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Utilization of black walnut trees and wood products by the invasive walnut twig beetle and the associated

fungal pathogen Geosmithia morbida: Components of the Thousand Cankers Disease complex, in eastern

Tennessee

Jackson Audley

M.S. Candidate

University of Tennessee

Department of Forestry, Wildlife and Fisheries

November 20th 2013 12:20 PM

160 Plant Biotechnology Building

What is Thousand Cankers Disease?

•  Walnut twig beetle (Pityophthorus juglandis) + fungal pathogen (Geosmithia morbida)

•  Invasive •  Native to the

southwest on Arizona walnut (Juglans major)

Tisserat et al. 2009; Cranshaw 2011; Moltzan 2011

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•  Cankers form around WTB galleries in the phloem

•  Not systemic •  Beetle feeding +

cankers = girdle branches

•  Top down dieback •  Symptoms: Yellowing

leaves in the crown; flagging; branch dieback

Tisserat et al. 2009

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A Brief History of TCD

•  Walnut decline across the west since 1990’s •  Walnut twig beetle (WTB) associated with decline in NM

in 2001

•  2007 unk. Geosmithia spp. Recovered in CO •  Thousand Cankers Disease (TCD) confirmed in J. nigra

native range – 2010 Knoxville, TN

•  Now confirmed in TN, NC, VA, PA, OH

Tisserat et al. 2009; Kolarik et al. 2011; Grant et al. 2011; Seybold et al. 2012, Ohio Department of Agriculture 2013, North Carolina Forest Service 2013

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Distribution of TCD as of June 2013 5

History continued

•  Black walnut and butternut (J. cinerea)- most susceptible

•  State quarantines: Wood; Firewood; Nursery Stock; Bark material

•  Required: Steam treated; fumigated; squared edge, kiln dried, bark-free lumber; and finished wood products

Utley et al. 2009; TN.gov

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Quarantine and Buffer Regulated Counties

TN.gov

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Why do we care?

•  Walnut: estimated >$500billion standing timber •  TN: $1.37 billion in urban areas and $1.47 billion in forest lands •  Veneer, furniture, gun stocks, cabinetry, and other specialty items •  Important ecological species; nuts = human and wildlife food

source

•  Invasive phytophagous insects on the rise •  Another iconic American hardwood species is threatened

Newton and Fowler 2009; Haun et al. 2010; Moltzan 2011; Aukema 2010

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Research Objectives

1.  Determine the ability of Pityophthorus juglandis to colonize treated black walnut wood – Post-treatment Colonization Study

2.  Determine the ability of Pityophthorus juglandis to colonize black walnut seedlings – Nursery Stock Study

3.  Determine the length of time Geosmithia morbida remains viable in cut black walnut wood – Geosmithia Persistence Study

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Post-treatment Colonization Study - Justification

•  Successful phytosanitation: Heat and Methyl Bromide fumigation treatment

•  Once treated, further protection?

•  Regulations – invasives still introduced •  Wood boring and bark beetles colonize slivers of bark left on WPM •  Costly for the walnut lumber industry •  Threat of further spread

Aukema et al. 2010; Haack and Petrice 2009; Mayfield et al. 2013; Myers et al.; Newton and Fowler 2009

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Post-treatment Colonization Study – Methods

´  n = 150, 70 cm(2ft) long pieces assigned to 1 of 5 treatments ´  90 walnut bolts (7-16cm diam):

´  30 heat treated; 30 fumigated; 30 no treatment ´  60 pieces of walnut lumber:

´ 30 bark on; 30 bark free ´  Bolts hung in 30 trees at 11 sites around Knoxville, TN with a WTB lure ´  Bolts hung for 30 days: from August 5th to September 5th 2013 ´  ½ placed into emergence barrel ´  ½ gallery length measured and WTB life stages recorded

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Post-treatment Colonization Study - Analysis

•  Response variables: •  Densities of beetles/treatment •  Gallery length/treatment •  WTB life stage count/treatment

•  Means compared independently using analysis of variance (ANOVA) in SAS

•  α = .05

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Nursery Stock Study - Justification

•  Are seedlings at risk of WTB attack?

•  Possible TCD pathway •  Seedlings are currently

restricted under quarantine

•  No method to treat

Aukema et al. 2010; Newton and Fowler 2009; thousandcankers.com

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Nursery Stock Study – Methods

No - Choice •  24 seedlings; 8 per size class:

•  = 2.5cm

•  50 beetles (25M and 25F) caged •  May 2014; 15 days in greenhouse •  Remaining adults and attack

holes counted

•  Emergence barrel •  Repeated August 2014

Choice •  24 seedlings by size class •  2 blocks of 12 seedlings – 3 clusters

of 4

•  1 infested bolt/cluster •  Set the groups out in May 2014 for

30 days

•  Attack holes will be counted •  ½ to emergence, ½ excavated for

galleries

•  Repeated in August 2014

Seybold et al. 2012

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Nursery Stock Study - Analysis

No-Choice Assay •  Response variables:

•  # adults in cage •  # of attack holes •  Emergence

•  Means compared independently using SAS ANOVA

•  α = .05

Choice Assay

•  Response variables: •  # of attack holes/seedling •  # of beetles emerged •  Gallery length

•  Means compared independently using SAS ANOVA

•  α = .05

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Geosmithia Persistence Study - Justification

•  How long will cut walnut wood remain a potential source?

•  Geosmithia morbida spores possibly “picked-up” by other species

•  Several ambrosia beetle spp on walnut – potential vectors

•  Recovery attained 12 weeks post cutting

Kolorik et al. 2011; Newton and Fowler 2009; Reed et al. 2013; Fraedrich personal communication

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Geosmithia Persistence Study – Methods

•  60, 80 cm walnut bolts (n=60) •  4 treatments (15/treatment): Inoculated

Outside; Inoculated Inside; Control Outside; Control Inside

•  4 wounds at 6 locations – agar plug •  Inoculation = Geosmithia morbida

culture and

•  Control bolts = ½ strength Potato Dextrose agar (PDA) plug

Utely et al. 2012; Klepzig et al. 1995

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4

3

2

110 cm

4 wk cut 24 wk cut

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Geosmithia Persistence Study –Methods •  The Outside bolts in mesh cages •  Inside in climate controlled room •  A cookie cut every 4 weeks •  Bark chipped and plated on ½

PDA

•  Incubated for 14 days •  Fungal growth assessed

Utely et al. 2012; Klepzig et al. 1995; Fraedrich personal communication

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Geosmithia Persistence Study - Analysis

•  Response variable: Proportion of plates with GM growth per treatment

•  Mean proportions analyzed using a Chi Squared test in SAS, •  Significant differences – pairwise comparisons using Fisher’s Exact

tests in SAS

•  α = .05

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References ´  Aukema, Juliann, Deborah G. McCullough, Betsy Von Holle, Andrew M. Liebhold, Kerry Britton and Susan J. Frankel. 2010. Historical Accumulation of Nonindigenous Forest Pests in the

Continental United States. BioScience, 60(11):886-897.

´  Cranshaw, W. 2011. Recently recognized range extensions of the walnut twig beetle, Pityophthorus juglandis Blackman (Coleoptera: Curculionidae: Scolytinae), in the western United States. The Coleopterists Bulletin 65: 48-49.

´  Fraedrich, Steven. Personal communication. Unpublished data from Geosmithia morbida persistence trials in 2012.

´  Grant, J.F., M.T. Windham, W.G. Haun, G.J. Wiggins, and P.L. Lambdin. 2011. Initial assessment of thousand cankers disease on black walnut, Juglans nigra, in eastern Tennessee. Forests 2: 741-748, doi:10.3390/f2030741.

´  Hack, Robert A. and Toby R. Petrice. 2009. Bark- and Wood-Borer Colonization of Logs and Lumber After Heat Treatment to ISPM 15 Specifications: The Role of Residual Bark. J. Econ. Entomol. 102(3): 1075Ð1084.

´  http://www.thousandcankers.com/media/docs/TN_TCD_Quarantine_9_2010.pdf

´  http://www.tn.gov/agriculture/publications/regulatory/TCD_regs_PlainLanguage.pdf 

´  Kolařík, M., E. Freeland, C. Utley, & N. Tisserat. 2011. Geosmithia morbida sp. nov. a new phytopathogenic species living in symbiosis with the walnut twig beetle (Pityophthorus juglandis) on Juglans in the USA. Mycologia 103: 325-332. 

´  Moltzan, B.D. 2011. National response framework for thousand cankers disease (TCD) on walnut. US Forest Service, Animal Plant Health Inspection Service, National Association of State Foresters, and the National Plant Board. October 2011. http://www.fs.fed.us/foresthealth/docs/TCD_National_Framework.pdf

´  Newton, L. and G. Fowler. 2009. Pathway Assessment: Geosmithia sp. and Pityophthorus juglandis Blackman movement from the western into the eastern United States. United States Department of Agriculture, Animal and Plant Health Inspection Service. 50 p.

´  North Carolina Forest Service 2013. Thousand cankers disease confirmed in North Carolina mountains. Forest Health Notes, Volume 201301-TCD, January 2013. http://ncforestservice.gov/forest_health/pdf/FHN/FHN1301TCD.pdf

´  Ohio Department of Agriculture 2013. Ohio Department of Agriculture encourages Ohio citizens to check their trees for signs of invasive pests: State taking measures against walnut twig beetle and Asian longhorned beetle; officials confirm thousand cankers disease for first time in Ohio. News release, 12 August 2013. http://www.agri.ohio.gov/public_docs/news/2013/08.12.13%20Tree%20Check%20Release.pdf

´  Reed, S., J. English, J. Juzwik, M. Ginzel. 2013. Bark and Ambrosia Beetles and Their Associated Fungi Colonizing Stressed Walnut in Missouri and Indiana. USDA Forest Service. http://www.nrs.fs.fed.us/pubs/jrnl/2013/nrs_2013_reed_001.pdf.

´  Seybold, S.J., T.W. Coleman, P.L. Dallara, N.L. Dart, A.D. Graves, L.A. Pederson, and S. Spichiger. 2012. Recent collecting reveals new state records and geographic extremes in the distribution of the walnut twig beetle, Pityophthorus juglandis Blackman (Coleoptera: Scolytidae), in the United States. Pan-Pacific Entomologist 88: 277-280.

´  Tisserat, N., W. Cranshaw, D. Leatherman, C. Utley, and K. Alexander. 2009. Black walnut mortality in Colorado caused by the walnut twig beetle and thousand cankers disease. Plant Health Progress, doi: 10.1094/PHP-2009-0811-01-RS.

´  Utley, C., W. Cranshaw, S. Seybold, A. Graves, C. Leslie, W. Jacobi, and N. Tisserat. 2009. Susceptibility of Juglans and Carya species to Geosmithia; a cause of thousand cankers disease. Poster presentation, American Phytopathological Society Meeting, August 1-5, 2009, Portland, Oregon

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Photo References ´  http://abernathyforestresources.com/wp-content/gallery/veneer- ´  logs/Black%20Walnut%20Veneer%20Logs%202.jpg ´  http://dnr.wi.gov/topic/foresthealth/thousandcankers.html ´  http://thousandcankers.com ´  http://www.1stdibs.com/furniture/tables/dining-room-tables/burled-walnut-veneer-table/id-f_612023/ ´  http://www.agnr.umd.edu/news/article.cfm?id=fbe34d590a5a5a8f00985fc6077a013a ´  http://www.daff.gov.au/biosecurity/import/timber/timber-packaging-and-dunnage-import-

requirements/bark

´  http://www.forestryimages.org/browse/detail.cfm?imgnum=5406097 ´  http://www.insectimages.org ´  http://www.tn.gov/agriculture/publications/regulatory/tcd_map.pdf ´  Jackson Audley personal photos. 2013. ´  Jim LaBonte, Oregon Department of Agriculture ´  mda.mo.gov ´  Scott Myers personal photos. 2012. ´  UC Davis Department of Entomology http://169.237.77.3/news/walnuttwigbeetle.html

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Acknowledgements

Committee: Adam Taylor (UT) Albert “Bud” Mayfield (USFS) Scott Myers (APHIS) Bill Klingeman (UT)

Funding Sources: USDA Forest Service USDA APHIS

Field and Lab assistance: Andy Tait Robert Camp Steven Fraedrich

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Questions?

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Or email me @ Jaudley@utk.edu