Live Microbial Bionematicides

IR-4 Western Region Biopesticide Training Workshop

Fort Collins, Colorado April 25 - 26, 2017

Michael B. Dimock Certis USA

Bacteria • Bacillus firmus • Pasteuria spp.

Nematophagous Fungi • Purpureocillium lilacinum

(formerly Paecilomyces lilacinus)

Live Microbial Bionematicides

Live Microbial Bionematicides Bacillus firmus

Modes of Action: PGPR that protects from nematodes through:

• Colonization of rhizoplane (physical barrier) • Degradation of root exudates (interferes with host finding) • Producing phytohormones (growth promotion) • Enzymatic degradation of eggs and J1-J2 larvae

Juvenile Meloidogyne damaged by B. firmus.

Bayer CropScience • B. firmus I-1582

From AgroGreen (Israel)

• VOTiVO FS (seed treatment) Broad crop labeling for many nematodes Also available as premix with insecticide

• Nortica 10WP (turf) Cyst, lance, lesion, ring, root knot sheath, spiral, sting & stunt nematodes

Products (Bayer):

Pasteuria spp. Live Microbial Bionematicides

Mode of Action: • Specific, obligate endoparasites

of nematodes. • Endospores attach to cuticle,

form “infection peg,” proliferates within host.

• New endospores released into soil when host cadaver disintegrates.

Pasteuria penetrans attached to and germinating on Meloidogyne. Bird et al. (2003). Int. J. Parasitol. 33: 1269 - 1276.

Products (Syngenta):

• Clariva pn - seed treatment Pasteuria nishizawae Pn1 Soybean, sugar beet cyst nematodes Premix with fungicide also available

• Naviva ST - seed treatment Pasteuria sp. Ph3 Reniform nematode in cotton, soy, vegetables, cucurbits, & floriculture

• NewPro - spray/chemigation Pasteuria usgae Bl1 + Pasteuria sp. Ph3 Lance & sting nematodes in turf (Bermudagrass & St. Augustine grass)

Purpureocillium lilacinum Live Microbial Bionematicides

Mode of Action: • Generalist mycoparasite of nematodes. • Fungal infection (mycosis), production of chitinases and proteases. • Particularly effective vs. eggs in cysts and galls.

P. Lüth (Prophyta/Bayer CropScience) R. Holland, Macquarie Univ (Australia)

(Enlarged below)

Purified conidia

Spore germination

Lesion nematode killed by PL251

Hyphae penetrating Meloidogyne egg

Products (Bayer): • MeloCon, BioAct, NemOut

P. lilacinus strain 251 (from Prophyta) Formulations: WG, WP; liquid (DC) in development Broad crop and nematode labeling

Live Microbial Bionematicides Effect on Beneficial Nematodes?

3rd stage infective juvenile (dauer stage) of Steinernema carpocapsae

R. Gaugler (Rutgers University)

Bacillus firmus • Close association with plant roots

provides niche separation from free-living nematodes.

Pasteuria spp. • Extreme host specificity - Spores

don’t attach to nonhost nematodes.

Purpureocillium lilacinum • Studies show no effect on EPNs • Protective sheath - barrier to infection. • Other mechanisms (e.g. avoidance)?

Background: Entomopathogenic nematodes • Steinernema & Heterorhabditis

• Free living IJ is only stage that occurs outside the insect host.

• Cuticle from previous molt retained as protective sheath.

Protective sheath

Live Microbial Bionematicides

Active ingredients Products

Fungicides Bacillus subtilis QST713 Serenade Fosetyl-Al Aliette Metalaxyl Ridomil PCNB Terrachlor Propamocarb-HCL Tattoo, Previcur Trichoderma harzianum RootShield

Insecticides & nematicides Aldicarb Temik Bifenthrin Talstar, Capture Cadusafos Rugby Carbofuran Furadan Ethoprop Mocap Fenamiphos Nemacur Fipronil Regent Fosthiazate Nemathorin Furfural Multiguard Imidacloprid Admire, others Oxamyl Vydate Terbufos Counter

Herbicides EPTC Eptam Metolachlor Dual, Bicep Metribuzin Sencor

MeloCon/BioAct Compatibility No significant effect on spore germination or colony growth:

Active Ingredients Products

Azoxystrobin Amistar, Heritage, Quadris

Chlorothalonil Bravo, Daconil, Echo, etc.

Mancozeb Dithane

Propiconazole Tilt, Banner MAXX

Tebuconazole Folicur

Fungicides that inhibit spore germination and/or colony growth:

Application efficiency may be enhanced by inclusion of a soil wetting agent to help the spores penetrate soil into the root zone.

Live Microbial Bionematicides Storage Recommendations

Active Product Storage

B. firmus VOTiVo FS Cool & dry

Nortica WP Cool & dry

Pasteuria spp.

Clariva pn 32 -100°F

Naviva ST Refrigerate (4°C)

NewPro 20 - 22°C

P. lilacinus MeloCon/BioAct Refrigerate (35-40°F) or freeze

Seed treatment slurries - do not hold > 72 hours Spray/chemigation - do not hold mix > 24 hours

Considerations for Use

Influenced by of mode of action • Association with plant roots • Parasitism of nematodes

Annual vs. perennial crops

Persistence in soil/rhizosphere

Effects of soil temperature

Effects of soil type

Live Microbial Bionematicides

Effect of soil temperature on persistence of PL251 spores in loamy sand (Rumbos 2005).

Timing of applications

Considerations for Use Live Microbial Bionematicides

Seed treatment

Banded vs. broadcast

Method of incorporation • Tillage/shank injection • Drip chemigation • “Watering in” • Soil penetrant/wetting agent?

Delivery to site of activity Iowa State Univ.

Considerations for Use

Nematode biology Ecto- vs. endoparasite

Migratory vs. sedentary

Cysts, root galls

Live Microbial Bionematicides

Univ. of Kentucky

USDA-ARS

Female

Egg mass

Soybean cyst nematode (Heterodera glycines)

J. Eisenback, Virginia Tech (Bugwood.org)

Lesion nematode (Pratylenchus sp.)

Considerations for Use

Evaluation methods Soil nematode counts Root galls or other damage Plant growth & vigor Yield effects

Live Microbial Bionematicides