Dead Microbial Insecticides

IR-4 Western Regional

Workshop

EPA Registered Bio-insecticides that are Dead

© 2016 Marrone Bio Innovations -

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Dead = heat-killed or no viable cells in end product Marrone Bio Innovations Grandevo® - 30% Chromobacterium subtsugae and spent fermentation media (no viable cells), DF and WDG formulations. Venerate® - 94.46% heat-killed Burkholderia rinojensis strain A396 cells and spent fermentation media, aqueous-based formulation sold as Venerate or Venerate XC.

GRANDEVO® Bioinsecticide

New species of Chromobacterium (subtsugae) isolated from forest soil from Maryland by Dr. Phyllis Martin of the USDA-ARS

C. subtsugae strain PRAA4-1T

– Commercial product is dead bacteria plus associated compounds produced in the cells and spent fermentation media

– DF (WP) and WDG formulations

Page 3 Photos courtesy of: Lygus Entomart; Beet armyworm Clemson Univ. USDA Cooperative Extension Slide Series Bugwood.org; Western flower thrip Frank Peairs, CO St. Univ. Bugwood.org; citrus leaf miner

Center for Invasive Species Research, UC Riverside; : cabbage Looper RJ Reynolds Tobacco Company, Bugwood.org; twospotted spider mite Clemson EDU

GRANDEVO® Bioinsecticide

Registered in the U.S. and in Mexico

Residues are exempt from tolerances

4 hour REI, 0 days to harvest Easy on pollinators and

beneficials NOP compliant, OMRI listed

Page 4 Photos courtesy of: Lygus Entomart; Beet armyworm Clemson Univ. USDA Cooperative Extension Slide Series Bugwood.org; Western flower thrip Frank Peairs, CO St. Univ. Bugwood.org; citrus leaf miner

Center for Invasive Species Research, UC Riverside; : cabbage Looper RJ Reynolds Tobacco Company, Bugwood.org; twospotted spider mite Clemson EDU

VENERATE® XC Bioinsecticide

New bacterial species of Burkholderia (rinojensis strain A396) – Discovered in MBI’s discovery

screen; isolated from soil; not related to pathogenic species

– Commercial product contains heat-killed cells, spent fermentation media and associated compounds

– Aqueous formulation

Page 5 Photos courtesy of: Pepper weevil Alton N Sparks, Univ of GA, Boxwood.org; Beet armyworm Clemson Univ. USDA Cooperative Extension Slide Series Bugwood.org; Western flower thrip Frank Peairs, CO St.

Univ. Bugwood.org; Cabbage Looper RJ Reynolds Tobacco Company, Bugwood.org; Twospotted spider mite Clemson EDU

VENERATE® Bioinsecticide

Registered in the U.S. and Mexico

Residues are exempt from tolerances

4 hour REI, 0 days to harvest Easy on pollinators and

beneficials NOP compliant, OMRI listed

Page 6 Photos courtesy of: Pepper weevil Alton N Sparks, Univ of GA, Boxwood.org; Beet armyworm Clemson Univ. USDA Cooperative Extension Slide Series Bugwood.org; Western flower thrip Frank Peairs, CO St.

Univ. Bugwood.org; Cabbage Looper RJ Reynolds Tobacco Company, Bugwood.org; Twospotted spider mite Clemson EDU

Some Active Compounds Produced by Chromobacterium subtsugae

CH3O

HO

OCH3

N

O

H

ONH

H

O

NH

H3C

ON

N

O

O

H O

H3C CH3

NO

CH3

CH3

HO H

H

Chromamide A Chromobacterium subtsugae

N

NOH

H

NOH

HO

Deoxyviolacein

Chromobacterium subtsugae

N

NOH

H

NOH

HO

Violacein Chromobacterium subtsugae

Some Active Compounds Produced by Burkholderia rinojensis strain A396

O

N

CH3H3C

H2NO

Templazole B Burkholderia rinojensis

N CH3 HO OH

OH3CO

O

H

OH3CCH3

O

HO

CH3OH

OH

CH3

Templamide A Burkholderia rinojensis

N CH3 HO OH

OH3C

O

O

H

OH3CCH3

O

O

CH3OH

CH3

Templamide B Burkholderia rinojensis

N

O

N

H

OOH3C

CH3H3C

Templazole A Burkholderia rinojensis

Modes of Action for C. subtsugae and B. rinojensis

Both Grandevo and Venerate function as stomach poisons in laboratory bioassays with species such as Trichoplusia ni and Spodoptera exigua and resemble Bt-based insecticides; rapid cessation of feeding and death in 3-5 days. Both products will also kill piercing sucking insects such as Lygus in sachet feeding assays. The mode-of-action at the cellular level is unknown. Both products also have biological activity via contact or exposure and possibly through ingestion of active compounds that have moved into a treated plant through translaminar movement.

Mode-of-action - Grandevo

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Mode-of-action - Venerate

A B

C D

Larvae are stunted

Problems molting

Problems molting, pupating, and developing

Liquefied frass

Spectrum of Activity – both products

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Certain Lepidoptera, especially Torticidae Certain leaf-feeding beetles and larvae (Chrysomelidae) Certain weevils (Curculionidae) Certain grubs (Scarabaeidae) Certain Heteroptera (Hemiptera and Homoptera) Thysanoptera Certain Diptera Mites

Compatibility With Beneficials – 3X Maximum Field Rate 24 hour Acute Test

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Beneficial species

Grandevo Venerate

Bombus sp. No effect No effect Orius insidiosus – minute pirate bug*

No effect* No effect*

Atheta coriaria – rove beetle*

No effect* No effect*

Aphidoletes apidimyza – predatory midge

No effect No effect

Aphidius colemani – aphid parasitoid

No effect No effect

*No effects on reproduction

Compatibility With Beneficials – 3X Maximum Field Rate 24 hour Acute Test

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Beneficial species

Grandevo Venerate

Cryptoplaimus montrouezi – mealybug destroyer

No effect No effect

Delphastus catalinae – whitefly predator

No effect No effect

Encarsia formosa – parasitic wasp

No effect No effect

Green lacewing No effect No effect Ladybird beetle No effect No effect Amblysius swirskii – predatory mite

LD50 4.3X max label rate

LD50 1.5x max label rate

Primary use crops – both products

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• Tree fruit • Tree nuts • Fruiting vegetables • Small fruit and berries • Alfalfa • Potatoes • Corn, cotton, soybean (seed treatment) –

Venerate only

Cabbage Looper Bioassay Test Results

Grandevo DF: Effect of Adjuvants

0

2

4

6

8

10

12

GVO DF BondMaxx

Biolink LI 700 Tactic ContactXcel

Activator90

Phase(MSO)

Attach Franchise

LC50

0.27

0.39 0.32

0.96

0.55

0.19

0.49 0.48

0.16 0.20

0.37

0.56

0.39

0.00

0.20

0.40

0.60

0.80

1.00

1.20

Cabbage Looper Bioassay Test Results

Venerate : Effect of Adjuvants

LC50

Adjuvants Evaluated with Grandevo DF

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Adjuvant LC 50 Description Function Rate per 100gal 1. Bond Maxx 0.79 Synthetic latex, 1, 2 propanediol and alcohol

Ethoxylate Spreader Sticker Deposition aid

12-24oz/100gal

2. Biolink- spreader And sticker.

4.48 Soap bark alkyl phenol ethoxylate, polysaccharide

Spreader sticker 2-4oz/100gal

Biolink- surfactant and penetrant.

0 Yucca and Garlic extract Surfactant and penetrant

05-2quarts/100gal

4. LI- 700 7.34 Phosphatidylcholine, methylacetic acid and alkyl polyoxyethlene ether

Soy oil derived 1-4pints/100gal

5. Tactic 9.76 Synthetic latex, 1,2 propanediol, alcohol ethoxylate and silicone polyether copolymer

Sticker organosilicone surfactant deposition aid

8-16oz/100gal

6. Contact Xcel 2.77 Linear alcohol ethoxylate and propylene glycol 25ml/100L

7. Activator 90 2.05 Alkylphenol ethoxylate, alcohol ethoxylate and tall oil fatty acids

1-4pints/100gal

8. Phase 2.32 Methyl esters of fatty acids, alkyl polyoxyethylene ether and polyether modified polysiloxane

Blend of methylated seed oil

1-4pints/100gal

9. Attach 1.72 Pinene(terpene) polymers, petrolatum, alpha(p- dodecylphenyl) omega- hydorxypolys (oxyethelene)

Pinene based 4-16oz/100gal

10 Franchise 2.91 Lecithin, methyl esters of fatty acids and alcohol ethoxylate

Lecithin based 3-4oz/100gal

Impact of Grandevo DF on SWD Mortality After Contact with Treated Blueberries (48 hours)

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Personal communication, Andrew Cuthbertson, Fera Science Ltd, UK.

Impact of Grandevo on SWD Oviposition After Contact Sprays

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Laboratory of Behavioral and Development Genetics, University of Leuven, Belgium, 2016

DF WDG DF

WDG

Impact of Grandevo on SWD Oviposition After Feeding on Treated Food

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Laboratory of Behavioral and Development Genetics, University of Leuven, Belgium, 2016

DF WDG

San Jose Scale Control on Apple – New York 2016

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0123456789

VEN XC 1 qt2C-5C

VEN XC 2 qt2C-5C

GVO WDG 1lb. 2C-5C

GVO WDG 2lb. 2C-5C

SivantoPrime SL @14 fl. oz. PK

SivantoPrime f/b

Movento oz.PK, 1C

LorsbanAdvanced

EC 64 fl. oz.TC

Imidan 3 lb.PF, 1C-6C

UTC

% SJS Damage at Harvest

Dr. Art Agnello, Cornell University. Sivanto and Movento applications included LI-700. No adjuvant included with Venerate or Grandevo applications. P=0.05

b ab ab b b b b b a

San Jose Scale Control on Peaches – New Jersey 2016

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05

10152025303540

VEN XC 2qt CSx2

GVO WDG2 lb. CSx2

Damoil 4gal. DD

Movento 9fl. oz. CS

Sivanto @10.5 fl. oz.

CS

Sivanto @14 fl. oz. CS

Closer 5.75oz. CS

UTC

% Fruit Injury

Dr. Anne Nielsen, Rutgers, Bridgeton, NJ. Season total of crawlers/5 cm tape in UTC = 657. Movento included LI-700. P=0.05.

ab b b b b b b a

Venerate Rotations with Radiant® for Control of Western Flower Thrips on Strawberry-2016

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05

101520253035404550

3DA-A 7DA-A 3DA-B 9DA-B 3DA-C 7DA-C 3DA-D 8DA-D

UTCRadiant @ 10 fl. oz.VEN XC @ 1 QTVEN XC @ 2 QTVEN XC @ 4 QTRAD/VEN XC @ 2 QT

Better Crops LLC, Dr. John Curtis – Florida 2016

Application dates: 3/26, 4/2, 4/11, 4/18 in 60 GPA.

# la

rvae

/25

flow

ers

Venerate XC Rotations with Radiant® for Control of Western Flower Thrips on Strawberry-2016

25

0

10

20

30

40

50

60

70

80

3DA-A 7DA-A 3DA-B 9DA-B 3DA-C 7DA-C 3DA-D 8DA-D

UTCRadiant @ 10 fl. oz.VEN XC @ 1 QTVEN XC @ 2 QTVEN XC @ 4 QTRAD/VEN XC @ 2 QT

Better Crops LLC, Dr. John Curtis – Florida, 2016

Application dates: 3/26, 4/2, 4/11, 4/18 in 60 GPA.

# ad

ults

/25

flow

ers

Rosy Apple Aphid Control in Apples– Michigan 2016

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0

5

10

15

20

25

30

UTC GVO WDG 2lb

GVO WDG 3lb

VEN XC 1 qt VDN XC 2 qt

# RAA and GAA colonies per 100 terminals 4DAT

Dr. John Wise, MSU. Treatments applied June 23, counts taken June 27. P=0.05.

A a B b B b B b B b

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Corn Root Protection With Venerate XC – 2015 Trials ISU 0-3 Nodal Root Damage Ratings & Meta-analysis

1.9

1.2 1.1

0.8 0.9

0.0

0.5

1.0

1.5

2.0

2.5

Untreated Venerate XC @ 170ml/100 m-row

Venerate XC @ 340ml/100 m-row

Venerate XC @ 680ml/100 m-row

Capture LFR @ 47.6ml/100 m-row

15-008NWV (IA) 15-009NWV (MN) 15-010NWV (WI) 15-012NWV (OH) Meta Avg

Trial design was a randomized complete block with 6 replications. A replication was 4 rows on 0.76 meter spacing x 9-15 meters in length. Treatments applied with planter-equipped units delivering materials in an in-furrow band over the open seed furrow before row closure. Means followed by different letters are statistically different from one another according to LSD test; actual P value = 0.0001 in all cases. Meta-analysis data was analyzed by Friedman Two-Way Nonparametric AOV with means separated by Dunn's All-Pairwise Comparisons Test, P value = 0.0001.

a a a a A bc b a b AB bc b b c BC c c c e BC b c d d BC

Treat now Not now

Neither Grandevo or Venerate are Knockdown Insecticides, Both Must be Applied Early

Best Use and Testing Recommendations for Grandevo DF/WDG and Venerate XC

Grandevo DF/WDG Venerate XC Adjuvant Yes (avoid LI-700) Not required (avoid LI-

700) Sticker Yes if rainfall expected Yes if rainfall expected pH 6.5 – 7.5 Yes Yes Adjust water hardness Yes for DF if >1000

ppm No

Spray interval 7 days (SWD may be 5 days)

7 days

Typical use rate 1-3 pounds/acre 1-4 quarts/acre

Best Use and Testing Recommendations for Grandevo DF/WDG and Venerate XC

Grandevo DF/WDG Venerate XC Alternate row middles Not at this time Not at this time Sensitive to water volume

Yes and more isn’t better

Not that we know of

Tank-mix compatible Yes (but not with VEN) Yes (but not with GVO) Shelf-life 3 years minimum 3 years minimum Timing Treat early for most

pests Treat early for most pests

Rotation Generally recommended

Generally recommended

Best Use and Testing Recommendations for Grandevo DF/WDG and Venerate XC

1.Treat early in the pest population life cycle. Established treatment thresholds might not be relevant.

2.Use a quality adjuvant with both Grandevo DF/WDG and with Venerate XC (avoid LI-700).

3.For lepidopteran pests apply at egg hatch

4.Plot size and untreated buffers

Questions?

Tim Johnson, PhD

VP Field Development and Technical Services

IR-4 Western Regional

Workshop