IR-4 Western Regional Workshop. Dead Microbial Insecticides

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1 IR-4 Western Regional Workshop Dead Microbial Insecticides

2 EPA Registered Bio-insecticides that are Dead 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 % heat-killed Burkholderia rinojensis strain A396 cells and spent fermentation media, aqueous-based formulation sold as Venerate or Venerate XC Marrone

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

3 GRANDEV Bioinsecticide New species of Chromobacterium (subtsugae) isolated from forest soil from Maryland by Dr. Phyllis Martin of the USDA-ARS C. subtsugae strain PRAA4-1 T Commercial product is dead bacteria plus associated compounds produced in the cells and spent fermentation media DF (WP) and WDG formulations Photos courtesy of: Lygus Entomart; Beet armyworm Clemson Univ. USDA Cooperative Extension Slide Series Bugwood.org; Western flower thrip Frank Peairs, C St. Univ. Bugwood.org; citrus leaf miner Page 3 Center for Invasive Species Research, UC Riverside; : cabbage Looper RJ Reynolds Tobacco Company, Bugwood.org; twospotted spider mite Clemson EDU

4 GRANDEV 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 NP compliant, MRI listed Photos courtesy of: Lygus Entomart; Beet armyworm Clemson Univ. USDA Cooperative Extension Slide Series Bugwood.org; Western flower thrip Frank Peairs, C St. Univ. Bugwood.org; citrus leaf miner Page 4 Center for Invasive Species Research, UC Riverside; : cabbage Looper RJ Reynolds Tobacco Company, Bugwood.org; twospotted spider mite Clemson EDU

5 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 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, C St. Page 5 Univ. Bugwood.org; Cabbage Looper RJ Reynolds Tobacco Company, Bugwood.org; Twospotted spider mite Clemson EDU

6 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 NP compliant, MRI listed 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, C St. Page 6 Univ. Bugwood.org; Cabbage Looper RJ Reynolds Tobacco Company, Bugwood.org; Twospotted spider mite Clemson EDU

7 Some Active Compounds Produced by Chromobacterium subtsugae H N H 3 C H H H N H N N H CH 3 CH 3 H H 3 C N H H N CH 3 N CH 3 CH 3 H H N N H H Deoxyviolacein Chromobacterium subtsugae H N Chromamide A Chromobacterium subtsugae H N H N H Violacein Chromobacterium subtsugae

8 Some Active Compounds Produced by Burkholderia rinojensis strain A396 H 3 C N H N H 3 C CH 3 H 2 N N H 3 C CH 3 Templazole A Burkholderia rinojensis Templazole B Burkholderia rinojensis CH 3 CH H 3 C H 3 C 3 CH 3 H N CH 3 H H H H H Templamide A Burkholderia rinojensis H 3 C CH 3 CH H 3 C 3 CH 3 H N CH 3 H H H Templamide B Burkholderia rinojensis

9 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.

10 Mode-of-action - Grandevo 10

11 Mode-of-action - Venerate A B Larvae are stunted Liquefied frass C D Problems molting Problems molting, pupating, and developing

12 Spectrum of Activity both products 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 12

13 Compatibility With Beneficials 3X Maximum Field Rate 24 hour Acute Test Beneficial Grandevo Venerate species Bombus sp. No effect No effect rius insidiosus No effect* No effect* minute pirate bug* Atheta coriaria No effect* No effect* rove beetle* Aphidoletes No effect No effect apidimyza predatory midge Aphidius colemani aphid parasitoid No effect No effect 13 *No effects on reproduction

14 Compatibility With Beneficials 3X Maximum Field Rate 24 hour Acute Test Beneficial Grandevo Venerate species Cryptoplaimus No effect No effect montrouezi mealybug destroyer Delphastus catalinae No effect No effect whitefly predator Encarsia formosa No effect No effect parasitic wasp Green lacewing No effect No effect Ladybird beetle No effect No effect 14 Amblysius swirskii predatory mite LD50 4.3X max label rate LD50 1.5x max label rate

15 Primary use crops both products Tree fruit Tree nuts Fruiting vegetables Small fruit and berries Alfalfa Potatoes Corn, cotton, soybean (seed treatment) Venerate only 15

16 Grandevo DF: Effect of Adjuvants Cabbage Looper Bioassay Test Results 12 LC GV DF Bond Maxx Biolink LI 700 Tactic Contact Xcel Activator 90 Phase (MS) Attach Franchise

17 Venerate : Effect of Adjuvants Cabbage Looper Bioassay Test Results 0.96 LC

18 Adjuvants Evaluated with Grandevo DF 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 Soap bark alkyl phenol ethoxylate, polysaccharide Spreader sticker 2-4oz/100gal Biolink- surfactant 0 Yucca and Garlic extract Surfactant and 05-2quarts/100gal and penetrant. penetrant 4. LI Phosphatidylcholine, methylacetic acid and Soy oil derived 1-4pints/100gal alkyl polyoxyethlene ether 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 Alkylphenol ethoxylate, alcohol ethoxylate and tall oil fatty acids 8. Phase 2.32 Methyl esters of fatty acids, alkyl polyoxyethylene ether and polyether modified polysiloxane 9. Attach 1.72 Pinene(terpene) polymers, petrolatum, alpha(pdodecylphenyl) omegahydorxypolys (oxyethelene) 10 Franchise 2.91 Lecithin, methyl esters of fatty acids and alcohol ethoxylate Blend of methylated seed oil Pinene based Lecithin based 1-4pints/100gal 1-4pints/100gal 4-16oz/100gal 3-4oz/100gal Page 18

19 Impact of Grandevo DF on SWD Mortality After Contact with Treated Blueberries (48 hours) Personal communication, Andrew Cuthbertson, Fera Science Ltd, UK. 19

20 Impact of Grandevo on SWD viposition After Contact Sprays DF WDG DF WDG Laboratory of Behavioral and Development Genetics, University of Leuven, Belgium,

21 Impact of Grandevo on SWD viposition After Feeding on Treated Food DF WDG Laboratory of Behavioral and Development Genetics, University of Leuven, Belgium,

22 San Jose Scale Control on Apple New York VEN XC 1 qt 2C-5C VEN XC 2 qt 2C-5C % SJS Damage at Harvest b ab ab b b b b b a GV WDG 1 lb. 2C-5C GV WDG 2 lb. 2C-5C Sivanto Prime 14 fl. oz. PK Sivanto Prime f/b Movento oz. PK, 1C Lorsban Advanced EC 64 fl. oz. TC Imidan 3 lb. PF, 1C-6C UTC Dr. Art Agnello, Cornell University. Sivanto and Movento applications included LI-700. No adjuvant included with Venerate or Grandevo applications. P=

23 San Jose Scale Control on Peaches New Jersey VEN XC 2 qt CSx2 GV WDG 2 lb. CSx2 Damoil 4 gal. DD % Fruit Injury ab b b b b b b a Movento 9 fl. oz. CS 10.5 fl. oz. CS Closer fl. oz. CS oz. CS UTC Dr. Anne Nielsen, Rutgers, Bridgeton, NJ. Season total of crawlers/5 cm tape in UTC = 657. Movento included LI-700. P=

24 Venerate Rotations with Radiant for Control of Western Flower Thrips on Strawberry-2016 # larvae/25 flowers Better Crops LLC, Dr. John Curtis Florida 2016 UTC 10 fl. oz. VEN 1 QT VEN 2 QT VEN 4 QT RAD/VEN 2 QT 3DA-A 7DA-A 3DA-B 9DA-B 3DA-C 7DA-C 3DA-D 8DA-D Application dates: 3/26, 4/2, 4/11, 4/18 in 60 GPA. 24

25 Venerate XC Rotations with Radiant for Control of Western Flower Thrips on Strawberry-2016 Better Crops LLC, Dr. John Curtis Florida, # adults/25 flowers UTC 10 fl. oz. VEN 1 QT VEN 2 QT VEN 4 QT RAD/VEN 2 QT 0 3DA-A 7DA-A 3DA-B 9DA-B 3DA-C 7DA-C 3DA-D 8DA-D Application dates: 3/26, 4/2, 4/11, 4/18 in 60 GPA. 25

26 Rosy Apple Aphid Control in Apples Michigan # RAA and GAA colonies per 100 terminals 4DAT 10 5 A a B b B b B b B b 0 UTC GV WDG 2 lb GV WDG 3 lb VEN XC 1 qt VDN XC 2 qt Dr. John Wise, MSU. Treatments applied June 23, counts taken June 27. P=

27 Corn Root Protection With Venerate XC 2015 Trials ISU 0-3 Nodal Root Damage Ratings & Meta-analysis 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 0.0 Untreated Venerate 170 ml/100 m-row Venerate 340 ml/100 m-row Venerate 680 ml/100 m-row Capture 47.6 ml/100 m-row NWV (IA) NWV (MN) NWV (WI) NWV (H) 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 = in all cases. Meta-analysis data was analyzed by Friedman Two-Way Nonparametric AV with means separated by Dunn's All-Pairwise Comparisons Test, P value =

28 Neither Grandevo or Venerate are Knockdown Insecticides, Both Must be Applied Early Treat now Not now

29 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 Yes Yes Adjust water hardness Yes for DF if >1000 ppm Spray interval 7 days (SWD may be 5 days) No 7 days Typical use rate 1-3 pounds/acre 1-4 quarts/acre

30 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 Tank-mix compatible Yes and more isn t better Not that we know of Yes (but not with VEN) Yes (but not with GV) Shelf-life 3 years minimum 3 years minimum Timing Rotation Treat early for most pests Generally recommended Treat early for most pests Generally recommended

31 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

32 Questions? Tim Johnson, PhD VP Field Development and Technical Services IR-4 Western Regional Workshop

IR-4 Western Regional Workshop. Dead Microbial Nematicides

IR-4 Western Regional Workshop Dead Microbial Nematicides EPA Registered Bionematicides that are Dead Dead = heat-killed or no viable cells in end product Marrone Bio Innovations Majestene - 94.46% heat-killed