WO2020006555A1 - Compositions comprenant des bactéries et procédés de lutte contre des organismes nuisibles de plantes et d'amélioration de la santé des plantes - Google Patents

Compositions comprenant des bactéries et procédés de lutte contre des organismes nuisibles de plantes et d'amélioration de la santé des plantes Download PDF

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WO2020006555A1
WO2020006555A1 PCT/US2019/040107 US2019040107W WO2020006555A1 WO 2020006555 A1 WO2020006555 A1 WO 2020006555A1 US 2019040107 W US2019040107 W US 2019040107W WO 2020006555 A1 WO2020006555 A1 WO 2020006555A1
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plant
cfu
active variant
bacterial strain
pest
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PCT/US2019/040107
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English (en)
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Narendra PALEKAR
Chad Alton KEYSER
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AgBiome, Inc.
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Publication of WO2020006555A1 publication Critical patent/WO2020006555A1/fr

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    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
    • A01N63/20—Bacteria; Substances produced thereby or obtained therefrom
    • A01N63/27—Pseudomonas
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
    • A01N63/20—Bacteria; Substances produced thereby or obtained therefrom

Definitions

  • the invention relates to bacterial strains and populations for controlling plant pests and/or improving an agronomic trait of interest in a plant.
  • Damage and diseases caused by plant pests are responsible for significant agricultural losses. Effects can range from mild symptoms to catastrophic plant damage, which can lead to major economic and social consequences. Methods are needed to effectively control plant pests.
  • compositions and methods for controlling plant pests and/or for improving at least one agronomic trait of interest in a plant comprise bacterial strains that control one or more plant pests, and/or improve at least one agronomic trait of interest.
  • the bacterial strains can be used as an inoculant for plants.
  • methods for growing a plant susceptible to a plant pest or plant disease caused by a plant pest and for treating or preventing a plant disease or damage caused by a plant pest are also provided. Further provided are methods and compositions for making a modified bacterial strain having resistance to a biocide of interest.
  • compositions and methods for controlling one or more plant pests and/or improving at least one agronomic trait of interest are provided.
  • a biological agent, biocontrol agent, bacterial strain, modified bacterial strain, modified biological agent, or modified biocontrol agent or active variant thereof are used herein to describe a microorganism that is used to control plant pests and/or improve at least one agronomic trait of interest.
  • biocontrol agents or bacterial strains are provided which can be used to control one or more plant pest and/or improve at least one agronomic trait of interest.
  • Such bacterial strains include AIP031898 (a Pseudomonas chlororaphis FB24 strain), AIP023234 (a Lonsdalea quercina strain), AIP024552 (an Agrobacterium tumefaciens strain), AIP035573 (a Lonsdalea quercina strain), AIP071234 (a Pseudarthrobacter phenanthrenivoram strain), AIP080021 (a Pseudomonas libanensis strain), AIP001237 ⁇ a Pseudomonas brassicaceanm strain), AIP050674 (a Pseudomonas chlororaphis FB24 strain), AIP023234 (a Lonsdalea quercina strain), AIP024552 (an Agrobacterium tumefacien
  • AIP071546 a Pseudomonas brassicacearum strain
  • AIP049805 a Pseudomonas Uni strain
  • AIPO 16229 a Pseudomonas brassicacearum strain
  • AIP081435 a Pseudomonas poae strain
  • AIP082140 an Agrohacierium tumefaciens strain
  • AIP000817 a Mucilaginibacter gossypii strain
  • AIP060333 a Pseudomonas fluorescens strain
  • AIP070494 a Mycolicibacterium frederiksbergense strain
  • AIPO 15104 a Pseudomonas graminis strain
  • AIP011586 an Agrobacterium tumefaciens strain
  • AIP010884 an Agrohacierium tumefaciens strain
  • AIP082287 a Pseudomonas brass
  • Ochrobaclrum intermedium strain AIP065969 (a Microbacterium arbor escens strain), AIPO 18514 (an Exiguobacterium indicum strain), AIP033041 (a Variovorax ginsengisoii strain), AIP092281 (a Microbacterium testaceum strain), AIP081114 (a Microbacterium hominis strain), AIP046403 (a Microbacterium oleivorans strain), AIP022635 (a Paenarthrobacter ureafaciens strain),
  • AIP070925 (a Pseudomonas fluorescens strain), and AIP039063 (a Klebsiella aerogenes strain).
  • Cell populations comprising one or more of, AIP031898, AIP023234, AIP024552, AIP035573, AIP071234, AIP080021, AIP001237, AIP050674, AIP071546, AIP049805, AIP016229,
  • AIP081114, AIP046403, AIP022635, AIP070925, and AIP039063 are provided, as well as, populations of spores derived from each of these strains, or any preparation thereof.
  • various bacterial strains and/or the pesticidal compositions provided herein comprise as an active ingredient a cell population comprising one or more of AIP031898, AIP023234, AIP024552, AIP035573, AIP071234, AIP080021, AIP001237, AIP050674, AIP071546,
  • AIP049805 AIP016229, AIP081435, AIP082140, AIP000817, AIP060333, AIP070494,
  • AIP015104 AIP011586, AIP010884, AIP082287, AIP088026, AIP065969, AIP018514,
  • AIP031898 was deposited with the Patent Depository of the National Center for
  • AIP023234 was deposited with the Patent Depository of the National Center for
  • AIP024552 was deposited with the Patent Depository of the National Center for
  • AIP035573 was deposited with the Patent Depository of the National Center for
  • AIP071234 was deposited with the Patent Depository of the National Center for
  • AIP080021 was deposited with the Patent Depository of the National Center for
  • AIP001237 was deposited with the Patent Depository of the National Center for
  • AIP050674 was deposited with the Patent Depository of the National Center for
  • AIP049805 was deposited with the Patent Depository of the National Center for
  • AIP081435 was deposited with the Patent Depository of the National Center for
  • AIP082140 was deposited with the Patent Depository of the National Center for
  • AIP000817 was deposited with the Patent Depository of the National Center for
  • AIP060333 was deposited with the Patent Depository of the National Center for
  • AIP082287 was deposited with the Patent Depository of the National Center for
  • AIP088026 was deposited with the Patent Depository of the National Center for
  • AIP065969 was deposited with the Patent Depository of the National Center for
  • AIP018514 was deposited with the Patent Depository of the National Center for
  • AIP033041 was deposited with the Patent Depository of the National Center for
  • AIP092281 was deposited with the Patent Depository of the National Center for
  • AIP081114 was deposited with the Patent Depository of the National Center for
  • AIP046403 was deposited with the Patent Depository of the National Center for
  • AIP070925 was deposited with the Patent Depository of the National Center for
  • AIP039063 was deposited with the Patent Depository of the National Center for
  • isolated encompasses a bacterium, spore, or other entity or substance, that has been (1) separated from at least some of the components with which it was associated when initially produced (whether in nature or in an experimental setting), and/or (2) produced, prepared, purified, and/or manufactured by the hand of man.
  • Isolated bacteria may be separated from at least about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, or more of the other components with which they were initially associated.
  • a substance is “pure” if it is substantially free of other components.
  • the terms “purify,” “purifying” and “purified” refer to a bacterium, spore, or other material that has been separated from at least some of the components with which it was associated either when initially produced or generated (e.g., whether in nature or in an experimental setting), or during any time after its initial production.
  • a bacterium or spore or a bacterial population or a spore population may be considered purified if it is isolated at or after production, such as from a material or environment containing the bacterium or bacterial population or spore, and a purified bacterium or bacterial population or spore may contain other materials up to about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, or above about 90% and still be considered purified.
  • purified bacteria or spores and bacterial populations or spore populations are more than about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or more than about 99% pure.
  • a culture of bacteria contains no other bacterial species in quantities to be detected by normal bacteriological techniques.
  • compositions that comprise a population of at least one bacterial strain or a mixed population of individuals from more than one bacterial strain.
  • the population of at least one of a bacterial strain i.e., cells of AIP031898, AIP023234, AIP024552, AIP035573, AIP071234, AIP080021, AIP001237, AIP050674, AIP071546, AIP049805,
  • AIP092281, AIP081114, AIP046403, AIP022635, AIP070925, and AIP039063 or an active variant of any thereof, or spores or forespores or a combination of cells, forespores and/or spores, formed from one or more of AIP031898, AIP023234, AIP024552, AIP035573, AIP071234, AIP080021, AIP001237, AIP050674, AIP071546, AIP049805, AIP016229, AIP081435,
  • AIP046403, AIP022635, AIP070925, and AIP039063, or an active variant of any thereof) comprises a concentration of at least about 10 5 CFU/ml to about 10 u CFU/ml, about 10 5 CFU/ml to about 10 10 CFU/ml, about 10 5 CFU/ml to about 10 12 CFU/ml, about 10 5 CFU/ml to about 10 6 CFU/ml, about 10 6 CFU/ml to about 10 7 CFU/ml, about 10 7 CFU/ml to about 10 8 CFU/ml, about 10 8 CFU/ml to about 10 9 CFU/ml, about 10 9 CFU/ml to about 10 10 CFU/ml, about 10 10 CFU/ml to about 10 11 CFU/ml, about 10 11 CFU/ml to about 10 12 CFU/ml.
  • the concentration of the bacterial strain provided herein or active variant thereof comprises at least about 10 5 CFU/ml, at least about 10 6 CFU/ml, at least about 10 7 CFU/ml, at least about 10 8 CFU/ml, at least about 10 9 CFU/ml, at least about 10 10 CFU/ml, at least about 10 11 CFU/ml, or at least about 10 12 CFU/ml.
  • A“spore” refers to at least one dormant (at application) but viable reproductive unit of a bacterial species.
  • AIP018514, AIP033041, AIP092281, AIP081114, AIP046403, AIP022635, AIP070925, and AIP039063 are disclosed elsewhere herein. It is further recognized the populations disclosed herein can comprise a combination of vegetative cells and forepores (cells in an intermediate stage of spore formation); a combination of forespores and spores; or a combination of forespores, vegetative cells and/or spores.
  • compositions comprising a cell of a bacterial strain (i.e., at least one of AGR031898, AIP023234, AIP024552, AIP035573, AIP071234, AIP080021, AIP001237, AIP050674,
  • AIP081114, AIP046403, AIP022635, AIP070925, and AIP039063, or an active variant of any thereof) can further comprise an agriculturally acceptable carrier.
  • the term "agriculturally acceptable carrier" is intended to include any material that facilitates application of a composition to the intended subject (i.e, a plant or plant part susceptible to damage or disease caused by a plant pest or a plant or plant part for improving an agronomic trait of interest).
  • Carriers used in compositions for application to plants and plant parts are preferably non-phytotoxic or only mildly phytotoxic.
  • a suitable carrier may be a solid, liquid or gas depending on the desired formulation.
  • carriers include polar or non-polar liquid carriers such as water, mineral oils and vegetable oils. Additional carriers are disclosed elsewhere herein.
  • variants will retain the ability to control one or more plant pests or improve one or more agronomic traits of interest in a plant.
  • the active variants of the bacterial strains provided herein will retain pesticidal activity against a plant pest.
  • “pesticidal activity” refers to activity against one or more pests, including insects, fungi, bacteria, nematodes, viruses or viroids, protozoan pathogens, and the like, such that the pest is killed or controlled.
  • variants will retain the ability to control one or more insect pests or nematode pests.
  • variants will retain the ability to control Coleopteran insect pests, including com rootworms (e.g., Western corn rootworm), Colorado potato beetle, weevils (e.g., sweetpotato weevil), or Hemipteran insect pests.
  • com rootworms e.g., Western corn rootworm
  • Colorado potato beetle e.g., Colorado potato beetle
  • weevils e.g., sweetpotato weevil
  • Hemipteran insect pests including com rootworms (e.g., Western corn rootworm), Colorado potato beetle, weevils (e.g., sweetpotato weevil), or Hemipteran insect pests.
  • Active variants of the various bacterial strains provided herein include, for example, any isolate or mutant of AIP031898, AIP023234, AIP024552, AIP035573, AIP071234, AIP080021, AIP001237, AIP050674, AIP071546, AIP049805, AIP016229, AIP081435, AIP082140,
  • AIP022635, AIP070925, and AIP039063 are examples of IP
  • the bacterial strain is compatible with a biocide.
  • a biocide is a chemical substance that can exert a controlling effect on an organism by chemical or biological means.
  • Biocides include pesticides, such as fungicides or insecticides; herbicides; other crop protection chemicals, and the like. Such compounds are discussed in detail elsewhere herein.
  • a bacterial strain is compatible with a biocide when the bacterial strain is able to survive and/or reproduce in the presence of an effective amount of a biocide of interest.
  • methods can be undertaken to modify the bacterial strain to impart the compatibility of interest. Such methods to produce modified bacterial strains include both selection techniques and/or transformation techniques.
  • modified bacterial strain is intended a population wherein the strain has been modified (by selection and/or transformation) to have one or more additional traits of interest.
  • the modified bacterial strain comprises any one of AIP031898, AIP023234, AIP024552,
  • the modified bacterial strain is compatible with a biocide of interest, including but not limited to, resistance to a herbicide, fungicide, pesticide, or other crop protection chemical.
  • the modified biocide-resistant strains have the same identification characteristics as the original sensitive strain except they are significantly more resistant to the particular herbicide, fungicide, pesticide, or other crop protection chemical. Their identification is readily possible by comparison with characteristics of the known sensitive strain. Thus, isolated populations of modified bacterial strains are provided.
  • An increase in resistance to a biocide refers to the ability of an organism (e.g., bacterial cell or spore) to survive and reproduce following exposure to a dose of the biocide (e g, herbicide, insecticide, fungicide, pesticide, or other crop protection chemical) that would normally be lethal to the unmodified organism or would substantially reduce growth of the unmodified organism.
  • the increase in resistance to a biocide is demonstrated in the presence of an agriculturally effective amount of the biocide.
  • compositions provided herein include selected or engineered bacterial strains and modified populations of bacterial strains. These bacterial strains or modified bacterial strains can be used as an inoculant for plants. They can also be applied as a spray application directly to the aerial parts of plants or can be applied as a seed coating, and can be mixed with the herbicide or other chemical to which they have been modified to become tolerant.
  • active variants of the bacterial strains disclosed herein include for example, a modified strain, such that the active variant controls a plant pest and further is able to grow in the presence of at least one biocide.
  • Recombinant bacterial strains having resistance to an herbicide, insecticide, fungicide, pesticide, or other crop protection chemical can be made through genetic engineering techniques and such engineered or recombinant bacterial strains grown to produce a modified population of bacterial strains.
  • a recombinant bacterial strain is produced by introducing polynucleotides into the bacterial host cell by transformation. Methods for transforming microorganisms are known and available in the art. See, generally, Hanahan, D.
  • Transformation may occur by natural uptake of naked DNA by competent cells from their environment in the laboratory.
  • cells can be made competent by exposure to divalent cations under cold conditions, by electroporation, by exposure to polyethylene glycol, by treatment with fibrous nanoparticles, or other methods well known in the art.
  • Herbicide resistance genes for use in transforming a recombinant bacterial strain include, but are not limited to, fumonisin detoxification genes (U.S. Patent No. 5,792,931); acetolactate synthase (ALS) mutants that lead to herbicide resistance, in particular the sulfonylurea-type herbicides, such as the S4 and/or Hra mutations; inhibitors of glutamine synthase such as phosphinothricin or basta (e.g., bar gene); and glyphosate resistance (EPSPS gene); gluphosinate, and HPPD resistance (WO 96/38576, U.S. Patent Nos.
  • the bar gene encodes resistance to the herbicide basta
  • the nptll gene encodes resistance to the antibiotics kanamycin and geneticin
  • the ALS-gene mutants encode resistance to the sulfonylurea herbicides including chlorsulfuron, metsulfuron, sulfometuron, nicosulfuron, rimsulfuron, flazasulfuron, sulfosulfuron, and triasulfuron
  • the imadizolinone herbicides including imazethapyr, imazaquin, imazapyr, and imazamethabenz.
  • the bacterial strains are grown in the presence of the herbicide, insecticide, fungicide, pesticide, or other crop protection chemical as the selection pressure. Susceptible agents are killed while resistant agents survive to reproduce without competition. As the bacterial strains are grown in the presence of the herbicide, insecticide, fungicide, pesticide, or other crop protection chemical, resistant bacterial strains successfully reproduce and become dominant in the population, becoming a modified population of bacterial strains. Methods for selecting resistant strains are known and include U.S. Patent Nos. 4,306,027 and 4,094,097, herein incorporated by reference.
  • the active variant of the bacterial strain comprising a modified population of bacterial strains will have the same identification characteristics as the original sensitive strain except they are significantly more tolerant to the particular herbicide, insecticide, fungicide, pesticide, or other crop protection chemical. Thus, their identification is readily possible by comparison with characteristics of the known sensitive strain.
  • AIP049805 AIP016229, AIP081435, AIP082140, AIP000817, AIP060333, AIP070494,
  • AIP015104 AIP011586, AIP010884, AIP082287, AIP088026, AIP065969, AIP018514,
  • AIP033041, AIP092281, AIP081114, AIP046403, AIP022635, AIP070925, and AIP039063 include strains that are closely related to any of the disclosed strains by employing the Bishop MLST method of organism classification as defined in Bishop et al. (2009) BMC Biology
  • an active variant of a bacterial strain disclosed herein includes a bacterial strain that falls within at least a 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%. 94%, 95%, 96%, 97%, 98%, 98.5%, 98.8%, 99%, 99.1%, 99.2%, 99.3%, 99.4%, 99.5%, 99.6%, 99.7%, 99.8%, or 99.9% sequence cut off employing the Bishop method of organism classification as set forth in Bishop et al. (2009) BMC Biology 7(1)1741-7007-7-3, which is herein incorporated by reference in its entirety.
  • Active variants of the bacteria identified by such methods will retain the ability to control at least one plant pest and/or to improve at least one agronomic trait when applied in an effective amount to a plant, plant part, or an area of cultivation, including for example reducing plant pests, reducing infestations of plant pests, and/or increasing pest resistance including insect pest resistance (e.g., Coleoptera insects such as Western corn rootworm, Colorado potato beetle, and/or sweet potato weevil).
  • insect pest resistance e.g., Coleoptera insects such as Western corn rootworm, Colorado potato beetle, and/or sweet potato weevil.
  • the active variant of the bacterial strain(s) disclosed herein include strains that are closely related to any of the disclosed strains on the basis of the Average Nucleotide Identity (ANI) method of organism classification.
  • ANI Average Nucleotide Identity
  • ANI see, for example, Konstantinidis, K.T., et al. , (2005) PNAS USA 102(7):2567 72; and Richter, M., et al, (2009) PNAS 106(45): 19126 31
  • variants see, for example, Varghese, N.J., et al, Nucleic Acids Research (July 6, 2015):
  • gkv657 are based on summarizing the average nucleotides shared between the genomes of strains that align in WGAs.
  • an active variant of bacterial strain is based on summarizing the average nucleotides shared between the genomes of strains that align in WGAs.
  • AIP070925, and AIP039063 disclosed herein includes a bacterial strain that falls within at least a 90%, 95%, 96%, 97%, 97.5%, 98%, 98.5%, 98.8%, 99%, 99.5%, or 99.8% sequence cut off employing the ANI method of organism classification as set forth in Konstantinidis, K.T., et al, (2005) PNAS USA 102(7):2567-72, which is herein incorporated by reference in its entirety.
  • Active variants of the bacteria identified by such methods will retain the ability to control at least one plant pest and/or to improve at least one agronomic trait when applied in an effective amount to a plant, plant part, or an area of cultivation, including for example, reducing plant pests, reducing infestations of plant pests, and/or increasing pest resistance including insect pest resistance (e.g., Coleoptera insects such as Western corn rootworm, Colorado potato beetle, and/or sweet potato weevil).
  • insect pest resistance e.g., Coleoptera insects such as Western corn rootworm, Colorado potato beetle, and/or sweet potato weevil.
  • the active variants of the isolated bacterial strain(s) disclosed herein include strain(s) that are closely related to any of the above strains (for example, closely related to AIP031898, AIP023234, AIP024552, AIP035573, AIP071234, AIP080021, AIP001237, AIP050674, AIP071546, AIP049805, AIP016229, AIP081435, AIP082140, AIP000817,
  • the active variant is at least 95% identical to any of the above strains on the basis of 16S rDNA sequence identity, at least 96% identical to any of the above strains on the basis of 16S rDNA sequence identity, at least 97% identical to any of the above strains on the basis of 16S rDNA sequence identity', at least 98% to any of the above strains on the basis of 16S rDNA sequence identity, at least 98.5% identical to any of the above strains on the basis of 16S rDNA sequence identity, at least 99% identical to any of the above strains on the basis of 16S rDNA sequence identity, at least 99.5% to any of the above strains on the basis of 16S rDNA sequence identity or at least 100% to any of the above strains on the basis of 16S rDNA sequence identity.
  • Active variants of the bacteria identified by such methods will retain the ability to control at least one plant pest and/or to improve at least one agronomic trait when applied in an effective amount to a plant, plant part, or an area of cultivation, including for example, reducing plant pests, reducing infestations of plant pests, and/or increasing pest resistance including insect pest resistance (e.g., Coleoptera insects such as Western corn rootworm, Colorado potato beetle, and/or sweet potato weevil).
  • insect pest resistance e.g., Coleoptera insects such as Western corn rootworm, Colorado potato beetle, and/or sweet potato weevil.
  • the MinHash (Mash) distance metric is a comparison method that defines thresholds for hierarchical classification of microorganisms at high resolution and requires few parameters and steps (Ondov et al. (2016) Genome Biology 17: 132). Mash distance strongly corresponds to Average Nucleotide Identity method (ANI) for hierarchical classification (See, Konstantinidis, K.T. et al. (2005) PNAS USA 102(7):2567-72, herein incorporated by reference in its entirety). That is, an ANI of 97% is approximately equal to a Mash distance of 0.03, such that values put forth as useful classification thresholds in the ANI literature can be directly applied with the Mash distance.
  • ANI Nucleotide Identity
  • Active variants of the bacterial strain(s) disclosed herein include strains that are closely related to AIP031898, AIP023234, AIP024552, AIP035573, AIP071234, AIP080021, AIP001237, AIP050674, AIP071546, AIP049805, AIP016229, AIP081435, AIP082140, AIP000817,
  • an active variant of a bacterial strain disclosed herein includes bacterial strains having a genome within a Mash distance of less than about 0.015 to the disclosed strains.
  • an active variant of a bacterial strain disclosed herein includes a distance metric of less than about 0.005, 0.010, 0.015, 0.020, 0.025, or 0.030.
  • a genome, as it relates to the Mash distance includes both bacterial chromosomal DNA and bacterial plasmid DNA.
  • the active variant of a bacterial strain has a genome that is above a Mash distance threshold to the disclosed strains that is greater than dissimilarity caused by technical variance. In further instances, the active variant of a bacterial strain has a genome that is above a Mash distance threshold to the disclosed strains that is greater than dissimilarity caused by technical variance and has a Mash distance of less than about 0.015. In other instances, the active variant of a bacterial strain has a genome that is above a Mash distance threshold to the disclosed strains that is greater than dissimilarity caused by technical variance and has a Mash distance of less than about 0.005, 0.010, 0.015, 0.020, 0.025, or 0.030.
  • “above technical variation” means above the Mash distance between two strains caused by errors in the genome assemblies provided the genomes being compared were each DNA sequenced with at least 20X coverage with the Mumina HiSeq 2500 DNA sequencing technology and the genomes are at least 99% complete with evidence for contamination of less than 2%. While 20X coverage is an art recognized term, for clarity, an example of 20X coverage is as follows: for a genome size of 5 megabases (MB), 100 MB of DNA sequencing from the given genome is required to have 20X sequencing coverage on average at each position along the genome. There are many suitable collections of marker genes to use for genome completeness calculations including the sets found in Campbell et al. (2013) PNAS USA 110(14):5540-45,
  • Contamination is defined as the percentage of typically single copy marker genes that are found in multiple copies in the given genome sequence (e.g. Parks et al. (2015) Genome Research 25: 1043- 1055); each of these references is herein incorporated in their entirety. Completeness and contamination are calculated using the same collection of marker genes. Unless otherwise stated, the set of collection markers employed in the completeness and contamination assay is those set forth in Campbell et al. (2013) PNAS USA 110(14):5540-45, herein incorporated by reference.
  • Genomes of sufficient quality for comparison must be produced.
  • a genome of sufficient quality is defined as a genome assembly created with enough DNA sequence to amount to at least 20X genome coverage using Ulumina HiSeq 2500 technology. The genome must be at least 99% complete with contamination of less than 2% to be compared to the claimed microbe’s genome.
  • Genomes are to be compared using the Minhash workflow as demonstrated in Ondov et al. (2016) Genome Biology 17: 132, herein incorporated by reference in its entirety. Unless otherwise stated, parameters employed are as follows:“sketch” size of 1000, and“k-mer length” of 21.
  • Active variants of the bacteria identified by such methods will retain the ability to control at least one plant pest and/or to improve at least one agronomic trait when applied in an effective amount to a plant, plant part, or an area of cultivation, including for example, reducing plant pests, reducing infestations of plant pests, and/or increasing pest resistance including insect pest resistance (e.g., Coleoptera insects such as Western corn rootworm, Colorado potato beetle, and/or sweet potato weevil).
  • insect pest resistance e.g., Coleoptera insects such as Western corn rootworm, Colorado potato beetle, and/or sweet potato weevil.
  • the bacterial strains provided herein i.e., cells of AIP031898, AIP023234, AGR024552, AIP035573, AIP071234, AIP080021, AIP001237, AIP050674, AIP071546, AIP049805,
  • AIP070925, and AIP039063, or an active variant of any thereof can be formulated as a cell paste, wettable powders, a cell pellet, dusts, granules, a slurry, a dry powder, aqueous or oil based liquid products, and the like.
  • Such formulations will comprise the bacteria provided herein or an active variant thereof in addition to carriers and other agents.
  • the formulations can be used in a variety of methods as disclosed elsewhere herein.
  • the bacterial strains disclosed herein and the active variants thereof can be formulated to include at least one or more of an extender, a solvent, spontaneity promoter, carrier, emulsifier, dispersant, frost protectant, thickener, and/or adjuvant.
  • the extender, solvent, spontaneity promoter, carrier, emulsifier, dispersant, frost protectant, thickener, and/or adjuvant is a non-natural or synthetic extender, a solvent, spontaneity promoters, carriers, emulsifiers, dispersants, frost protectants, thickeners, and/or adjuvants.
  • the bacterial strains disclosed herein and the active variants thereof can be formulated to include at least one or more natural extender, a solvent, spontaneity promoter, carrier, emulsifier, dispersant, frost protectant, thickener, and/or adjuvant.
  • Examples of typical formulations include water-soluble liquids (SL), emulsifiable concentrates (EC), emulsions in water (EW), suspension concentrates (SC, SE, FS, OD), water- dispersible granules (WG), granules (GR) and capsule concentrates (CS); WG; GR; BB; SG; ZC these and other possible types of formulation are described, for example, by Crop Life International and in Pesticide Specifications, Manual on development and use of FAO and WHO specifications for pesticides, FAO Plant Production and Protection Papers - 173, prepared by the FAO/WHO Joint Meeting on Pesticide Specifications, 2004, ISBN: 9251048576.
  • the formulations may comprise active agrochemical compounds other than one or more active compounds of the invention.
  • the formulations or application forms of the various bacterial strains or active variants thereof can comprise, but are not limited to, auxiliaries, such as extenders, solvents, spontaneity promoters, carriers, emulsifiers, dispersants, frost protectants, biocides, solid carriers, surfactants, thickeners and/or other auxiliaries, such as adjuvants.
  • auxiliaries such as extenders, solvents, spontaneity promoters, carriers, emulsifiers, dispersants, frost protectants, biocides, solid carriers, surfactants, thickeners and/or other auxiliaries, such as adjuvants.
  • An adjuvant in this context is a component which enhances the biological effect of the formulation, without the component itself having a biological effect.
  • adjuvants are agents which promote the retention, spreading, attachment to the leaf surface, or penetration.
  • Non-limiting extenders are, for example, water, polar and nonpolar organic chemical liquids, for example from the classes of the aromatic and non-aromatic hydrocarbons (such as paraffins, alkyl benzenes, alkylnaphthalenes, chlorobenzenes), the alcohols and polyols (which, if appropriate, may also be substituted, etherified and/or esterified), the ketones (such as acetone, cyclohexanone), esters (including fats and oils) and (poly)ethers, the unsubstituted and substituted amines, amides, lactams (such as N-alkylpyrrolidones) and lactones, the sulphones and sulphoxides (such as dimethyl sulphoxide).
  • the extender used is water, it is also possible to employ, for example, organic solvents as auxiliary solvents.
  • organic solvents are:
  • aromatics such as xylene, toluene or alkylnaphthalenes, chlorinated aromatics and chlorinated aliphatic hydrocarbons such as chlorobenzenes, chloroethylenes or methylene chloride, aliphatic hydrocarbons such as cyclohexane or paraffins, for example petroleum fractions, mineral and vegetable oils, alcohols such as butanol or glycol and also their ethers and esters, ketones such as acetone, methyl ethyl ketone, methyl isobutyl ketone or cyclohexanone, strongly polar solvents such as dimethylformamide and dimethyl sulphoxide, and also water. In principle it is possible to use any suitable solvent.
  • Non-limiting solvents are, for example, aromatic hydrocarbons, such as xylene, toluene or alkylnaphthalenes, for example, chlorinated aromatic or aliphatic hydrocarbons, such as chlorobenzene, chloroethylene or methylene chloride, for example, aliphatic hydrocarbons, such as cyclohexane, for example, paraffins, petroleum fractions, mineral and vegetable oils, alcohols, such as methanol, ethanol, isopropanol, butanol or glycol, for example, and also their ethers and esters, ketones such as acetone, methyl ethyl ketone, methyl isobutyl ketone or cyclohexanone, for example, strongly polar solvents, such as dimethyl sulphoxide, and water.
  • aromatic hydrocarbons such as xylene, toluene or alkylnaphthalenes
  • chlorinated aromatic or aliphatic hydrocarbons such as chlor
  • Non-limiting examples of suitable carriers include, for example, ammonium salts and ground natural minerals such as kaolins, clays, talc, chalk, quartz, attapulgite, montmorillonite or diatomaceous earth, and ground synthetic minerals, such as finely divided silica, alumina and natural or synthetic silicates, resins, waxes and/or solid fertilizers. Mixtures of such carriers may likewise be used.
  • ground natural minerals such as kaolins, clays, talc, chalk, quartz, attapulgite, montmorillonite or diatomaceous earth
  • ground synthetic minerals such as finely divided silica, alumina and natural or synthetic silicates, resins, waxes and/or solid fertilizers. Mixtures of such carriers may likewise be used.
  • Carriers suitable for granules include the following: for example, crushed and fractionated natural minerals such as calcite, marble, pumice, sepiolite, dolomite, and also synthetic granules of inorganic and organic meals, and also granules of organic material such as sawdust, paper, coconut shells, maize cobs, and tobacco stalks.
  • Liquefied gaseous extenders or solvents may also be used.
  • extenders or carriers which at standard temperature and under standard pressure are gaseous, examples being aerosol propellants, such as halogenated hydrocarbons, and also butane, propane, nitrogen and carbon dioxide.
  • aerosol propellants such as halogenated hydrocarbons, and also butane, propane, nitrogen and carbon dioxide.
  • emulsifiers and/or foam-formers, dispersants or wetting agents having ionic or nonionic properties, or mixtures of these surface-active substances are salts of polyacrylic acid, salts of lignosulphonic acid, salts of phenolsulphonic acid or
  • naphthalenesulphonic acid polycondensates of ethylene oxide with fatty alcohols or with fatty acids or with fatty amines, with substituted phenols (preferably alkylphenols or arylphenols), salts of sulphosuccinic esters, taurine derivatives (preferably alkylta urates), phosphoric esters of polyethoxylated alcohols or phenols, fatty acid esters of polyols, and derivatives of the compounds containing sulphates, sulphonates and phosphates, examples being alkylaryl polyglycol ethers, alkyl sulphonates, alkyl sulphates, aryl sulphonates, protein hydrolysates, lignin-sulphite waste liquors and methylcellulose.
  • the presence of a surface-active substance is advantageous if one of the active compounds and/or one of the inert carriers is not soluble in water and if application takes place in water.
  • auxiliaries that may be present in the formulations and in the application forms derived from them include colorants such as inorganic pigments, examples being iron oxide, titanium oxide, Prussian Blue, and organic dyes, such as alizarin dyes, azo dyes and metal phthalocyanine dyes, and nutrients and trace nutrients, such as salts of iron, manganese, boron, copper, cobalt, molybdenum, and zinc.
  • colorants such as inorganic pigments, examples being iron oxide, titanium oxide, Prussian Blue, and organic dyes, such as alizarin dyes, azo dyes and metal phthalocyanine dyes, and nutrients and trace nutrients, such as salts of iron, manganese, boron, copper, cobalt, molybdenum, and zinc.
  • Stabilizers such as low-temperature stabilizers, preservatives, antioxidants, light stabilizers or other agents which improve chemical and/or physical stability may also be present. Additionally present may be foam-formers or defoamers.
  • formulations and application forms derived from them may also comprise, as additional auxiliaries, stickers such as carboxymethylcellulose, natural and synthetic polymers in powder, granule or latex form, such as gum arabic, polyvinyl alcohol, polyvinyl acetate, and also natural phospholipids, such as cephalins and lecithins, and synthetic phospholipids.
  • additional auxiliaries include mineral and vegetable oils.
  • auxiliaries present in the formulations and the application forms derived from them.
  • additives include fragrances, protective colloids, binders, adhesives, thickeners, thixotropic substances, penetrants, retention promoters, stabilizers, sequestrants, complexing agents, humectants and spreaders.
  • the active compounds may be combined with any solid or liquid additive commonly used for formulation purposes.
  • Suitable retention promoters include all those substances which reduce the dynamic surface tension, such as dioctyl sulphosuccinate, or increase the viscoelasticity, such as hydroxypropylguar polymers, for example.
  • Suitable penetrants in the present context include all those substances which are typically used in order to enhance the penetration of active agrochemical compounds into plants.
  • Penetrants in this context are defined in that, from the (generally aqueous) application liquor and/or from the spray coating, they are able to penetrate the cuticle of the plant and thereby increase the mobility of the active compounds in the cuticle. This property can be determined using the method described in the literature (Baur et ah, 1997, Pesticide Science 51 : 131-152).
  • Examples include alcohol alkoxylates such as coconut fatty ethoxylate (10) or isotridecyl ethoxylate (12), fatty acid esters such as rapeseed or soybean oil methyl esters, fatty amine alkoxylates such as tallowamine ethoxylate (15), or ammonium and/or phosphonium salts such as ammonium sulphate or diammonium hydrogen phosphate, for example.
  • alcohol alkoxylates such as coconut fatty ethoxylate (10) or isotridecyl ethoxylate (12)
  • fatty acid esters such as rapeseed or soybean oil methyl esters
  • fatty amine alkoxylates such as tallowamine ethoxylate (15)
  • ammonium and/or phosphonium salts such as ammonium sulphate or diammonium hydrogen phosphate, for example.
  • compositions and formulations disclosed herein can comprise an amount of a cell of a bacterial strain, such as AIP031898, AGR023234, AIP024552, AIP035573, AIP071234, AIP080021, AIP001237, AIP050674, AIP071546, AIP049805, AIP016229, AIP081435,
  • Such an amount can comprise a concentration of the bacterial strain of at least about 10 4 to about 10 11 , at least about 10 5 CFU/gram to about 10 11 CFU/gram, about 10 5 CFU/gram to about 10 10 CFU/gram, about 10 5 CFU/gram to about 10 12 CFU/gram, about 10 5 CFU/gram to about 10 6 CFU/gram, about 10 6 CFU/gram to about 10 7 CFU/gram, about 10 7 CFU/gram to about 10 8 CFU/gram, about 10 8 CFU/gram to about 10 9 CFU/gram, about 10 9 CFU/gram to about 10 10 CFU/gram, about 10 10 CFU/gram to about 10 11 CFU/gram, or about 10 u CFU/gram to about 10 12 CFU/gram.
  • the concentration of the bacterial strain comprises at least about 10 4 CFU/gram, at least about 10 5 CFU/gram, at least about 10 6 CFU/gram, at least about 10 7 CFU/gram, at least about 10 8
  • concentrations of the bacterial strain can occur in any formulation type of interest, including, for example in a liquid formulation, wettable power, spray dried formulation, in a cell paste, wettable granule, or freeze dried formulation.
  • the bacterial strain can occur in a liquid forumulation.
  • Liquid formulations can comprise an amount of a cell of a bacterial strain, such as AIP031898,
  • the amount of bacterial strain, or active variant thereof, disclosed herein can comprise a concentration of at least about 10 4 to about 10 11 CFU/mL, at least about 10 5 CFU/mL to about 10 11 CFU/ mL, about 10 5 CFU/ mL to about 10 10 CFU/ mL, about 10 5 CFU/ mL to about 10 12 CFU/ mL, about 10 5 CFU/ mL to about 10 6 CFU/ mL, about 10 6 CFU/ mL to about 10 7 CFU/ mL, about 10 7 CFU/ mL to about 10 8 CFU/ mL, about 10 8 CFU/ mL to about 10 9 CFU/ mL, about 10 9 CFU/ mL to about 10 10 CFU/ mL, about 10 10 CFU/ mL to about 10 11 CFU/
  • Dry formulations such as cell pastes, wettable powders, and spray dried formulations can comprise a cell of a bacterial strain, such as AIP031898, AIP023234, AIP024552, AIP035573, AIP071234, AIP080021, AIP001237, AIP050674, AIP071546, AIP049805, AIP016229,
  • the amount of the bacterial strain in the dry formulation can comprise a concentration of the bacterial strain of at least about 10 5 CFU/gram to about 10 11 CFU/gram, about 10 7 CFU/gram to about 10 10 CFU/gram, about 10 7 CFU/gram to about 10 11 CFU/gram, about 10 6 CFU/gram to about 10 10 CFU/gram, about 10 6 CFU/gram to about 10 11 CFU/gram, about 10 11 CFU/gram to about 10 12 CFU/gram, about 10 5 CFU/gram to about 10 10 CFU/gram, about 10 5 CFU/gram to about 10 12 CFU/gram, about 10 5 CFU/gram to about 10 6 CFU/gram, about 10 6 CFU/gram to about 10 7 CFU/gram, about 10 7 CFU/gram to about 10 8 CFU/gram, about 10 8 CFU/gram to about 10
  • the concentration of the bacterial strain comprises at least about 10 5 CFU/gram, at least about 10 6 CFU/gram, at least about 10 7 CFU/gram, at least about 10 8 CFU/gram, at least about 10 9 CFU/gram, at least about 10 10 CFU/gram, at least about 10 11
  • CFU/gram at least about 10 12 CFU/gram, or at least about 10 13 CFU/gram.
  • a“cell paste” comprises a population of cells that has been centrifuged and/or filtered or otherwise concentrated.
  • a coated seed which comprises a seed and a coating on the seed, wherein the coating comprises a cell of at least one bacterial strain, such as AIP031898, AIP023234, AIP024552, AIP035573, AIP071234, AIP080021, AIP001237, AIP050674, AIP071546,
  • AIP049805 AIP016229, AIP081435, AIP082140, AIP000817, AIP060333, AIP070494,
  • AIP015104 AIP011586, AIP010884, AIP082287, AIP088026, AIP065969, AIP018514,
  • Various plants of interest are disclosed elsewhere herein.
  • seeds are provided which comprise a heterolous coating on the seed, wherein the heterologous coating comprises a cell of at least one bacterial strain, such as AIP031898, AIP023234, AIP024552, AIP035573, AIP071234, AIP080021, AIP001237,
  • AIP070925, and AIP039063 or an active variant of any thereof, or a spore or a forespore or a combination of cells, forespores and/or spores, from any one of AIP031898, AIP023234,
  • AIP049805 AIP016229, AIP081435, AIP082140, AIP000817, AIP060333, AIP070494,
  • AIP015104 AIP011586, AIP010884, AIP082287, AIP088026, AIP065969, AIP018514,
  • “heterologous” in reference to a coating can refer to a seed coating comprising a bacterial strain that is not found in nature on the seed, or, if found in nature on the seed, is substantially modified from its native form in composition and/or concentration by deliberate human intervention.
  • “heterologous” in reference to a coating can refer to a seed coating comprising a bacterial strain suspended in a solution in which the bacterial strain is not naturally found.
  • the suspension solution for heterologous coatings can be natural or non-natural and can provide the bacterial strain with properties that the strain would not normally possess.
  • the suspension solution of a heterologous coating can permit the bacterial strain to adhere to the seed in such as a manner that the bacteria retains activity during seed storage and germination.
  • a seed coating can further comprise at least one nutrient, at least one biocide (e.g., herbicide or pesticide). See, for example, US App Pub. 20040336049, 20140173979, and 2015003381 1.
  • biocide e.g., herbicide or pesticide.
  • compositions disclosed herein can be stable for at least 30, 40, 50, 60, 70, 80,
  • Biological activity refers to the ability of the formulation to improve an agronomic trait of interest or control a plant pest.
  • the stable formulation retains at least about 1%, about 10%, about 20%, about 30% about 40%, about 50%, about 60%, about 70%, about 80%, or about 90% of CFU/gram in the formulation at a given storage time point when compared to the CFU/gram produced after immediate preparation of the formulation.
  • the stable formulation retains at least about 30% to 80%, about 50% to about 80%, about 60% to about 70%, about 70% to about 80%, about 40% to about 50%, about 50% to about 60%, about 60% to about 70% of biological activity in the formulation at a given storage time point when compared to the biological activity found in the formulation immediately after production.
  • the stable formulation at a given storage time point retains at least about 30%, 45%, 50%, 60%, 70%, 80%, 90% of biological activity when compared to the biological activity found in the formulation immediately after production.
  • the stable formation retains any combination of the viability and biological activity noted above.
  • the formulations preferably comprise between 0.00000001 % and 98% by weight of active compound or, with particular preference, between 0.01 % and 95% by weight of active compound, more preferably between 0.5% and 90% by weight of active compound, based on the weight of the formulation.
  • the active compound content of the application forms prepared from the formulations may vary within wide ranges.
  • the active compound concentration of the application forms may be situated typically between 0.00000001 % and 95% by weight of active compound, preferably deliberate human intervention.
  • “heterologous” in reference to a coating can refer to a seed coating comprising a bacterial strain suspended in a solution in which the bacterial strain is not naturally found.
  • the suspension solution for heterologous coatings can be natural or non-natural and can provide the bacterial strain with properties that the strain would not normally possess.
  • the suspension solution of a heterologous coating can permit the bacterial strain to adhere to the seed in such as a manner that the bacteria retains activity during seed storage and germination.
  • a seed coating can further comprise at least one nutrient, at least one biocide (e.g., herbicide or pesticide). See, for example, US App Pub. 20040336049, 20140173979, and 20150033811.
  • biocide e.g., herbicide or pesticide
  • compositions disclosed herein can be stable for at least 30, 40, 50, 60, 70, 80,
  • Biological activity refers to the ability of the formulation to improve an agronomic trait of interest or control a plant pest.
  • the stable formulation retains at least about 1%, about 10%, about 20%, about 30% about 40%, about 50%, about 60%, about 70%, about 80%, or about 90% of CFU/gram in the formulation at a given storage time point when compared to the CFU/gram produced after immediate preparation of the formulation.
  • the stable formulation retains at least about 30% to 80%, about 50% to about 80%, about 60% to about 70%, about 70% to about 80%, about 40% to about 50%, about 50% to about 60%, about 60% to about 70% of biological activity in the formulation at a given storage time point when compared to the biological activity found in the formulation immediately after production.
  • the stable formulation at a given storage time point retains at least about 30%, 45%, 50%, 60%, 70%, 80%, 90% of biological activity when compared to the biological activity found in the formulation immediately after production.
  • the stable formation retains any combination of the viability and biological activity noted above.
  • the formulations preferably comprise between 0.00000001 % and 98% by weight of active compound or, with particular preference, between 0.01 % and 95% by weight of active compound, more preferably between 0.5% and 90% by weight of active compound, based on the weight of the formulation.
  • the active compound content of the application forms prepared from the formulations may vary within wide ranges.
  • the active compound concentration of the application forms may be situated typically between 0.00000001 % and 95% by weight of active compound, preferably between 0.00001 % and 1 % by weight, based on the weight of the application form.
  • Application takes place in a customary manner adapted to the application forms.
  • the bacterial strain provided herein or an active vari nt thereof can be mixed with a biocide, such as a fungicide, insecticide, or herbicide to enhance its activity or the activity of the chemical to which it has been added.
  • a biocide such as a fungicide, insecticide, or herbicide
  • the combination of the bacterial strain and chemical may show synergistic activity where the mixture of the two exceeds that expected from their simple additive effect.
  • the biocontrol agents described herein can be mixed with other biocontrol agents.
  • the bacterial strain or active variant thereof is compatible with agricultural chemicals used to improve performance of biocides.
  • agricultural chemicals include safeners, surfactants, stickers, spreaders, UV protectants, and suspension and dispersal aids.
  • Safeners are chemicals that improve or modify the performance of herbicides.
  • Surfactants, spreaders, and stickers are chemicals included in agricultural spray preparations that change the mechanical properties of the spray (for example, by altering surface tension or improving leaf cuticle penetration).
  • UV protectants improve the performance of agricultural biocides by reducing degradation by ultraviolet light.
  • Suspension and dispersal aids improve the performance of biocides by altering their behavior in a spray tank.
  • methods can be undertaken to modify the bacterial strain to impart the compatibility of interest. Such methods to produce modified bacterial strains include both selection techniques and/or transformation techniques.
  • the bacterial strain provided herein can be used to significantly improve at least one agronomic trait of interest (e.g., reduce susceptibility to plant pests, such as insect and nematode pests).
  • the bacterial strain provided herein can be used with other pesticides for an effective integrated pest management program.
  • the biocontrol populations can be mixed with known pesticides in a manner described in WO 94/10845, herein incorporated by reference.
  • Non-limiting examples of compounds and compositions that can be added to the formulation include but are not limited to, Acetyl tributyl citrate [Citric acid, 2-(acetyloxy)-, tributyl ester]; Agar; Almond hulls; Almond shells; alpha-Cyclodextrin; Aluminatesilicate;
  • Aluminum magnesium silicate [Silicic acid, aluminum magnesium salt]; Aluminum potassium sodium silicate [Silicic acid, aluminum potassium sodium salt]; Aluminum silicate; Aluminum sodium silicate [Silicic acid, aluminum sodium salt]; Aluminum sodium silicate (1:1 : l)[Silicic acid (H4Si04), aluminum sodium salt (1 : 1 : 1)]; Ammonium benzoate [Benzoic acid, ammonium salt]; Ammonium stearate [Octadecanoic acid, ammonium salt]; Amylopectin, acid-hydrolyzed, 1- octenylbutanedioate; Amylopectin, hydrogen 1 -octadecenylbutanedioate; Animal glue; Ascorbyl palmitate; Attapulgite-type clay; Beeswax; Bentonite; Bentonite, sodian; beta-Cyclodextrin; Bone meal; Bran; Bread crumbs; (+)-Butyl lactate;
  • Glycerin [l,2,3-Propanetriol]; Glycerol monooleate [9-Octadecenoic acid (Z)-, 2,3-dihydroxypropyl ester]; Glyceryl dicaprylate [Octanoic acid, diester with 1,2,3-propanetriol]; Glyceryl dimyristate [Tetradecanoic acid, diester with 1,2,3- propanetriol]; Glyceryl dioleate [9-Octadecenoic acid (9Z)-, diester with l,2,3-propanetriol];
  • Glyceryl distearate Glyceryl monomyristate [Tetradecanoic acid, monoester with 1,2,3- propanetriol]; Glyceryl monooctanoate [Octanoic acid, monoester with l,2,3-propanetriol];
  • Glyceryl monooleate [9-Octadecenoic acid (9Z)-, monoester with l,2,3-propanetriol]
  • Glyceryl monostearate [Octadecanoic acid, monoester with l,2,3-propanetriol]
  • Glyceryl stearate [Octadecanoic acid, monoester with l,2,3-propanetriol]
  • Hydrogenated rapeseed oil Hydrogenated soybean oil; Hydroxyethyl cellulose [Cellulose, 2- hydroxyethyl ether]; Hydroxypropyl cellulose [Cellulose, 2-hydroxypropyl ether]; Hydroxypropyl methyl cellulose [Cellulose, 2-hydroxypropyl methyl ether]; Iron magnesium oxide (Fe 2 Mg0 4 );
  • Magnesium carbonate [Carbonic acid, magnesium salt (1 : 1); Magnesium benzoate; Magnesium oxide; Magnesium oxide silicate (Mg 3 0(Si 2 05)2), monohydrate; Magnesium silicate; Magnesium silicate hydrate; Magnesium silicon oxide (Mg 2 Si 3 0 8 ); Magnesium stearate [Octadecanoic acid, magnesium salt]; Magnesium sulfate; Magnesium sulfate heptahydrate; Malic acid; Malt extract; Malt flavor; Maltodextrin; Methylcellulose [Cellulose, methyl ether]; Mica; Mica-group minerals; Milk; N/A Millet seed; Mineral oil (U.S.P.); 1-Monolaurin [Dodecanoic acid, 2,3-dihydroxypropyl ester]; 1-Monomyristin [Tetradecanoic acid, 2,3-dihydroxypropy
  • Pigment Blue 29 ; Urea; Vanillia; Vermiculite; Vinegar (maximum 8% acetic acid in solution); Vitamin C [L-Ascorbic acid]; Vitamin; Walnut flour; Walnut shells; Wheat; Wheat flour; Wheat germ oil; Whey; White mineral oil (petroleum); Wintergreen oil; Wollastonite (Ca(Si03)); Wool; Xanthan gum; Yeast; Zeolites (excluding erionite (CAS Reg. No. 66733-21-9)); Zeolites, NaA; Zinc iron oxide; Zinc oxide (ZnO); and Zinc stearate [Octadecanoic acid, zinc salt]
  • the bacterial strains or modified bacterial strains or active variants thereof provided herein can be employed with any plant species to control a plant pest or improve an agronomic trait of interest.
  • Agronomic traits of interest include any trait that improves plant health or commercial value.
  • Non limiting examples of agronomic traits of interest including increase in biomass, increase in drought tolerance, thermal tolerance, herbicide tolerance, drought resistance, pest resistance (e.g., nematode resistance, insect resistance, fungus resistance, virus resistance, bacteria resistance), male sterility, cold tolerance, salt tolerance, increased yield, enhanced nutrient use efficiency, increased nitrogen use efficiency, increased tolerance to nitrogen stress, increased fermentable carbohydrate content, reduced lignin content, increased antioxidant content, enhanced water use efficiency, increased vigor, increased germination efficiency, earlier or increased flowering, increased biomass, altered root-to-shoot biomass ratio, enhanced soil water retention, or a combination thereof.
  • the agronomic trait of interest includes an altered oil content, altered protein content, altered seed carbohydrate composition, altered seed oil composition, and altered seed protein composition, chemical tolerance, cold tolerance, delayed senescence, disease resistance, drought tolerance, ear weight, growth improvement, health enhancement, heat tolerance, herbicide tolerance, herbivore resistance, improved nitrogen fixation, improved nitrogen utilization, improved root architecture, improved water use efficiency, increased biomass, increased root length, increased seed weight, increased shoot length, increased yield, increased yield under water-limited conditions, kernel mass, kernel moisture content, metal tolerance, number of ears, number of kernels per ear, number of pods, nutrition enhancement, photosynthetic capability improvement, salinity tolerance, stay-green, vigor improvement, increased dry weight of mature seeds, increased fresh weight of mature seeds, increased number of mature seeds per plant, increased chlorophyll content, increased number of pods per plant, increased length of pods per plant, reduced number of wilted leaves per plant, reduced number of severely wilted leaves per plant, and increased number of non-wilted leaves
  • the bacterial strain or active variant thereof provided herein can be employed with any plant species susceptible to a plant pest or at risk of developing a plant disease or damage caused by a plant pest.
  • pest resistance is intended that the bacterial strain or active variant thereof provided herein can inhibit (inhibit growth, feeding, fecundity, or viability), suppress (suppressing growth, feeding, fecundity, or viability), reduce (reduce the pest infestation, reduce the pest feeding activities on a particular plant) or kill (cause the morbidity, mortality, or reduced fecundity of) a pest, such as an insect pest.
  • a plant susceptible to a pest is meant that a pest is able to infect or damage the plant.
  • a plant susceptible to a pest can be susceptible to damage caused by a fungal, insect, or nematode pest as disclosed elsewhere herein.
  • plant species of interest include, but are not limited to, com (Zea mays), Brassica sp. (e.g., B. napus, B. rapa, B.juncea ), particularly those Brassica species useful as sources of seed oil, alfalfa ( Medicago sativa ), rice ( Oryza sativa ), rye ( Secale cereale), sorghum ( Sorghum bicolor, Sorghum vulgare), millet (e.g., pearl millet ⁇ Pennisetum glaucum), proso millet ( Panicum miliaceum), foxtail millet ( Setaria italica), finger millet ( Eleusine coracana )), sunflower ( Helianthus annum), safflower ( Carthamus tinctorius ), wheat ( Triticum aestivum), soybean ( Glycine max), tobacco ( Nicotiana tabacum), potato ⁇ Solanum tuberosum), peanuts ( Arachis hypogae,
  • Vegetables include tomatoes ( Lycopersicon esculentum), lettuce (e.g., Lactuca sativa), green beans ⁇ Phaseolus vulgaris ), lima beans ( Phaseolus limensis ), peas ( Lathyrus spp ), and members of the genus Cucumis such as cucumber (C. sativus), cantaloupe (C. cantalupensis ), and musk melon (C. melo).
  • tomatoes Lycopersicon esculentum
  • lettuce e.g., Lactuca sativa
  • green beans ⁇ Phaseolus vulgaris lima beans
  • Phaseolus limensis Phaseolus limensis
  • peas Lathyrus spp
  • members of the genus Cucumis such as cucumber (C. sativus), cantaloupe (C. cantalupensis ), and musk melon (C. melo).
  • Ornamentals include azalea ⁇ Rhododendron spp.), hydrangea ⁇ Macrophylla hydrangea), hibiscus ⁇ Hibiscus rosasanensis ), roses ⁇ Rosa spp.), tulips ⁇ Tulipa spp.), daffodils ⁇ Narcissus spp.), petunias ⁇ Petunia hybrida), carnation ⁇ Dianthus caryophyllus), poinsettia ⁇ Euphorbia pulcherrima), and chrysanthemum.
  • Conifers that may be employed in practicing the present invention include, for example, pines such as loblolly pine ⁇ Pinus taeda ), slash pine ⁇ Pinus elliotii), ponderosa pine ⁇ Pinus ponderosa), lodgepole pine ⁇ Pinus contorta), and Monterey pine ⁇ Pinus radiata), Douglas-fir ⁇ Pseudotsuga menziesii ); Western hemlock ⁇ Tsuga canadensis ); Sitka spruce ⁇ Picea glauca); redwood ⁇ Sequoia sempervirens), true firs such as silver fir ⁇ Abies amabilis) and balsam fir ⁇ Abies balsamea), and cedars such as Western red cedar ⁇ Thuja plicata) and Alaska yellow-cedar (C hamaecyparis nootkatensis).
  • pines such as loblolly pine
  • plants of the present invention are crop plants (for example, com, alfalfa, sunflower, Brassica, soybean, cotton, safflower, peanut, sorghum, wheat, millet, tobacco, etc.).
  • com and soybean plants are optimal, and in yet other embodiments com plants are optimal.
  • plants of interest include grain plants that provide seeds of interest, oil-seed plants, and leguminous plants.
  • Seeds of interest include grain seeds, such as corn, wheat, barley, rice, sorghum, rye, etc.
  • Oil-seed plants include cotton, soybean, safflower, sunflower, Brassica, maize, alfalfa, palm, coconut, etc.
  • Leguminous plants include beans, peas, and dry pulses. Beans include guar, locust bean, fenugreek, soybean, garden beans, cowpea, mungbean, lima bean, fava bean, lentils, chickpea, etc.
  • the bacterial strains provided herein are those that target one or more plant pests.
  • the term“pests” includes but is not limited to, insects, fungi, bacteria, nematodes, viruses or viroids, protozoan pathogens, and the like.
  • the bacterial strains provided herein are those that target one or more insect or insect pests.
  • insects or“insect pests” as used herein refers to insects and other similar pests such as, for example, those of the order Coleoptera including, but not limited to Agriotes spp., Anthonomus spp., Atomaria linearis, Chaetocnema tibialis, Cosmopolites spp., Curculio spp., Dermestes spp., Epilachna spp., Eremnus spp., Leptinotarsa decemlineata, Lissorhoptrus spp., Melolontha spp., Orycaephilus spp., Otiorhynchus spp., Phlyctinus spp., Popillia spp., Psylliodes spp., Rhizopertha spp., Scarabeidae, Sitophilus spp., Sitotroga s
  • Coleoptera insects include, but are not limited to weevils from the families Anthribidae, Bruchidae, and Curculionidae (e.g., sweetpotato weevil ( Cylas formicarius (Fabricius)), boll weevil ( Anthonomus grandis Boheman), rice water weevil ⁇ Lissorhoptrus oryzophilus Kuschel), rice weevil ⁇ Sitophilus oryzae L.)); flea beetles, cucumber beetles, rootworms, leaf beetles, potato beetles, and leafminers in the family Chrysomelidae (e.g., Colorado potato beetle ⁇ Leptinotarsa decemlineata Say), western com rootworm ⁇ Diabrotica virgifera virgifera LeConte)); chafers and other beetles from the family Scaribaei
  • insect pests include Coleoptera pests of the corn rootworm complex: Western corn rootworm, Diabrotica virgifera virgifera ; northern com rootworm, I) barberi ;
  • the insect pest is Western com rootworm, Diabrotica virgifera virgifera.
  • the methods and compositions provided herein can also be used against Hemiptera such as Lygus hesperus , Lygus lineolaris , Lygus pratensis , Lygus rugulipennis Popp , Lygus pabulums.
  • Calocoris norvegicus Orthops compestris, Plesiocoris rugicollis , Cyrtopeltis modestus , Cyrtopeltis notatus, Spanagonicus albofasciatus , Diaphnocoris chlorinonis, Labopidicola allii,
  • Insect pests of interest also include Araecerus fasciculatus, coffee bean weevil;
  • Insect pests that can be controlled with the compositions and methods disclosed herein further include insects of the order Lepidoptera, e.g. Achoroia grisella, Acleris glover ana, Acleris van ana, Adoxophyes orana, Agrotis ipsilon , Alabama argillacea, Alsophila pometaria , Amyelois transitella, Anagasta kuehniella, Anarsia lineatella, Anisota senatoria, Antheraea pernyi,
  • insects of the order Lepidoptera e.g. Achoroia grisella, Acleris glover ana, Acleris van ana, Adoxophyes orana, Agrotis ipsilon , Alabama argillacea, Alsophila pometaria , Amyelois transitella, Anagasta kuehniella, Anarsia lineatella, Anisota senatoria, Antheraea pernyi,
  • Thaurnstopoea pityocampa Tinsola bisselliella, Trichoplusia hi, Tuta absoluta , Udea rubigalis, Xylomyges curiails , and Yponomeuta padella.
  • Insect pests also include insects selected from the orders Diptera, Hymenoptera,
  • Insect pests of the present invention can further include those of the order Acari including, but not limited to, mites and ticks.
  • Coleoptera pests or Coleopteran pests include Western corn rootworm, Colorado potato beetle, and/or sweet potato weevil.
  • Insect pests that can be controlled with the compositions and methods of the invention for the major crops include, but are not limited to: Maize: Ostrinia nubilalis , European corn borer; Agrotis ipsilon , black cutworm; Helicoverpa zeae , com earworm; Spodoptera frugiperda , fall armyworm; Diatraea grandiosella , southwestern com borer; Elasmopalpus lignosellus , lesser cornstalk borer; Diatraea saccharalis, surgarcane borer; western corn rootworm, e.g., Diabrotica virgifera virgifera, northern corn rootworm, e.g., Diabrotica longicornis barbery southern corn rootworm, e.g., Diabrotica nndecimpunctata howardi ; Melanotus spp., wireworms; Cyclocephala borealis, northern masked chafer (white gru
  • Sphenophorus maidis maize billbug; Rhopalosiphum maidis , corn leaf aphid; Anuraphis maidiradicis, corn root aphid; Myzu persicae, green peach aphid; Nezara viridula, southern green stink bug; Blissus leucopterus leucopterus, chinch bug; Melanoplus femurrubrum, redlegged grasshopper; Melanoplus sanguinipes , migratory grasshopper; Hylemya platura , seedcorn maggot; Agromyza parvicornis, com blotch leafminer; Anaphothrips obscrurus, grass thrips; Solenopsis milesta, thief ant; Tetranychus urticae, two spotted spider mite; Sorghum: Chilo partellus, sorghum borer; Spodoptera frugiperda , fall armyworm; Helicoverpa zea, corn earworm
  • Rhopalosiphum maidis corn leaf aphid; Sipha flava, yellow sugarcane aphid; chinch bug, e.g., Blissus leucopterus leucopterus, Contarinia sorghicola, sorghum midge; Tetranychus
  • Melanoplus femurrubrum redlegged grasshopper
  • Melanoplns differ entialis, differential grasshopper
  • Thrips /abaci onion thrips
  • Frankliniella fusca tobacco thrips
  • Tetranychus cinnabarinus carmine spider mite
  • Tetranychus urticae two-spotted spider mite
  • Rice Diatraea saccharalis, sugarcane borer; Spodoptera frugiperda, fall armyworm; Helicoverpa zea, com earworm; Colaspis brunnea, grape colaspis; Lissorhoptrus oryzophilus, rice water weevil;
  • insects encompasses eggs, larvae, juvenile and mature forms of insects. Insects can be targeted at any stage of development. For example, insects can be targeted after the first instar, during the second instar, third instar, fourth instar, fifth instar, or any other developmental or adult growth stage. As used herein, the term“instar” is used to denote the developmental stage of the larval or nymphal forms of insects.
  • Nematodes include parasitic nematodes such as root-knot, cyst, and lesion nematodes, including of the species Meloidogyne such as the Southern Root-Knot nematode (Meloidogyne incognita), Javanese Root-Knot nematode (Meloidogyne javanica), Northern Root-Knot Nematode (Meloidogyne hapla) and Peanut Root-Knot Nematode (Meloidogyne arenaria ); nematodes of the species Ditylenchus such as Ditylenchus destructor and Ditylenchus dipsaci; nematodes of the species Pratylenchus such as the Cob Root-Lesion Nematode (Pratylenchus penetrans),
  • Meloidogyne such as the Southern Root-Knot nematode (Meloidogyne incognita), Javanese Root-Knot
  • Nematodes of the species Globodera such as Globodera rostochiensis and Globodera pallida
  • Nematodes of the species Heterodera such as Heterodera glycines (soybean cyst nematode); Heterodera schachtii (beet cyst nematode); Heterodera avenae (cereal cyst nematode)
  • Nematodes of the species Aphelenchoides such as the Rice White-tip Nematode (Aphelenchoides besseyi), Aphelenchoides ritzemabosi and Aphelenchoides fragariae ; Nematodes of the species Aphelench
  • Globodera rostochiensis and Globodera pailida potato cyst nematodes
  • Lesion nematodes include Pratylenchus spp.
  • the term "nematode” encompasses eggs, larvae, juvenile and mature forms of nematodes.
  • a cell of the bacterial strain AIP031898, AIP023234, AIP024552, AIP035573, AIP071234, AIP080021, AIP001237, AIP050674, AIP071546, AIP049805,
  • the plant pest disclosed herein is an insect pest from the order Coleoptera.
  • a cell of the bacterial strain AIP031898, AIP023234, AIP024552, AIP035573, AIP071234, AIP080021, AIP001237, AIP050674,
  • a cell of the bacterial strain AIP031898, AIP023234, AGR024552,
  • AIP092281, AIP081114, AIP046403, AIP022635, AIP070925, and AIP039063, or an active variant of any thereof, can have activity against Western com rootworm, Colorado potato beetle, and/or sweet potato weevil.
  • AIP092281, AIP081114, AIP046403, AIP022635, AIP070925, and AIP039063, or an active variant of any thereof, has activity against root-knot nematodes and Southern green stink bug.
  • Bacterial strains or active variants thereof can be tested for pesticidal activity against a plant pest (e.g., an insect pest) in early developmental stages, e.g., as larvae or other immature forms.
  • the insects may be reared in total darkness at from about 20° C to about 30° C and from about 30% to about 70% relative humidity.
  • Bioassays may be performed as described in Czapla and Lang (1990) J Econ. Entomol. 83 (6): 2480-2485. Methods of rearing insect larvae and performing bioassays are well known to one of ordinary skill in the art.
  • a fungal pest can be, but is not limited to, a fungus selected from the group consisting of Botrytis spp., Botrytis ciner ea, Cersospora spp, Cercospora sojina, Cercospora beticola, Alternaria spp., Alternaria solani, Rhizoctonia spp., Rhizoctonia solani, Blumeria graminis f sp.
  • Tritici Erysiphe necator, Podosphaera xanthii, Golovinomyces cichoracearum, Erysiphe lager stroemiae, Sphaerotheca pannosa, Colletotrichum cereale, Apiognomonia errabunda, Apiognomonia veneta, Colletotrichum spp, Colletotrichum gloeosporiodes, Discula fraxinea, Mycosphaerella spp., Phomopsis spp., Plasmopara viticola, Pseudoperonospora cubensis, Peronospora belbahrii, Bremia lactucae, Peronospora lamii, Plasmopara obduscens, Pythium spp., Pythium cryptoirregulare, Pythium aphanidermatum, Pythium irregulare, Pythium
  • the fungal pest is selected from the group consisting of Botrytis cinerea, Cercospora sojina, Alternaria solani, Rhizoctonia solani, Erysiphe necator, Podosphaera xanthii, Colletotrichum cereal, Plasmopara viticola, Peronospora belbahrii, Pythium
  • the fungal pathogen is Phakopsora sp., including Phakopsora pachyrhizi and/ or Phakopsora meibomiae.
  • Examples of fungal plant conditions and diseases caused by fungal pests include, but are not limited to, Asian Soybean Rust (ASR), gray mold, leaf spot, Frogeye Leaf Spot, Early Blight, Damping off complex, Brown Patch, black scurf, root rot, belly rot, sheath blight, Powdery Mildew, Anthracnose leaf spot, Downy Mildew, Pythium Blight, Late Blight, Fusarium Head Blight, sudden death syndrome (SDS), Fusarium Wilt, Com Stalk Rot, Brown Rust, Black Rust, Yellow Rust, Wheat Rust, Rust, Apple Scab, Verticillium Wilt, Fire Blight, and Brown Rot.
  • ASR Asian Soybean Rust
  • kits for controlling plant pests comprising applying to a plant an effective amount of at least one bacterial strain provided herein or an active variant thereof wherein the bacterial strain controls the plant pest, such as an insect pest.
  • methods of reducing susceptibility to a plant pest and/or increasing resistance to a plant pest comprising applying to a plant having a plant pest, a plant disease or damage caused by a plant pest or damage or at risk of developing a plant disease or damage caused by a plant pest an effective amount of at least one bacterial strain provided herein or an active variant thereof wherein the bacterial strain controls the plant pest.
  • a plant disease or damage comprising applying to a plant having a plant disease or damage or at risk of developing a plant disease or damage an effective amount of at least one bacterial strain provided herein or an active variant thereof wherein the bacterial strain controls a plant pest that causes the plant disease or damage.
  • the plant disease is a disease caused by an insect pest, such as a coleopteran pest.
  • the bacterial strain provided herein or active variant thereof may comprise a cell of at least one of AIP031898, AIP023234, AIP024552, AIP035573, AIP071234, AIP080021, AIP001237, AIP050674, AIP071546, AIP049805, AIP016229,
  • the effective amount of the bacterial strain or active variant thereof comprises at least about 10 12 to 10 16 CFU per hectare or least about 10 4 to 10 16 CFU per hectare, or least about 10 5 to 10 u CFU per hectare.
  • any of the bacterial strains provided herein or active variants thereof can control one, two, three, four, five, or more plant pests described herein.
  • the bacterial strain controls one, two, three, four, five or more insect pests, such as Coleoptera pests.
  • any of the bacterial strains provided herein or active variants thereof can have activity against a combination of insect pests and other plant pests, including fungi, viruses or viroids, bacteria, insects, nematodes, and protozoa pests.
  • the bacterial strain provided herein or an active variant thereof can be employed with any plant species susceptible to a plant pest of interest.
  • Examples of diseases causes by exemplary plant pests are provided in Table 1. Also provided are non-limiting exemplary crop species that are susceptible to the plant diseases caused by the pests. For example, Table 1 shows that Bortrytis cinerea causes gray mold on all flowering crops. Therefore, a bacterial strain provided herein or active variant thereof that controls Bortrytis cinerea can be applied to a plant having gray mold or at risk of developing gray mold in order to treat or prevent gray mold in the plant. Similarly, Table 1 shows that Rhizoctonia solani causes Damping off complex in com, Damping off complex in soybean, Brown Patch in turf, and Damping off complex in ornamentals.
  • a bacterial strain provided herein or active variant thereof that controls Rhizoctonia solani can be applied to a plant having Damping off complex and/or brown patch or at risk of developing Damping off complex and/or brown patch in order to treat or prevent Damping off complex and/or brown patch in the plant.
  • Table 1 shows that Colletotrichum cereal, Apiognomonia errabunda, Apiognomonia veneta , Colletotrichum gloeosporiodes , Discula fraxinea cause Anthracnose leaf spot.
  • a bacterial strain provided herein or active variant thereof that controls one or more of Colletotrichum cereal, Apiognomonia errabunda, Apiognomonia veneta, Colletotrichum gloeosporiodes, Discula fraxinea can be applied to a plant having Anthracnose leaf spot or at risk of developing
  • Anthracnose leaf spot in order to treat or prevent Anthracnose leaf spot in the plant.
  • the bacterial strain provided herein or active variants thereof controls one or more nematode pests.
  • the bacterial strain or active variants thereof can control or treat root knot nematodes, (Meloidogyne spp ).
  • Plant parasitic nematodes may attack the roots, stem, foliage and flowers of plants. All plant parasitic nematodes have piercing mouthparts called stylets. The presence of a stylet is the key diagnostic sign differentiating plant parasitic nematodes from all other types of nematodes.
  • Typical root symptoms indicating nematode attack are root knots or galls, root lesions, excessive root branching, injured root tips and stunted root systems.
  • Symptoms on the above-ground plant parts indicating root infection are a slow decline of the entire plant, wilting even with ample soil moisture, foliage yellowing and fewer and smaller leaves. These are, in fact, the symptoms that would appear in plants deprived of a properly functioning root system. Bulb and stem nematodes produce stem swellings and shortened internodes. Bud and leaf nematodes distort and kill bud and leaf tissue. In some cases, such as with SCN, yield loss may take place with no visible symptoms.
  • “treat” or“treating” or its derivatives includes substantially inhibiting, slowing, or reversing the progression of a condition, substantially ameliorating symptoms of a condition or substantially preventing the appearance of symptoms or conditions brought about by the insect pest, or the pathogen or pest that causes the plant disease.
  • controlling refers to one or more of inhibiting or reducing the growth, feeding, fecundity, reproduction, and/or proliferation of a plant pest or killing ( e.g ., causing the morbidity or mortality, or reduced fecundity) of a plant pest.
  • a plant treated with the bacterial strain provided herein may show a reduced infestation of pests, or reduced damage caused by pests by a statistically significant amount.
  • “controlling” and “protecting” a plant from a pest refers to one or more of inhibiting or reducing the growth, germination, reproduction, and/or proliferation of a pest; and/or killing, removing, destroying, or otherwise diminishing the occurrence, and/or activity of a pest.
  • a plant treated with the bacterial strain provided herein may show a reduced severity or reduced development of disease or damage in the presence of plant pests by a statistically significant amount.
  • prevent and its variations means the countering in advance of bacterial, fungal, viral, insect or other pest growth, proliferation, infestation, spore germination, and hyphae growth.
  • the composition is applied before exposure to the plant pests.
  • ameliorate and“amelioration” relate to the improvement in the treated plant condition brought about by the compositions and methods provided herein.
  • the improvement can be manifested in the forms of a decrease in pest growth and/or an improvement in the damaged or diseased plant height, weight, number of leaves, root system, or yield.
  • the term refers to the improvement in a damaged or diseased plant’s physiological state.
  • inhibitor and all variations of this term is intended to encompass the restriction or prohibition of bacterial, fungal, viral, nematode, insect, or any other pest growth, as well as spore germination.
  • eliminate relates to the substantial eradication or removal of bacteria, fungi, viruses, nematodes, insects, or any other pests by contacting them with the composition of the invention, optionally, according to the methods of the invention described below.
  • delay means the slowing of the progress of bacterial, fungal, viral, nematode, insect, or any other pest growth, and spore germination.
  • the expression “delaying the onset” is interpreted as preventing or slowing the progression of bacterial, fungal, viral, nematodes, insect, or any other pest growth, infestation, infection, spore germination and hyphae growth for a period of time, such that said bacterial, fungal, viral, nematode, insect, or any other pest growth, infestation, infection, spore germination and hyphae growth do not progress as far along in development, or appear later than in the absence of the treatment according to the invention.
  • a plant, plant part, or area of cultivation treated with the bacterial strain provided herein or an active variant thereof may show a reduced severity or reduced development of disease or damage in the presence of plant pests by a statistically significant amount.
  • a reduced severity or reduced development of disease or damage can be a reduction of about 10% to about 20%, about 20% to about 30%, about 30% to about 40%, about 40% to about 50%, about 50% to about 60%, about 60% to about 70%, about 70% to about 80%, about 80% to about 90%, or about 90% to about 100% when compared to non-treated control plants.
  • the plant treated with a bacterial strain provided herein or an active variant thereof may show a reduced severity or reduced development of disease or damage in the presence of a plant pest of at least about 10%,
  • Methods for assessing plant damage or disease severity include, measuring percentage of damaged or diseased leaf area (Godoy el al. (2006) Fitopatol. Bras. 31(1) 63-68 or by measuring uredinia counts.
  • a plant, plant part, or area of cultivation treated with the bacterial strain provided herein or an active variant thereof may show a reduction of plant pests, including insect and/or nematode pests.
  • a reduction of plant pests can be a reduction of about 10% to about 20%, about 20% to about 30%, about 30% to about 40%, about 40% to about 50%, about 50% to about 60%, about 60% to about 70%, about 70% to about 80%, about 80% to about 90%, or about 90% to about 100% when compared to non-treated control plants.
  • the plant treated with a bacterial strain provided herein or an active variant thereof may show a reduction of plant pests of at least about 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%,
  • Methods for measuring the number of plant pests include, counting the number of pests, or contacting plants with one or more pests and determining the plant's ability to survive and/or cause the death of the pests. See, for example, Czapla and Lang, (1990) J. Econ. Entomol. 83:2480-2485; Andrews, et al., (1988) Biochem. J. 252: 199-206; Marrone, et al., (1985)
  • the bacterial strains and active variants thereof provided herein have pesticidal activity against an insect pest (i.e., insecticidal activity).
  • insecticidal activity is activity against a Coleopteran species.
  • insecticidal activity is against a Lepidoptera insect.
  • insecticidal activity is against a Hemiptera species.
  • insecticidal activity is against one or more insect pests, such as the Western com rootworm, Southern com rootworm, Northern corn rootworm, Mexican corn rootworm, the Colorado potato beetle, the sweet potato weevil, or the Southern green stink bug.
  • the bacterial strains or active variants thereof provided herein reduce the damage or disease symptoms resulting from a plant pest by a statistically significant amount, including for example, at least about 10% to at least about 20%, at least about 20% to about 50%, at least about 10% to about 60%, at least about 30% to about 70%, at least about 40% to about 80%, or at least about 50% to about 90% or greater.
  • the methods of the invention can be utilized to protect plants from disease or damage caused by plant pests.
  • Assays that quantitate damage or disease resistance following pest infestation are commonly known in the art. See, for example, U.S. Patent No. 5,614,395, herein incorporated by reference. Such techniques include, measuring over time, the average lesion diameter, the pest biomass, and the overall percentage of decayed plant tissues. For example, a plant either expressing a pesticidal polypeptide or having a pesticidal composition applied to its surface shows a decrease in tissue necrosis (i.e., lesion diameter) or a decrease in plant death following challenge with a pest when compared to a control plant that was not exposed to the pesticidal composition. Alternatively, pesticidal activity can be measured by a decrease in pest biomass.
  • a plant expressing a pesticidal polypeptide or exposed to a pesticidal composition is challenged with a pest of interest.
  • tissue samples from the pest-infested tissues are obtained and RNA is extracted.
  • the percent of a specific pest RNA transcript relative to the level of a plant specific transcript allows the level of pest biomass to be determined. See, for example, Thomma et al. (1998) Plant Biology 95: 15107-15111, herein incorporated by reference.
  • in vitro pesticidal assays include, for example, the addition of varying concentrations of the pesticidal composition to paper disks and placing the disks on agar containing a suspension of the pest of interest. Following incubation, clear inhibition zones develop around the discs that contain an effective concentration of the pesticidal composition (Liu et al. (1994) Plant Biology 91 : 1888-1892, herein incorporated by reference). Additionally,
  • microspectrophotometrical analysis can be used to measure the in vitro pesticidal properties of a composition (Flu et al. (1997) Plant Mol. Biol. 34:949-959 and Cammue et al. (1992) J. Biol.
  • compositions and methods for inducing pest and/or disease resistance in a plant, wherein the disease is caused by a plant pest are also provided. Accordingly, the compositions and methods are also useful in protecting plants against any type of plant pest, including fimgal pests, viruses, nematodes, and insects.
  • methods of inducing resistance against a plant pest comprising applying to a plant that is susceptible to infection or infestation by a plant pest or a plant disease caused by the plant pest an effective amount of at least one bacterial strain provided herein or active variant thereof.
  • the bacterial strain provided herein or active variant thereof may comprise a cell of at least one of AIP031898, AIP023234, AGR024552, AIP035573, AIP071234, AIP080021, AIP001237, AIP050674, AIP071546, AIP049805,
  • AIP022635, AIP070925, and AIP039063 or an active variant of any thereof.
  • AIP022635 AIP070925, and AIP039063, or an active variant of any thereof.
  • the bacterial strain provided herein or active variant thereof promotes a defensive response to the pest that causes the plant disease or damage.
  • the effective amount of the bacterial strain provided herein or active variant thereof comprises at least about 10 12 to 10 16 CFU per hectare.
  • a defensive response in the plant can be triggered after applying the bacterial strain provided herein or active variant thereof to the plant, but prior to pest challenge and/or after pest challenge of the plant treated with the bacterial strain provided herein or active variant thereof.
  • the bacterial strain provided herein or active variant thereof induces resistance to one, two, three, four, five or more plant pests described herein. In other methods, the bacterial strain provided herein or active variant thereof induces resistance to one, two, three, four, five or more insect pests, fungal plant pests, or nematode pests described herein.
  • disease resistance is intended that the plants avoid the disease symptoms that result from plant-pest interactions. That is, pests are prevented from causing plant diseases and the associated disease symptoms, or alternatively, the disease symptoms caused by the pest are minimized or lessened as compared to a control.
  • pest resistance can be intended that the plants avoid the symptoms that result from infection or infestation of a plant by a pest. That is, pests are prevented from causing plant diseases and the associated disease symptoms, or alternatively, the disease symptoms caused by the pest are minimized or lessened as compared to a control.
  • the bacterial strain provided herein or active variant thereof may comprise a cell of at least one of AIP031898, AIP023234, AIP024552, AIP035573, AIP071234, AIP080021, AIP001237,
  • AIP070925, and AIP039063 or an active variant of any thereof; or a spore, or a forespore or a combination of cells, forespores and/or spores from any one of AIP031898, AIP023234,
  • AIP049805 AIP016229, AIP081435, AIP082140, AIP000817, AIP060333, AIP070494,
  • AIP015104 AIP011586, AIP010884, AIP082287, AIP088026, AIP065969, AIP018514,
  • the effective amount of the bacterial strain provided herein or active variant thereof comprises at least about 10 12 to 10 16 CFU per hectare.
  • the agronomic trait of interest that is improved by the bacterial strains or active variants thereof described herein is improved plant health.
  • improved plant health is meant increased growth and/or yield of a plant, increased stress tolerance and/or decreased herbicide resistance, to name a few.
  • Increased stress tolerance refers to an increase in the ability of a plant to decrease or prevent symptoms associated with one or more stresses.
  • the stress can be a biotic stress that occurs as a result of damage done to plants by other living organisms such as a pest (for example, bacteria, viruses, fungi, parasites), insects, nematodes, weeds, cultivated or native plants).
  • the stress can also be an abiotic stress such as extreme temperatures (high or low), high winds, drought, salinity, chemical toxicity, oxidative stress, flood, tornadoes, wildfires, radiation and exposure to heavy metals.
  • abiotic stress such as extreme temperatures (high or low), high winds, drought, salinity, chemical toxicity, oxidative stress, flood, tornadoes, wildfires, radiation and exposure to heavy metals.
  • improved agronomic traits are disclosed elsewhere herein.
  • an effective amount of the bacterial strain or active variant thereof improves plant health or improves an agronomic trait of interest by a statistically significant amount, including for example, at least about 10% to at least about 20%, at least about 20% to about 50%, at least about 10% to about 60%, at least about 30% to about 70%, at least about 40% to about 80%, or at least about 50% to about 90% or greater.
  • the bacterial strains provided herein or active variant thereof are applied in an effective amount.
  • An effective amount of a bacterial strain provided herein or active variant thereof is an amount sufficient to control, treat, prevent, inhibit the pest, such as an insect pest, and/or improve an agronomic trait of interest.
  • an effective amount of a bacterial strain provided herein or active variant thereof is an amount sufficient to control, treat, prevent, or inhibit a pest that causes plant disease or damage and/or reduce plant disease severity or reduce plant disease development.
  • the effective amount of the bacterial strain provided herein or active variant thereof is an amount sufficient to improve an agronomic trait of interest and/or to promote or increase plant health, growth or yield of a plant susceptible to a disease and/or infection by a plant pest or infestation by a plant pest, such as an insect pest.
  • the rate of application of the bacterial strain provided herein or active variant thereof may vary according to the pest being targeted, the crop to be protected, the efficacy of the bacterial strain provided herein or active variant thereof, the severity of the disease, the climate conditions, the agronomic trait of interest to improve, and the like.
  • the methods provided herein can comprise a single application of at least one bacterial strain provided herein or an active variant thereof to a plant, plant part, or area of cultivation or multiple applications of at least one bacterial strain provided herein or an active variant thereof to a plant, plant part, or area of cultivation.
  • the rate of bacterial strain provided herein or active variant thereof is 10 7 to 10 16 colony forming units (CFU) per hectare.
  • the rate of bacterial strain provided herein or active variant thereof application is 3 x 10 7 to 1 x 10 11 colony forming units (CFU) per hectare. (This corresponds to about 1 Kg to 10kg of formulated material per hectare).
  • the rate of bacterial strain provided herein or active variant thereof application is 3 x 10 7 to 1 x 10 16 colony forming units (CFU) per hectare; about lxlO 12 to about lxlO 13 colony forming units (CFU) per hectare, about lxlO 13 to about lxlO 14 colony forming units (CFU) per hectare, about lxlO 14 to about lxlO 15 colony forming units (CFU) per hectare, about lxlO 15 to about lxlO 16 colony forming units (CFU) per hectare, about lxlO 16 to about lxlO 17 colony forming units (CFU) per hectare; about lxlO 4 to about lxlO 14 colony forming units (CFU) per hectare; about lxlO 5 to about lxlO 13 colony forming units (CFU) per hectare; about lxlO
  • the rate of bacterial strain provided herein or active variant thereof application is at least about lxlO 4 , about lxlO 5 , about lxlO 6 , about lxlO 7 , about lxlO 8 , about lxlO 9 , about lxlO 10 , about lxlO 11 , about
  • the rate of bacterial strain provided herein or active variant thereof application is at least lxlO 7 to at least about lxlO 12 CFU/hectare.
  • the bacterial strain provided herein or active variant thereof applied comprises the strain deposited as AIP031898, AIP023234, AIP024552, AIP035573, AIP071234, AIP080021, AIP001237, AIP050674, AIP071546, AIP049805, AIP016229, AIP081435, AIP082140,
  • Any appropriate agricultural application rate for a biocide can be applied in combination with the bacterial strain provided herein or active variant thereof disclosed herein.
  • Methods to assay for the effective amount of the bacterial strain provided herein or active variant thereof include, for example, any statistically significant increase in the control of the pest targeted by the bacterial strain or active variant thereof. Methods to assay for such control are known.
  • a statistically significant increase in plant health, yield and/or growth can occur upon application of an effective amount of the bacterial strain provided herein or active variant thereof when compared to the plant health, yield and/or growth that occurs when no bacterial strain provided herein or active variant thereof is applied.
  • compositions comprising a bacterial strain provided herein or active variant thereof provided herein (i .e., a cell of at least one of AIP031898,
  • AIP081114, AIP046403, AIP022635, AIP070925, and AIP039063, or an active variant of any thereof By“applying” is intended contacting an effective amount of the bacterial strain provided herein or active variant thereof to a plant, area of cultivation, and/or seed with one or more of the bacterial strains provided herein or active variant thereof so that a desired effect is achieved.
  • the application of the bacterial strain provided herein or active variant thereof can occur prior to the planting of the crop (for example, to the soil, the seed, or the plant).
  • the application of the bacterial strain provided herein or active variant thereof is a foliar application. Therefore, a further embodiment of the invention provides a method for controlling or inhibiting the growth of a plant pest by applying the population of bacterial strain provided herein or active variant thereof to an environment in which the plant pest may grow.
  • the application may be to the plant, to parts of the plant, to the seeds of the plants to be protected, or to the soil in which the plant to be protected are growing or will grow.
  • Application to the plant or plant parts may be before or after harvest. Application to the seeds will be prior to planting of the seeds.
  • an effective amount of at least one bacterial strain provided herein or active variant thereof provided herein is used as a foliar application to control or inhibit growth of one or more nematode pathogens from the group consisting of Southern Root-Knot nematode ⁇ Meloidogyne incognita), Javanese Root-Knot nematode ( Meloidogyne javanica ), Northern Root- Knot Nematode (Meloidogyne hapla ) and Peanut Root-Knot Nematode.
  • an effective amount of at least one bacterial strain provided herein or active variant thereof provided herein is used as a foliar or soil or seed application to control or inhibit growth of one or more insect pests.
  • an effective amount of at least one bacterial strain provided herein, or active variant thereof can be used as a foliar application to control or inhibit growth of Coleopteran insects including com rootworms, Western com rootworm, Colorado potato beetle, weevils, and the sweetpotato weevil.
  • an effective amount of at least one bacterial strain provided herein or active variant thereof provided herein is applied to the soil in which the plants to be protected are growing or will grow to control or inhibit growth of one or more nematode or nematode pest.
  • an effective amount of at least one bacterial strain provided herein or active variant thereof provided herein is applied to plant seed for inhibiting (inhibiting growth, feeding, fecundity, or viability), suppressing
  • a plant pest e.g., an insect pest, such as a Coleoptera pest.
  • an effective amount of at least one bacterial strain provided herein or active variant thereof provided herein is applied to the plant propagule (i.e. seed, slip, stem cutting, com etc.) from which the plant to be protected are growing or will grow to control or inhibit growth of one or more plant pests.
  • an effective amount of at least one bacterial strain provided herein, or active variant thereof can be applied to the plant propagule to control or inhibit growth of insect pests (e.g., Coleopteran insects including corn rootworms, Western corn rootworm, Colorado potato beetle, weevils, and the sweetpotato weevil).
  • an effective amount of at least one bacterial strain provided herein, or active variant thereof can be applied to the plant tissue (including fruit) before or after harvest to control or inhibit growth of a plant pest (e.g., insect pest, such as Coleopteran insects including corn rootworms, Western corn rootworm, Colorado potato beetle, weevils, and the sweetpotato weevil).
  • a plant pest e.g., insect pest, such as Coleopteran insects including corn rootworms, Western corn rootworm, Colorado potato beetle, weevils, and the sweetpotato weevil.
  • an effective amount of a bacterial strain provided herein or active variant thereof provided herein is applied to the plant tissue (including fruit) after harvest to control or inhibit growth of one or more nematode pests.
  • an effective amount of at least one bacterial strain provided herein or active variant thereof provided herein is applied to the soil in which the plant to be protected are growing or will grow to control or inhibit growth of one or more pests selected from the group consisting of Southern Root-Knot nematode ( Meloidogyne incognita ), Javanese Root-Knot nematode ( Meloidogyne javanica ), Northern Root-Knot Nematode (Meloidogyne hapla ) and Peanut Root-Knot Nematode.
  • an effective amount of a bacterial strain provided herein or active variant thereof provided herein is applied to the plant after harvest to control or inhibit growth of one or more pests selected from the group consisting of Southern Root-Knot nematode
  • the term plant includes plant cells, plant protoplasts, plant cell tissue cultures from which plants can be regenerated, plant calli, plant clumps, and plant cells that are intact in plants or parts of plants such as embryos, pollen, ovules, seeds, leaves, flowers, branches, fruit, kernels, ears, cobs, husks, stalks, roots, root tips, anthers, and the like.
  • Grain is intended to mean the mature seed produced by commercial growers for purposes other than growing or reproducing the species.
  • the application of the bacterial strain provided herein or active variant thereof i.e., a cell of at least one of AIP031898, AIP023234, AIP024552, AIP035573, AIP071234, AIP080021, AIP001237, AIP050674, AIP071546, AIP049805, AIP016229,
  • AIP070925, and AIP039063, or an active variant of any thereof) is applied to the seeds of a plant, such as the seeds of a corn (maize) plant.
  • Application of the bacterial strain, or an active variant thereof, to com seed can comprise a concentration of aboutlO 5 CFU/gram to about 10 11 CFU/gram, about 10 7 CFU/gram to about 10 10 CFU/gram, about 10 7 CFU/gram to about 10 11 CFU/gram, about 10 6 CFU/gram to about 10 10 CFU/gram, about 10 6 CFU/gram to about 10 11 CFU/gram, about 10 11 CFU/gram to about 10 12 CFU/gram, about 10 5 CFU/gram to about 10 10 CFU/gram, about 10 5 CFU/gram to about 10 12 CFU/gram, about 10 5 CFU/gram to about 10 6 CFU/gram, about 10 6 CFU/gram to about 10 7 CFU/gram, about 10 7 CFU/gram to about 10 8 CFU/gram, about 10 8 CFU/
  • the concentration of the bacterial strain comprises at least about 10 5 CFU/gram, at least about 10 6 CFU/gram, at least about 10 7 CFU/gram, at least about 10 8 CFU/gram, at least about 10 9 CFU/gram, at least about 10 10 CFU/gram, at least about 10 11 CFU/gram, at least about 10 12 CFU/gram, or at least about 10 13 CFU/gram.
  • the bacterial strain, or active variant thereof, applied to the corn seed is applied in the form of a heterologous seed coating as described elsewhere herein. The concentration and timing of application can vary depending on the conditions and geographical location.
  • the application of the bacterial strain provided herein or active variant thereof i.e., a cell of at least one of AIP031898, AIP023234, AIP024552, AIP035573, AIP071234, AIP080021, AIP001237, AIP050674, AIP071546, AIP049805, AIP016229,
  • AIP070925, and AIP039063, or an active variant of any thereof is applied to the leaves of a soybean plant.
  • the timing of application can vary depending on the conditions and geographical location.
  • a plant pest such as one that causes a plant disease, in an area of cultivation containing a plant susceptible to the plant pest or a plant disease caused by a plant pest.
  • the method comprises planting the area of cultivation with seeds or plants susceptible to the plant disease or pest; and applying to the plant susceptible to the disease or pest, the seed or the area of cultivation of the plant susceptible to the plant disease or pest an effective amount of at least one bacterial strain provided herein or active variant thereof (i.e., a cell of at least one of AIP031898, AIP023234, AIP024552, AIP035573, AIP071234, AIP080021, AIP001237, AIP050674, AIP071546, AIP049805, AIP016229, AIP081435, AIP082140, AIP000817,
  • AIP070925, and AIP039063 or an active derivative of any thereof, or a spore, or a forespore or a combination of cells, forespores and/or spores from any one of AIP031898, AIP023234,
  • AIP049805 AIP016229, AIP081435, AIP082140, AIP000817, AIP060333, AIP070494,
  • AIP015104 AIP011586, AIP010884, AIP082287, AIP088026, AIP065969, AIP018514,
  • the effective amount of the bacterial strain provided herein or active variant thereof controls the plant pest without significantly affecting the plant.
  • the effective amount comprises at least about 10 12 to 10 16 colony forming units (CFU) per hectare.
  • a method for growing a plant susceptible to a plant pest or a plant disease caused by a plant pest comprises applying to a plant susceptible to the disease or pest, a seed, or an area of cultivation of the plant susceptible to the disease or pest an effective amount of a composition comprising at least one bacterial strain provided herein or active variant thereof.
  • the bacterial strain provided herein or active variant thereof may comprise a cell of at least one of AIP031898, AIP023234, AIP024552, AIP035573, AIP071234, AIP080021, AIP001237, AIP050674, AIP071546, AIP049805, AIP016229,
  • AIP070925, and AIP039063 or an active variant of any thereof.
  • Various effective amounts of at least one bacterial strain provided herein or active variant thereof are disclosed elsewhere herein and in one, non-limiting example, the effective amount of the bacterial strain provided herein or active variant thereof comprises at least about 10 12 to 10 16 colony forming units (CFU) per hectare.
  • the “yield” of the plant refers to the quality and/or quantity of biomass produced by the plant.
  • biomass is intended any measured plant product.
  • An increase in biomass production is any improvement in the yield of the measured plant product.
  • An increase in yield can comprise any statistically significant increase including, but not limited to, at least a 1% increase, at least a 3% increase, at least a 5% increase, at least a 10% increase, at least a 20% increase, at least a 30%, at least a 50%, at least a 70%, at least a 100% or a greater increase in yield compared to a plant not exposed to the bacterial strain provided herein or active variant thereof.
  • a method for increasing yield in a plant comprises applying to a crop or an area of cultivation an effective amount of a composition comprising at least one bacterial strain comprising AIP031898, AIP023234, AIP024552, AIP035573, AIP071234, AIP080021, AIP001237, AIP050674, AIP071546, AIP049805, AIP016229, AIP081435,
  • AIP046403, AIP022635, AIP070925, and AIP039063 or an active variant of any thereof, a spore or a forespore or a combination of cells, forespores and/or spores from any one of AGR031898, AIP023234, AIP024552, AIP035573, AIP071234, AIP080021, AIP001237, AIP050674,
  • CFU colony forming units
  • an“area of cultivation” comprises any region in which one desires to grow a plant.
  • Such areas of cultivations include, but are not limited to, a field in which a plant is cultivated (such as a crop field, a sod field, a tree field, a managed forest, a field for culturing fruits and vegetables, etc.), a greenhouse, a growth chamber, etc.
  • a plant of interest i.e., plant susceptible to a plant pest or plant disease caused by a plant pest
  • the area of cultivation comprising the plant can be treated with a combination of an effective amount of the bacterial strain provided herein or active variant thereof and an effective amount of a biocide or other biocontrol agent.
  • a bacterial strain provided herein or active variant thereof and a biocide or other biocontrol agent By“treated with a combination of’ or“applying a combination of’ a bacterial strain provided herein or active variant thereof and a biocide or other biocontrol agent to a plant, area of cultivation or field it is intended that one or more of a particular field, plant, and/or weed is treated with an effective amount of one or more of the bacterial strains provided herein or active variant thereof and one or more biocide or other biocontrol agent so that a desired effect is achieved.
  • the application of one of the bacterial strains provided herein or active variant thereof and the biocide or other biocontrol agent can occur prior to the planting of the crop (for example, to the soil, or the plant).
  • the application of the bacterial strains provided herein or active variant thereof and the biocide or other biocontrol agent may be simultaneous or the applications may be at different times
  • the active variant comprises a bacterial strain provided herein that is resistant to one or more biocide.
  • the bacterial strain provided herein or active variant thereof i.e., a cell of at least one of AIP031898, AIP023234, AIP024552, AIP035573, AIP071234, AIP080021, AIP001237, AIP050674, AIP071546,
  • AIP049805 AIP016229, AIP081435, AIP082140, AIP000817, AIP060333, AIP070494,
  • AIP015104 AIP011586, AIP010884, AIP082287, AIP088026, AIP065969, AIP018514,
  • AIP022635, AIP070925, and AIP039063, or an active variant of any thereof) is resistant to glyphosate.
  • a plant, crop, or area of cultivation is treated with a combination of an effective amount of the bacterial strain provided herein or active variant thereof that is resistant to glyphosate and an effective amount of glyphosate, wherein the effective amount of glyphosate is such as to selectively control weeds while the crop is not significantly damaged.
  • the active variant comprises a bacterial strain provided herein that is resistant to glufosinate.
  • a plant, crop, or area of cultivation is treated with a combination of an effective amount of the bacterial strain provided herein or active variant thereof that is resistant to glufosinate and an effective amount of glufosinate, wherein the effective amount of glufosinate is such as to selectively control weeds while the crop is not significantly damaged.
  • the effective amount of the bacterial strain provided herein or active variant thereof is sufficient to result in a statistically significant increase in plant health, yield, and/or growth when compared to the plant health, yield, and/or growth that occurs when the same concentration of a bacterial strain provided herein or active variant thereof that was not modified to be resistant to glufosinate is applied in combination with the effective amount of the glufosinate or active derivative thereof.
  • the bacterial strain provided herein or active variant thereof comprises an effective amount of a cell of at least one of
  • AIP070925, and AIP039063 or an active variant of any thereof, or a spore, or a forespore or a combination of cells, forespores and/or spores from any one of AIP031898, AIP023234,
  • AIP049805 AIP016229, AIP081435, AIP082140, AIP000817, AIP060333, AIP070494,
  • AIP015104 AIP011586, AIP010884, AIP082287, AIP088026, AIP065969, AIP018514,
  • the bacterial strain provided herein or active variant thereof can be used in combination with a biocide (i.e., an herbicide, insecticide, fungicide, pesticide, or other crop protection chemical).
  • a biocide i.e., an herbicide, insecticide, fungicide, pesticide, or other crop protection chemical.
  • the bacterial strain provided herein or active variant thereof is compatible with the biocide of interest.
  • herbicide fungicide, pesticide, insecticide or other crop protection chemical tolerance or herbicide, fungicide, pesticide, insecticide or other crop protection chemical resistance
  • an organism i.e., the plant and/or the bacterial strain provided herein or active variant thereof etc.
  • Herbicides that can be used in the various methods and compositions discloses herein include glyphosate, ACCase inhibitors (Arloxyphenoxy propionate (FOPS)); ALS inhibitors (Sulfonylurea (SU)), Imidazonlinone (IMI), Pyrimidines (PM)); microtubule protein inhibitor (Dinitroaniline (DNA)); synthetic auxins (Phenoxy (P)), Benzoic Acid (BA), Carboxylic acid (CA)); Photosystem II inhibitor (Triazine (TZ)), Triazinone (TN), Nitriles (NT),
  • Benzothiadiazinones BZ
  • Ureas US
  • EPSP Synthase inhibitor glycines
  • Glutamine Synthesis inhibitor Phosphinic Acid (PA)
  • DOXP synthase inhibitor Isoxazolidinone (IA)
  • HPPD inhibitor Pyrazole (PA)
  • NP N- phenylphthalimide
  • PPO inhibitors Diphenyl ether (DE), N- phenylphthalimide (NP) (Ary triazinone (AT)
  • VLFA inhibitors chloroacetamide (CA)), Oxyacetamide (OA)
  • Photosystem I inhibitor Bipyridyliums (BP)
  • BP Photosystem I inhibitor
  • Pesticides that can be used in the various methods and compositions disclosed herein include imidacloprid clothianidin, arylpyrazole compounds (W02007103076); organophosphates, phenyl pyrazole, pyrethoids caramoyloximes, pyrazoles, amidines, halogenated hydrocarbons, carbamates and derivatives thereof, terbufos, chloropyrifos, fipronil, chlorethoxyfos, telfuthrin, carbofuran, imidacloprid, tebupirimfos (U S. Patent No. 5,849,320).
  • Insecticides that can be used in the various methods and compositions disclosed herein include imidacloprid, beta-cyfluthrin, cyantraniliprole, diazinon, lambda-cyhalothrin, methiocarb, pymetrozine, pyrifluquinazon, spinetoram, spirotetramat, thiodicarb, and Ti-435, carbamates, sodium channel modulators/voltage dependent sodium channel blockers, pyrethroids such as DDT, oxadiazines such as indoxacarb, acetylcholine-receptor agonists/antagonists, acetylcholine- receptor-modulators, nicotine, bensultap, cartap, chloronicotyinyls such as acetamiprid, bifenthrin, clothianidin, dinotefuran, imidac loprid, nitenpyram, nithiazine, thiacloprid
  • Nematicides that can be used in the various methods and compositions disclosed herein include, but are not limited to, acibenzolar-S-methyl, an avermectin (e.g., abamectin), carbamate nematicides (e.g., aldicarb, thiadicarb, carbofuran, carbosulfan, oxamyl, aldoxycarb, ethoprop, methomyl, benomyl, alanycarb), organophosphorus nematicides (e.g., phenamiphos (fenamiphos), fensulfothion, terbufos, fosthiazate, dimethoate, phosphocarb, dichlofenthion, isamidofos, fosthietan, isazofos ethoprophos, cadusafos, terbufos, chlorpyrifos, dichlofenthion, heterophos, isa
  • Fungicides that can be used in the various methods and compositions disclosed herein include aliphatic nitrogen fungicides (butylamine, cymoxanil, dodicin, dodine, guazatine, iminoctadine); amide fungicides (benzovindiflupyr, carpropamid, chloraniformethan, cyflufenamid, diclocymet, diclocymet, dimoxystrobin, fenaminstrobin, fenoxanil, flumetover, furametpyr, isofetamid, isopyrazam, mandestrobin, mandipropamid, metominostrobin, orysastrobin, penthiopyrad, prochloraz, quinazamid, silthiofam, triforine); acylamino acid fungicides (benalaxyl, benalaxyl-M, furalaxyl, metalaxyl, metalaxyl-M, pe
  • antibiotic fungicides aureofungin, blasticidin-S, cycloheximide, griseofulvin, kasugamycin, moroxydine, natamycin, polyoxins, polyoxorim, streptomycin, validamycin
  • strobilurin fungicides fluoxastrobin, mandestrobin
  • methoxyacrylate strobilurin fungicides azoxystrobin, bifujunzhi, coumoxystrobin, enoxastrobin, flufenoxystrobin, jiaxiangjunzhi, picoxystrobin, pyraoxystrobin
  • methoxycarbanilate strobilurin fungicides pyraclostrobin, pyrametostrobin, triclopyricarb
  • methoxyiminoacetamide strobilurin fungicides (dimoxystrobin, fenaminstrobin, metominostrobin, orysastrobin); methoxyiminoacetate strobilurin fungicides (kresoxim-methyl, trifloxystrobin); aromatic fungicides (biphenyl, chlorodinitronaphthalenes, chloroneb, chlorothalonil, cresol, dicloran, fenjuntong, hexachlorobenzene, pentachlorophenol, quintozene, sodium
  • conazole fungicides triazoles
  • polysulfide fungicides barium polysulfide, calcium polysulfide, potassium polysulfide, sodium polysulfide
  • pyrazole fungicides benzovindiflupyr, bixafen, fenpyrazamine, fluxapyroxad, furametpyr, isopyrazam, oxathiapiprolin, penflufen, penthiopyrad, pyraclostrobin, pyrametostrobin, pyraoxystrobin, rabenzazole, sedaxane
  • pyridine fungicides (boscalid, buthiobate, dipyrithione, fluazinam, fluopicolide, fluopyram, parinol, picarbutrazox, pyribencarb, pyridinitril, pyrifenox, pyrisoxazole, pyroxychlor, pyroxyfur, triclopyricarb); pyrimidine fung
  • anilinopyrimidine fungicides cyprodinil, mepanipyrim, pyrimethanil
  • pyrrole fungicides cyprodinil, mepanipyrim, pyrimethanil
  • fungicides dimetachlone, fenpiclonil, fludioxonil, fluoroimide
  • quaternary ammonium fungicides berberine, sanguinarine
  • quinoline fungicides ethoxyquin, halacrinate, 8-hydroxyquinoline sulfate, quinacetol, quinoxyfen, tebufloquin
  • quinone fungicides chloranil, dichlone, dithianon
  • quinoxaline fungicides chinomethionat, chlorquinox, thioquinox
  • thiazole fungicides ethaboxam, isotianil, metsulfovax, octhilinone, oxathiapiprolin, thiabendazole, thifluzamide
  • thiazolidine fungicides flutianil, thiadifluor
  • thiocarbamate fungicides metalhasulfocarb, prothiocarb
  • thiophene fungicides ethaboxam, isofetamid, silthiofam
  • triazine fungicides anilazine
  • triazole fungicides amisulbrom, bitertanol, fluotrimazole, triazbutil
  • conazole fungicides triazoles
  • a composition comprising a cell, a spore, a forespore, or a combination of cells, spores, and/or forespores of at least one of bacterial strain AIP031898, AIP023234, AIP024552,
  • AIP092281, AIP081114, AIP046403, AIP022635, AIP070925, and AIP039063 or an active variant of any thereof, wherein the active variant comprises a cell, spore, forespore, or a combination of cells, spores and/or forespores having a genome within a Mash distance of about 0.015;
  • said cell, spore, forespore, or combination of cells, spores and/or forespores or the active variant of any thereof is present at about 10 5 CFU/gram to about 10 12 CFU/gram or at about 10 5 CFU/ml to about 10 12 CFU/ml, and wherein an effective amount of said composition controls a plant pest or improves an agronomic trait of interest of a plant.
  • composition of embodiment 1, wherein said plant pest comprises an insect pest.
  • composition of embodiment 2, wherein said insect pest comprises a Coleoptera insect or Hemiptera insect.
  • composition of embodiment 3, wherein said Coleoptera insect is a corn rootworm, Colorado potato beetle, or a weevil.
  • composition of embodiment 3, wherein said Coleoptera insect is a Western com rootworm, Colorado potato beetle, or sweetpotato weevil.
  • composition of any of embodiments 1-5 wherein said cell, spore, forespore, or combination of cells, spores and/or forespores or the active variant of any thereof is present at about 10 5 CFU/gram to about 10 10 CFU/gram or at about 10 5 CFU/ml to about 10 10 CFU/ml.
  • composition comprises a cell paste.
  • a composition comprising a cell paste comprising a cell, a spore, a forespore, or a combination of cells, spores and/or forespores of at least one of bacterial strain AIP031898, AIP023234, AIP024552, AIP035573, AIP071234, AIP080021, AIP001237, AIP050674,
  • an effective amount of said composition controls a plant pest or improves an agronomic trait of interest of a plant.
  • composition of embodiment 9, wherein said plant pest comprises an insect pest.
  • composition of embodiment 10, wherein said insect pest comprises a Coleoptera insect or Hemiptera insect.
  • composition of embodiment 11, wherein said Coleoptera insect is a corn rootworm, Colorado potato beetle, or a weevil.
  • composition of embodiment 13, wherein said Coleoptera insect is a Western com rootworm, Colorado potato beetle, or sweetpotato weevil.
  • a composition comprising a wettable power comprising a cell, a spore, a forespore, or a combination of cells, spores and/or forespores of at least one of bacterial strain AIP031898, AIP023234, AIP024552, AIP035573, AIP071234, AIP080021, AIP001237, AIP050674,
  • composition of embodiment 14 wherein an effective amount of said composition controls a plant pest or improves an agronomic trait of interest of a plant.
  • plant pest comprises at least one insect pest.
  • composition of embodiment 15, wherein said insect pest comprises a Coleoptera insect or Hemiptera insect.
  • composition of embodiment 16, wherein said Coleoptera insect is a corn rootworm, Colorado potato beetle, or a weevil.
  • composition of embodiment 16, wherein said Coleoptera insect is a Western com rootworm, Colorado potato beetle, or sweetpotato weevil.
  • composition of embodiment 19, wherein said active variant is selected under herbicide, fungicide, pesticide, insecticide, or other crop protection chemical pressure and is resistant to said herbicide, fungicide, pesticide, insecticide, or other crop protection chemical.
  • composition of embodiment 21, wherein said herbicide is selected from the group consisting of glyphosate, glufosinate (glutamine synthase inhibitor), sulfonylurea and
  • imidazolinone herbicides branched chain amino acid synthesis inhibitors.
  • An isolated biologically pure culture of a bacterial strain comprising a cell, a spore, a forespore, or a combination of cells, spores and/or forespores of AIP031898, AIP023234,
  • AIP049805 AIP016229, AIP081435, AIP082140, AIP000817, AIP060333, AIP070494,
  • AIP015104 AIP011586, AIP010884, AIP082287, AIP088026, AIP065969, AIP018514,
  • an effective amount of said cell, spore, forespore, or combination of cells, spores and/or forespores controls a plant pest or improves an agronomic trait of interest of a plant.
  • AIP022635, AIP070925, and AIP039063 or an active variant of any thereof, wherein the active variant comprises a cell, spore, forespore, or a combination of cells, spores and/or forespores having a genome within a Mash distance of about 0.015;
  • said bacterial culture is able to grow in the presence of glufosinate
  • an effective amount of said bacterial culture controls a plant pest or improves an agronomic trait of interest of a plant.
  • a method for controlling a plant pest population comprising contacting said population with an effective amount of the composition of any one of embodiments 1-22, the isolated biologically pure culture of any one of embodiments 23-29, or the bacterial culture of any one of embodiments 30-34, wherein said bacterial strain controls said plant pest.
  • a method for growing a plant susceptible to a plant pest or plant disease caused by a plant pest or improving an agronomic trait of interest in a plant comprising applying to the plant an effective amount of a cell, a spore, a forespore, or a combination of cells, spores and/or forespores of at least one of bacterial strain AIP031898, AIP023234, AIP024552, AIP035573, AIP071234, AIP080021, AIP001237, AIP050674, AIP071546, AIP049805, AIP016229, AIP081435,
  • AIP046403, AIP022635, AIP070925, and AIP039063 or an active variant of any thereof wherein the active variant comprises a cell, spore, forespore, or a combination of cells, spores and/or forespores having a genome within a Mash distance of about 0.015;
  • said effective amount comprises at least about 10 12 to 10 16 colony forming units (CFU) per hectare;
  • said effective amount controls said plant pest or improves the agronomic trait of interest.
  • Colorado potato beetle or a weevil.
  • a method of controlling a plant pest in an area of cultivation comprising:
  • AIP022635, AIP070925, and AIP039063 or an active variant of any thereof, wherein the active variant comprises a cell, spore, forespore, or combination of cells, spores and/or forespores having a genome within a Mash distance of about 0.015; and wherein said effective amount comprises at least about 10 12 to 10 16 colony forming units (CFU) per hectare.
  • CFU colony forming units
  • a method of making a modified bacterial strain comprising:
  • AIP092281, AIP081114, AIP046403, AIP022635, AIP070925, and AIP039063 or an active variant of any thereof, wherein the active variant comprises a cell, spore, forespore, or a combination of cells, spores and/or forespores having a genome within a Mash distance of about 0.015, wherein said population is susceptible to a biocide of interest;
  • a method of treating or preventing a plant disease or damage caused by a plant pest comprising applying to a plant having said plant pest, said plant disease, said damage, susceptible to said plant pest, or at risk of developing said plant disease or damage an effective amount of a cell, a spore, a forespore, or a combination of cells, spores and/or forespores of at least one of bacterial strain AIP031898, AIP023234, AIP024552, AIP035573, AIP071234, AIP080021, AIP001237, AIP050674, AIP071546, AIP049805, AIP016229, AIP081435, AIP082140,
  • said effective amount comprises at least about 10 12 to 10 16 CFU per hectare, and wherein said cell, spore, forespore, or a combination of cells, spores and/or forespores controls the plant pest that causes the plant disease.
  • Colorado potato beetle or a weevil.
  • Table 2 A bacterial strain selected for evaluation of inhibition of pest activity.
  • the bacterial strains set forth in Table 2 were cultured in TB medium.
  • Table 3 summarizes the incubation time. Following incubation, each strain was at a concentration of at least 10 7 CFU/ml, at least 10 8 CFU/ml, at least 10 9 CFU/ml, at least 10 10 CFU/ml, at least 10 11 CFU/ml, or at least 10 12 CFU/ml.
  • Plant material The susceptible soybean cultivar Williams 82 was used in strain evaluation using the detached-leaf technique (Twizeyimana and Hartman (2010) Plant Dis 94: 1453-1460) and using whole plant in growth chambers. Briefly, soybean plants were planted every two weeks and placed inside a growth chamber (Percival Scientific, Inc., Boone, IA) maintained at 75% relative humidity (RH) with a daily cycle of 14 h of light (350 mitio ⁇ m V PAR) and 10 h of darkness at 24° and 23°C, respectively, for a constant supply of 2- to 3-week-old rust-free leaves. A mixture of P. pachyrhizi urediniospores obtained from infected soybean leaves collected from Gadsden County, Florida in 2015 and 2016 was used in this experiment.
  • Percival Scientific, Inc., Boone, IA maintained at 75% relative humidity (RH) with a daily cycle of 14 h of light (350 mitio ⁇ m V PAR) and 10 h of darkness at 24° and 23
  • leaf disks (3.5-cm diameter each) were sprayed with 120 pl of each bacterial strain of interest (1 x 10 8 CFEl/ml of sterile distilled water) using a fingertip sprayer (Container & Packaging Supply, Eagle, ID) fitted to a 15 mL conical centrifuge tube (Fisher Scientific, Cat. No. 14-59-53A).
  • Fingertip sprayer Container & Packaging Supply, Eagle, ID
  • 15 mL conical centrifuge tube (Fisher Scientific, Cat. No. 14-59-53A).
  • Leaf disks were placed adaxial side down on saturated 20 x 20 cm filter paper (Whatman International Ltd., Kent, England) in a plastic container (Blister Box 20 x 20 cm, Placon, Madison, WI); two filter papers were used per box.
  • Leaf disks 20 per box were incubated at room temperature in the dark for 24 h.
  • the leaf disks were than inoculated with a spore suspension of P. pachyrhizi urediniospores (120 pL per leaf disk at 5 x 10 4 urediniospores/mL of sterile distilled water) using an atomizer attached to an air compressor (Twizeyimana and Hartman, 2010).
  • the boxes with leaf disks were incubated in the dark for a period of 12 h followed by a cycle of 13 hours of light (40-60 nmol m V ' ) at 22.5°C and 11 h of darkness at 22°C in a growth chamber (Percival Scientific, Inc.) maintained at 78% RH. Prior to placing in a growth chamber, boxes were placed inside zip bags (Webster Industries, Peabody, MA).
  • Rust severity was scored by counting the number of sporulating uredinia in two arbitrarily selected l-cm diameter circle of leaf tissue from an inoculated leaf disk (Table 4). Data was analyzed using analysis of variance (ANOVA) in PROC GLM of SAS (version 9.4; SAS Institute Inc., Cary, NC) and significant differences (P ⁇ 0.05) were observed among treatments.
  • Rhizoctonia solani infested grain 11-14 day old Rhizoctonia solani infested grain is ground.
  • the ground inoculum is screened through a #10 screen to remove any grain that is not ground well.
  • the ground, screened infected grains are added to Fafard Superfine Germination media at 1.5 grams of ground inoculum to 1 liter of soil mix by volume.
  • Germination mix, inoculum, and 1 liter of water per 75 liters of germination media are added to a cement mixer and mix until everything is well incorporated.
  • the well incorporated media-inoculum material is placed into a secondary holding container with a lid and held at 20°C for 18 hours before using in the assay.
  • 606-cell planting trays are filled with inoculated germination media making sure to not pack the media too firmly.
  • One soybean seed is sown per 606 cell, planting at a depth of 1.5 to 2cm leaving the planting holes open if applying treatments as a liquid formulation
  • Individual planting cells are treated with the re-suspended strain set forth in Table 2 at 3ml per cell/seed. The seed treatment is directly over the top of the seed.
  • the shake flats is shaken lightly shake to close planting holes.
  • the planting trays are lightly watered and placed in a humidity dome on the flat. After 3-4 days, flats are checked for moisture and lightly watered as needed to ensure cells are evenly moist. The humidity dome is replaced after watering.
  • a starter culture was prepared by filling a 96-well block with l-ml (per well) LB media. From a freezer-stock screenmate, using pipet tips or an inoculation loop each well of the block was inoculated with a bacterial strain. This starter culture was grown at 30°C shaking at 225 rpms for 24 h. Assay cultures were prepared by filling two 48-well blocks with ⁇ 1.7 ml (per well) media.
  • Assay block were grown at 30 °C for either 24, 48 or 72 h at 225 rpms. After growth, the assay blocks were removed from the incubator/shaker and centrifuged for 20 min at 4000 rpms to pellet the microbial content. The supernatant was then poured off so that only the pellet remained. Pellets were then re-suspended in 0.5 ml buffer and placed on ice until they were used in the assay. All microbial preparations were applied within 12 h of preparation.
  • Fall armyworm (FAW) and com earworm (CEW) eggs were purchased from a commercial insectary (Benzon Research Inc., Carlisle, PA). The FAW and CEW eggs were incubated to the point that eclosion occurred within 12 hrs of the assay setup. Assays were carried out in 24-well trays containing multispecies lepidopteran diet (SOUTHLAND PRODUCTS INCORPORATED, Lake Village, AR). For most strains, whole culture bacterial suspensions were applied to the surface of the diet (diet overlay) and allowed to evaporate and soak into the diet. For some strains, bacterial cultures were centrifuged and the supernatant was removed.
  • FAW Fall armyworm
  • CEW com earworm
  • the remaining cell material was resuspended in lysate buffer and subsequently sonicated to lyse the cells.
  • the cell lysate was then applied to the surface of the diet (diet overlay) and allowed to evaporate and soak into the diet.
  • the bioassay plates were sealed with a plate sealing film vented with pin holes. The plates were incubated at 26C at 65%RH on a 16:8 daymight cycle in a Percival for 5 days.
  • the assays were assessed for level of mortality, growth inhibition and feeding inhibition. A microbe is considered active on CEW or FAW when mortality, growth inhibition and/or feeding inhibition is greater than the controls in three or more independent repetitions. Results are provided in Table 5.
  • the strains for which a lysed pellet was used are indicated in Table 5.
  • Leaf disc assay Cork bore size #8 leaf disks were excised from potato leaves, dipped into a whole culture bacterial suspension (at least two replicates per treatment) until thoroughly wet and then allowed to dry before being placed on top of a filter disk (Millipore, glass fiber filter, 13 mm). Each filter disk was pre-moistened with 60 m. 1 distilled water and the disks were placed in wells of a 24-well plate (Cellstar, 24-well, Greiner Bio One). Five second instar CPB larvae were introduced into each well using a fine tip paint brush. Plates were covered with membrane (Viewseal, Greiner Bio One), and a small hole was punctured in the membrane for each well.
  • the bacterial cultures were evaluated in an insect diet overlay bioassay in wells of 24-well plates (Cellstar, 24-well, Greiner Bio One. 60 m ⁇ volume of a 1 :6 dilution of whole culture microbial suspension (three replicates each) was inoculated on the top surface of diet and allowed to dry. Each well contained 500 m ⁇ diet (modified from Marrone et al. (1985) Journal of Economic Entomology 78:290-293). Fifteen to twenty neonate larvae were introduced in each well using a fine tip paint brush and the plate was covered with membrane (Viewseal, Greiner Bio One). The bioassay was stored at ambient temperature and scored for mortality, and/or growth/feeding inhibition at day 4.
  • Microbes were considered to have pesticidal activity if 80% mortality of the larvae and growth inhibition was observed.
  • the negative control was AFS094370, a bacterium that consistently shows negative activity against WCR
  • the positive control was AFS037424, a bacterium that consistently shows activity against WCR.
  • Table 5 The results are set forth in Table 5.
  • sucrose suspension was added to 4 wells of a 24 well plate, which was then covered with stretched parafilm that served as a feeding membrane to expose the SGSB to the diet/sample mixture.
  • the plates were incubated at 25 C:2lC, 16:8 daymight cycle at 65%RH for 5 days.
  • Treatments are applied at 16.8 Gallons/Acre with treatments applied to achieve uniform plant coverage per general treatment guidelines for ASR treatment.
  • the first treatment is applied at R1 with a follow up treatment applied at 14 days and 28 days after first treatment.
  • the specific treatments are outlined below.
  • the seed treatment formulation is made by mixing lOg formulated strain plus 30ml water plus 15ml Unicoat Polymer. The weighed out seed is placed in a sterilized mason jar. An appropriate amount of seed treatment solution based off of seed weight ( 05ml/25g seed), the mixture is shaken for 60 seconds or until the seeds were visually well coated. The seeds are placed into a single layer in a foil roasting pan and placed under a laminar flow hood for 1 hour or until seeds are dry. Once the seeds dry, they are placed in an air tight container and stored at RT.
  • One hundred grams of cell paste from each of the strains denoted in Table 2 is mixed with 5 g of glycerol and 20 g of synthetic calcium silicate using a food processor. This material is dried at 40°C to a water activity of less than 0.30. The dried powder formulation is stored in vacuum sealed mylar pouches at 22 C. The dried powder formulation retains antifungal activity.
  • the bacterial strains set forth in Table 2 are applied as seed treatments to Soybean variety W3103.
  • the bacterial strains are formulated as a wettable powder as described in Example 10 and then turned into seed treatments by combining lOg of formulated bacterial strain with 30ml water and l5ml Seed Coating Polymer (Unicoat) and then shaking until a uniform solution was made.
  • the finished solution was applied to lkg of soybean seed and allowed to dry under a laminar flow hood for 12 hours
  • Pythium inoculum was grown on millet grain and applied via in-furrow application at l .25g/ft and was applied at planting with treated soybeans seeded at 130,000 seeds per acre on day 1. Whole row stand counts were taken 17 days later. The specific treatments are outlined below. Treatments: Example 12 Rhizoctonia solani Field Trials.
  • the bacterial strains set forth in Table 2 are applied as seed treatments to Soybean variety W3103.
  • the bacterial strains are each formulated as a wettable powder as noted in Example 10 and then turned into seed treatments by combining lOg of formulated bacterial strain with 30ml water and l5ml Seed Coating Polymer (Unicoat) and then shaking until a uniform solution is made.
  • the finished solution is applied to lkg of soybean seed and allowed to dry under a laminar flow hood for 12 hours.
  • Rhizoctonia solani inoculum is grown on sorghum grain and applied via in-furrow application at l.25g/ft and is applied at planting with treated soybeans seeded at 130,000 seeds per acre on day 1. Whole row stand counts were taken 17 days later. The specific treatments are outlined below:
  • Example 13 On-plant assays for evaluation of bacterial strains for nematicidal activity.
  • a starter culture is prepared by filling 10 ml culture tubes with 3 ml of LB media and inoculating with a bacterial strain of Table 2. This starter culture is grown at 30 °C with shaking at 225 rpms for 18 h. After 18 hours 1 ml from each starter culture is passaged through 10 ml of LB media. These cultures are incubated with agitation at 30 °C with shaking at 225 rpms for 5-6 hrs. Assay cultures are prepared by filling 1 liter flasks with 350ml of media. 1.5 ml of the 10 ml culture is added to the assay culture flask. Flasks are grown at 30 C for either 24, 48, or 27 h at 175 rpms.
  • the flasks are removed from the incubator/shaker and culture poured into 500mL centrifuge tubes and centrifuged for 15 min at 8000 rpms to pellet the microbial content.
  • Cucumber plants are assessed for infectivity with Root-Knot Nematodes in the presence of a bacterial strain of interest.
  • Cucumber seeds are planted in a 4 inch plastic pot with 90% sand and 10% soil. Following germinating, plants are allowed to grow for one week.
  • the cucumber plants received a drench application with 10X microbial concentration or 3X microbial concentration, prepared as described above.
  • some cucumber plants are treated with a lOx or 3x negative microbe control (using Escherichia coli, strain BL21) or with the nematicide Velum ® as a positive control.
  • a lOx or 3x negative microbe control using Escherichia coli, strain BL21
  • the nematicide Velum ® as a positive control.

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Abstract

L'invention concerne des compositions et des procédés de lutte contre des organismes nuisibles de plantes et/ou d'amélioration d'au moins un caractère agronomique d'intérêt dans une plante. De telles compositions et procédés comprennent une souche bactérienne qui peut être utilisée comme inoculant pour des plantes. Par conséquent, l'invention concerne des procédés de culture d'une plante sensible à un organisme nuisible de plante et/ou à une maladie provoquée par un organisme nuisible de plante et des procédés pour lutter contre des organismes nuisibles de plante et/ou une maladie de plante sur une plante sensible à l'organisme nuisible de plante et/ou à la maladie de plante.
PCT/US2019/040107 2018-06-29 2019-07-01 Compositions comprenant des bactéries et procédés de lutte contre des organismes nuisibles de plantes et d'amélioration de la santé des plantes WO2020006555A1 (fr)

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WO2022225926A1 (fr) 2021-04-19 2022-10-27 AgBiome, Inc. Compositions et procédés d'amélioration de la santé de plantes et de lutte contre les maladies de plantes
WO2022245786A1 (fr) 2021-05-18 2022-11-24 AgBiome, Inc. Compositions et méthodes pour améliorer la santé des plantes et lutter contre des maladies de plantes
WO2024026305A1 (fr) 2022-07-26 2024-02-01 AgBiome, Inc. Compositions et procédés basés sur pseudomonas chlororaphis pour améliorer la santé des plantes et lutter contre les maladies des plantes

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WO2022225925A1 (fr) 2021-04-19 2022-10-27 AgBiome, Inc. Compositions et méthodes pour améliorer la santé de plantes et lutter contre des maladies des plantes
WO2022245786A1 (fr) 2021-05-18 2022-11-24 AgBiome, Inc. Compositions et méthodes pour améliorer la santé des plantes et lutter contre des maladies de plantes
WO2024026305A1 (fr) 2022-07-26 2024-02-01 AgBiome, Inc. Compositions et procédés basés sur pseudomonas chlororaphis pour améliorer la santé des plantes et lutter contre les maladies des plantes

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