wanatah, and Vincennes.

Each accession was propagatedby tillers from the mother plant so thatplants for each individual accession usedin different locations are genetically identi-cal. Screening of perennial ryegrass popu-lations in three locations over multipleyears will generate more reliable data forevaluating drought response and tolerance.

A total of 105 simple sequence

repeat (SSR) markers were used to screen192 populations of diploid perennial rye-grass for assessing genetic diversity.

The cluster analysis by unweight-ed pair-group methods using arithmeticaverages (UPGMA) and principal compo-nent analysis (PCA) showed that perennialryegrass accessions chosen for this studywas diverse without forming the majorpopulation groups, except for 5 commer-cial cultivars that clustered together, sug-gesting that genetic diversity in commer-cial cultivars is limited.

The first three principle compo-nents only explained 11.4 % (7.0 %, 2.3 %,and 2.1 %) of total variation. The pair-wiseplots of first three principle componentsexhibited some levels of separation, butnot clearly identifiable groups. The resultindicated that this perennial ryegrass col-lection would be an ideal population forfurther association mapping of candidategenes for drought tolerance.

Perennial ryegrass is a self-incompatible species that can potentiallyprovide high-resolution association map-ping. The project will sample naturaldiverse populations to evaluate the physio-logical adaptations of perennial ryegrass todrought stress and to identify, through theassociation mapping approach, genes thatplay an important role in drought toleranceand adaptation.

Linking Drought Tolerance Traits and Candidate Genes inPerennial Ryegrass through Association Mapping


Perennial ryegrass accessions were planted in the fieldand drought tolerant traits will be evaluated atVincennes (A), West Lafayette (B), and Wanatah (C).

Yiwei JiangPurdue University

Objectives:1. Evaluate the natural diversity of drought tolerance traits in perennial ryegrass.2. Estimate the genetic population structure of perennial ryegrass.3. Choose candidate genes involved in drought tolerance and genotyping the population for candidate genes.4. Identify the correlation between the candidate gene and plant performance for target traits.


Summary PointsThe diploid perennial ryegrass was

planted and drought tolerance traits will beevaluated.

The mapping population showed alarge genetic diversity.

Genetic population structure will beestimated and candidate genes will be con-tinuously chosen for future associationanalysis.

43

A

B

C

A research site was assigned and

planted using greenhouse-grown bermuda-grass plugs on June 22, 2007 at theOklahoma State University TurfgrassResearch Center, Stillwater, OK. The sitewas specifically constructed to host thisand future shade selection projects. Fundsfor development of the site were obtainedfrom the Oklahoma AgriculturalExperiment Station, Stillwater, OK.Additional research funding was securedfor this project through the OklahomaTurfgrass Research Foundation and fromthe Huffine Endowed Professorship.Additional sources of funding are underconsideration.

The research site receives mid- tolate afternoon shade, depending on season,from a dense, mature evergreen canopy onthe west side of the site. The site meets themost important parameters for effectiveshade research. Late afternoon vegetativeshade is provided by conifers on the westside of the plots. These conifers also pro-vide root competition and reduce the pre-dominately westerly airflow.

Maple trees have been plantedalong the south side of the site and redbudtrees along the east side. We attempted toincrease the duration of shade at the site byplanting vines along a hoop structure in2008, but we had limited success.Consequently, in the next few years, as thedeciduous shade trees mature, we willdesign neutral shade canopies that limitphotosynthetic efficiency and adjust to theduration that we desire. Using canopies,we can increase the shade duration toincrease photosynthetic stress or decreaseduration to limit the loss of potentialselections.

The study consists of 45bermudagrass selections and four stan-dards ('Celebration', 'Patriot', 'Tifton 4', and'Tifton 10'), replicated five times on theshade site and on an adjacent site that is infull sun for about 90% of each day. Visualturf quality (TQ) and NDVI (NormalizedDifference Vegetation Index) wereassessed every two weeks in 2008 andresults are reported for six rating datesfrom May 19 to August 15, 2008.

In 2008, shade stress occurred onthe shade site for 12% longer each daythan on the sun site. However, this shortduration of shade stress was presumablysufficient to cause an average 4.9% declinein TQ and a 3.4% decline in NDVI whenselections were individually comparedbetween the shade site and the sun site.

The top performing selectionsranked by LSD mean separation (P=0.05)of TQ ratings indicate that there is signifi-cant variation among selections in both sunand shade. Visual TQ ratings change withrating date. They are accurate for rankingselections on any given day but are not

accurate for comparisons among ratingdates. The NDVI is consistent and can beaccurately compared among dates.Consequently, it is a good tool for accu-rately evaluating seasonal changes or forindividual comparisons over multiple rat-ing dates.

According to NDVI, the selec-tions that had the best TQ in sun declinedfrom 0.4% to 10.3% in shade suggestingthat some of these selections tolerate shadebetter than others. As we increase shadestress, we are confident that we can deter-mine selections with potential for shadetolerance.

Selection of Bermudagrass Germplasm that Exhibits PotentialShade Tolerance and Identification of Techniques for Rapid

Selection of Potential Shade Tolerant Cultivars


Gregory E. Bell and Yanqi WuOklahoma State University

Objectives:1. Screen bermudagrass germplasm collections and selections for their performance in shaded environments.2. Determine turfgrass characteristics that may useful for screening future selections for potential shade tolerance.3. Create 1 or 2 genetic populations by physiological and molecular selections of shade tolerant and susceptibleparents for future research.

Top 24 bermudagrass selections determined by visual quality means collected on six rating dates from May 19 toAugust 15, 2008.


Summary PointsIn 2008, the shade site received 88% of

the solar irradiance received on the sunsite.

Turfgrass quality ratings and NDVIindicated significant diversity amongselections.

The mean visual turf quality declinebetween like selections in full sun to shadewas 4.9% and the mean decline in NDVIquality was 3.4%.

44

Bermudagrass Sun Shade Shade/sunVisualTQ Visual Rank* NDVI** VisualTQ Visual Rank NDVI Decline***

1-9=best ---- LSD ---- 1-9=best ---- LSD ---- ---- % ----

Patriot 7.6 A 0.7646 7.1 A 0.7365 -3.7C19 7.2 B 0.7681 6.7 BC 0.7479 -2.6C23 7.0 BC 0.7772 6.3 EFGHIJKL 0.7333 -5.6Tifton10 7.0 BCD 0.7300 6.5 BCDEFG 0.7261 -0.5Tifton4 7.0 BCDE 0.7579 5.8 QRSTUV 0.6796 -10.3C17 6.9 BCDEF 0.7515 6.6 BCD 0.7462 -0.7C20 6.9 BCDEFG 0.7703 6.3 DEFGHIJK 0.7460 -3.2C29 6.8 BCDEFGH 0.7463 6.4 CDEFGHI 0.7099 -4.5C28 6.8 CDEFGHI 0.7443 6.7 B 0.7183 -3.5C116 6.8 CDEFGHI 0.7532 6.1 JKLMNOP 0.7154 -5.0C34 6.8 CDEFGHIJ 0.7329 6.0 KLMNOPQ 0.6775 -7.6C35 6.7 CDEFGHIJK 0.7419 6.3 DEFGHIJ 0.7022 -5.4C12 6.7 CDEFGHIJK 0.7338 5.8 PQRST 0.7168 -2.3C13 6.7 DEFGHIJKL 0.7582 6.6 BCD 0.7464 -1.6C27 6.7 DEFGHIJKL 0.7466 6.4 BCDEFGH 0.7295 -2.3PRC-06-5 6.6 EFGHIJKLM 0.7394 6.4 CDEFGHI 0.7140 -3.4Celebration 6.6 EFGHIJKLM 0.7537 6.3 DEFGHIJ 0.7387 -2.0C72 6.6 EFGHIJKLM 0.7402 6.3 EFGHIJKL 0.7182 -3.0C16 6.6 EFGHIJKLM 0.7401 6.2 HIJKLMNO 0.7114 -3.9C4 6.6 FGHIJKLMN 0.7554 6.5 BCDE 0.7524 -0.4C73 6.6 FGHIJKLMN 0.7270 6.0 KLMNOPQ 0.7055 -3.0

A creeping bentgrass mapping popu-

lation (L93-10 x 7418-3) segregating fordollar spot resistance was evaluated forvariation in drought tolerance in this study.We examined both parents and 100 F2progenies. An intra-specific pseudo F2mapping population (180 individuals) wasgenerated from a cross between a dollarspot resistant (L93-10) and a susceptible(7418-3) genotype in the spring of 2003.All individual clones (104 from L93-10 x7418-3) were planted in one large plasticcontainer (60 cm x 120 cm x 30 cm deep)filled with a mixture of sand and soil andreplicated five times.

Plants were exposed to droughtstress by withholding irrigation. Duringthe dry-down period, several drought-stress indicators were evaluated to deter-mine the genotypic variation and pheno-typic traits for drought tolerance in thesetwo mapping populations. Turf quality wasrated visually based on the scale of 1-9 (9= best) as the indicator of general turf per-formance. Several most commonly usedparameters for phenotyping drought toler-ance, including relative water content,osmotic adjustment, cell membrane stabil-

ity, and water use efficiency wereanalyzed.

The phenotypic data with the ‘L-93’ x 7418 population demonstrated thatturf quality for two parents and 102 proge-nies exhibited large variation, with visualrating ranging from 3 to 6 following 7 daysof drought stress. Leaf relative water con-tent (RWC) and osmotic adjustment (OA),two commonly used stress indicators, aresignificantly correlated to turf qualityunder drought stress, with correlation coef-ficients being 0.73 and 0.68, respectively.Both traits exhibited a wide range of vari-ation, with RWC varying from 33% to94% and OA ranging from 0 to 0.50.

A field study was conducted insummer and fall 2008 in a fully automatedmobile shelter (35' x 60') at RutgersUniversity in North Brunswick. The L93-10 x 7418-3 mapping population wasestablished in a mowed spaced-plant eval-uation trial in the rainout shelter in April2008. The individual clones were arrangedin a randomized complete block designwith 3 replications.

From July to September, allplants were maintained under well-wateredconditions and examined for turf quality,canopy green leaf biomass, leaf area index,and cell membrane stability to analyze forvariation and phenotypic traits for heat tol-erance. During October and November,

plants were exposed to drought stress bywithholding irrigation to examine geneticvariation in drought tolerance. Data for thefield study will be analyzed and summa-rized in December.

QTL analysis was conducted onseveral phenotypic traits including elec-trolyte leakage (EL), relative water content(RWC), and osmotic adjustment in the‘L93’ x 7418-3 mapping population. Nosignificant QTLs were found on the moredrought sensitive parent (‘L-93’). SevenQTLs were found on the more drought tol-erant parent (7418-3). Four significantQTLs (1 RWC; 2 EL; 1 TQ) and three sug-gestive QTLs (1 RWC; 1 WUE; 1 OA)were detected. These QTLs were found infour chromosome regions, with co-loca-tion of RWC and TQ, RWC and EL, andWUE and OA.

QTLs markers for TQ, RWC, andEL were highly correlated to drought toler-ance. These initial QTLs from the green-house study will be compared to phenotyp-ic traits associated with drought toleranceevaluated under field conditions.

Identification of Quantitative Trait Loci (QTL) Associated withDrought and Heat Tolerance in Creeping Bentgrass


A field study was conducted in summer and fall 2008 in a fully automated mobile shelter (35' x 60') at RutgersUniversity in North Brunswick, NJ.

Bingru Huang, Stacy Bonos, and Faith BelangerRutgers University

Objectives:1. To evaluate variations in drought and heat tolerance for two mapping populations of bentgrass segregating for disease resistance.

2. To identify phenotypic traits associated with drought and heat tolerance.3. To identify QTL markers associated with drought and heat tolerance utilizing the available linkage maps.


Summary PointsPhenotypic variations in drought toler-

ance exist in the creeping bentgrass map-ping population developed for dollar spotresistance.

Significant QTLs were identified forTQ, RWC, and EL, and those QTL mark-ers are highly correlated to drought toler-ance.

QTL markers for drought tolerancewere located in four regions of the geneticlinkage map for the drought tolerant par-ent.

Major QTLs located in the same chro-mosomal region for turf quality, relativewater content, cell membrane stability,osmotic adjustment, and water use effi-ciency may be useful as molecular markersfor marker-assisted selection.

45

The New England Velvet Bentgrass

germplasm collection is a collaborativeeffort by researchers at the University ofRhode Island and the University ofMassachusetts. More then 250 accessionshave been collected from old golf coursesthroughout New England. In 2008, morethen 750 individuals were evaluated in thefield at UMass for turf quality and resist-ance to brown patch and dollar spot. Theentire collection was screened for resist-ance to copper spot in the greenhouse atURI.

Velvet bentgrass (Agrostiscanina) has excellent tolerance to lowerlevels of sunlight, nitrogen, and water.Velvet bentgrass is considered native toNew England and coastal regions as farsouth as Maryland. Many of the old velvetgreens have been regrassed with moderncreeping bentgrass varieties due to overmanagement, high thatch layer production,and lack of available velvet bentgrass com-mercial seed. The stress tolerance genesfound in velvet bentgrass need to be pre-served as potentially irreplaceable geneticresources.

The identification and geneticdiversity of the collection are being meas-ured with molecular markers and flowcytometry to confirm that accessions are

diploid Agrostis canina. Fingerprintingthe accessions will permit them to begrouped based on genetic identity, subdi-viding the collection into groups which canbe converted to seed populations forgenebank storage. The groups can be rep-resented by a subset of the clones for moreefficient screening.

Screening for copper spot resist-ance was done in the greenhouse usingclones grown in a sand-peat mix, clippedto fairway height, and fertilized withexcess nitrogen. The plants were inoculat-ed using copper spot (Gloeocercosporasorghi) isolates and maintained underappropriate conditions to promote disease.Disease severity was evaluated on a 0-10scale with 0 indicating complete loss offoliage and 10 indicating no damage. Thevarieties 'SR7200' and 'Greenwich' wereused as standards for comparison.

Sixty-two percent of the acces-sions were effectively defoliated by copperspot. Both 'SR7200' and 'Greenwich' showsome resistance to copper spot; 30% of theaccessions tested were at least as resistant.The germplasm collection is being evaluat-ed in the field for turfgrass quality andgenetic color. Velvet bentgrass has a ten-dency to be lighter green than other bent-grasses. The identification of velvet bent-

grass which is genetically darker greenmay make turfgrass managers more com-fortable applying less nitrogen.

The core collection is being eval-uated for wear tolerance, a common abiot-ic stress on highly utilized turfgrass. Golfcourse putting greens, tees, and fairwaysare particularly susceptible to wear injurydue to the funneling of intense and fre-quent traffic onto a relatively small surfacearea.

Evaluation of the New England Velvet Bentgrass Collection


Rebecca Nelson Brown and Cynthia PercivalleUniversity of Rhode Island

Objectives:1. Collect velvet bentgrass (Agrostis canina L.) germplasm accessions.2. Identify and group accessions based on their genetic similarities for analysis and breeding.3. Evaluate accessions for improved resistance to biotic and abiotic stresses.

More then 250 accessions have been collected from old golf courses throughout New England.


Summary PointsCollaborative studies continue with

several researchers in New England.

Significant progress has been made toidentify germplasm that have enhancedresistance to biotic and abiotic stresses.

46

Geunhwa Jung and Jason M DowgiewiczUniversity of Massachusetts

Planting plugs of velvet bentgrass.

The velvet bentgrass collection at the University ofRhode Island.

Poa annua is widely recognized to

provide a large portion of high quality put-ting surfaces in many regions of the U.S.,Canada, Australasia, and Scandinavia.However, despite repeated attempts todevelop improved cultivars of greens-typePoa annua for the golf course industry,there currently are no commercially avail-able sources suitable for use in new con-struction or renovation. Our previousgenetic research suggests that the perenni-al greens-type phenotypes result from theaction of mowing which causes a repres-sion of the plant growth hormone gibberel-lic acid (GA) signaling pathway through anon-Mendelian epigenetic mechanism.

A somewhat analogous situationoccurs in the wild, weedy annual-types ofPoa annua from applications of plantgrowth regulators, such as Primo.However, in the absence of the mowingstimulus, the GA pathway progressivelybecomes unsilenced resulting in reversionof the greens-type plants back to the annu-

al type. The annual type is undesirable as aputting surface and requires years of mow-ing in order to develop a perennial greens-type form. We believe that the genetic sta-bility of perennial greens-type Poa annuais capable of being maintained throughvegetative propagation in combinationwith mowing.

We established vegetative plotsusing different source plant materialsincluding aerification cores, solid sod, andshredded sod/plugs. Planting densities ofthese materials were varied in order tomaximize the limited amount of our plant-ing stock while, at the same time, attempt-ing to achieve full coverage in a reasonableamount of time.

We also began evaluating vegeta-tive establishment in the greenhouse incombination with the exogenous applica-tions of gibberellic acid in an attempt tofurther reduce the time required to achievefull coverage. All greenhouse and fieldplots are being mowing to a height of 1 cm(approx. 3/8 inch) as soon as possible afterplant materials have rooted in order tomaintain the greens-type phenotype.

This year, we were fortunateenough to supply vegetative planting mate-rial of greens-type Poa annua to turfgrassresearchers at 5 universities participatingin the Northeast 1025 Project titled:"Biology, Ecology, and Management ofEmerging Pests of Annual Bluegrass onGolf Courses".

Future expectations of thisresearch project will be to extend the mostsuccessful techniques identified for thevegetative propagation of greens-type Poaannua to a larger scale approaching that ofcommercial sod production. Vegetativematerial from the previous vegetativepropagation studies will provide the neces-sary planting material for us to workdirectly with interested sod producers for

an evaluation of scaled-up production.Ultimately, our research will enable thecommercial availability of greens-typePoa annua cultivars that possess high turfquality, disease resistance, and stress toler-ance which will significantly reduce inputsfor the management of Poa greens and willincrease the supply of greens-type Poaannua for turfgrass scientists throughoutthe USA to conduct valuable collaborativeresearch in other areas of Poa annua golfcourse management.

Evaluating Methods for Vegetative Propagation and Enhancementof Seed Production of Greens-type Poa annua Cultivars


Poa annua is widely recognized to provide a large por-tion of high quality putting surfaces in many regions ofthe U.S., Canada, Australasia, and Scandinavia.

David R. HuffPennsylvania State University

Objectives:1. Establish efficient vegetative propagation methods of greens-type Poa annua for sod production and for establishing/renovating golf course putting greens.

2. To scale-up our results for greens-type cultivar production to a larger commercial level by collaborating with sodproducers.

3. To release a genetically stable, vegetative greens-type cultivar exhibiting superior putting green quality and stresstolerance.

vegetative plots were established using different sourceplant materials including aerification cores, solid sodand shredded sod/plugs.


Summary PointsOur research continues to indicate that

the dwarf nature of perennial greens-typePoa annua results from an epigenetic inhi-bition of the biosynthetic pathway for theplant growth hormone gibberellic acid(GA).

We demonstrated that greens-type Poaannua is responsive to exogenous applica-tions of gibberellic acid which may aid inits vegetative establishment.

This year, we supplied vegetative plant-ing material of greens-type Poa annua toturfgrass researchers at 5 universities par-ticipating in the Northeast 1025 Projecttitled: "Biology, Ecology, andManagement of Emerging Pests of Annual

Bluegrass on Golf Courses".47

It is widely known that one of thegreatest challenges confronting the turfindustry today is water. Golf course super-intendents and other landscape managersare being asked to use less irrigation water,use lower quality water sources, and allowmore use of the turfgrass areas. Turfgrasswith high turfgrass quality and greater salttolerance is essential.

Beginning in 2005, the USDA-ARS Forage and Range Research Lab(FRRL) in Logan, Utah and Utah StateUniversity started work in the area of saltand drought tolerance evaluation in Poa. Acooperative agreement has also been estab-lished with Utah State University. As partof this cooperative effort, we are evaluat-ing a large number of National PlantGermplasm accessions, collections fromaround the world, somaclonal lines of P.pratensis from Dr. Nancy Ehlke at theUniversityof Minnesota, and numerouscurrently available varieties.

These lines are being evaluated infield conditions for turf and seed produc-tion characteristics, drought tolerance, andother traits and will be used to combinestress tolerance and turf quality traits ingermplasm that can be used by the seedindustry.

Many Poa species,and most of the availablegermplasm within Poapratensis, has not been rigor-ously evaluated for salt toler-ance and turf characteristicsin the arid West climate.Similarly, there is not a goodunderstanding of the geneticcontrol of salt tolerance inPoa. We are expanding abreeding and genetics pro-gram to develop improvedsalt-tolerant bluegrass (Poa) germplasm.

Using our large and growing col-lection of germplasm from a variety ofsources, we have and will continue to con-duct a rigorous program of screening forsalt tolerance and will identify candidategenes associated with salt tolerance in Poausing suppressive-subtractive hybridizationmethods.

We completed an initial evalua-tion of salinity tolerance of Poa germplasmwith a proven method developed at theFRRL. In a greenhouse, seedings weregrown in sand and when at the 3-tillerstage, were subjected to increasing salt con-centrations (increased by 3.0 dS m-1 incre-ments every 2 weeks) and evaluated untilplant death.

Based on two runs of the experi-ment, several germplasm sources wereidentified with consistently high salinitytolerance during both runs of the study.The salinity tolerance of some of thegermplasm sources was similar to that ofthe tall fescue and perennial ryegrasscheck entries. Results suggest that selec-tion and hybridization among the bettergermplasm sources will potentially resultin bluegrass cultivars with increased salin-ity tolerance. The results of this study willbe submitted for publication in a refereedjournal in the coming months.

A Ph.D. graduate student, Mr.Shyam Shridhar, is currently conductingthe gene expression work, specifically toidentify candidate genes turned on during.This process involves imposing salt stress

on salt-tolerant and salt-sensitiveKentucky bluegrass accessions. Theseaccessions were identified in the screeningprocedure described above. RNA, theresults of gene expression, was extractedfrom roots and shoots, purified, and sub-jected to suppression-subtractivehybridization (SSH) to detect differentiallyexpressed sequences of DNA.

This procedure will allow us toidentify genes expressed at a higher levelin the salt-tolerant germplasm shoots androots than sensitive germplasm shoots androots. Currently the SSH procedure is com-pleted, and the differentially-expressedgenes are being detected and sequenced. Inthe coming months, we will be comparingthe expressed gene sequences to a publiclyavailable plant sequence database to assignpossible gene function.

Evaluation and Development of Poa Germplasm for Salt Tolerance


Seedings were grown in sand and when at the 3-tillerstage, were subjected to increasing salt concentrations(increased by 3.0 dS m-1 increments every 2 weeks)and evaluated until plant death.

Paul JohnsonUtah State University

Objectives:1. To identify salt-tolerant Poa germplasm that can be incorporated into breeding and genetics efforts.2. To identify genes whose RNA transcript levels vary between control and high salinity treatments.

Poa germplasm was emersed in increasingly saline water and evaluat-ed in the greenhouse for salinity tolerance.


Summary PointsWe have observed significant variation

in salt tolerance among the National PlantGermplasm accessions.

The most salt-tolerant accessionsexceeded the tolerance of Kentucky blue-grass check varieties and approached orexceeded the tolerance of perennial rye-grass and tall fescue check varieties.

These encouraging results will result inthe evaluation of a wider selection of Poapratensis and Poa spp. germplasm.

Genes expressed at higher levels insalt-tolerant germplasm will be identifiedand considered for future DNA markers.

48

Joseph Robins and B. Shaun BushmanUSDA-ARS

In order to improve the stress tol-erance of turfgrass, we have undertakenthe following five projects:

Development of Danthonia spicata as aLow Maintenance, Native TurfgrassSpecies

A key aspect to the developmentof Danthonia spicata (Poverty grass) as alow maintenance turfgrass species isobtaining a better understanding of thebiology of the species. Therefore, we havespent time collecting plant material andobserving the growth and reproductivecharacteristics of the species. Initial obser-vations indicate that there is variation pres-ent in the species, and there may be morethan one species present in natural stands.

Several new locations for the col-lection of germplasm were identified. Aseeding rate trial was established at theUniversity of Maryland turfgrass centerusing three different seeding rates. Theresults of the first year indicate no signifi-cant difference in turf quality between thethree seeding rates.

Field Screening of BentgrassGermplasm for Resistance to ImportantTurfgrass Diseases

A field trial containing clonallypropagated plants from a bentgrass map-ping population, developed by Dr.Geunhwa Jung at the University ofMassachusetts, was established at theUniversity of Maryland turfgrass center.The study involves approximately 300entries replicated three times, plugged intoa ryegrass turf.

The first rating of this material fordisease resistance was conducted in thesummer of 2008. The artificial inoculationwith Sclerotinia homoeocarpa was not aseffective as we would have liked. Slowfungal growth during inoculum develop-ment resulted in a late inoculation of thefield trial. However, disease developmentdid occur, and our first rating of the mate-rial was conducted with differences clearlypresent.

Development of Koeleria macrantha as aLow-input Turfgrass

The goal of this research is toincrease the genetic potential of prairiejunegrass (Koleria macrantha Ledeb.) foruse as a low-input turfgrass. Research onKoeleria is all being conducted by theUniversity of Minnesota and has initiallybeen focused on germplasm collection.Future research may involve the establish-ment of research plots in Maryland todetermine the tolerance of Koeleriagermplasm to the stressful growing condi-tions often present during Marylandsummers.

Identification of Brown Patch ResistantTall Fescue

The objective of this study was touse digital image analysis (DIA) to evalu-ate tall fescue plant introductions (PIs) forresistance to both Rhizoctonia solani andR. zeae. This study included 15 PIs select-ed from the USDA germplasm databaseand three commercial cultivars with vary-ing brown patch resistance.

The commercial cultivars had thelowest mean disease severity in eachexperiment. Mean disease severity rangedfrom 59-93% for R. solani and from 32-64% for R. zeae. Current work involvesscreening the best PIs and using 20 seeds

sampled from each PI to evaluate thediversity of resistance to both R. solani andR. zeae that may be present in each PI.

Bentgrass Breeding Consortium:Molecular Breeding for Dollar Spot andSnow Mold Resistances

The objective of this study was toidentify candidate Miniature Inverted-repeat Transposable Elements (MITEs), aclass of transposable elements that has notbeen previously described in turfgrasses,from Agrostis and assess their value as amolecular-marker tool.

DNA sequences were screenedusing the FindMITE program to identifycandidate MITE sequences. FindMITEidentified 202 MITE-like sequences, or1.26% of the 16,064 sequences. The MITEdisplay markers had a significantly higherpolymorphism rate (0.42) between themapping population parents thanAmplified Fragment LengthPolymorphism (AFLP) markers at 0.28.The MITE display protocol would be aneffective tool for diversity analyses andmapping in Agrostis.

Genetic Enhancement of TurfgrassGermplasm for Reduced-input Sustainability


Kevin MorrisNational Turfgrass Federation, Inc.

Objectives:1. The objective of this research, conducted at the USDA, ARS Beltsville Agricultural Research Center in Beltsville,MD, is to use genetic and biotechnology approaches to identify and develop turfgrass germplasm with improvedbiotic and abiotic stress resistance. Efforts will be made to identify molecular markers associated with desirabletraits and to combine useful traits into germplasm able to grow with reduced inputs.


Summary PointsThere is variation present in natural-

ized stands of the low maintenance speciesDanthonia spicata (Poverty grass), allow-ing for potential selection of superior planttypes.

A field trial of bentgrass germplasm,planted in Maryland, exhibited differencesin response to dollar spot (Sclerotiniahomoeocarpa) infection.

Tall fescue plant introductions (PIs)inoculated with Rhizoctonia solani and R.zeae showed varying degrees of toleranceto both diseases.

Miniature Inverted-repeat TransposableElements (MITEs) show promise as aneffective tool for diversity analyses andmapping in Agrostis species.

49

Scott WarnkeUSDA-ARS

Research findings from this project are

a continuation of earlier research from pastyears. Screening germplasm and cultivarsfor salt tolerance is an important aspect ofthe ryegrass turf breeding program inTexas. Results indicate a good correlationbetween field screening and greenhousescreening. Methods studied were plantingseed in the field in a high salinity soil andirrigating with high salinity water at Pecos,Texas.

Each entry was planted in 5 ftrows at a seeding rate of 2 g/5 ft. Plantswere grown in sand in cone-tainers (4-cmwide and 20-cm tall) which were placed ina salt tank where salt concentration wasgradually increased over time. Plants were

immersed every 3to 4 days in saltwater. Seed wereplanted in rows insand in flats onDecember 14,2007. The firstimmersion (4000ppm salt water, or6.4 dS m-1) of flatsoccurred onJanuary 25 whenplants were in 3-leaf stage.Thereafter, saltwater concentrationwas graduallyincreased as flatswere immersed for2 minutes every 3days.

Each entrywas rated on sever-al dates on a 1 to 9scale, where 9 =dead plants. Resultsfrom the first yearof experimentationindicated that thefield trial and thegreenhouse immer-sion technique arecorrelated and are agood indication ofsalinity tolerance ofgenotypes tested.

We will continue to screen geno-types using both of these techniques in thefuture. We will also screen segregatingpopulations for salt tolerance with the goalof developing and releasing a salt-tolerantcultivar in the future. We will also increaseseed from the latest and hopefully salt-tol-erant populations for further testing.

Breeding Turf-type Annual Ryegrass for Salinity Tolerance


Methods studied were planting seed in the field in a highsalinity soil and irrigating with high salinity water atPecos, Texas.

L. R. NelsonTexas AgriLife Research

Objectives:1. To identify improved salt-tolerant genotypes and introgress the tolerance into adapted turf-type genotypes of

annual ryegrass and increase seed of these populations for experimental testing.2. To select plants of turf-type annual ryegrass which are infected with a fungal endophyte and determine if pres-

ence of the endophyte provides this population with increased tolerance to high salt concentrations.3. To test the experimental lines of annual ryegrass under high salinity growing conditions in hydroponic and field

conditions and release the line as a salt-tolerant variety.

Screening germplasm and cultivars for salt tolerance is an important aspect of the rye-grass turf breeding program in Texas. Results indicate a good correlation between fieldscreening and greenhouse screening.


Summary PointsField and greenhouse screening of

genotypes for salinity tolerance ratingswere correlated.

A greenhouse screening techniquewhere plants in flats were immersed in saltwater for 2 minutes every 3 days holdspromise for future research.

50

While sting nematodes are found

most commonly in sandy coastal areas, theUniversity of Florida NematodeAssay Labhas diagnosed this nematode on bermuda-grass from inland areas of Alabama,Mississippi, Arkansas, Louisiana, Georgia,Tennessee, Texas, Oklahoma, Kansas, andCalifornia.

Recent cancellation offenamiphos (Nemacur, BayerCropscience) has resulted in the need ofbetter nematode management tactics.Currently, Curfew Soil Fumigant (DowAgrosciences) is the most effective man-agement for sting nematodes. However,Curfew is not cost effective and environ-mental restrictions highlight the need foralternative options.

Utilization of resistant or tolerantcultivars is the most efficient, least costlypractice for nematode management on turf.Although a range in responses of bermuda-grass to sting nematodes has been found,most cultivars were reported as suscepti-ble. Information about the responses ofseveral newer bermudagrass and seashorepaspalum cultivars is lacking.

Separate glasshouse experimentsfor bermudagrass and seashore paspalumwill be conducted to assess the range ofthese species for response to sting nema-todes. In 2008, two experiments were con-ducted using six cultivars of bermudagrassin one experiment and three cultivars ofseashore paspalum in the second.

All cultivars were propagated bymeans of nematode free aerial stolons intoclay pots (1,500 cm3) with a USGA sand.Once established, inoculations were doneon June 21, 2008 using 200 nematodes per

pot. The experiments were main-tained at a temperature range of24o C to 34o C under natural day-light. They were trimmed at 1.0-cmmowing height and fertilized onceevery other week using 18-3-6 (N-P-K) at a rate of 2 kg / 100 m2 (4 lb/ 1000 sq. ft) per year.

Experiments were har-vested 90 days after inoculation byremoving a 5-cm diameter corefrom the center of each pot.Nematodes were extracted fromthe soil cores and counted under amicroscope. Roots were analyzedusing WinRhizo root scanningsoftware to determine root lengthsand surface areas.

Reproduction and meannematode counts were greatest on'Champion' bermudagrass with no differ-ences between the remaining cultivars.Significant differences were not foundbetween the seashore paspalum cultivars.However; biologically, 'SeaDwarf' was thebest host for sting nematodes.

Mean total root length compar-isons were made within a given cultivar forits uninoculated and inoculated treatments.For all cultivars except 'TifEagle', the inoc-ulated treatments led to reductions in totalroot length. Differences between uninocu-lated controls and inoculated treatmentswere significant only for ‘Champion’bermudagrass, and ‘Aloha’ and ‘Sea Isle 1’seashore paspalum.

The percent reduction of‘Champion’ was similar to all bermuda-grass cultivars except ‘TifEagle’.However, the uninoculated root length of‘Champion’ was approximately one-thirdof the other greens-type uninoculated rootlengths. This may be the basis for the dif-ference observed for its inoculatedtreatment.

Similar reductions between‘Champion’ and other cultivars will bemore detrimental to ‘Champion’ due to itssmaller root system. Because ‘Champion’was a better host for sting nematode repro-duction (least resistant) and its root systemwas more heavily damaged (poor toler-ance), it was considered the most suscepti-ble cultivar to sting nematode damage inthis study. ‘SeaDwarf’ supported highernematode reproduction than other seashorepaspalum cultivars.

Bermudagrass (Cynodon spp.) and Seashore Paspalum(Paspalum vaginatum) Cultivar Response to the Sting Nematode

(Belonolaimus longicaudatus)


Wenjing Pang, William T. (Billy) Crow, and Kevin E. KenworthyUniversity of Florida

Objectives:1. Determine the range of response (resistance or tolerance) of bermudagrass and seashore paspalum cultivars to thesting nematode and identify the best performing cultivars.

2. Investigate if a proposed alternative method for assessing sting nematode response is as effective or more effi-cient than traditional methods.

In 2008, two experiments were conducted using six cultivars ofbermudagrass in one experiment and three cultivars of seashorepaspalum in the second.


Summary PointsThe sting nematode caused damage to

‘Champion’, ‘Sea Isle 1’, and ‘Aloha’.

Sting nematode damage was not as sig-nificant to ‘TifSport’, ‘Celebration’,‘Floradwarf’, ‘TifEagle’, ‘Tifgreen’, or‘SeaDwarf’.

‘Champion’ was the most susceptiblebermudagrass cultivar exhibiting poorresistance and tolerance.

‘SeaDwarf’ exhibited better tolerancethan other seashore paspalum cultivarsstudied.

51

There is a nationwide effort to use

native grass species in turf systems, asmost species used now are typically non-native. This study involves initial collec-tion and description of growth charactersof native range grasses as mowed and un-mowed plants. The research may result inpopulations of native grasses that are suit-able for use in turf plantings in the south-western U.S. Procedures we perfect in thisresearch may be used in projects to evalu-ate native plants for potential turf develop-ment.

From July to September 2007,almost 300 clones from seven species ofperennial range grasses were collectedfrom a 150-mile radius of Tucson, Arizonafrom rangelands with a known history ofintensive livestock grazing. Collectionsincluded the sod-forming grasses, curlymesquite, and false grama, and the bunch-grasses sprucetop grama, wolftail, bluegrama, black grama, and hairy grama.These clones were propagated and grownunder near-optimal conditions in a green-house.

In early January 2008, and againsix weeks after a "grazing" (severe defoli-ation) event, data were collected fromplants actively growing in the greenhouse.These included (1) plant height (2) plantdiameter in two directions, and visualscores for (3) innate plant density, and (4)overall turf quality (1-5, 5 = best for thelatter traits). During this phase of ourresearch, clones noted by mere subjectiveobservations were identified as "flagged"clones.

Height/width ratios (lower valuessuggest a more "wider" than "taller"growth habit) before and after defoliationwere relatively consistent up to a value of

5, for pre- and post-defoliation measure-ments, respectively. Interestingly, subjec-tively "flagged" clones were very close tothe 1:1 line of best fit.

Stem and box plot data show thatthere may be considerable useful variationfor growth habit in curly mesquite, wolf-tail, and sprucetop grama. These specieshad a low H/W ratio before defoliation.

After a single defoliation event,most clones exhibited increased height,with some variation remaining for lowH/W ratio growth response for curlymesquite, wolftail, and sprucetop grama.Other species showed very little variationfor H/W ratios following defoliation.

When plotted against the averageof turfgrass quality and density scores afterdefoliation, clones with the lowerheight/width ratio values had the greatestnumerical mean quality-density averages.In this case again, the "flagged" cloneswere again close in fit to the 1:1 line ofH/W vs. the mean quality and densityscores. This data set shows that that plantheight alone is not related to turf quality ordensity, but the height/width ratio is a bet-ter predictor of turf quality.

The next phase of researchincludes the establishment of clonallyreplicated spaced-plant nurseries (accom-plished inAugust 2008) that include clonesselected based on greenhouse perform-ance. Field evaluations will involve thesame measurements taken in the green-house, but also the response of these clonesto regular mowing events. In addition tothe identification of clones appropriate foruse in turf plantings, results from this com-ponent of the projects should allow fordetermination of the utility of greenhouse-based measurements for predicting actualturf quality in the field.

Collection and Evaluation of Native Grasses fromGrazed Arid Environments for Turfgrass Development


D.M. Kopec, S.E. Smith, and M. PessarakliUniversity of Arizona

Objectives:1. Collect native grasses from rangelands with long history of livestock grazing in southern Arizona.2. Evaluate primary growth habit-related characteristics of collected clones.3. Evaluate relationships among characteristics in clones evaluated and select clones for field evaluation based on a

simple index involving independent culling levels.

From July to September 2007, almost 300 clones from seven species of perennial range grasses were collectedfrom a 150-mile radius of Tucson. Species included curly mesquite, false grama, sprucetop grama, wolftail bluegrama, black grama, and hairy grama.


Summary PointsFrom July to September 2007, almost

300 clones from seven species of perenni-al range grasses were collected from a150-mile radius of Tucson.

There may be considerable useful vari-ation for growth habit in curly mesquite,wolftail, and sprucetop grama.

Plant height alone is not related to turfquality or density. The height/width ratiois a better predictor of turf quality.

52

Environmental ImpactThe public is concerned about the effects of golf courses on the environment.

In response to this concern, the USGA has conducted research examining the fate ofpesticides and fertilizers since 1991. The USGA continues to support scientificallybased investigations on the environmental impact of golf courses. The focus remains onresearch to understand the effects of turfgrass pest management and fertilization onwater quality and the environment.

Research on best management practices evaluates pesticide and fertilizer pro-grams for golf courses in order to make turfgrass management recommendations thatprotect environmental quality. The research is conducted on university experiment sta-tions and golf courses. The projects evaluate pesticides or nutrients that pose an envi-ronmental risk, and identify cultural practices that minimize volatilization, surfacerunoff, and groundwater contamination.

Pesticide and nutrient fate models are used to predict the environmental impactof turfgrass pesticides and fertilizers. From 1991 through 1997, research sponsored bythe USGA demonstrated:

1. Measured nitrogen and pesticide leaching was minimal and that surfacetransport (runoff) posed a greater problem for golf courses, especially onheavy textured soils in high rainfall areas of the country.

2. The turf/soil ecosystem enhances pesticide adsorption and degradation thatgreatly reduces the amount of chemical that moves below the rootzone.

3. Current agricultural fate models need modification to predict the fate ofpesticides and fertilizers applied to turfgrasses grown under golf courseconditions.

The results of USGA-sponsored pesticide and fertilizer fate research is beingused to calibrate and validate existing pesticide fate models for turfgrasses managedunder golf course conditions.

Locations of the following projects funded in 2008 by the USGA Turfgrass andEnvironmental Research Program under the category of Environmental Impact

53

Golf course superintendents consider

many factors when selecting a pesticide fora specific use, including cost, efficacy, andturf safety. However, it is currently muchmore difficult for a superintendent toassess environmental risk and its relevanceto their golf course. What is the risk togroundwater supplies when a particularpesticide is applied? What is the risk tosurface water, fish, or bees?

These are complex questionsrequiring not only data, but also a methodto integrate the data into a form that allowsmeaningful conclusions. The first step ofthis project is to collect relevant data onenvironmental fate, toxicology, and envi-ronmental endpoints from publicly avail-able databases. To date, we have collectedthe majority of the data needed.

The second part is to create amodel or software program that calculatesthe relative risk to specific environmentalfeatures from the application of a specificpesticide active ingredient. Consider a golfcourse with a stream flowing through the

property. The golfcourse superintendentmay want to know theprobability of a pesti-cide intended for useto reach the stream,and if it does, what isthe likelihood that itwill cause problemsfor fish in the stream?A rudimentary riskassessment deter-mines the likely con-centration of the pesti-cide in the stream and whether this con-centration is high enough for concern.

Integral to our process of buildingthis resource has been the solicitation offeedback from the following groups:

End-users of the resource, golf coursesuperintendents;

EPA staff with expertise in pesticidefate assessment and modeling, regulatoryenforcement and economic impact;

The scientific community via presenta-tions at scientific meetings; and

Scientists from pesticide manufacturersrepresented by Crop Life America andResponsible Industry for a Sound

Environment.The challenge of this project is to

develop a tool that is easy to use, whileretaining a sound scientific basis for esti-mating potential environmental risks ofusing a particular pesticide. At this point,several components of the final modelhave been selected. We expect the model toyield information on risk to ground water,surface water, birds, and non-target inver-tebrates for each of the over 100 pesticideactive ingredients in our database. The riskdetermination will be based on risk ratiosand presented in a format that is easy tointerpret.

Recommendations for best man-agement practices to minimize environ-mental risk of an application and maximizeenvironmental stewardship will be provid-ed. The resource will help superintendentsmake more informed environmental deci-sions on the pesticides they choose to use.

The Pesticide Matrix Project: Developing a Data Base Tool toGuide Environmentally Responsible Pesticide Selection


The development of a pesticide matrix involves obtaining pesticide environmental fate and toxicological endpointsand calculating toxicological reference points when necessary.

Bruce Branham and Thomas Fermanian Stuart Cohen Jennifer GrantUniversity of Illinois Environmental and Turf Services Cornell University

Objectives:1. To create a resource that is available electronically to help superintendents better understand the environmental

characteristics of golf course pesticides and optimize their environmental stewardship.

Categories of risk factors included in the pesticide matrix model include risks toground water, surface water, birds, and non-target invertebrates.


Summary PointsA database of pesticide properties

needed for risk assessment is beingcompiled.

The database will serve as the founda-tion to predict potential environmental riskof a pesticide’s active ingredient.

The end result of this research will be aresource that will help superintendentsmake more informed environmental deci-sions regarding the pesticides they apply.

54

This study seeks to determine actu-

al levels of golfer exposure to "reduced-risk" pesticides following application toturfgrass. The fate of pesticides after appli-cation largely determines how much of it isavailable for potential human exposure.

We have analyzed pesticideresidues in the air and on turfgrass (dis-lodgeable foliar residues, DFR) in 43 pes-ticide applications using either chlorpyri-fos, carbaryl, cyfluthrin, chlorothalonil,2,4-D, MCPP-p, dicamba, imidacloprid,and carfentrazone. This season, threeapplications of the reduced-risk fungicideazoxystrobin were made. Analyses of thesesamples are in progress.

This study also evaluates bestmanagement practices for reducing golferexposure to reduced-risk turfgrass pesti-cides. While many "standard" pesticideshave been removed from use, newreduced-risk pesticides have been added tothe IPM practitioner's toolbox. To date,there is no dosimetry or biomonitoringdata on these reduced-risk pesticides,which exhibit low mammalian and envi-ronmental toxicity, low potential forground water contamination, low pestresistance potential, and are compatiblewith IPM.

To determine precisely how muchof the environmental residues is actuallytransferred to golfers during a round ofgolf, we measure exposure to volunteergolfers using dosimetry (measuring pesti-cide residues on full-body cotton suits andpersonal air samplers) and biomonitoring(measuring urinary metabolites). Thiswork is being done in cooperation with the

New England Regional TurfgrassFoundation.

Dosimetry and biomonitoring, togetherwith concurrently collected dislodgeablefoliar and airborne residue data, provides aunique database on golfer exposure andhas allowed us to develop a golfer expo-sure model. The central predictor of expo-sure in the model is the transfer factor(TF), which is ratio between the pesticideresidues measured in the environment ver-sus the amount that actually ends up trans-ferring to the golfer (as measured bydosimetry).

A transfer factor of 1,800 hasbeen calculated for carfentrazone. We willcompare the biomonitoring and dosimetryresults for these reduced risk compoundswith those previously determined forchlorpyrifos, carbaryl, cyfluthrin, 2,4-D,MCPP, dicamba, chlorothalonil and imida-cloprid . This season (2008) we determinedexposure in 24 rounds of golf followingapplication of azoxystrobin without post-application irrigation.

Hazard quotients (HQs) less than orequal to 1.0 indicate that the exposureresulted in a pesticide dose at whichadverse effects are unlikely. To date, HQsdetermined for chlorpyrifos, carbaryl,

cyfluthrin, 2,4-D, dicamba, MCPP,chlorothalonil, imidacloprid, and carfen-trazone have been 20- to 1.25 million-foldbelow 1.0, indicating safe exposure levelsusing the EPA Hazard Quotient criteria.

The hazard quotient calculatedfor carfentrazone was 0.0000008, 1.25-million fold lower than a hazard quotientof 1.0. This HQ was significantly lowerthan those calculated for 2,4-D, MCPP,and dicamba (0.001 - 0.01). It is importantto note that carfentrazone is considered areduced-risk replacement for these phe-noxy acid herbicides. The TF model willstill allow us to calculate a hazard quotientin pesticides that are not amenable to bio-monitoring.

Utilizing Reduced-risk Pesticides and IPM Strategiesto Mitigate Golfer Exposure and Hazard


J. Marshall Clark, Raymond Putnam, and Jeffery DohertyUniversity of Massachusetts

Objectives:1. Determine the level of hazard of volatile and foliar dislodgeable residues of the reduced-risk pesticides carfentra-zone, halofenozide, and azoxystrobin following total course and full-rate applications.

2. Determine the effect of partial course application strategies (e.g. tees and greens only) and post application irriga-tion on volatile and foliar dislodgeable pesticide residues following full-rate applications of carfentrazone,halofenozide, and azoxystrobin.

3. Model the relationship of volatile and dislodgeable foliar residues vs. actual golfer exposure using urinary biologi-cal monitoring techniques or, for pesticides that are not amenable to biomonitoring, dosimetry techniques.

Summary PointsResearchers have evaluated exposure in

24 rounds of golf following the applicationof azoxystrobin (2008) and will comparethis with future results from halofenozide(2009) and with those results of previousexperiments on chlorpyrifos, cyfluthrin,carbaryl, chlorothalonil, 2,4-D, MCPP-p,dicamba, imidacloprid, and carfentrazone.Determination of golfer exposure to

reduced-risk pesticides will provide anovel dataset for these IPM-friendly com-pounds.


To determine precisely how much of the environmental residues is actually transferred to golfers during a round ofgolf, volunteers are used for dosimetry (measuring pesticide residues on full-body cotton suits and personal air sam-plers) and biomonitoring (measuring urinary metabolites).

The loss of pesticides and nutrients

into surrounding bodies of water and theresulting decreases in water quality has ledto the use of best management practices ongolf courses. One such practice is the useof vegetative filter strips (VFS) to interceptrunoff water and thus prevent its loss andthe loss of any associated pesticides andnutrients to surrounding water bodies.

Joint greenhouse and field studieshave been implemented to evaluate select-ed plants for their effectiveness in remov-ing pesticides and nutrients from turfgrassrunoff waters that enter vegetative filterstrips (VFS). Ten plant species were evalu-ated in a greenhouse pot study to deter-mine which species most effectivelyremove six pesticides (2 fungicides, 2 her-bicides, and 2 insecticides) from a silt loamsoil. Five species (big blue stem, blue flagiris, eastern gama grass, prairie cord grass,and woolgrass) were determined to bemost effective.

A run-on plot, consisting of 12VFS, was established. Each VFS had a 5%slope and was lined with an impermeableliner. A manifold was placed on the front(top) edge of each VFS to evenly applyrun-on water using a solvent pump deliv-

ery system. At the bottom of each VFS,runoff water was collected. Lysimetersnear the bottom of each strip sampled sub-surface water. A bromide tracer studydetermined any hydraulic differencesbetween VFS prior to planting. VFS werethen established in replicates of three(unvegetated, random mixture of plants,succession of plants, and turfgrass cut tothree heights).

The six pesticides used in thegreenhouse study, plus cyfluthrin, will beused in the VFS run-on field trials (grow-ing seasons 2009 and 2010). Pesticideswill be applied with a water volume thatwould be generated for both a 1-year and a5-year rain event. Bromide will also beadded to the pesticide containing water at 4g/L as a tracer.

Several storm/run-on scenarioson the bare (pre-planted) VFS were evalu-ated. The volume of runoff water appliedas runon to each VFS was based on a 1-year rain event. The runoff water generat-ed during a 1-year rain event was calculat-ed to be 25.4 gallons over the course of 24hours from an turfgrass area 3 feet by 20feet with a 5% slope. This water volume isthen applied to the top of the VFS as run-on water. Soil presaturation was achievedprior to the initiation of the storm event.

An artificial rainfall system hasbeen constructed similar to those used inprevious USGA-funded runoff studies in

Minnesota. This system provides uniformdroplet size with low kinetic energy. Thestorm scenario selected for the 1-year rainevent and the initial bromide tracer studieswas as follows: Artificial rain for 6 hourstotal (6 am - 12 noon, for ~2.4 inches totalrainfall, 0.4 inches/hr) and run-on for 2hours (11 am - 1 pm @ 12.7 gal/hour). Inthe case of the 5-year rain event, thiswould involve adding 3.8 inches of wateras rain over 24 hours and 62.1 gallons ofwater as runoff over 24 hours.

The mass of pesticide lost will beevaluated using the concentration of thepesticide and the volume of water collect-ed during runoff. In addition to pesticides,runoff water will be monitored for lossesof nitrogen and phosphorus from fertilizerinputs. Soil, soil water, and plants withinthe VFS will be analyzed to determine ifthe pesticides lost from the runoff waterare sorbing to the soil, being degraded inthe soil, taken up by the plants, or lost toleaching or subsurface flow. Values will becompared against the bromide tracer,which will move freely with the run-onwater.

Soil sampling will be conductedat three different depths at 3 locationswithin the VFS (0.3 m, 1.83 m, and 3.35 mfrom the top of each strip). Even chemi-cals that sorb tightly to the soil can befound deeper in the soil profile than wouldbe expected based on their physical andchemical properties because of preferentialflow pathways.

Optimization of Vegetative Filter Strips forMitigation of Runoff from Golf Course Turf


J. Marshall Clark, Jeffery J. Doherty, Guy R. Lanza, and Om ParkashUniversity of Massachusetts

Objectives:1. Use selected plant species in a field study to evaluate the efficacy of vegetative filter strips (VFS) and their mosteffective arrangement.

2. Determine the fate of pesticides retained in VFS and the major mechanisms of degradation.


Summary PointsA pump-driven delivery system has

been fabricated to deliver run-on wateronto the VFS.

Nine VFS have been planted (3 VFSwill remain unvegetated) with either turf-grass or the five plant species selectedfrom the greenhouse study and allowed tomature.

An artificial rainfall simulator systemhas been fabricated and installed at therun-on plot.

56

Soil, soil water, and plants within the VFS will be analyzed to determine if the pesticides lost from the runoff waterare sorbing to the soil, being degraded in the soil, taken up by the plants, or lost to leaching or subsurface flow.

USGA initially funded research at

Michigan State University to determinenitrogen fate and leaching from aKentucky bluegrass turf in 1991. Similarto previous research, the initial research atMSU conducted from 1991 through 1993indicated that there was minimal risk ofnitrate-nitrogen leaching from Kentuckybluegrass (Poa pratensis L.) turfgrass.

Since the summer of 1998, perco-late samples have been collected from thesame monolith lysimeters and analyzed fornitrate-nitrogen (NO3-N). As of 2008, the

turfgrass area has now been under continu-al fertilization practices for 18 years withpercolate collection for the last 10 yearsconsecutively.

From July 1998 through 2002,lysimeters were treated annually with ureaat a low N rate 98 kg N ha-1 (4 applica-tions) and a high N rate of 245 kg N ha-1 (4applications). From 1998-2002 for thehigh N rate, there was a dramatic increasein NO3-N leaching from 5 mg L-1 in 1998

to 25 mg L-1 in 2002. During the sametime frame, there was a modest increase inNO3-N leaching from 3 mg L-1 in 1998 to

5 mg L-1 in 2002. In 2003, the N rate wasreduced to 196 kg N ha-1 for the high Nrate, while the low N rate remained at 98kg N ha-1.

Since 2003, phosphorus fromtriple superphosphate (20% P) has beenapplied at two rates, 49 and 98 kg P ha-1

split over two applications. The phospho-rus application dates coincide with nitro-gen application dates in the spring andautumn.

In 2003, the concentration ofNO3-N leaching from the high N rate treat-

ment did not decline from the previousyears. The average NO3-N concentration

leached from the low and high N rate treat-ments was 6.3 and 31.6 mg L-1. In 2004,the concentration of NO3-N leaching from

the high N rate treatment declined drasti-

cally from previous years. The averageconcentration of NO3-N in leachate for the

high N rate was 8.5 mg L-1. This was adecrease in NO3-N concentration of 23.1

mg L-1 from 2003. For the low N rate theaverage concentration of NO3-N in

leachate for the low N rate was 1.2 mg L-1.From 2004 through 2008, the mean NO3-N

concentration for the low and high N rateswas 2.9 and 9.4 mg L-1.

The concentration of phosphorusdetected in leachate remains very lowregardless of treatment. The mean concen-tration of phosphorus detected in leachatesince initiating phosphorus treatments in2003 has been less than 0.02 mg L-1.

Nitrogen and Phosphorus Fate in a10+ Year Old Kentucky Bluegrass Turf

Start Date: 2003Project Duration: five yearsTotal Funding: $68,886

Kevin W. FrankMichigan State University

Objectives:1. Determine nitrate-nitrogen and phosphorus leaching from a turfgrass stand that has been continually fertilized for18 years.

2. Continue data collection from the Long Term Nutrient Fate Research Area at MSU; currently we have data collec-tion for 10 years consecutively.

Dr. Kevin Frank (above right) explains to members of USGA’s Research Committee the efficiency of nitrogen useby mature Kentucky bluegrass when fertilized at a low (2 lb./1000 ft2/year) and a high rate (5 lb./1000 ft2/year).Results indicate that the high rate of nitrogen fertilization is much more than the turf needs and can result in unac-ceptable levels of nitrate-nitrogen in leachate.


Summary PointsThe mean NO3-N concentration from

2004 through 2008 is less than 10 mg L-1.

Results continue to indicate lowamounts of phosphorus leaching.

57

Since the summer of 1998, percolate samples havebeen collected from the same monolith lysimeters andanalyzed for nitrate-nitrogen (NO3-N).

Little is known about the fate and

transport of emerging contaminants suchas pharmaceuticals, personal care prod-ucts, and endocrine-disrupting compounds(PPCP/EDCs) from reuse water in land-scape systems receiving reuse waterthrough irrigation. This project will inves-tigate the fate and transport of PPCP/EDCsin turf with the goal to evaluate the effec-tiveness of turf in mitigating their verticalmovement (i.e., leaching).

Studies to determine the stabilityof the target compounds in wastewaterwere initiated. Samples were taken at t=0and regular intervals thereafter to be ana-lyzed for the target compounds.Simvastatin hydroxy acid and estrone wereremoved from the target list due to their

historically low concentrations in post-UVwastewater effluent. In addition, sul-famethoxazole, phenytoin, primidone,TCPP, and TCEP were added due to theirhigh concentrations in post-UV waste-water effluent.

It was found that all compoundswere stable for the first 10 days. However,by day 20, atenolol, fluoxetine, diclofenac,

and naproxen began to show decreases inconcentration, indicating degradation hadbegun to occur. Therefore, it is recom-mended that the wastewater be sampledduring the initial filling of the wastewaterstorage containers to determine analyteconcentrations and then replaced every 2to 3 weeks to avoid changes in analyteloading.

Studies were also undertaken toidentify any effects that the lysimeter com-ponents would have on the concentrationof the target analytes. Deionized water thathad been spiked with the target analyteswas poured into a beaker containing ~400

g of diatomaceous earth and allowed tostand for three days. Samples were takendaily and analyzed to determine the degreeto which the target analytes adsorbed ontothe diatomaceous earth. Only fluoxetinewas found to be affected by the diatoma-ceous earth, decreasing from 52 ng/L to 3.1ng/L over the course of three days.Therefore, any fluoxetine degradationobserved in the study should be viewedaccordingly.

Attenuation of Pharmaceuticals, Personal CareProducts, and Endocrine-disrupting Compounds

by Golf Courses Using Reuse Water


Setting the lysimeters into the holes at UCR.Researchers will check leachate collection equipmentfrom each plot and identify 8 plots for use in theresearch commencing in the summer of 2009.

Mike McCulloughNorthern California Golf Association

Objectives:1. To understand the fate and transport of PPCP/EDCs in golf courses when reuse water is used as the sole sourceof irrigation water.

2. To evaluate the capacity of turfgrass as a biofilter to remove PPCP/EDCs in reuse water.3. To extend knowledge to stakeholders and reduce the uncertainty of how long-term use of reuse water for irriga-tion may impact groundwater quality.


Summary PointsWe have purchased non-labeled and

13C-labeled standards for all the candidatecompounds listed in the proposal and areclose to establishing analytical conditionsfor these compounds.

We are developing sample preparationmethods and evaluating methodperformance.

We have received two soils that will beused for the laboratory-scale experiments.We plan to develop and finalize the exper-imental design for these laboratory-scaleexperiments in the second quarter.

Efforts were devoted to assisting in thecollection of soil in Eldorado Valley forthe meso-scale experiments.

The lysimeter plots at UCR are cur-rently being renovated. New sod will beinstalled before spring of 2009. We plan tocheck leachate collection equipment fromeach plot and identify 8 plots for use in theresearch commencing in the summer of2009.

Meters were ordered from the vendorand shipped to Las Vegas. Equipmentchecked out correctly and installation atWildhorse Golf Course is scheduled forOctober 2008. Other flux meters are beingshipped in mid-October to golf courses inSan Jose and southern California withinstallation to follow.

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Compound ConcentrationSulfamethoxazole 1,500Atenolol 500Trimethoprim 16Fluoxetine 16Meprobamate 480Phenytoin 120Carbamazepine 170Diazepam 2.3Atorvastatin 7.7Primidone 130TCPP 1,900TCEP 470Gemfibrozil 2.3Bisphenol A <5.0Diclofenac 34Naproxen 14Triclosan 16

Analyte concentrations (ng/L) at t=0 in post-UV waste-water effluent

Audubon InternationalAudubon International strives to educate, assist, and inspire millions of people from all walks of life to protect

and sustain the land, water, wildlife, and natural resources around them. In 1991, Audubon International launched theAudubon Cooperative Sanctuary Program for Golf Courses (ACSP), an environmental education program designed tohelp golf courses play a significant role in enhancing and protecting wildlife habitat and natural resources. The ACSPprovides an advisory information service to help golf courses conduct environmental projects and achieve positive recog-nition for their efforts.

In addition, the Audubon Signature Program works closely with planners, architects, managers, and key stake-holders of new golf course developments to merge wildlife conservation, habitat enhancement, resource conservation,and environmental improvement with the economic agenda associated with the development. Involvement in the devel-opment stages of a project enables Audubon International to ensure that biodiversity conservation, environmental quali-ty, and sustainable management are built into the project and continue after construction is completed.

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The redesigned and greatly improved wash pad at PrairieDunes Country Club provides adequate collection and fil-tering of wash water. Identifying problems, choosing themost environmentally responsible treatment, and imple-menting a solution are a key focus area for AudubonCooperative Sanctuary Program for Golf Course members.

The Green Golfer Challenge gave the staff at PecanHollow in Plano TX an opportunity to introduce golfers toour efforts to become a Certified Audubon CooperativeSanctuary and to encourage them to be good stewardsof the environment as they played golf. Outreach proj-ects like this are simple way to get golfers involved andbuild support for the game, the environment, and theprogram.

Signage provides students and the general public witheducation about the inhabitants of the golf course. Signslike this one at Washington Reservation in Ohio, whenplaced strategically around the nine holes, provide infor-mation about birds and other wildlife that may be seen.Washington Reservation is the first Certified GolfSignature Sanctuary in Ohio.

Jennifer and Philip Nola were the 2007 Grand Prize win-ners of the Green Golfer Challenge, receiving 2 tickets tothe 2008 US Open at Torrey Pines Golf Course in LaJolla,California. Their home base is Farmingdale NY andhome course is Bethpage State Park.


Audubon International is a non-profit

environmental organization that envisionscommunities becoming more sustainablethrough good stewardship of the naturalenvironment where people live, work, andrecreate through responsible managementof land, water, wildlife, and naturalresources. Since 1991, it has worked inpartnership with the USGA to offer theAudubon Cooperative Sanctuary Programfor Golf Courses (ACSP), an award-win-ning education and certification programthat promotes ecologically-sound landmanagement and the conservation of natu-ral resources on golf courses. AudubonSignature Programs provide comprehen-sive environmental education and planningassistance to new developments.

Today, more than 2,200 golfcourses in 30 countries participate in theACSP for Golf Courses. More than half ofthose enrolled have developed an environ-mental plan to guide management of thegolf course and 723 have achieved certifi-cation for their outstanding best practices.We also awarded 664 "Certificates ofAchievement" to recognize golf coursesfor outstanding accomplishments toimprove wildlife habitat, save water, con-serve energy, and reduce waste.

Audubon International providedenvironmental planning services to 161projects (126 golf-related) developmentprojects in 36 U.S. states and 10 countries,covering 75,000 acres of land, through theAudubon Signature Programs in 2007/08.Seventeen projects achieved certification,bringing the total number of golf coursesthat have been designated as CertifiedAudubon Signature Sanctuaries to 70 in 22US states and 3 countries.

An analysis of environmental

improvements on certified golf coursescompared with golf courses that areenrolled but not yet certified revealed sig-nificant differences. Certified courses are2-4 times more likely to be involved inenvironmental projects, such as exoticplant removal; prairie, wetland, and streamrestoration; and endangered or threatenedspecies reintroduction.

Efforts to enhance wildlife habi-tats on golf courses are done on a regionaland site-specific basis. In addition to par-ticipation in the program's environmentalmanagement and certification activities,golf courses are invited to participate inwildlife monitoring events and activities.Birdwatching teams from 72 golf courseseach identified an average of 46 birdspecies during the 2008 North AmericanBirdwatching Open, a 24-hour bird countconducted at the height of bird migrationin March, April, and May. (Start date basedon location.)

Audubon International encour-ages active participation in conservationprograms by golfers, golf course superin-tendents, golf officials, and the generalpublic through a variety of outreach andeducation activities. In 2005, AudubonInternational, the United States Golf

Association, and The PGA of Americalaunched the Golf and the EnvironmentInitiative to foster environmental aware-ness, action, and positive results through-out the golf industry. A Web site,www.golfandenvironment.org serves as aclearinghouse of golf and environmentinformation. In addition, a Green Golferpledge drive to engage golfers has beenlaunched. Currently there are over 6,400Green Golfers who have taken the pledgeand over 105 Golf Courses involved in theGreen Golf Challenge.

Over 2000 people took part inseminars and field training conducted byAudubon International staff in 2007/08.More than 500 conservation organizationsare directly involved with golf courses as aresult of their participation in the ACSP.

The Audubon Cooperative Sanctuary Program for Golf Courses

Start Date: 1991Project Duration: ongoingTotal Funding: $100,000 per year

Ron DodsonAudubon International

Objectives:1. Enhance wildlife habitats on existing golf courses by working with golf course superintendents and providingadvice for ecologically sound course management.

2. Encourage active participation in conservation programs by golfers, golf course superintendents, golf officials,and the general public.

3. Recognize the people who are actively participating in environmentally responsible projects.4. Educate the public and golfing community about the benefits of golf courses and the role they play relative to theenvironment and wildlife.

Summary PointsNumber of golf courses enrolled in the

Audubon Cooperative Sanctuary Program(ACSP): 2,205Number of golf courses enrolled in the

Audubon Signature Program: 237Total number of acres registered in

ACSP for Golf Courses: 519,474Number of conservation organizations

directly involved with golf courses as aresult of their participation in the ACSP:>500Average number of bird species sighted

on 72 golf courses participating in the2008 North American Birdwatching Open:46

Number of Certified AudubonCooperative Sanctuary Golf Courses: 80Total number of golf courses certified

in Environmental Planning: 1,780Number of golf course personnel and

others educated by Audubon Internationalin 2007/08 through seminars, conferencepresentations, and site visits: >2,000


Southern Chorus Frog. Out of Golf Digest America's100 Greatest Golf Courses, 56% are members of theAudubon Cooperative Sanctuary Program for GolfCourses and undertaking voluntary action to protectand enhance wildlife habitats. Photo Credit: MarvinBouknight, Oldfield Naturalist, SC

Grant-in-Aid Research Program

USGA’s Turfgrass and Environmental Research Program relies on science to attainanswers that will help ensure the long-term success of the golf course management industry.Frequently projects may span several years. Many times, however, golf course superintend-ents need answers to very applied problems to help them meet the many challenges of prop-erly managing golf courses. The Grant-in-Aid Research Program was created to address thisneed. This program allows directors of all eight USGA Green Section regions to identifyapplied problems and the appropriate researchers in their regions to solve those problems.Research projects funded under this program most often include cultural aspects of golfcourse management. Examples include what fungicides work best on a particular disease, orthe management of new turfgrass cultivars, renovation techniques, safe and effective use ofherbicides, insecticides, or fertilizers. These projects are usually short duration (one to threeyears), but can offer golf course superintendents answers to practical, management-orientedchallenges that they can put into use quickly.

Location of current projects funded in 2008 by the USGA Turfgrass andEnvironmental Research Program under the Grant-in-Aid Program

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Physical water conditioning units such

as AquaPhyd, Carefree, Fre-Flo,Magnawet, and Zeta Core have been usedin the golf course industry because of man-ufacturers' claims that they improve turfquality and reduce scaling, compaction,the effect of salts on soil physical andchemical characteristics and plant growth,and reduce the amount of irrigation waternecessary for adequate turfgrass growthand quality. These products are gainingpopularity, particularly in the Southwest,despite the lack of scientific evidence tosupport the manufacturers' claims.

A study conducted at NewMexico State University in spring andsummer of 2008 investigated the effect ofphysical conditioning systems on turfcolor, quality, and cover, and chemicalrootzone composition of a perennial rye-grass stand irrigated with either saline orpotable water. The soil at the site is aTorriorthent, a sandy entisol typical of aridregions.

Treatments consisted of 2 levelsof irrigation water quality and 5 condition-ing products. During 2008, irrigation was

applied daily with either potable water(TDS 397 ppm, SAR 1.55, pH 8) atapproximately 90% ETo or saline water

(TDS 3,110 ppm, SAR 8.94, pH 7.5) atapproximately 125% ETo. Conditioning

treatments included two catalytic waterconditioners (Zetacore and FreFlo), a mag-netic water conditioner (Magnawet), ahydro-electrical conditioner (Aqua-PhyD),and a control treatment.

The area was established in 2007with ‘IG2’ perennial ryegrass. Data col-lection included turfgrass color and cover(measured by digital image analysis), visu-al assessment of turfgrass quality, salinitybuild-up in the rootzone, and turfgrassstress as measured by NormalizedDifference Vegetation Index (NDVI).

The water conditioning units hadno effect on turfgrass quality and onlysaline water reduced quality significantly.When plot coverage and turf color datawere averaged for water qualities,Magnawet and Zetacore plots had lowercoverage and lighter green color than con-trol plots or plots irrigated with Fre Flo-treated water at the end of the growing

period (November 2008). Aqua-PhyDplots did not differ from control plots forcoverage or color.

There was an obvious and expect-ed difference in soil salinity between plotsirrigated with potable water and those irri-gated with saline water. All the treatmentsthat received saline water showed higherlevels of salts compared to treatmentsreceiving potable water. However, waterconditioning had no effect on the chemicalcomposition of the rootzone.

Assessing the Usefulness of Physical Water Conditioning Productsto Improve Turfgrass Quality and Reduce Irrigation Water Use


A study conducted at New Mexico State University in spring and summer of 2008 investigated the effect of physi-cal conditioning systems on turf color, quality, and cover, and chemical rootzone composition of a perennial rye-grass stand irrigated with either saline or potable water.

Bernd Leinauer, Ty Barrick, and Cody RobertsonNew Mexico State University

Objectives:1. To assess the effect of three water conditioning systems on perennial ryegrass establishment, overall turfgrass

performance, and root distribution under potable and saline irrigation.2. To study the impact of water conditioning units on salinity build-up in the rootzone.


Summary PointsStudies were conducted at New

Mexico State University to assess physicalwater conditioning products under potableand saline irrigation.

Products tested had no effect on thequality of a perennial ryegrass stand.

At the end of the 2008 growing period,Zetacore- and Magnawet- treated plots hadlower coverage and lighter green colorthan control plots.

Physical water conditioning had noeffect on the chemical composition ofeither saline or potable irrigated rootzones.

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Conditioning treatments included two catalytic waterconditioners (Zetacore and FreFlo), a magnetic waterconditioner (Magnawet), a hydro-electrical conditioner(Aqua-PhyD), and a control treatment.

Turfgrass quality on golf course put-

ting greens receiving few hours of sunlightor exposed to prolonged periods of exces-sive moisture is often reduced for a num-ber of reasons. Shaded environments gen-erally are associated with poor air move-ment, excessive soil moisture, and extend-ed periods of leaf wetness. Even greenslocated in excellent growing environ-ments, but built with no or poor internaldrainage, are susceptible to thinning dur-ing the stressful summer months. Stress-related injury to the turfgrass often leads toan increased susceptibility to other turf-grass disorders including algae and mossinvasion, as well as infection from variousturfgrass pathogens.

Algae infestation on closelymown putting greens continues to be diffi-cult to manage. While applications of var-ious fungicides such as chlorothalonil,maneb, and mancozeb have been shown tosuppress algae, these products must beapplied on relatively short intervals priorto the appearance of symptoms and theirefficacy varies. Further complicating theuse of fungicides is the exclusion ofchlorothalonil from use in specific regionsof New England. In areas wherechlorothalonil use remains legal, new labelrestrictions on the total amount of productthat can be applied and increased timebetween application intervals may drasti-cally limit the ability to control algae overthe course of an entire season.

Previous studies revealed a reduc-tion in algae growth from phosphite-basedfungicides and select wetting agents.Although the main purpose of these studiesdid not involve an investigation of algae,products suppressed algae between 83 and100% when compared to the untreatedcontrol. Little information is availableregarding the influence of phosphite prod-ucts and wetting agents for controlling

algae. The objective of this study will beto determine the ability of various phos-phites and wetting agents to suppressalgae; and 2) assess the interaction amongeffective algae control products and fungi-cides commonly used on golf course put-ting greens.

Studies were initiated at theUniversity of Connecticut Plant ScienceResearch and Education Facility located inStorrs, CT. When applied preventively,most phosphite treatments, as well asDaconil, reduced algae populations whencompared to the untreated control. Whensingle applications of phosphites andfungicides were applied as a curative treat-ment, algae development was not reducedor slowed and no differences wereobserved among treatments.

In addition to the impact of phos-phites, the interaction among fungicidesand wetting agents was assessed. The leastamount of algae was observed within plotstreated with Insignia + Magnus, Daconil +Revolution, and Daconil alone. Moderatesuppression of algae was exhibited withinplots treated with Insignia + Revolution,Protect + Dispatch, Trinity + Magnus,Daconil + Duplex, and Protect + Duplex.No other treatment provided a reduction ofalgae compared to the untreated controlplots.

Although various treatments pro-vided suppression of algae, it appeared that

applications of each product alone provid-ed varying results than when the productswere tank-mixed with different wettingagents. A final greenhouse study investi-gated the influence of nitrogen source onalgae development. In this study, only potsreceived ammonium sulfate (21-0-0), cal-cium nitrate (15.5-0-0), and ammoniumnitrate (34-0-0) significantly reduced algaewhen compared to the untreated control orother nitrogen sources.

Additional field and greenhousestudies are planned for 2009 to furtherinvestigate the preventive and curativesuppression of algae, as well as any inter-actions with other commonly applied prod-ucts. Laboratory studies will also bedeveloped to assess the direct inhibition ofalgae from phosphorous acid and variouscommercially available phosphiteproducts.

Preventive and Curative Control of Algae onPutting Greens Using Products Other than Daconil


John E. KaminskiUniversity of Connecticut

Objectives:1. To determine the influence of various products on algae control of golf course putting greens.

The influence of fungicides on algae suppression may be influenced by other tank-mixed partners such as wettingagents.


Summary PointsAlgae growth is a chronic problem on

thinning golf course putting greens duringthe summer months.

Current data suggest that most phos-phite products can reduce algae whenapplied preventively, but curative controlmay be more difficult.

The influence of fungicides on algaesuppression may be influenced by othertank-mixed partners such as wettingagents.

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Common bermudagrass is a trouble-

some weed in cool-season roughs and fair-ways. Several programs have been recom-mended for common bermudagrass sup-pression in cool-season turfgrass, howevercomplete control is rarely achieved andhighly variable in field situations.

Four treatments of fenoxaprop-Pplus triclopyr applied monthly providedacceptable suppression of commonbermudagrass in trials in North Carolinaand California. Researchers in Georgiasuppressed bermudagrass in tall fescuewith fenoxaprop and ethofumesate combi-nations. None of these programs resultedin complete control, only suppression.

Research at Virginia Tech foundthat concentrating herbicide applications inspring and fall at three-week intervals con-trolled bermudagrass and avoided herbi-cide treatment during summer when injuryto cool-season turf is more likely. This

research also incorporated mesotrione as atank mix partner to fenoxaprop-P and tri-clopyr. The addition of mesotrioneimproved bermudagrass control over theproducts alone, and all three products tank-mixed controlled bermudagrass 98%.

The limitation of Virginia Tech'sfindings is that better performing programsexceed labeled maximum annual productuse restrictions. Combinations that maxi-mize bermudagrass control and do notexceed these annual use limits are needed.There are only five herbicides that canselectively suppress bermudagrass inKentucky bluegrass. These include

fenoxaprop (Acclaim Extra), ethofumesate(Prograss), siduron (Tupersan), triclopyr(Turflon), and mesotrione (Tenacity).

The experimental location isKentucky bluegrass rough that was infest-ed with 'Vamont' common bermudagrasssod 4 years ago. Experimental design israndomized complete block with 3 replica-tions and 6 by 10 ft. plots. Data collectedwill include bermudagrass and other weedcontrol and cover, turfgrass injury, color,and quality. The duration of this trial willbe from spring 2008 to summer 2009 toevaluate treatment effects 1.5 years aftertreatment initiation.

Preliminary results indicate thatseveral treatment scenarios have signifi-cantly reduced bermudagrass cover whileincreasing Kentucky bluegrass cover. Veryfew treatments significantly injuredKentucky bluegrass. However, treatment 3controlled bermudagrass greater than 97%;this treatment exceeds label restrictions forannual usage rates. The modified VirginiaTech programs (Treatments 6 and 7) havealso controlled bermudagrass equivalent totreatment 5. Treatments 5, 6, and 7 all con-trol bermudagrass significantly better thanthe industry standard (treatment 4).

Bermudagrass Control Programs in Kentucky Bluegrass

Start Date: 2008Project Duration: one yearTotal Funding: $3,000

Herbicide treatments used in the bermudagrass control study at Virginia Tech.

J. B. Willis and S. D. AskewVirginia Polytechnic and State University

Objectives:1. Develop effective bermudagrass control programs utilizing multiple active ingredients, avoiding summer stress,and targeting bermudagrass throughout its growing season.

Treatment combinations containing ethofumesate and triclopyr applied in summer significantly injured Kentuckybluegrass.


Summary PointsTreatment combinations containing

ethofumesate and triclopyr applied in sum-mer significantly injured Kentucky blue-grass.

Several treatment combinations wereshown to control bermudagrass better thanthe current industry standard.

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Herbicide Treatments1) Nontreated control no herbicides applied.2) Negative control for bermudagrass industry standard treatments for broadleaf and summer annual grassweed control, three-way plus crabgrass preemergence control product both at labeled rates and timings.3) Industry Standard Renovation fall renovation using glyphosate spot-treatment according to label recommen-dations (2 apps. at 5 qt/A each at 3 wk interval and reseed).4) North and South Carolina recommendation 4 applications of fenoxaprop-P at 28 fl oz/A plus triclopyr at 32 floz/A applied at monthly intervals starting June.5) Virginia Tech top 3-Way 3 applications of mesotrione at 4 fl oz/A plus triclopyr at 32 fl oz/A plus fenoxaprop-P at 28 fl oz/A at 3 week intervals both spring and fall.6) VT modified 1 2 applications of fenoxaprop-P at 28 fl oz/A plus triclopyr at 32 fl oz/A applied at 3 week inter-vals both spring and fall, with mesotrione at 4 fl oz/A at 3 week intervals throughout summer (not to overlap withfenoxaprop-P plus triclopyr applications)7) VT modified 2 2 applications of fenoxaprop-P at 28 fl oz/A applied at 3 week intervals both spring and fall,with mesotrione at 4 fl oz/A plus triclopyr at 16 fl oz/A at 3 week intervals throughout summer (not to overlapspring and fall applications).8) 5 herbicide combo fenoxaprop-P at 28 fl oz/A plus triclopyr at 32 fl oz/A plus ethofumesate at 64 fl oz/A plussiduron at 24 lbs/A plus mesotrione at 4 fl oz/A applied 2 times spring and fall at three week intervals.9) 5 herbicide sequence 1 fenoxaprop-P at 28 fl oz/A plus ethofumesate at 64 fl oz/A plus siduron at 24 lbs/Aapplied 2 times spring and fall at three week intervals, and triclopyr at 32 fl oz/A plus mesotrione at 4 fl oz/Aapplied 4 times in summer at 3 week intervals (not to overlap spring and fall applications).10) 5 herbicide sequence 2 fenoxaprop-P at 28 fl oz/A plus ethofumesate at 64 fl oz/A plus siduron at 24 lbs/Aapplied 4 times in summer at three week intervals, and triclopyr at 32 fl oz/A plus mesotrione at 4 fl oz/A applied2 times spring and fall at 3 week intervals.11) 4 herbicide combo same as #6 but drop siduron to save costs.12) 4 herbicide sequence 1 same as #7 but drop siduron.13) 4 herbicide sequence 2 same as #8 but drop siduron.

Aggressive organic matter dilution

programs are intended to slow loss of aer-ation porosity and subsequent infiltrationrates thereby allowing superintendents tomore easily manage their putting greensand lessen the effects of summer bentgrassdecline. Our research compared variouscultivation approaches that removed from10 to 27% surface area to determine theeffects on agronomic performance of amature putting green in east-centralVirgina.

We used a mix of tine sizes andverticutting to achieve a range of surfaceremoval. Our research was done on mature(8-yr-old) ‘Penn A4’ greens that are inplay. Our treatments were imposed on twopractice putting greens at theIndependence Golf Club, the home courseof the Virginia State Golf Association nearRichmond.

Prior to initiation of the study,analysis of 4 randomly-selected cup cuttercores revealed a mat layer with 5.8%organic matter and an infiltration rate of 11inches/hr. Various combinations of smalltines (0.25"), big tines (0.50"), and verti-cutting (3-mm blade) were imposed in lateMarch and early September to provide a

range of seasonal surface removal from0% to 26.6% (Table 1). Verticut bladespacing and depth was 1 inch, while tinespacing was 1.33" X 1.5", with a coringdepth of 2".

Heavy sand topdressing ofapproximately 12 ft3 (1,200 lbs/1000 sq.ft.) was applied on both days of cultiva-tion, supplemented by six light topdress-ings of 2 ft3 (200 lbs/1000 sq. ft) every 3weeks between cultivations for a seasonaltotal of about 36 ft3. Cultural managementof these greens were identical to all otherson the golf course, receiving preventivepesticide applications, daily mowing at0.115", andApril through September fertil-ization of 1.6 lbs N/1000 sq. ft.

Cultivation treatment had noeffect on soil temperature, soil moisture, orball roll distance throughout the summerseason. Close-up pictures were taken fordigital image analysis of recovery rate, butanalysis is still in process. Visual qualityobservations indicated almost completerecovery of cover at 2 weeks after coringin the spring for treatment 2, where 0.25"tines were used in 2 passes to remove 5%surface area. All other spring treatmentswere more aggressive, removing 10 to14%; these treatments required 8 to 10weeks for recovery.

Fastest fall recovery was againseen with the small tines (0.25").Interestingly, fill-in was just as fast

when 2 passes were madewith the ¼-inch tines aswhen only 1 pass wasmade. This observationagrees with others whoobserved that recoverywas only affected by tinediameter and not by tinespacing. Cup-cutter plugsfor quantification of matlayer percent organic mat-ter were taken in lateOctober and have yet tobe processed.

Organic Matter Dilution Programs for Sand-based Putting Greens


Table 1. Cutivation treatments and precent surface area removed.

Erik Ervin and Brandon HorvathVirginia Polytechnic Institute and State University

Objectives:To compare various cultivation approaches that remove from 10 to 27% surface area and determine treatmenteffects on agronomic performance of a mature putting green in east-central Virginia. The ultimate goal is to deter-mine which organic matter dilution program maintains mat layer organic matter at less than 4% while providing thefewest days of putting quality disruption each year.

Cultivation treatments being applied on Sept 4. Leftlane shows a 0.25" tine treatment; middle lane shows a0.5" treatment; right lane shows a 0.25" coring + verti-cutting treatment.


Summary PointsPrevious transition zone research at the

University of Arkansas on a relativelyyoung (2-yr-old) ‘Penn G2’ sand-basedgreen indicated that aggressive annualremoval of 15-23% surface area via verti-cutting was required to keep mat layerorganic matter below 4%. However,recovery of putting green uniformity tookat least twice as long as coring.

Our transition zone study in Richmond,Virginia is being conducted on 8-yr-old‘Penn A4’ sand-based putting greens thatare in-play, and began the trial at 5.8%organic matter (above the USGA recom-mended mat layer limit of 4%).

Closely spaced core aeration, with orwithout verticutting treatments, wereimposed spring and fall so that annual sur-face area removal of plots ranged from10% to 27%.

No overall bentgrass decline was notedduring the 2008 growing season and noneof the treatments affected soil temperature,soil moisture, or ball roll distance.

Faster recovery was seen on those plotswhere surface area removal was the least.That is, where ¼-inch tines were used atclose spacing (1.33" x 1.5"), and the greenwas either cored once or twice.

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Jason HendersonUniversity of Connecticut

Surface Area Removed (%)Treatment March 28 Sept 4 Total

1. Untreated check 0 0 02. 0.25” tine core aerations X2 5 5 103. 3-mm blade verticultting 11.8 11.8 23.64. 0.25” tine core aeration + 2.5 2.53-mm blade verticultting 11.8 16.8

5. 0.5” tine core aeration + 9.80.25” tine core aeration X2 5 14.8

6. 0.5” tine core aeration 9.8 9.8 19.67. 0.5” tine core aeration + 9.8

3-mm blade verticultting + 11.80.25” tine core aerations X2 5 26.6

Fairway topdressing is a relatively new

cultural practice that is being adopted byseveral golf course superintendentsthroughout the United States to improveplaying conditions. Some of the benefitsreported have been improved drainage,less disease, and firmer fairways. The ben-efits to fairway topdressing seem unani-mous, but the practice requires a signifi-cant budget, considerable labor, time, andcommitment to implement properly.Additionally, many questions remainunanswered with regards to topdressingmaterial selection, application rates, andthe turfgrass management implications asthe topdressing layer accumulates.

Sands used in USGA puttinggreen construction and subsequent top-dressing have been thoroughly researchedto optimize macroporosity while maintain-ing sufficient water-holding capacity.However, due to the strict specifications,these sands are prohibitively expensivewhen considered for use on larger fairwayacreage. Therefore, recommendations forfairway sands are often very general.

The cost of a USGA sand doesnot improve the practicality of implement-ing this program for many golf courses.The impact of using sand that does notmeet USGA specifications, however, hasnot been thoroughly investigated. Particlesize distribution will likely affect infiltra-tion and water retention at the playing sur-face. Topdressing materials that are toofine may retain excess moisture, whereas,a sand that is too coarse may predispose alarge portion of the course to moisturestress. The short- and long-term impact oftopdressing native soils is unknown.

This study was initiated on an ‘L-93’ creeping bentgrass (Agrostisstolonifera) stand managed as a golfcourse fairway at the University ofConnecticut Plant Science Education andResearch Facility in the summer of 2007.The first factor is sand type; fine, medium,and coarse.. The second factor is applica-tion rate; 4 ft3/1000 ft2, 8 ft3/1000 ft2, and12 ft3/1000 ft2. A control is included thatreceives no topdressing applications.

Topdressing applications are keptconstant and are applied once per monthstarting in May and ending in November.This design allows the comparisons ofeach sand type applied at each of the threerates. The three different rates will also

enable the development of three differentdepths of topdressing over time.

Weekly data collection includesvolumetric soil moisture, soil penetrationresistance, turfgrass cover, turfgrass color,and turfgrass quality. Soil physical pro-prties will be determined on an annualbasis.

Impact of Sand Type and Application Rate ofFairway Topdressing on Soil Physical Properties,

Turfgrass Quality, Disease Severity, and Earthworm Castings


Topdressing applications are kept constant and are applied once per month starting in May and ending inNovember. This design allows the comparisons of each sand type applied at each of the three rates.

Jason J. HendersonUniversity of Connecticut

Objectives:1. Determine whether particle size distribution and/or application rate will affect turfgrass quality, disease incidenceand earthworm activity.

2. Quantify the effects of particle size distribution and topdressing layer depth on soil physical properties.3. Use the resultant data to make recommendations to improve the practice of fairway topdressing.


Summary PointsTopdressed plots showed a faster

green-up response than the untreated con-trol plots in mid-April regardless of sandtype. Plots that received higher rates ofapplication exhibited a greater greeningresponse than plots receiving lighter ratesof application.

Topdressed plots exhibited less dollarspot incidence than untreated plots. Plotsthat received higher rates of topdressinghad less dollar spots than plots thatreceived lower rates of topdressing,regardless of sand type.

Earthworm pressure was not sufficientto determine treatment differences.

Topdressed treatments had higherresistance to penetration than the untreatedcontrol plots demonstrating a firmer sur-face than the untreated controls. The finesand had the greatest resistance to penetra-tion, followed by the medium sand and thecoarse sand, respectively. Plots receivinghigher rates of topdressing exhibitedgreater firmness than plots receiving thelower rates.

Untreated controls had the highest vol-umetric soil moisture content in the top 2"of the playing surface compared to all top-dressing treatments. The fine and mediumsand treatments hold more water than thecoarse sand treatments. Regardless of sandtype, the higher the rates of application theless water is held in the top 2" of the play-ing surface.

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Growing concern over the sufficiency

and variability of present water supplies inwestern Texas and other areas of the aridsouthwest led to the examination of sever-al reduced-input turfgrass species for waterconservation. Buffalograss [Buchloedactyloides (Nutt) Engelm.] is a warm-sea-son turfgrass that shows excellent drought,cold, and salinity tolerance. Increasingawareness and acceptance of buffalograssas a viable turfgrass option in arid environ-ments requires investigation into conver-sion techniques for its establishment.

Research was conducted at theTexas Tech Turfgrass Research Stationduring the summer of 2008 on an estab-lished bermudagrass rough. The parame-ters evaluated included four seedbedpreparation treatments and two buffalo-grass seeding rates. Bermudagrass was

sprayed with glyphosate at 2.4 kg ai/ hausing small-plot spray equipment twoweeks prior to seedbed preparation. Allplots were scalped following herbicideapplication and subsequent desiccation ofbermudagrass.

Seedbed preparation treatmentsconsisted of verticutting in two directions,verticutting plus topdressing (0.6-cmlayer), verticutting plus topdressing plusaeration (hollow-tine), or no seedbedpreparation. 'Texoka' buffalograss wasexamined at seeding rates of 101 or 146kg/ha and was planted on June 17, 2008. Astarter fertilizer was applied at seeding andall plots were lightly brushed to ensuregood seed-to-soil contact. Plots were irri-gated daily by an automated irrigation sys-tem that applied approximately 3.8 cm ofwater per week. Plots were mowed once aweek to a height of 5.1 cm with a rotarymower.

Treatments were arranged in arandomized complete block design withfour replications of treatments.

Buffalograss conver-sion was visually eval-uated weekly for thefirst 2 months andmonthly thereafterusing a scale of 0 (nocover) to 100% (com-pletecover). Buffalo-grass turfgrass qualitywas assessed on a sim-ilar time frame using ascale of 0 to 9, where 9is considered to beoptimal turf qualityand 6.5 the minimumacceptable level.

Buffa lograssgermination was mini-mal regardless oftreatment and compe-tition from bermuda-grass regrowth be-came evident one

month after seeding. Bermudagrass hadrecovered fully from glyphosate applica-tions by the conclusion of the trial(October 15, 2008) and occupied a majori-ty of each plot. Differences in buffalograssestablishment did not exist between thetwo seeding rates regardless of seedbedpreparation treatment. However, verticut-ting, aeration, and topdressing treatmentsincreased buffalograss cover by 10% 2months after seeding compared to noseedbed preparation.

Percent buffalograss cover neverexceeded 15% regardless of seedbedpreparation or seeding rate treatments atthe conclusion of the trial. Due to low buf-falograss conversation, turfgrass qualityratings were not taken. Further research isnecessary to identify chemical treatmentsto reduce the regrowth potential ofbermudagrass and increase the establish-ment of buffalograss from seed.

Best Management Practices for the Conversionof Established Bermudagrass to Buffalograss


Seedbed preparation treatments consisted of verticutting in two directions, verti-cutting plus topdressing (0.6-cm layer), verticutting plus topdressing plus aeration(hollow-tine), or no seedbed preparation.

Gerald HenryTexas Tech University

Objectives:1. Examine the effect of seeding rate on the establishment of seeded buffalograss.2. Examine the effect of various seedbed preparation techniques (verticutting, aeration, and topdressing) on theestablishment of seeded buffalograss.


Summary PointsBermudagrass was difficult to control

chemically and grew back fromglyphosate treatments 3 to 4 weeks afterbuffalograss seeding.

Bermudagrass germination was mini-mal regardless of treatment and competi-tion from bermudagrass regrowth becameevident one month after seeding.

Differences in buffalograss establish-ment did not exist between the two seedingrates regardless of seedbed preparationtreatment.

Verticutting, aeration, and topdressingtreatments increased buffalograss cover by10% 2 months after seeding compared tono seedbed preparation.

Percent buffalograss cover neverexceeded 15% regardless of seedbedpreparation or seeding rate treatments atthe conclusion of the trial (October 15,2008).

Due to low buffalograss conversation,turfgrass quality ratings were not taken.

67

Chlorothalonil, propiconazole, and

iprodione are among the most popular andeffective fungicides for controlling foliardiseases of turfgrasses. Three fungicide tri-als were conducted on bentgrass main-tained as a golf course fairway at two dif-ferent locations in Pennsylvania (a golfcourse in the southeastern portion of thestate and a research facility in centralPennsylvania). Treatments included threeor four different products applied at thesame rate and timing for each active ingre-dient (chlorothalonil, propiconazole, andiprodione).

Results show differences in dis-ease control among active ingredients intwo of the three trials and a few differencesamong products with the same activeingredient in all three trials. At the south-eastern Pennsylvania site, all fungicidetreatments provided good to excellent con-trol of dollar spot. No differences in dollarspot incidence were observed betweenchorothalonil and propiconazole fungi-cides, or among the three chlorothalonilproducts (Daconil Ultrex, Echo Ultimate,and Chlorothalonil DF), or the four propi-conazole products (Banner MAXX,Propiconazole 14.3, Propensity 1.3ME,and Spectator Ultra 1.3).

Two of the three iprodione prod-ucts (Chipco 26GT and Ipro 2SE) per-formed similarly to the chlorothalonil andpropiconazole products with respect todollar spot control (no differences occur-ring on any rating date). Only one iprodi-one product, Raven, showed reduced effi-cacy compared to Chipco 26GT and Ipro2SE on two of the seven rating dates.

Area under the disease progresscurve (AUDPC) is a calculated value usedto assess disease epidemics for an entiretest period. The only fungicide treatmentwith a higher AUDPC value ( than all oth-

ers) was the Raven treatment.Two tests were conducted at the

central Pennsylvania site. Differences indollar spot severity were observed amongchorothalonil, propiconazole, and iprodi-one treatments, with iprodione generallyshowing better control than chlorothaloniland propiconazole over the test period. Nodifferences in control were observedamong the three chlorothalonil products(Daconil Ultrex, Echo Ultimate, and

Chlorothalonil DF) or the four propicona-zole products (Banner MAXX,Propiconazole 14.3, Propensity 1.3ME,and Spectator Ultra 1.3).

As in the southeasternPennsylvania trial, two of the three iprodi-one products (Chipco 26GT and Ipro 2SE)performed similarly with respect to dollarspot control. However, Raven showedreduced dollar spot control compared toChipco 26GT and Ipro 2SE on three of thenine rating dates. Area under the diseaseprogress curve values indicated the bestdollar spot control was achieved with theiprodione products, followed by propi-conazole and chlorothalonil products.

In the colonial bentgrass trial, allfungicide treatments were applied in Juneprior to brown patch symptom develop-ment. By mid-summer, differences inbrown patch severity were observedamong chorothalonil, propiconazole, andiprodione fungicides, with chlorothaloniland iprodione generally showing better

control than propiconazole. No differ-ences in brown patch control wereobserved among the three chlorothalonilproducts (Daconil Ultrex, Echo Ultimate,and Chlorothalonil DF) or three iprodioneproducts (Chipco 26GT, Ipro 2SE, andRaven) on any rating date or amongAUDPC values. Area under the diseaseprogress curve values indicate the bestbrown patch control was achieved with thechlorothalonil and iprodione treatments,followed by propiconazole treatments in2008.

Comparison of Chlorothalonil, Propiconazole, and IprodioneProducts for Control of Dollar Spot and Brown Patch Diseases


Peter Landschoot and Michael FidanzaPennsylvania State University

Objectives:1. To determine if different fungicide products containing chlorothalonil, propiconazole, or iprodione provide differ-ences in control of dollar spot and brown patch diseases of bentgrass maintained as a golf course fairway.

Comparative area under the disease progress curve(AUDPC) to control dollar spot for various fungicides


Summary PointsWith respect to dollar spot control,

iprodione products generally showed bet-ter control than chlorothalonil and propi-conazole products at the centralPennsylvania location. However, in thesoutheastern Pennsylvania trial, the threedifferent active ingredients were very sim-ilar in controlling dollar spot.

Generally, chlorothalonil and iprodioneproducts provided better brown patch con-trol than propiconazole products.

All chlorothalonil products included inthis study (Daconil Ultrex, Echo Ultimate,and Chlorothalonil DF) performed similar-ly in both dollar spot trials and the brownpatch trial.

No differences among propiconazole-containing products (Banner MAXX,Propiconazole 14.3, Propensity 1.3ME,and Spectator Ultra 1.3) were observedwith respect to dollar spot control in eitherdollar spot trial. However, AUDPC valuesindicated better brown patch efficacy forPropensity 1.3ME compared withSpectator Ultra 1.3 when evaluated overthe entire 2008 test period.

Of the three iprodione products(Chipco 26GT, Ipro 2SE, and Raven)Chipco 26GT and Ipro 2SE provided themost consistent dollar spot control. Ravenshowed less dollar spot control comparedwith Chipco 26GT and Ipro 2SE on sever-al rating dates and in AUDPC values inboth trials.

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In recent years there has been anincreasing frequency of occurrence of 'leafand sheath blight' caused by Rhizoctoniazeae (or related fungi) on bermudagrassputting greens. In South Carolina, R. zeaeis a well documented pathogen of creepingbentgrass putting greens, causing brown-patch like symptoms in the heat of sum-mer. The fungus has been identified andpathogenicity documented as well on St.Augustinegrass, centipedegrass, andseashore paspalum. In all cases, the diseasehas not been controlled with benzimida-zole fungicides, as R. zeae is essentiallyimmune to that chemistry.

Recent outbreaks have occurredon all of the common ultradwarf bermuda-grass cultivars on putting greens(‘TifEagle’, ‘Champion’, and ‘Mini-Verde’) as well as ‘TifDwarf’ and even‘TifGreen’ in the southeastern UnitedStates. Generally, the disease is firstnoticed in late August to early fall months.Initial symptoms are bronze patches of afew cm up to 100 or more cm in diameter.If not controlled quickly with fungicides,the pathogen blights and bleaches thelower leaves that results in a persistent dis-tinct patch symptom. Disease has been

known to recur in spring months after turfemerges from dormancy and presumablycan detrimentally affect spring transition inoverseeded systems.

In several instances, stressinduced by aggressive verticutting hasinduced severe outbreaks of persistent dis-ease symptoms. Low nutrition or reducedrecuperative potential by any of the rea-sons outlined would be expected toincrease disease severity. In the summerand fall of 2008, the disease again occurredon several golf courses in South Carolinaand throughout portions of the southeast-ern United States. Cultivars were‘TifEagle’, ‘Champion’, ‘TifDwarf’,‘MiniVerde’, and ‘TifGreen’.

Identification of R. zeae has beenbased solely on cultural characteristics,based primarily on sclerotia size and abun-dance. Recently, however, molecularmethods have been used to distinguishamong R. zeae, R. oryzae, and R. circinata.Cultures have been recovered from bermu-da patches that resemble (culturally) allthree of these fungi. Molecular identifica-tion of these isolates will be conductedduring the winter of 2008/09.

In 2007, three trials were con-ducted but went out curatively after symp-toms became well established. Trials wereplaced in late September (Experiment 1)and late October (Experiment 2) whensymptoms were severe and day length andtemperatures were not favorable forbermudagrass growth. In 2008, trials wereconducted on a chipping green in

Columbia, SC and a putting green nearFlorence. Intentions were to initiate thetrials in July prior to symptom expression,but symptoms already were present whenthe trials were established.

Only Heritage gave some controlin the single component fungicide trial. Inanother experiment, however, we looked ata 'program' approach of rotation of fungi-cides and fungicide mixtures with one setof treatments watered-in before spraysdried, and the other set left on the leaf. Wedid observe that watering-in the fungicidesmade a positive difference, and these trialsgave us some evidence that preventivetreatments have to be initiated much earli-er than we first supposed.

Identification, Pathogenicity, and Control of Leafand Sheath Blight of Bermudagrass Putting Greens


Bruce Martin and Dara ParkClemson University

Objectives:1. Collect isolates of R. zeae from symptomatic bermudagrass and complete Koch's postulates in greenhouse tests.2. Determine the influence of Primo and thiophanate-methyl and Heritage treatments on disease severity.3. Determine the effects of various rates of N and K on disease severity and control.


Summary PointsLeaf and sheath blight caused by

Rhizoctonia zeae, and related fungi, onbermudagrass putting greens has not beencontrolled with benzimidazole fungicides.

Molecular methods have been used todistinguish among R. zeae, R. oryzae, andR. circinata and will be conducted duringthe winter of 2008.

Fungicide trials were conducted in2007 and 2008. Only Heritage gave somecontrol in the single component fungicide

trial.

Watering-in fungicides made a positivedifference and suggested preventive treat-ments have to be initiated much earlierthan first supposed.

69

Fungicide Exp No. Rate Spray Dates Disease Severity (%)

Heritage TL 1 1.0 fl oz/1000 ft2 July 15, 29 and August 12 66.8 aEndorse 1 4 oz/1000 ft2 July 15, 29 and August 12 62.8 aTartan 1 2 fl oz/1000 ft2 July 15, 29 and August 12 51.8 abTrinity 2 2 fl oz/1000 ft2 July 15, 29 and August 12 35 bDisArm 1 0.27 fl oz/1000 ft2 July 15, 29 and August 12 55.8 abBanner Maxx 1.3 ME 1 1 fl oz/1000 ft2 July 15, 29 and August 12 45.5 abInsignia 1 0.9 oz/1000 ft2 July 15, 29 and August 12 60.5 abFore + 1 8 + 4 oz/1000 ft2 July 15, 29 and August 12 59.5 abChipco Signature

Untreated 1 61.3 a

Table 1. Single component fungicides, rates and timings for early curative control of Rhizoctonia leaf and sheathspot in bermudagrass putting greens.

Generally, the disease is first noticed in late August toearly fall months. Initial symptoms are bronze patchesof a few cm up to 100 or more cm in diameter.

Saltgrass (Distichlis spicata) is a

warm-season potential turfgrass speciesthat has the ability to grow under highlysaline, poor soil conditions, and with limit-ed available water and nutrient (i.e. nitro-gen) sources. This characteristic couldprove to be beneficial in certain turfgrassareas requiring low maintenance in aridregions with saline soils and limited wateror low fertility/limited nutrient contentsand poor soil conditions. Growth of thisspecies could be important to aid in con-servation of water and nutrients and use ofsalt-affected soils. Test of the species' abil-ity to grow under highly saline conditionsand limited nutrients, as well as low avail-able water sources, are needed before itcan be applied to a turfgrass system.

Twelve saltgrass genotypes (A37,A49, A50, A60, 72, A86, A107, A126,A136, A138, 239 and 240) collected fromseveral southwestern states of the UnitedStates were studied in a greenhouse toevaluate their growth in terms of shoot androot lengths and dry mater (DM) weights(shoot clipping weight) and nitrogen (15N)uptake under salt stress condition. Thegrasses were grown under saline (NaCl)conditions at EC of 20 dSm-1 using ahydroponic technique using Hoaglandsolution. A randomized complete block(RCB) design trial was used with fourreplications of each genotype and eachtreatment. Nitrogen (15N) was used todetermine the exact amount of N taken upby the plants.

Each week, the plant shoots (clip-pings) were harvested for the evaluation ofthe dry matter (DM) production. At eachharvest, both the shoot and root lengthswere measured and recorded. The harvest-ed plant materials were oven-dried at 60ºCand DM weights were measured andrecorded. Six harvests were included in theexperiment. The 15N samples for only one

harvest were analyzed. At the terminationof the experiment, plant roots were alsoharvested, oven-dried at 60º C and DMweights were measured and recorded.

The results of the average shootlength per week showed a decrease in allvarieties of plants grown under saline con-dition compared to the control. The rootlength for the duration of the experimentshowed increased root growth of eight ofthe twelve varieties grown under salinecondition when compared to the control.These eight varieties include A60, A86,A107, A126, A136, A138, 239 and 240.

All varieties, except A86, pro-duced a higher weekly average freshweight for control plants than those grownunder saline conditions. The fresh weightof the roots showed an increase in six often of the varieties (A49, A86, A107,A126, A136 and 240) for the plants grownunder saline conditions. Two of the vari-eties, A138 and 239, did not producerecordable root fresh weight.

The grass DM weight showed anincrease in ten of the twelve varieties(A37, A50, A60, 72, A107, A126, A136,A138, 239, 240) and in the control plantsin comparison to those grown under salineconditions. However, the percent DMweight of the shoots (average dryweight/average fresh weight) showed an

increase in nine out of thetwelve varieties (A49,A50,A60, 72, A86, A107, A126,239 and 240) in the plantsgrown under saline condi-tions compared to the con-trol, with variety A136being equal under the bothsalt conditions.Root DM weight

showed an increase in eightof the ten varieties (A49,A50, 72, A86, A107, A126,A136 and 240) grownunder saline conditions.The percent root DM (aver-age dry weight/averagefresh weight) showed

increased DM production in seven of theten varieties (A37, A50, A60, 72, A86,A126 and 240) grown under salineconditions.

The partial results of the 15Nanalysis show a substantial increase innitrogen concentration of the plant tissues.This indicates that saltgrass is a true halo-phyte and accumulates the extra N in itstissues which can be gradually available tothe growing parts of the grass.

Nitrogen Nutrition of Distichlis (Saltgrass) underNormal and Salinity Stress Conditions Using 15N


Grasses were grown under saline (NaCl) conditions at EC of 20 dSm-1using a hydroponic technique using Hoagland solution.

Mohammad PessarakliUniversity of Arizona

Objectives:1. To determine nitrogen uptake of saltgrass under salt stress conditions using 15N.


Summary PointsThe results of this experiment con-

firmed that saltgrass has a high salt toler-ance and is a true halophyte.

Root length, root fresh wt., root DMwt., percent shoot, and root DM wts. inplants grown under saline conditionincreased.

The amount of DM produced is themost significant result as it is a direct rep-resentation of saltgrass' ability to growunder highly saline conditions.

Based on the results of this experiment,each of the tested varieties had a highdegree of salt tolerance.

Based on the results of the 15N analy-sis, all the clones accumulated significantamounts of nitrogen in their tissues undersalinity stress condition.

70

Foliar fertilization refers to theprocess of nutrient uptake through thefoliage or other aerial plant parts. As asupplement to traditional root-feeding pro-grams, foliar fertilization has beenobserved to be an increasingly commonpractice in today's golf course manage-ment. Recent surveys of Arkansas golfcourse superintendents indicate that nearlyall golf course superintendents use foliarfertilization on some area of their golfcourse, and this method of nutrient appli-cation often comprises a major portion ofannual nitrogen (N) inputs to puttinggreens.

Although previous research hasdocumented uptake of N by turfgrassleaves in controlled-environment studies,there have been no studies which demon-strate its potential effectiveness in a fieldenvironment. It is known from previousagricultural research that environmentalfactors and seasonal dynamics of leaf cuti-cle characteristics can influence the foliarabsorption of N solutions. Therefore, theaim of this project is to increase scientificunderstanding of the turfgrass leaf cuticle,while assessing foliar uptake of N during atwo-year putting green research trial.

Experimental areas of 'Penn A1'

creeping bentgrass and 'Tifeagle' ultra-dwarf bermudagrass were developed on anestablished sand-based putting green inFayetteville, Arkansas. The greens weremaintained according to typical manage-ment practices for the region. Foliar uptakeof N was studied each month from May toSeptember to determine if foliar uptakewas consistent across the growing season.

An isotopic tracer technique thatallows for positive identification and directmeasurement of fertilizer N in the plant tis-sue was used in the study. 15N-labeledurea solutions were sprayed at two differ-ent rates to represent a low and high rate(0.10 or 0.25 lb. N / 1000 ft2) common tofoliar fertilization rates of golf coursesuperintendents. For a 24-hour periodafter treatment, plots received no irrigationor rainfall in order to track only foliarabsorption of N. Plant tissues were sam-pled at 1, 4, 8, and 24 hours after applica-tion in order to develop a time-courseanalysis of foliar N uptake. In addition, anenclosed chamber with an internal acidtrap was installed within the plots aftertreatment to estimate the volatilization ofN as ammonia from the plots.

Results from the first year ofmonthly foliar urea-N applications indicatethat both species proved receptive to foliaruptake and ammonia volatilization losseswere minimal (averaged < 2% of appliedN). A range of 24-57% of the fertilizer Napplied was recovered in leaves/shoots at 1hour after treatment, while peak foliar

absorption was normally observed around4 hours after treatment.

Foliar uptake (measured as a % ofN applied) was significantly reduced athigher application rates. The highest maxi-mum absorption of N applied observedover 24 hours, was in the month of May onbentgrass (76%). Absorption of N by bent-grass leaves was affected by month of year,while ultradwarf bermudagrass was notaffected.

On bentgrass, the significantdecrease in N recovered within plant tissueas the season progressed (May averaged59% of applied N across all samplingtimes, while September averaged 37%), iscurrently believed to be attributed to leafcuticle changes that made leaves morehydrophobic and possibly less receptive tonutrient absorption. Continued laboratoryinvestigations are underway to betterunderstand this observed trend.

Leaf Cuticle Characteristics and Foliar NutrientUptake by a Cool-season and Warm-season Turfgrass


Golf course superintendents often use foliar fertilizationas a primary means of feeding putting greens.

Michael D. Richardson, James C. Stiegler, Douglas E. Karcher, and Aaron J. PattonUniversity of Arkansas

Objectives:1. Determine the seasonal dynamics of the turfgrass cuticle in a cool-season and warm-season turfgrass speciesmanaged under putting green conditions.

2. Compare the seasonal uptake of foliar nitrogen by a cool-season and warm-season turfgrass species managedunder putting green conditions.

Foliar nitrogen is applied to the putting green turf and an ammonia volatilization trap is installed after application.


Summary PointsBoth creeping bentgrass and ultradwarf

bermudagrass greens are receptive tofoliar uptake of urea nitrogen.

Most of the nitrogen applied to puttinggreen turfgrass foliage is absorbed in thefirst 4 hours after application.

Foliar uptake in creeping bentgrass wasreduced during warmer months, suggest-ing a change in the composition of the leafcuticle.

Loss of foliar-applied nitrogen toammonia volatilization appears to beminimal.

71

Wildlife Links Program

Golf courses offer excellent opportunities to provide important wildlifehabitat in urban areas. With more than 17,000 golf course in the United States com-prising in excess of two million acres, great potential exists for golf courses tobecome an important part of the conservation landscape.

Wildlife Links is a cooperative program with the National Fish andWildlife Foundation that funds innovative research, management, and educationprojects that help golf courses become an important part of the conservation land-scape. The United States Golf Association is providing through the Wildlife LinksProgram which began in 1996. The objectives of Wildlife Links are to:

1. Facilitate research on wildlife issues of importance to the golf industry.2. Provide scientifically credible information on wildlife management to

the golf industry.3. Develop wildlife conservation education materials for the golf industry

and golfers.4. Implement wildlife monitoring programs that will improve manage-

ment on golf courses.

Location of the following project funded in 2008 through the Wildlife LinksProgram administered by the National Fish and Wildlife Federation

72

The eastern population of the Painted

Bunting (Passerina ciris) breeds in arestricted range of the Atlantic CoastalPlain, from North Carolina to Florida. ThePainted Bunting is considered a species atrisk by various conservation organizationsdue in part to its restricted distribution andlow population size.

Within the coastal counties ofSouth Carolina, golf courses and golfcourse communities are common andbecoming more prevalent. Recent researchfunded by the Wildlife Links Programshowed that while Painted Buntings werefound on numerous courses in HorryCounty, SC, they did not occur on courseswith highly altered landscapes.

We are assessing the conserva-tion value of golf courses in the BeaufortCounty area of SC for Painted Buntings.The Beaufort County region represents anarea of high risk for the species due toongoing development, but also potentiallyhigh conservation reward. If courses in thisarea can provide habitat that will supportsuccessful breeding populations of PaintedBuntings, then a substantial portion of thebreeding range can be managed in such away as to take advantage of the extensiveprivate land holdings in the region.

A pilot season of field researchwas initiated in May 2008. Searching forPainted Bunting nests is an pain-stakingand tedious endeavor, and, for that reason,we chose to focus our efforts on only twogolf courses. The field research teamsearched the Chechessee Creek Club andOld Tabby Links on Spring Island inBeaufort County, South Carolina, through-out May, June, and July 2008.

Researchers located individualbuntings during the day and over thecourse of several days and carefully plot-ted their locations and activities. This tech-nique often resulted not only in the loca-tion of a nest, but also in a home range mapand a better understanding of the habitatsbeing used during the nesting period.Locations were plotted with a handheldGPS and developed into a GIS data layer.This data layer can be overlaid with GIScoverage of the courses to examine habitatuse at the nest, patch, and territory scale.

Using the territory-mappingapproach, we located 14 nests of PaintedBunting on the two study courses. Fivenests were located on the Old Tabby Linkson Spring Island. Three of these nests werelocated in wax myrtle bushes, while theother two nests were located in thickets ofmixed shrubs. Nine nests were located atthe Chechessee Creek Club. Four of thesewere located in wax myrtles, while theremaining nests were located in saplings orlarge trees.

We also were able to locate and

track six groups of Painted Bunting fledg-lings on Spring Island and eight groups onChechessee Creek. These data will be usedto develop habitat use models for fledg-lings and hence improve our ability todevelop habitat management guidelines.

Field research will be renewed inMay of 2009. We will focus our efforts onthe same two courses and begin mappingterritories as soon as possible during thebreeding season. An early start to territorymapping will improve our ability to findnests and track fledgling groups. Duringthe 2009 breeding season, we also willbegin to investigate the value of mowedversus unmowed grass plots for PaintedBunting foraging activity.

Reproductive Success and Habitat Use of PaintedBuntings on Golf Courses in Coastal South Carolina


Patrick G.R. Jodice, Mark Freeman, and Jessica GorzoClemson University

Objectives:1. Locate nests of Painted Buntings on study courses and relate various metrics of reproductive success (clutch size,daily nest survival, hatching success, brood size at fledging) to characteristics of nesting habitat at the nest, patch,and territory scale .

2. Locate fledgling groups of Painted Buntings on study courses and determine home ranges and habitat use.3. Determine use of grass, out of play areas for foraging by Painted Buntings and compare its availability amongcourses.

4. Compare reproductive success and nesting habitat to previously measured values in the southeastern US.

By locating individual buntings over the course of sev-eral days and carefully plotted their locations and activ-ities, a home range map and a better understanding ofthe habitats being used during the nesting period wasachieved.


Summary PointsSearches for nests of Painted Buntings

were conducted on the Chechessee CreekClub and Old Tabby Links in BeaufortCounty, South Carolina, from May - July2008. Areas searched included roughs andshrub/scrub habitat throughout the cours-es.

We located 14 nests on the two cours-es. Most nests were found in wax myrtlesor shrub/scrub habitat, although a fewnests also were located in large hardwoodtrees. We also located and mapped thehabitat use of 14 groups of Pained Buntingfledglings. Data from both the nest loca-tions and fledgling groups are being incor-porated into a GIS data layer that will beused to assess habitat use.

Data collection in 2009 will focus onadditional nest searching and territorymapping and may use the assistance ofvolunteers in an effort to increase the sam-ple size of nests and territories that can bemonitored. We also will initiate research toexamine the use of mowed versusunmowed grass plots as feeding areas forPainted Buntings.

73

Chris MarshLow Country Institute

The southern Appalachian Mountain

region harbors an exceptional amphibiandiversity which is dominated by salaman-ders. These salamanders are integral eco-logical components of the headwaterecosystems they inhabit where they oftenaccount for the majority of vertebrate bio-mass. The majority of these salamandersare found in close association with head-water stream habitats, forming communi-ties that are comprised of five to ninespecies from the genera Desmognathus,Eurycea, Gyrinophilus, and Pseudotriton.These salamanders are stream salamandersof the family Plethodontidae and all havebiphasic life cycles consisting of an aquat-ic larval stage that is followed by a terres-trial adult stage.

Stream salamanders are depend-ent upon both the stream and the surround-ing riparian habitat for foraging, breeding,and dispersal. Though less conspicuousand least studied, the larval stage is essen-tial to the persistence of or the reestablish-ment of adult salamanders in the surround-ing riparian habitat.

Managing landscapes with an eyefor both human use and preservation ofbiodiversity can create a win-win situationfor stakeholders and wildlife. Consideringthat the average golf course consist ofmore than 150 acres of green space (70%is rough, non-play areas) and there are

more than 17,000 golf courses in the U.S.that total over 2.2 million acres, we sug-gest there is great potential for golf cours-es to serve as sanctuaries for many wildlifespecies if the habitat needs of species arepresent.

Our project will measure streamsalamander abundance and diversity inorder to make biologically relevant man-agement suggestions to improve the quali-ty of golf courses for stream salamanders.We will sample 10 golf courses in the mon-tane region of the southern Appalachianswithin a 25-mile radius of Highlands, NCto compare to adjacent (upstream, down-stream) control areas and to previous workwe have conducted on national forest land.

During the 2008 summer field

season, Mark Mackey (M.S. GraduateStudent) and Grant Connette (FieldResearch Technician) were able to estab-lish contact and participation agreementswith 10 golf courses in the Highlands, NCarea. They were also able to delineatestreams on each course to establish six 25-meter transects (2 upstream, 2 in-course, 2down stream) for intensive sampling dur-ing 2009. Further, they conducted a pilotproject to determine the effectiveness ofsampling protocols and leaf-bag samplingin different habitat types.

They found that leaf bags werehighly effective for larval salamanders andthat the four primary species found in

national forest streams were also presentin some streams on golf courses. Intensivesampling of larval and adult salamanderson all 10 golf courses will begin next yearin May through August 2009.

By comparing the abundance anddiversity of larval salamanders in streamsand adult salamanders in the adjacent ter-restrial habitat on golf courses relative toupstream and downstream areas, we willbe able to assess the adequacy of currentcourse management. We will then developrecommendations on the management ofin-stream and riparian habitats on eachgolf course to maintain or improve habitatfor salamanders.

Abundance and Diversity of Stream Salamanders on Golf Courses


Salamanders are integral ecological components of theheadwater ecosystems they inhabit.

Raymond D. SemlitschUniversity of Missouri

Objectives:1. Measure stream salamander abundance and diversity on 10 golf courses in western North Carolina to make bio-

logically relevant management suggestions to improve the quality of golf courses for amphibian biodiversity.

Four primary species of salamanders found in nationalforest streams were also present in some streams ongolf courses.


Summary PointsAgreements of participation were

obtained for 10 different golf courses in theHighlands-Cashiers area of NorthCarolina.

Six different 25-meter stream transectson each course were delineated for inten-sive sampling and comparison, yielding atotal of 60 different stream segments to besampled across all 10 golf courses.

Preliminary sampling of larval sala-manders was conducted to obtain the bestsampling methods and a preview ofspecies' presence on golf courses.

A pilot project was conducted examin-ing the efficiency of the leaf-litter bagtechnique as a method for sampling larvalsalamanders in different stream habitats.

74

Stream salamanders are dependent upon both thestream and the surrounding riparian habitat for forag-ing, breeding, and dispersal.

The brown-headed nuthatch (Sitta

pusilla) is a cooperatively breeding birdendemic to the southeastern United States.But for nearly half a century its numbershave been in decline. Habitat degradationis usually blamed. Brown-headednuthatches are said to be habitat specialists- dependent on old growth pine forests. Asdevelopment overtakes more of theSoutheast, there are fewer old pine standsusable by these birds.

Another cavity-nesting specieshas increased dramatically in number inthe same region - the Eastern Bluebird(Sialia sialis). Bluebirds tend to fare wellin a variety of human-altered habitats.Moreover, bluebirds have been the benefi-ciaries of nest box programs throughouttheir range. We hypothesized that the bur-geoning bluebird population in theSoutheast is negatively impacting brown-headed nuthatches.

My students and I have beenmonitoring nest boxes on six golf coursesnear Davidson, North Carolina since 2001.For each nest box, we measured the dis-tance to the nearest three pine trees (Pinusechinata). The distance to the third wasour measure of pine density. Boxes for

which the third closest pine was less than50 meters away were considered to be in"pine-rich" habitat; boxes for which thethird closest pine was > 50 meters awaywere considered to be in "pine-poor" habi-tat.

Standard 1.5" entrance holesaccommodate both bluebirds and nuthatch-es; 1.25" holes accommodate nuthatches,but are too small for bluebirds. We ran-domly assigned boxes on three golf cours-es to the 1.25" treatment. Pine densityhad no significant effect on nesting bynuthatches, contrary to the predictions ofthe conventional wisdom. Hole size, how-ever, had a highly significant effect onnuthatch settlement: nuthatches settlewhere competition with bluebirds is mini-mized, regardless of local pine density.

We recorded the number ofnuthatch nests on three courses with a sub-set of 1.25" entrance holes vs. the numberon three other courses on which all boxeshad standard "bluebird-friendly" 1.5"entrance holes. We monitored boxes in2004 (prior to the addition of "nuthatch-friendly" holes on our experimental cours-es) to ensure that all six courses were sim-ilar in their lack of nuthatches. For the2005, 2006, and 2007 breeding season, wemonitored the numbers of nuthatch nestson both experimental and control courses.Numbers of nuthatches increased in eachyear of the study, suggesting that bluebirdscompetitively exclude nuthatches fromavailable habitat.

These results clearly demonstratethat nuthatches flourish only where blue-birds are excluded. Prior to the 2008breeding season, we reversed the treat-ments on our six golf courses: our three"bluebird-friendly" courses became"nuthatch-friendly" and vice-versa. Of the32 boxes containing nuthatch nests in2007, 31 contained bluebird nests in 2008.The competitive superiority of bluebirds isunquestionable.

Brown-headed Nuthatch Enhancement Study


Mark StanbackDavidson College

Objectives:1. Assess the importance of pine density and competition with eastern bluebirds on the spatial distribution of

brown-headed nuthatch nests.2. Assess the numerical response of brown-headed nuthatches to the experimental exclusion of eastern bluebirds.3. Assess the ability of eastern bluebirds to usurp nest sites from brown-headed nuthatches.4. Provide golf course managers in the Southeast with recommendations to increase the numbers of brown-headed

nuthatches on their golf courses.

Brown-headed nuthatches breed in cooperative groupsin the southeastern US. (photo by Austin Mercadante)


Summary PointsBrown-headed nuthatch numbers have

declined throughout the Southeast. Theirsupposed dependence on old growth pineforests - and susceptibility to habitat alter-ation - is usually blamed for this decline.We offer an alternative hypothesis - thatnest site competition with a burgeoningeastern bluebird population is responsible.

We monitored nest box use by blue-birds and nuthatches on golf courses andfound that pine density had little effect onnest box use by nuthatches. Instead, theexclusion of bluebirds (via smallerentrance holes) was the best predictor ofnest box occupation by nuthatches.

Brown-headed nuthatch numbersincreased dramatically on three golf cours-es where bluebirds were excluded fromone-third of nest boxes. Control courseshad few, if any, breeding nuthatches overthe same period.

When bluebird-friendly holes werereturned to experimental boxes, bluebirdsquickly evicted resident nuthatches.

To prevent monopolization of nestboxes by bluebirds, golf courses in theSoutheast should provide smaller entranceholes on a subset of their nest boxes.

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1.5" entrance holes (left) accommodate both bluebirdsand nuthatches; smaller holes (right) exclude the larg-er bluebirds.

Several studies have illustrated theabundance and diversity of birds that canbe found at golf courses, but less is knownabout nesting success. In addition, thechemicals often employed by golf coursemanagers are known to negatively affectwildlife, including reproductive andbehavioral impairments, yet the extent ofavian exposure on golf courses remainslargely unknown.

Studies indicate that eastern blue-birds can breed successfully in nest boxesinstalled on golf courses, but there is alsoevidence that the nestling birds weigh lessthan same-aged counterparts at suburbansites with no pesticide inputs. Our researchobjectives were to determine the extent ofpesticide exposure in nestling bluebirdsand to determine if these insectivoroussongbirds are able to obtain sufficientinvertebrate food on golf courses treatedwith insecticides.

Experimental design consisted ofmonitoring and comparing breeding activ-ity in more than 500 nest boxes at 8 golfcourses and 7 non-golf course referencesites. Our reference sites consisted of sub-urban locales with some level of humanactivity (e.g. local parks), but which wereconfirmed to not apply pesticides.Participating golf courses shared their

detailed chemical application logs, allow-ing us to know specifically which type,where, and when pesticides were applied.

During the 2007 and 2008 breed-ing seasons, we collected and analyzed 290blood samples from nestling birds to assesscholinesterase enzyme activities, the stan-dard method for determining exposure tomany common insecticides. All nestlingswere measured and banded during sam-pling efforts. We collected more than 450prey items directly from nests through useof esophageal ligatures and an additional20 insects were collected from turf areas24 hours after insecticide application.These 20 insects and 30 insects from nestsafter pesticide applications were analyzedfor pesticide residues.

Lab analyses found no evidenceof enzyme inhibition in golf coursenestlings, suggesting they had not beenexposed to enough pesticide to detect anydirect effects. No pesticide residues weredetected in 50 prey items insects. Nestlingdiet and parental feeding rates were thesame at golf courses and reference sites.However, physical measurements ofnestlings revealed that, as previouslyreported, golf course chicks weighed lessthan birds on reference sites of the samedevelopmental age.

Much of the application of pesti-

cides occurred after nestlings fledged fromnests. Because the nestlings we sampledappear not to have been exposed to pesti-cides, additional research efforts are under-way to track survival of fledgling bluebirdsafter they leave the nest.

These young, inexperienced birdsmay be more likely to be exposed to pesti-cides through ingestion of dead or mori-bund insects. If these young birds have alow survival rate, it may be that the suc-cessful nesting efforts observed for adultbirds overestimate the habitat quality ofgolf courses. Only by measuring conditionand survivorship at every stage of the lifecycle can we gain confidence that golfcourses are not ecological traps.

Avian Pesticide Exposure on Golf Courses


During the 2007 and 2008 breeding seasons, we col-lected and analyzed 290 blood samples from nestlingbirds.

Daniel A. Cristol and Ryan B. BurdgeThe College of William and Mary

Objectives:1. To determine the extent of pesticide exposure in nestling bluebirds and to determine if these insectivorous song-birds are able to obtain sufficient invertebrate food on golf courses treated with insecticides.

Experimental design consisted of monitoring and com-paring breeding activity in more than 500 nest boxes at8 golf courses and 7 non-golf course reference sites.


Summary PointsNestling eastern bluebirds are at low

risk of pesticide exposure on golf courses.

Eastern bluebirds are able to foragecomparable type and amount of insect preyon golf courses compared to other subur-ban environments.

Nestling bluebirds tend to be lowerweight than reference birds, potentiallyaffecting post-fledging survivorship.

Continued research will track survivaland pesticide exposure in juvenile birds.

Additional work is needed to assesspesticide exposure and associated effectsin other golf course wildlife and at other

stages of the life cycle.

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Physical measurements of nestlings revealed that, aspreviously reported, golf course chicks weighed lessthan birds on reference sites of the same developmen-tal age.

Over the past several years,

Stewardship Partners has collaboratedwith the Oregon based Salmon-Safe Inc. todevelop a Salmon-Safe certification andlabeling program for Puget Sound areafarmers and other landowners who adoptfish-friendly farming practices and partici-pate in salmon habitat restoration projects.The project is proving to be successful,with over 40 farms certified to date, in-store retail campaigns with Seattle areagrocers, extensive media coverage, and avariety of promotional materials producedfor use by farmers in their direct marketingefforts.

As a follow-up to this successfulprogram, we are considering additionalopportunities to use the Salmon-Safe labeland evaluation tool to advance environ-mental management practices of othertypes of landowners and businesses. Forexample, the Salmon-Safe program hasbeen adapted for municipal park systemsas well as corporate and universitycampuses.

As a result of our promotion andoutreach in Puget Sound, we have beenapproached by the Western WashingtonGolf Course Superintendents Associationto explore opportunities to utilize theSalmon-Safe program as an evaluation toolto assess environmental performance and apublic education tool to promote exempla-ry golf course management.

This represents an importantopportunity because golf courses, many ofwhich are located along productive streamsystems, can have a large environmentalimpact from use of water, application ofchemicals, conversion of wetlands, andalteration of stream and riparian habitat.Golf courses are increasingly beingrequired to address these issues as severalspecies of salmon, such as Puget SoundChinook, have been listed under the feder-al Endangered Species Act.

Due to the increases in awarenessof these issues among the public and golfcourse managers themselves, there havebeen some initial efforts within the indus-try to improve management practices. Forexample, the Northwest Golf CourseEnvironmental Alliance is a newly formedeffort to pro-actively demonstrate positivetrends within the industry through theircommitment to a set of EnvironmentalStewardship Guidelines.

Another industry effort is theCooperative Sanctuary Program developedby the US Golf Association and AudubonInternational. Stewardship Partners, inconjunction with Salmon-Safe, Inc. willbuild on this momentum to establish ameaningful and regionally-based third-party certification program under the aus-pices of Salmon-Safe. We will adapt theSalmon-Safe Parks and Natural AreasCertification Guidelines to be more specif-ic for golf course management and conduct

independent pilot evaluations of three golfcourses based on these guidelines. We willalso help evaluate restoration opportunitiesand connect these golf courses with localorganizations to conduct on-the-groundrestoration work.

We have identified several golfcourses which would be good candidatesfor piloting such a program based on theirreputation for environmental performance,participation in habitat restoration alongsalmon bearing streams, and the interest ofthe managers. We will pilot the programwith these initial candidates to determinethe feasibility of more widescale imple-mentation of a Salmon-Safe golf certifica-tion program.

Puget Sound Salmon-Safe Golf Course Program


Salmon-Safe guidelines represent an important oppor-tunity because golf courses can have a large environ-mental impact from use of water, application of chemi-cals, conversion of wetlands, and alteration of streamand riparian habitat.

David BurgerStewardship Partners

Objectives:1. Conduct independent evaluations of area golf courses based on an adapted version of the Salmon-Safe Parks and

Natural Areas Certification Guidelines to determine how management addresses water quality and habitat conser-vation.

2. Identify habitat restoration opportunities with participating golf courses and connect them with local resources toconduct on-the-ground improvements.

3. Develop an education and outreach campaign to build support for Salmon-Safe golf course management.4. Test the opportunities for wide-scale implementation of Salmon-Safe golf course certification.5. Demonstrate leadership within the golf industry in voluntary restoration and conservation efforts to recover nativesalmon and other fish and wildlife.

Golf courses are increasingly being required to addressthese issues as several species of salmon, such asPuget Sound Chinook, have been listed under the fed-eral Endangered Species Act.


Summary PointsStewardship Partners, in conjunction

with Salmon-Safe, Inc. is establishing ameaningful and regionally-based third-party certification program under the aus-pices of Salmon-Safe for golf courses inthe Pacific Northwest.

Several golf courses have been identi-fied which would be good candidates forpiloting such a program based on their rep-utation for environmental performance,participation in habitat restoration alongsalmon bearing streams, and the interest ofthe managers.

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Research was conducted to

investigate how the use of buffer zonesaround water resources could help bolteramphibian populations. We made contactwith three golf courses [Twin Run GolfCourse (Hamilton, OH), Hueston WoodsGolf Course (College Corner, OH), andOxford Country Club (Oxford, OH)] thateach had two ponds: one that they agreedto leave a 1-meter terrestrial grass bufferzone, and another that they mowedto the edge. Additionally, we usedtwo "reference" ponds at theEcology Research Center (Oxford,OH).

We spent approximately 2months collecting cricket frog pairsfor the golf course and related proj-ects. Eight enclosures were addedto each golf course pond. To half ofthe enclosures 40 cricket frog tad-poles were added, and to the otherhalf green frog tadpoles were added(so that we could examine effectson two species).

Golf courses werechecked 2-3 times per week forsigns of metamorphosed frogs, andwater quality measurements weretaken weekly to every other week.The aquatic experiments are stillunderway and will be terminated by theend of October, 2008.

Cricket frogs that reached meta-morphosis were given individual marksand released at the golf course ponds sothat subsequent terrestrial survival can beassessed. We will search for these cricketfrogs next spring when the breeding seasonensues.

In another study, we exposedcricket frog tadpoles and green frog tad-poles reared in cattle tank mesocosmponds at Miami University's EcologyResearch Center (Oxford, OH) to presenceor absence of a potential predator (twograss carp, two bluegill sunfish, two cray-fish) and to the presence or absence of a

sublethal level of the insecticide imidaclo-prid. This experiment was conducted overthe summer and completed at the begin-ning of September. Currently, we are ana-lyzing the data and preparing the manu-script for publication.

We also conducted a laboratorystudy with cricket frogs exposed to waterfrom each golf course pond and the pres-ence or absence of a sublethal exposure tothe insecticide imidacloprid (which at leastone of the golf courses was using). Thisstudy allowed us to see how individual tad-poles were affected by the golf coursewater relative to controls (ponds from the

reference site with aged tap water) in thepresence or absence of the insecticide.

This study will give us insightinto the effects that we observe inProject 1 located on the golf course.The student collected water fromeach of the six golf course pondsand one reference site each week,and then raised the tadpoles in thiswater. In this study, survival wasmeasured daily, and at the end ofthe study, we determined mass anddevelopmental stage.

Additionally, in a separatestudy, the student also examinedwhether the presence of the insecti-cide and the golf course wateraffected susceptibility to a crayfishpredator. Both studies have beencompleted and a manuscript is cur-rently being prepared from thiswork.

Using Buffer Zones to Promote Amphibian Populations


Cricket frogs were used both at the golf course pondsand reference ponds to test for the effects ofimidicloprid.

Michelle D. BooneMiami University

Objectives:1. To determine the effects of a 1-meter buffer zone around ponds to bolster cricket frog populations.2. To determine the effects of the presence or absence of a potential predator (two grass carp, two bluegill sunfish,

two crayfish) and to the presence or absence of a sublethal level of the insecticide imidacloprid.

Eliminating fish from ponds is a critical step becausepondswithout fish allow for greater abundance ofamphibians and more diverse communities.


Summary PointsResearch is underway to test for the

effects of a 1-meter grassy buffer aroundgolf course ponds on the population ofcricket frogs.

A laboratory study was also conductedwith cricket frogs exposed to water fromeach golf course pond and the presence orabsence of a sublethal exposure to theinsecticide imidacloprid.

In a separate study, it was also exam-ined whether the presence of the imidaclo-prid in the golf course water affected sus-ceptibility to a crayfish predator.

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Green frogs were also used to test for the effects presence or absence of apotential predator (two grass carp, two bluegill sunfish, two crayfish) and tothe presence or absence of a sublethal level of the insecticide imidacloprid.

The demand for golf course commu-

nities in Arizona has steadily increased asbaby boomers transition into retirement.Presently, there is a lack of information onthe potential role that golf courses can playin maintaining native reptile populationsand communities, especially in arid envi-ronments like the southwestern UnitedStates. The more information we have onhow to design and manage golf coursecommunities, the better we will be at max-imizing the benefits that golf courses canprovide for wildlife.

During the past year, we usedmark-recapture to determine relative abun-dance and distribution of reptiles usinggolf courses in Arizona. To date, we havedocumented literally thousands of tortois-es, snakes, and lizards using various fea-tures of golf courses. Using high-resolu-tion imagery, we are analyzing distributionand abundance data in the context of land-scape structure. We are finding that certainaspects of the golf course are used by cer-

tain species, while others are avoided. Forexample, irrigated vegetation along fair-ways and surrounding greens and teeboxes are used well out of proportion totheir availability, while open fairwaysappear to be avoided, especially by snakes.Areas of the golf course where naturaldesert has been incorporated into thedesign of the course correspond to areas ofgreater relative abundance of most species.

We have been radio tracking Gilamonsters to gain a better understanding ofhow they are responding to the golf courseand its surroundings. We have observed aclear pattern of use by Gila monsters char-acterized by general avoidance of openfairways and other areas of turf. However,Gila monsters tend to spend more time onthe edges of fairways and greens, presum-ably searching for prey in the dense, irri-gated vegetation.

We have observed individual Gilamonsters changing the location and use oftheir home ranges as newly constructedhomes become more common.Interestingly, the Gila monsters have start-ed to concentrate more of their activity inareas immediately adjacent to the golfcourse that are off limits to development.

Although radiotelemetry is anexcellent tool for tracking the movementsof Gila monsters, you can only obtaininformation on the animal's location byphysically following its signal. In 2008,we began using a new technique thatallows us to quantify the exact movementpath taken by a Gila monster. The tech-nique involves the use of florescent pow-der that we apply to the Gila monster bygluing rabbit fur onto its belly and loadingup the fur with the powder. After allowingthe Gila monsters some time to moveabout, we come back after dark with a UVlight and follow its exact track. This tech-

nique allows us to determine if Gila mon-sters are avoiding certain features of theirhabitat, such as fairways and roads.

Using radiotelemetry, we only getthe straight-line distance between succes-sive fixes, but florescent powder allows usto determine the exact distance moved. Sofar, we have found that Gila monsters typ-ically move two to three times farther thantheir straight-line distances would indicate.

Population and Community Responses of Reptiles to Golf Courses


The more information we have on how to design andmanage golf course communities, the better we will beat maximizing the benefits that golf courses can providefor wildlife.

Matt GoodeUniversity of Arizona

Objectives:1. Examine diversity, distribution, and relative abundance of reptiles living on three golf courses varying in age,

location, and surrounding housing density.2. Implant radio transmitters into Gila monsters to examine movement patterns, habitat use, and survival in response

to varying features of golf courses.3. Inform residents, golfers, and golf course personnel about local reptiles via interactions, presentations, and thedevelopment and distribution of a field guide booklet.

4. Engage golfers in monitoring reptiles by writing down wildlife observations in a logbook available in each club-house.

5. Develop recommendations for golf course designers and managers that can be used to retrofit existing coursesand design new courses to maximize benefits to reptiles.

University of Arizona scientists have been radio trackingGila monsters to gain a better understanding of howthey are responding to the golf course and its sur-roundings.


Summary PointsReptile species vary in their use of golf

course features, but most species tend toavoid open fairways.

We have observed a greater abundanceof reptiles using thick, irrigated vegetationadjacent to fairways and greens.

The placement and layout of the golfcourse helps to determine the distributionof reptiles, and certain species alter theirdistributions to take advantage of irrigatedvegetation and natural areas that are incor-porated into the golf course as hazards orrough.

Using radiotelemetry and florescentpowder tracking, we have determined thatGila monsters tend to avoid open fairways,and tend to utilize irrigated areas along theedges of the golf course out of proportion

to their availability.79

Product Testing Program

Locations of the following projects funded in 2008 by the USGA Turfgrass andEnvironmental Research Program under the category of Product Testing

80

Every year golf course superintendents are introduced to new products inthe marketplace. Without results from properly designed, objective research, super-intendents are asked to make buying decisions based on word-of-mouth, previousexperience from colleagues, or the representative of the product. Several surveysindicate that golf course superintendents desire side-by-side product evaluations toassist them in making product purchases. The result of this desire for this type ofinformation is the Product Testing Program section of USGA’s Turfgrass andEnvironmental Research Program. Currently, USGA is funding three projects thatfall into this category of USGA-supported research, including cooperators at sever-al sites across the country (see map below).

Summer bentgrass decline is a majorconcern of superintendents growing creep-ing bentgrass on putting greens across thecountry, especially in the southern statesand the transition zone. Some golf coursesapply various compounds on bentgrass,hoping to improve turf growth during sum-mer months.

Numerous organic products,claimed to have the functions as plantgrowth regulators or biostimulants, haveemerged in recent years. Some organicmaterials such as seaweed extracts andamino acids are being used as fertilizersupplements in commercial turfgrass man-agement. Among them, seaweed extractsare widely used in various biostimulantproduct formulations. Many of biostimu-lants promise better turf quality and stresstolerance.

The use of PGRs has recentlybeen expanded as a means to promote turf-grass performance under stressful condi-tions. We propose to investigate whetherthe application of selected PGRs and bios-timulants to a bentgrass putting greenwould enhance turf quality during summermonths and to determine whether these

products would help alleviate summerbentgrass decline by delaying leaf senes-cence and promoting root growth.

The study was conducted in 2007and 2008. This report summarizes the 2007results. We tested the following selectedproducts that are extensively used in thegolf course maintenance or pure chemicalswith great potential to be marketed asstress reducers:Primo Max (from Syngenta, 0.125 oz. /1000 sq. ft.): Inhibitor of gibberellic acidsynthesis and vertical shoot growth.TurfVigor (from Novazymes, 15 oz. /1000 sq. ft.): Combines patented highimpact microbial strains, and macro andmicronutrients.CPR (from Emeraled Isle Solutions, 6 oz./ 1000 sq. ft.): a blend of natural sea plantextracts, micronutrients, and a surfactant.Aminoplex (from Grigg Brothers, 2 oz./1000 sq. ft.): A proprietary mixture of 15plant based L-amino and organic acids,complex polysaccharides, and natural hor-mones.Proxy and StressGard (from Bayer, 5 oz.+ 0.38 oz. / 1000 sq.ft.): Combined treat-ment with a Type II plant growth regulatorthat suppresses seedhead of Poa annua andwhite clover while promoting lateralgrowth of cool-season turf and a fungicideproduct that provides long-lasting broad-spectrum disease control and improvedturf quality.Aminoethoxyvinylglycine (AVG, fromSigma, 25 uM): an ethylene synthesisinhibitor that suppresses leaf senescenceand helped to maintain greener turf for anextended period in a recent growth cham-ber study when creeping bentgrass was

exposed to 35o C.6-benzylamine (BA, from Fisher, 25 uM):a synthetic cytokinin that demonstratedfunctions in delaying leaf senescence andimproving heat tolerance in creeping bent-grass in controlled-environmentconditions.

We also included two controltreatments:Nutrient control (Hoagland's solution):complete nutrient solution including allkinds of macronutrients and micronutrientsto sustain plant growth.Water control: regular ground water.

All chemical treatments wereapplied following the manufacture recom-mended rate (carried in 2 gal. of water).We evaluated turf quality, chlorophyll con-tent, photosynthesis rate, leaf area index(indicator of canopy density), green leafbiomass (indicator of canopy color), androot growth.

Evaluation of Plant Growth Regulators and Biostimulantsfor Use in Managing Summer Bentgrass Decline


Summer bentgrass decline is a major concern of super-intendents growing creeping bentgrass on puttinggreens across the country, especially in the southernstates and the transitional zone.

Bingru Huang and Yan XuRutgers University

Objectives:1. To investigate whether the application of selected PGRs and biostimulants to a bentgrass putting green wouldenhance turf quality during summer months.

2. To determine whether these products would help alleviate summer bentgrass decline by delaying leaf senescenceand promoting root growth.


Summary PointsTurfVigor- and CPR-treated plots con-

sistently maintained significantly higherturf quality than both nutrient and watercontrol plots during the whole treatmentperiod. Turf quality of Primo Max-treatedplots was also higher than that of the nutri-ent and water control plots starting frommid-August. Other products, such as BA,AVG, and Aminoplex had some positiveeffects in August.

Shoot growth and physiological activi-ty of creeping bentgrass was promotedwhen treated with certain biostimulants orPGRs. The decline of canopy photosyn-thetic rate was less severe in TurfVigor-,Primo Max-, and BA-treated plots duringmid-July and/or early August. Canopydensity and color were maintained higherin CPR-, TurfVigor-, and Primo Max-treat-ed plots in July and August.

The only positive effects on root lengthand surface area were observed in theProxy- and StressGard-treated plots inearly September, indicating this productmay affect the recovery of creeping bent-grass from summer stress.

Our results suggested that proper use ofselected biostimulants and PGRs couldpromote turf growth and alleviate summerbentgrass decline in warm climates.

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Golf course superintendents often

limit nitrogen (N) fertility on greens for thepurpose of improving green speed. Tomaintain acceptable turf color and planthealth under conditions of limited N input,superintendents often apply liquid biostim-ulants in the summer. Numerous compa-nies market products containing cytokini-ans (usually from seaweed) and other plantextracts in liquid forms for use on golfgreens.

Cytokinians are considered bios-timulants, which have been shown toimprove shoot and root health of creepingbentgrass in controlled environment stud-ies. Biostimulant is a broad term andrefers to products that may contain one ormore active ingredients including:cytokinians, nutrients, humates, organicacids, hormones, vitamins, microbial inoc-ulants, and other plant extracts.

Often biostimulant products con-tain either iron (Fe) or N or both.Manufacturer labels state that these prod-

ucts improve turf color, root growth, plantdevelopment, and environmental stress tol-erance. While N and Fe are known toimprove turf color and quality, it remainsunclear if products containing cytokiniansand/or other biostimulants provide anyadditional benefits in the field.

Lessons learned in 2007 resultedin a change in the treatment structure in2008. In 2007, IronRoots, Knife, Lesco's12-0-0 Chelated Iron Plus Micronutrients(hereafter Lesco 12-0-0), PanaSea Plus,Ultraplex, urea and Daconil Ultex wereevaluated for their ability to suppress dol-lar spot. Except for Daconil Ultrex, noneof the other products claim disease sup-pression on their labels.

As expected, plots treated withDaconil Ultrex exhibited the best summerquality in both IL and MD since it effec-tively controlled the disease. Urea was thenext best treatment, but urea-treated plotseventually were severely damaged by dol-lar spot. Except as noted below, none ofthe other treatments reduced dollar spotand plots treated with those products weredamaged severely, resulting in poor turfquality in both IL and MD. In MD, how-ever, some slight reduction in dollar spot

was observed in plots treated with Knife,but the effect was temporary and did notresult in acceptable quality at any time.

Products containing Fe and Nimproved turf color, but due to the pres-ence of disease, the plots could not be eval-uated for environmental stress impact onsummer quality. Hence, Daconil Ultrexwas replaced by Roots Concentrate in2008, and both sites were treated withfungicides to prevent blighting bypathogens.

Since PanaSea Plus, RootsConcentrate, and IronRoots contain no N,they were compared with and without Nfrom urea. Therefore, urea was appliedalone or tank-mixed with IronRoots, RootsConcentrate, and PanaSea Plus at 0.15 lbN/ 1000 ft2. All treatments were appliedon a 14-day interval, except Knife whichwas evaluated at two rates applied oneither a 14-day or 28-day interval as spec-ified on the label. All products were testedat label rates and application intervals asspecified on product labels. Both studysites were treated with fungicides in 2008throughout the study period to control dol-lar spot and brown patch (Rhizoctoniasolani). (continued on next page)

Evaluation of Cytokinian Plant Extract Biostimulants,Iron and Nitrogen Products For Their Effect On

Creeping Bentgrass Summer Quality


Both studies were conducted on USGA constructedbentgrass greens that were either a ‘G-2’ / ‘L-93 ‘ blend(IL) or ‘Providence’ (MD).

Peter DernoedenUniversity of Maryland

Objectives:1. To determine if products containing biostimulants, iron and/or nitrogen would affect dollar spot (Sclerotinia

homoeocarpa) severity and summer quality of creeping bentgrass (Agrostis stolonifera) putting green turf inIllinois and Maryland.

2. To evaluate the impact of products containing biostimulants, iron and/or nitrogen on summer quality of creepingbentgrass putting green turf in Illinois and Maryland in the absence of disease.

NDVI measurements were taken to supplement visualcolor ratings at both sites by use of a small handheldmeter.


Derek SettleChicago District Golf Association

Iron darkened the turfgrass at both sites immediatelyafter application and appeared as a checkerboard pat-tern across the study area where applied, but the effectwas transient unlike urea.

Iron-treated

The studies were conducted onmature stands of either 'G-2' + 'L-93' blend(IL) or 'Providence' (MD) creeping bent-grass grown on a sand-based rootzone con-structed to USGA recommendations. Theresearch greens were mowed five to sixtimes weekly to a height of 0.156 inches.The MD site received 1.75 lb N/ 1000 ft2

between April 19 and May 30 2008, andan additional 0.25 lb N/1000 ft2 wasapplied on August 6, 2008. The IL sitereceived 0.5 lb N/1000 ft2 during May2008. Otherwise, both sites did not receiveany supplemental N. All treatments wereapplied in 50-87 gal. water/A using a CO2pressurized backpack sprayer equippedwith an 8004 Tee Jet nozzle. Plots werearranged in a randomized complete blockwith four replications.

Turfgrass color and quality wereassessed visually on a 0 to 10 scale where0 = entire plot area brown or dead; 7 =minimum acceptable color and quality and10 = optimum greenness and unifomity.Chlorophyll levels influence turf color andnormalized difference vegetation index(NDVI) readings also were obtained usinga Field Scout TCM 500 Color Meter.Scalping injury was evaluated in MD on a0 to 5 visual scale where 0 = no scalping;2.5 = objectionable levels of scalping; and5.0 = severe injury.

The performance of treatments in

IL and MD were very similar. When datawere averaged over the season, treatmentscontaining urea provided the best quality atboth sites. There were no significant dif-ferences in color or quality in plots treatedwith urea alone compared to plots treatedwith IronRoots + urea, Roots Concentrate+ urea, or PanaSea Plus + urea at eithersite. Hence, little or no benefit wasobserved by tank-mixing any of the bios-timulant products with urea.

Furthermore, urea alone providedequal or better color and quality whencompared to all other treatments on mostrating dates in late summer at both loca-tions. Only treatments containing ureamitigated the injurious effects of scalpingin MD. When data were averaged over theseason, Ultraplex (IL and MD) and Lesco12-0-0 (IL) improved quality compared tothe control. Knife, IronRoots, RootsConcentrate and PanaSea Plus improvedquality in early summer in MD.

In IL, PanaSea Plus consistentlyimproved quality over the control frommidsummer on, whereas plots treated withKnife, IronRoots, and Roots Concentratehad a level of quality equal to the control.When data were averaged over the season,the aforementioned biostimulants had pro-vided a level of quality equivalent to thecontrol at both locations.

Visual color ratings often wereimproved by products containing Fe andN. PanaSea Plus and Roots Concentrate

sometimes improved color, but they didnot improve color to an acceptable level onmost rating dates. Color ratings, as deter-mined by NDVI, were similar betweensites. In both IL and MD, highest NDVIratings were observed in plots treated withurea alone or mixed with IronRoots, RootsConcentrate, or PanaSea Plus.

In MD, there were no other treat-ments that consistently improved color asmeasured by NDVI. In IL, plots treatedwith Ultraplex and Lesco 12-0-0 registeredNDVI ratings equivalent to urea-treatedplots when monitored 3 days, but not 7days, after application. Since NDVImeasures color as influenced largely bychlorophyll, it can be concluded that the Feapplied did not impact chlorophyll levels.Applications of Fe improved turf color byeliciting a darkening of foliar color. That isto say, the canopy does not appear greener,but darker.

Panasea Plus and Roots Concentrate, which do notcontain Fe or N, did not improve visual quality overuntreated control at either site.

The first year, products were evaluted for ability tosuppress dollar spot disease but were unable to do soat both sites.


Summary PointsEarly in the season in MD, IronRoots, Knife, Lesco 12-0-0, Ultraplex, and treatments

containing urea improved turf color and quality compared to the control and often pro-vided for enhancement of color and quality to a high level. In IL, differences amongtreatments became evident in July. Both Ultraplex and Lesco 12-0-0 consistentlyimproved turf quality and color compared to the control, whereas Knife had provided noapparent beneficial effect in IL.

When data were averaged over the season in both IL and MD, urea alone and treat-ments containing urea generally provided for best summer quality, but there were no dif-ferences among plots treated with urea alone or mixed with biostimulants.

Urea alone and treatments containing urea helped to mitigate scalping injury.

When data were averaged over the 2008 season, Lesco 12-0-0 (IL) and Ultraplex (ILand MD) improved quality compared to the control; whereas, IronRoots, Knife, PanaSeaPlus, and Roots Concentrate did not improve quality at either location.

Any potential visual benefits from the biostimulants were masked by the presence ofFe or N in Knife and IronRoots or by the N in urea when tank-mixed with RootsConcentrate or PanaSea Plus.

PanaSea Plus and Roots Concentrate, which do not contain Fe or N, had only a smallimpact on turf color and quality, and did not improve quality above the minimum accept-able level on most rating dates after early July in MD and mid-August in IL.

NDVI color ratings were consistently highest in plots treated with urea, IronRoots +urea, Roots Concentrate + urea, and PanaSea Plus + urea in IL (2007 and 2008) and MD(2008). Iron generally had little or no effect on color as measured by NDVI.

Urea alone and treatments containing urea helped mit-igate injury from scalping on a ‘Providence’ bentgrassgreen in MD, 2008

Panasea Plus Panasea Plus + UreaUreaLesco 12-0-0 plus Micros

Buffalograss [Buchole dactyloides

(Nutt.) Engelm] is a low-growing, warm-season turfgrass native to the Great Plainsof the US. It has an outstanding combina-tion of drought, heat, and cold toleranceand is ideally suited for turfgrass usewhere low or minimum inputs of water,nutrients, pesticides, and energy arerequired. Breeding and developing geno-types that meet the future need of the turf-grass industry requires the testing of elitegenotypes across a wide range of environ-mental conditions. For this reason, a trialconsisting of 20 turf-type buffalograssgenotypes was initiated in 2007.

In 2008, a second trial consistingof nine seeded and seven vegetative geno-types was established at nine locations ineight states. The experimental design wasa randomized complete block design withthree replications. The plots were 5 ft by 5ft. Data collected at each location werereported except the two locations inKansas.

At Mead, NE, data were collectedboth on 20 genotypes established in 2007and 17 genotypes re-established in 2008.Significant differences were observedamong genotypes tested for establishmentrate, seedling vigor, lateral spread, andcolor in 2007; for turfgrass cover, springgreen up, and color in 2008, and turfgrassdensity and quality in both years.

The 2008 trial also had significantdifferences among vegetative genotypesfor percent cover at 2, 4, and 8 weeks afterplantings, density, and quality. Seededgenotypes differed for percent cover two

weeks after planting and for summer den-sity. Some new experimental lines weresuperior to the best commercial cultivarcheck.

In Colorado, data were reportedonly for vegetative genotypes in 2007.Results indicated significant differencesamong genotypes tested for turfgrass coverat 2, 4, 8, and 12 weeks after planting, aswell as turfgrass leaf texture, spring green-up, density, and quality. Two genotypesconsistently outperformed the best checkat this location.

At Logan, Utah, 2007 data analy-sis indicated significant differences amonggenotypes tested for each group for turf-grass cover, quality, color, and leaf texture.The result indicated significant differenceamong genotypes for turfgrass color andquality. Some experimental lines demon-strated excellent performance.

At Las Cruces, NM, significantdifferences were observed among geno-types evaluated for turfgrass cover at dif-ferent times and fall discoloration. Someexperimental lines performed similar to orbetter than the commercially available

cultivars.At Tucson, AZ, significant differ-

ences were observed among seeded geno-types tested for turfgrass cover, color, andquality. Vegetative genotypes differed sig-nificantly only for percent cover. Twoexperimental lines performed similar to orbetter than the best commercially availablecultivar.

At Blacksburg, VA, significantdifferences were found among vegetativegenotypes for turfgrass cover, texture,color, and quality, but seeded genotypesdiffered only in turfgrass cover. Someexperimental lines were as good as the bestcommercially available check.

2007 Buffalograss Experimental Line and Cultivar Evaluation


Robert (Bob) Shearman and Bekele Abeyo University of NebraskaDavid M. Kopec University of ArizonaPaul G. Johnson Utah State UniversityRyan Goss New Mexico State University

Anthony Koski Colorado State UniversityMichael J. Goatley Virginia TechRodney St. John Kansas Sate UniversityGwen Stahnke Washington State University

Objectives:1. Assess the turfgrass performance of elite buffalograss genotypes across several locations involving a wide range

of environmental conditions and possibly make recommendations.

Breeding and developing genotypes that meet the future need of the turfgrass industry requires the testing of elitegenotypes across a wide range of environmental conditions.


Summary PointsSignificant differences were observed

among genotypes tested for most traits.

A few genotypes performed similar toor more than the best check in mostlocations.

Quality data were slightly low.

Genotypes performed differently to therange of environments at which they weretested.

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