pwaisen nematology discussion

Management of Root-knot and Reniform Nematodes using Oil Radish and Mustard as Cover Crops

Committee MembersDr. Koon-Hui Wang (Chair)

Dr. Brent Sipes Dr. Zhiqiang Cheng

Dr. Joe DeFrankDr. James Leary

February 17th, 2017Time: 9:00-10:00 AMRoom: Gilmore 302

Philip WaisenDissertation Proposal Seminar

Economic Damage of Plant-parasitic Nematodes

•Worldwide there are > 4,100 species of plant-parasitic nematodes.

•$100 billion/yr loss worldwide.

•$10 billion/yr loss in USA. •Root-knot nematodes can cause 20-38% crop loss.

Okra

tomato

Beet

Root-knot Nematodes

Picture: Koon-Hui Wang

Split roots of carrot

Blemishes on cross section of a potato tuber

Cucurbit crops are most susceptible

Kona root-knot nematodes on coffeePicture: Society of Nematologists

Crops in Hawai’i most damaged by Reniform nematode

Pineapple

Papaya Cowpea

Reniform Nematodes

Sweet potato

Broad host range

http://www.google.com/imgres?imgurl=http://mossagrow.files.wordpress.com/2008/06/papaya1.jpg&imgrefurl=http://mossagrow.wordpress.com/2008/06/20/the-papaya/&usg=__2k5_qFBKh11r_GiM7nlgfh2CrIc=&h=480&w=640&sz=89&hl=en&start=5&um=1&itbs=1&tbnid=g9-pZ9hAJ1vUoM:&tbnh=103&tbnw=137&prev=/images?q=papaya&um=1&hl=en&sa=N&rlz=1T4WZPC_enUS360US361&tbs=isch:1

http://www.google.com/imgres?imgurl=http://mossagrow.files.wordpress.com/2008/06/papaya1.jpg&imgrefurl=http://mossagrow.wordpress.com/2008/06/20/the-papaya/&usg=__2k5_qFBKh11r_GiM7nlgfh2CrIc=&h=480&w=640&sz=89&hl=en&start=5&um=1&itbs=1&tbnid=g9-pZ9hAJ1vUoM:&tbnh=103&tbnw=137&prev=/images?q=papaya&um=1&hl=en&sa=N&rlz=1T4WZPC_enUS360US361&tbs=isch:1

http://www.all-creatures.org/recipes/images/i-blackeyedpeas.jpg

http://www.all-creatures.org/recipes/images/i-blackeyedpeas.jpg

Cover Crops Suppressive to Plant-parasitic Nematodes

Radish and mustard-- glucosinolate

Sunn hempCrotalaria juncea-- monocrotaline

French Marigold Tagetes patula

-- α-terthinyl

Sorghum-sudangrass-- Dhurrin

1) Non-host or poor host 2) Trap crop3) Allelopathy 4) Enhance nematode antagonistic

microorganisms 5) Enhance host tolerance/resistance

Mechanisms of cover crops against Plant-parasitic Nematodes

Cover crops

Root-knot nematode

Reniform nematode

Biofumigation effect

Trap crop effect Soil health

Oil radish Poor host Poor host Strong Slightly Tillage effect

Mustard Excellent host Non-host Stronger Good for root-knot Not much information

Oil radish and Mustard as Trap and Biofumation crops

Objective 1 Objectives 2 & 3 Objective 4

Objectives1) Screen oil radish and mustard cultivars/species for their potential to serve

as trap crops and biofumigation crops against M. javanica and R. reniformis.

2) Determine degree days required for M. javanica to complete a life cycle on mustard.

3) Examine best time to terminate oil radish for optimum nematode trapping.

4) To compare the effects of oil radish and mustards termination methods on biofumigation and soil health.

Objective 1Screen oil radish and mustard cultivars/species for their potential to serve as trap crops and biofumigation crops against M. javanica and R. reniformis.

Oil radish cultivars: AC = April Cross; AL = Alpine; DI = Discovery; MI = Miyashige; OS = Oshin; SC = Summer Cross; SB = Sodbuster; TR = Tillage Radish.

Mustard species: Brassica hirta, Brassica juncea ‘Pacific Gold’, B. hirta + B. juncea (‘Caliente 199’)

Objective 1Two greenhouse experiments were conducted at Magoon Research & Teaching Facility.

Pi = 1000 J2 of M. javanica

Biofumigation test

Susceptibility test

1% (dw/dw), Mj and Rr infested soil

Objective 1

1 month after initiation of biofumigation test

Green manure effect

Objective 1Experiment 1.1. Biofumigation effects of oil radish

NA = no amendment; AC = April Cross; AL = Alpine; MI = Miyashige; OS = Oshin; SB = Sodbuster; SC = Summer Cross; TR = Tillage Radish.

Initial populations:Meloidogyne spp. = 2130/250 cm3

Rotylenchulus reniformis = 2270//250 cm3

Objective 1Experiment 1.2. Susceptibility of oil radish cultivars to M. javanica

NA = no amendment; AC = April Cross; AL = Alpine; MI = Miyashige; OS = Oshin; SB = Sodbuster; SC = Summer Cross; TR = Tillage Radish.

1 month after inoculation

Seed Cost

Objective 1

Experiments 1.3. Screening mustards for susceptibility to M. javanica.

‘Caliente 199’ (Brassica juncea & Brassica hirta) ‘Pacific Gold’ brown mustard (Brassica juncea) Yellow mustard (Brassica hirta) ‘Orange Pixie’ tomato (susceptible control)

Objective 1Experiments 1.3. Screening mustards for biofumigation & susceptibility to M. javanica and R. reniformis

Biofumigation test

Susceptibility test

Root-knot Reniform0

2000

4000

6000

8000

10000

Egg

coun

ts/r

oot

syst

em

Preliminary ResultField soil 1% dw/dw

Pi = 1000 M. j‘Caliente 199’

• ‘Sodbuster’ oil radish has a potential to serve as trap crop for M. javanica and as biofumigation crop against Meloidogyne spp. and R. reniformis.

• ‘Caliente 199’ mustard mix is a very good host for M. javanica.

Objective 1 (Summary)

Objective 21) Screen oil radish and mustard cultivars/species for their

potential to serve as trap crops and biofumigation crops against M. javanica and R. reniformis.


3) Examine best time to terminate oil radish for optimum trap.


Cover crops

Root-knot nematode

Reniform nematode

Biofumigation effect

Trap crop effect (Degree days)

Soil health

Oil radish

Poor host Poor host Strong Slightly Tillage effect

Mustard Excellent host Non-host Stronger Good for root-knot None

Objective 2

Objective 1 Objectives 2 Objective 4

Dead-end trap crop

• Sugarbeet cyst nematode (Heterodera schachtii) on radish (Smith et al., 2004).

• Root-knot nematode (Meloidogyne hapla) (450-500 DD, base 10°C) on oil radish (Melakeberhan et al., 2008).

Conventional trap crop

Non-host Sink

Host Sink and source

Nematode Degree DaysPlant-parasitic

NematodesCalendar days

(CD)Degree days

(DD)Base temperature

(°C)Host Reference

M. javanica 19 334 10.6 Okra Dávila-Negrón & Dickson (2014)

M. incognita 15 300 9.8 Okra Dávila-Negrón & Dickson (2014)

M. arenaria 19 316 10.3 Okra Dávila-Negrón & Dickson (2014)

M. hapla 450-500 10 Radish Melakeberhan et al., 2008

DD = Σ{[(Tmax + Tmin)/2]-Tbase}. Mustard?

Objective 2: Degree Days Experiment• A greenhouse mustard experiment will be

conducted.• Treatments: Mustard inoculated with 200 Mj

J2 will be stained for nematode developmental stages every 4 days up to 40 days using Acid Fucshin method; 4 reps.

• WatchDog Temperature Data Logger will be buried in soil.

• J3, J4, egg-laying females will be counted.• Degree days (DD, base 10.6°C) to reach

egg-laying female will be determined using the formula, DD = Σ{[(Tmax + Tmin)/2]-Tbase}.

• Experiment will be repeated once.

Objective 31) Screen oil radish and mustard cultivars/species for their potential to serve as trap

crops and biofumigation crops against M. javanica and R. reniformis.


3) Examine best time to terminate oil radish for optimum nematode trapping.


8 weeks

6 weeks

4 weeks

2 weeks

0 week

Growing period of Oil Radish as a Cover Crop

Term

inati

on o

f oil

radi

sh

Oil radish was planted for different length of time (0, 2, 4, 6 and 8 weeks). Experiment was arranged in RCBD with 4 replications. Pumpkin was planted after oil radish (OR) termination and incoporation, nematodes were sampled at OR termination and at 4 weeks after pumpkin planting.

Objective 3: Termination time for oil radish

20lb seeds/acre

RGI = 0 RGI = 12RGI = 6 RGI = 9

Root Gall Index based on 0 – 12 scale

Severity of pumpkin root galls by root-knot nematodes

Nematodes/250 cm3 soilHerbivores 0 2 4 6 8Root-knot nemamtode 178 A 140 A 213 A 160 A 467 AReniform nematode 371 A 256 A 874 A 168 A 312 AStubby root nematode 36 A 20 A 32 A 22 A 33 A

Repeated measure over 4 sampling dates at monthly interval

Oil radish did not suppress PPN in the soil but reduce root galls on pumpkin

Soil Health

Enrichment indexChannel index

Structure index

Richness, diversity

Bacterivore Fungivore Herbivore Omnivore Predator

Free-living nematodes

Nematode community AnalysisBased on their feeding behaviour

Objective 3: Results Weeks of oil radish growth

Nematode 0 2 4 6 8-----------------------Nematodes/250 cm3 soil------------------------

Bacteriovore 632 BC 550C 748B 1423A 1416AFungivore 461 ABC 309C 358BC 613A 841ABOmnivore 6 B 13AB 27A 17AB 29APredator 0A 0A 0A 1A 4ARichness 13A 13A 13A 14A 15ADiversity 5.24A 5.87A 4.46A 5.67A 5.34AF/(F+B) 0.44A 0.40AB 0.31C 0.33BC 0.33BCEI (%) 58C 58C 68AB 71A 62BCSI (%) 4.75A 10.29A 13.6A 5.78A 10.79ACI (%) 43A 36AB 24C 23C 27BC

• Planting oil radish for > 6 weeks enhanced bacterial decomposition == enriched soil food web, green manure effect

Throughout the 3-month of pumpkin crop

Objective 3: Results Weeks of oil radish growth

Nematode indices 0 2 4 6 8

Nematodes/250 cm3 soil

------------------July 7, 2016---------------------

F/(F+B) 0.46 A 0.41 A 0.25 B 0.25 B 0.33 AB

EI (%) 45.53 B 48.51 B 68.75 A 81.01 A 74.25 A

------------------August 4, 2016-----------------

F/(F+B) 0.45 AB 0.52 A 0.24 C 0.32 BC 0.17 C

EI (%) 68.74 A 68.07 A 72.63 A 70.43 A 53.48 A

--------------- September 23, 2016--------------

F/(F+B) 0.40 A 0.31 A 0.36 A 0.40 A 0.45 A

EI (%) 57.28 A 63.24 A 67.18 A 69.28 A 66.66 A

---------------- October 25, 2016----------------

F/(F+B) 0.47 A 0.34 A 0.38 A 0.37 A 0.39 A

EI (%) 59.31 A 55.64 A 63.22 A 63.12 A 52.61 A

• However the enrichment effects of oil radish amendment was transient.

Objective 3 (Summary)• Terminating oil radish at 4 weeks after planting reduced root gall

index and increased total fruit weight of pumpkin.• Oil radish only enhanced bacterial decomposition 1 week to 1

month after soil incorporation.

More improvement is needed to use Brassica-based cover crop for nematode management.

Objective 41) Screen oil radish and mustard cultivars/species for their

potential to serve as trap crops and biofumigation crops against M. javanica and R. reniformis.


3) Examine best time to terminate oil radish for optimum trap.


Oil Radish and Mustard as Biofumigants

Cover with black plastic

(Ploeg and Stapleton, 2001)

Isot

hioc

yana

tes

Weed whackedSoil incorporated

(Rosa et al., 1999)(Ohtsuru et al., 1973)

Aspergillus, Rhizobacter

Objective 4. Enhance biofumigation effects by cover crop termination practices

1) ORT=oil radish + weed whack + till2) ORBP=oil radish + weed whack + till + black plastic 3) ORNT=oil radish + sickle + weed mat (=NT)

7 Treatments 4) MST=mustard + weed whack + till5) MSBP=mustard + weed whack + till + black plastic 6) MSNT=mustard+sickle + weed mat (NT)7) BG=Bare ground

Objective 4: Biomass production

1) ORT=oil radish + weed whack + till; 2) ORBP=oil radish + weed whack + till + black plastic; 3) ORNT=oil radish + sickle + weed mat; 4) MST=mustard + weed whack + till; 5) MSBP= mustard + weed whack + till + black plastic; 6) MSNT=mustard + sickle + weed mat; 7) Bare ground control

Objective 4: Contrast Analysis

Nematodes BG ORNT MSNT ORT MST ORBP MSBP T vs NT T(+BPvs -BP)

BP vs

BG

MSvs OR

NT vs T(-BP)

NT vsT+BP

------------------------------------------------------------Numbers/250 cm3 soil---------------------------------------------------

Root-knot 298 222 390 412 335 242 205 NS NS NS NS NS NS

Reniform 320 630 472 602 635 365 380 NS NS NS NS NS NS

% Bacterivores

33 20 26 37 43 41 44 ** NS NS NS ** **

% Fungivores 17 22 17 12 22 17 15 NS NS NS NS NS NS

% Herbivores 46 52 52 45 30 29 27 ** NS * NS * **

% Omnivores 0.15 0.27 0.61 0.23 0.15 0.49 0.36 @ ** * NS ** NS

% Predators 0 0 0 0 0 0.22 0 NS * @ @ NS *

Richness 14 15 18 14 17 19 17 NS * ** NS NS @

Diversity 7 4 6 6 7 9 10 ** * @ NS NS **

MI 1.91 1.96 2.15 1.75 1.69 1.82 1.7 ** NS @ NS ** **

EI (%) 54 61 52 70 71 73 74 ** NS * NS * *

SI (%) 81 85 85 83 74 76 75 * NS NS NS NS *

CI (%) 40 40 43 18 25 17 15 ** NS * NS * **

1 week after CC termination

Biofumigation effects of oil radish and mustard on nematodes


• Tilling brassica cover crops into the soil suppressed % herbivores better than NT (P < 0.05).

• Maceration of cover crops followed by covering with black plastic (BP) reduced % herbivores better than BG (P < 0.05) based on contrast analysis.


Biofumigation effects of oil radish and mustard on bacterivores


• Tilling brassica cover crops into the soil increased % bacterivores compared to NT (P < 0.05), indicating an enhancement of bacterial decomposition.

• Biofumigation effect did not suppress bacterivores (BP vs no BP were not different).


Biofumigation effects of oil radish and mustard on omnivores


• NT increased % omnivores compared to tilled plots (P < 0.01), indicating an improved soil food web structure.

• However, covering tilled plots with BP increased % omnivores better than no BP (P<0.05). In fact BP increased % omnivores compared to BG (P<0.05).


Objective 4 (Summary)

Enrichment indexChannel index

Structure index

Richness, diversity

Bacterivore Fungivore Herbivore Omnivore Predator

Free-living nematodes

Tilled + BPNTTilled + BP

Plant growth after incorporation of radish and mustard green manures

BG MST ORT MSNT ORNT ORBP MSBP30

32

33

35

36

38

Chlo

roph

yll (

SPAD

uni

ts)

D

AB

A

ABAB

BCCD

BG MST MSNT ORT ORBP ORNT MSBP0

20

40

60

80

Cano

py w

idth

(cm

)

C

A AAABBB

2 weeks after planting

Conclusions ‘Sodbuster' oil radish is a potential conventional trap crop and a good biofumigant

cultivar against root-knot and reniform nematodes. Terminating oil radish at 4 weeks by tilling only reduced galling by 27% on

pumpkin, but increased the total fruit weight by 74%. However, biofumigation (weed whacking + till + black plastic) reduced herbivorous

nematodes by 39%. None-the-less, oil radish/mustard cover cropping (weed whacking + till + black

plastic) increased bacterivorous nematodes compared to NT and BG. NT increased % omnivores compared to tilled plots. Covering tilled plots with BP increased % omnivores better than no BP. In fact BP

increased % omnivores compared to BG.

Acknowledgements

Adviser: Dr. Koon-Hui Wang

Dissertation committees: Dr. Brent Sipes, Dr. Zhiqiang Cheng, Dr. Joe DeFrank and Dr. James Leary

Technical support staff: Donna Meyer, Gareth Nagai and Steve Yoshida

Poamoho Experiment Station: Farm crew Jari Sugano

Extension Agents: Jari Sugano and Jensen Uyeda

Funding source

Sustainable Pest Management LabUniversity of Hawai’i at Mānoa,

College of Tropical Agriculture and Human Resources

Shelby ChingJosiah MarquezJonathan KamBishnu Bandari

Objective 4.

1 week after covering weed mat 1 week after covering black plastic

‘Felix’ zucchini transplanted at 3 ft

spacing

pwaisen nematology discussion

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