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Dead Microbial Nematicides
IR-4 Western Regional
Workshop
EPA Registered Bionematicides that are Dead
© 2016 Marrone Bio Innovations -
2
Dead = heat-killed or no viable cells in end product Marrone Bio Innovations Majestene® - 94.46% heat-killed Burkholderia rinojensis strain A396 cells and spent fermentation media MBI-304 - 30% Chromobacterium subtsugae and spent fermentation media (no viable cells), DF and WDG formulations, not commercially available as a nematicide. Valent BioSciences Corporation DiTera ® – 90% Myrothecium verucarria strain AARC-0255 fermentation solids and solubles. Non-viable, “killed” microbial composition
Majestene® XC Bionematicide
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 – QC for nematode activity
Page 3 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
MAJESTENE® Bionematicide
Registered in the U.S. Residues are exempt from tolerances 4 hour REI, 0 days to harvest NOP compliant, OMRI listed
Page 4 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
Majestene® Reduces Gall Formation and Egg Mass Development
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Inhibition of egg mass development & hatching
• Reduction of # of egg masses per gall • Direct suppression on egg hatching
Reduction of Gall Formation: • Reduces nematode feeding
• Prevents colony establishment • Prevents disruption of plant function
Williamson and Gleason, 2003; modified
Causes direct mortality: • Dose dependent
• Kills free living stages • Prevents colony establishment
Some Labeled Crops and Controlled Nematode Genera
Meloidogyne
Pratylenchus
Criconema
Tylenchorhynchus
Belonolaimus
Helicotylenchus
Rotylenchulus
Heterodera
Radopholus
Haplolaimus
Corn
Cotton
Cucurbits
Tomato/Pepper
Soybean
Strawberry
2ee labels on potato, onion, cane berries, blueberries, carrots, mint, non-bearing fruit and nut trees, turf
Micro-Well Nematode Lethality Study USDA Corvallis, OR
7
0
10
20
30
40
50
60
70
80
90
100
% a
live
Meloidogyne chitwoodi
72 hrs
Root-knot Nematode Counts vs Majestene Concentration via Drip Irrigation
0.00 0.05 0.10 0.15 0.20
Majestene concentrations (v/v)
0
10
20
30
40
50
60
Roo
t-kno
t nem
atod
e po
poul
atio
n (n
o/10
0 m
L ro
ot)
y||=||1.68x -0.79 r2 = 0.90
Yield
0.00 0.05 0.10 0.15 0.20
Majestene concentrations (v/v)
15.00
15.50
16.00
16.50
Frui
t wei
ght (
ton/
acre
)
y||=||16.99+0.38ln||x r2 = 0.82
MBI-304 Bionematicide
Page 10
New species of Chromobacterium (subtsugae) isolated from forest soil from Maryland by Dr. Phyllis Martin of the USDA-ARS
C. subtsugae strain PRAA4-1T
– 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, 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
MBI-304 Bionematicide
Page 11
Registered in the U.S. and in Mexico but not commercially available as a nematicide
Residues are exempt from tolerances
4 hour REI, 0 days to harvest Easy on pollinators and
beneficials NOP compliant, OMRI listed
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
Product Overview
Page 12
• 2 gal/acre in-furrow spray/drip irrigation at planting or transplanting
• 1 gal/acre re-applications 3-4 weeks later. Make one to two applications during season.
• Apply toward the end of a drip irrigation event. • Specific instructions for overhead irrigation to maintain
a lethal concentration.
Typical Use Recommendation for Fruiting Vegetables and Strawberries:
Product Overview – MBI-304
Page 13
Typical Use Recommendation for Fruiting Vegetables and Strawberries:
Tomato: Meloidogyne incognita
• Majestene through drip. • One or two applications (30 days apart). • Galling index taken at 3 weeks after last application.
Agrisol Consulting, 2015 - Florida
Tomato: Meloidogyne incognita
• Majestene through drip. • One or two applications (30 days apart). • Counts taken at 3 weeks after last application. .
Agrisol Consulting, 2015 - Florida
Strawberry Demo, Plant City, FL
16
Untreated following fumigation
Majestene @ 2GPA following fumigation
Root-knot Nematode Control on Squash – NC 2016
17
0
200
400
600
800
1000
1200
1400
UTC Melocon@ 3 lb. AB
Majestene@ 1 gal
AB
Majestene@ 2 gal
AB
MBI-304@ 2 lb. A
MBI-304@ 2 lb. AB
MBI-304@ 4 lb. A
MBI-304@ 4 lb. AB
# Nematodes/sample 79DAA
a
cd
a
c
d
b
A= at planting, B=28 days after planting Biological Applied Research, 16-028TBJ
cd
e
Root-knot Nematode Control on Tomatoes – NC 2016
18
0
200
400
600
800
1000
1200
UTC Melocon@ 3 lb. AB
Majestene@ 1 gal
AB
Majestene@ 2 gal
AB
MBI-304WDG @ 2
lb. A
MBI-304WDG @ 2
lb. AB
MBI-304WDG @ 4
lb. A
MBI-304WDG @ 4
lb. AB
# Nematodes/sample 90DA-A
a
c
b
c c
ab
A= at planting, B=28 days after planting Biological Applied Research, 16-029TBJ
c c
Best Use and Testing Recommendations for Majestene and MBI-304
Majestene MBI-304 Adjuvant Soil adjuvants are under
investigation Soil adjuvants are under investigation
pH sensitivity ? ? Adjust water hardness ? Yes for DF if >1000 ppm Application interval Varies by crop Varies by crop Sensitive to water volume
Yes and more isn’t better. 0.05% v/v or greater concentration
Needs confirmation, likely an important variable
Tank-mix compatible Generally yes Under investigation Rates per acre (drip) 1-2 gallon/acre 2-4 lbs./acre Rates in-furrow 14-28 fl. oz./1000 row-ft. 2-4 oz./1000 row-ft. Rates for overhead chemigation
2.6 – 4 gallon/acre in 0.1” irrigation
TBD
Best Use and Testing Recommendations for Majestene and MBI-304
Majestene MBI-304 Shelf-life 3 years minimum 3 years minimum Timing At plant and in-season At plant and in-season Plant back restrictions None None Rotation No recommendation but
compatible with other nematicides
No recommendation but compatible with other nematicides
Questions?
Tim Johnson, PhD
VP Field Development and Technical Services
IR-4 Western Regional
Workshop