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WO2020006555A1 - Compositions comprising bacteria and methods for controlling plant pests and improving plant health - Google Patents

Compositions comprising bacteria and methods for controlling plant pests and improving plant health Download PDF

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Publication number
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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WIPO (PCT)
Prior art keywords
plant
cfu
active variant
bacterial strain
pest
Prior art date
Application number
PCT/US2019/040107
Other languages
French (fr)
Inventor
Narendra PALEKAR
Chad Alton KEYSER
Original Assignee
AgBiome, Inc.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
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Application filed by AgBiome, Inc. filed Critical AgBiome, Inc.
Publication of WO2020006555A1 publication Critical patent/WO2020006555A1/en

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Classifications

    • 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

Compositions and methods for controlling plant pests and/or improving at least one agronomic trait of interest in a plant are provided. Such compositions and methods comprise a bacterial strain that can be used as an inoculant for plants. Therefore, methods for growing a plant susceptible to a plant pest and/or plant disease caused by a plant pest and methods for controlling plant pests and/or plant disease on a plant susceptible to the plant pest and/or plant disease are also provided.

Description

COMPOSITIONS COMPRISING BACTERIA AND METHODS FOR CONTROLLING PLANT PESTS AND IMPROVING PLANT HEALTH
FIELD OF THE INVENTION
The invention relates to bacterial strains and populations for controlling plant pests and/or improving an agronomic trait of interest in a plant.
BACKGROUND
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.
SUMMARY
Compositions and methods for controlling plant pests and/or for improving at least one agronomic trait of interest in a plant are provided. Such compositions and methods 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. Also provided herein are 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. Further provided are methods and compositions for making a modified bacterial strain having resistance to a biocide of interest.
DETAILED DESCRIPTION
I. Overview
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.
II. Bacterial Strains
Various 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 brassicaceanm strain), 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 oryzihabitans strain), AIP088026 (an
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,
AIP081435, AIP082140, AIP000817, AIP060333, AIP070494, AIP015104, AIP011586,
AIP010884, AIP082287, AIP088026, AIP065969, AIP018514, AIP033041, AIP092281,
AIP081114, AIP046403, AIP022635, AIP070925, and AIP039063 are provided, as well as, populations of spores derived from each of these strains, or any preparation thereof.
Thus, 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,
AIP033041, AIP092281, AIP081114, AIP046403, AIP022635, AIP070925, and AIP039063, or an active variant of any thereof.
AIP031898 was deposited with the Patent Depository of the National Center for
Agricultural Utilization Research Agricultural Research Service, U S. Department of Agriculture, 1815 North University Street, Peoria, Illinois 61604 U.S.A. on June 4, 2019 and assigned NRRL No.B-67799.
AIP023234 was deposited with the Patent Depository of the National Center for
Agricultural Utilization Research Agricultural Research Service, U S. Department of Agriculture, 1815 North University Street, Peoria, Illinois 61604 U.S.A. on June 4, 2019 and assigned NRRL No.B-67784.
AIP024552 was deposited with the Patent Depository of the National Center for
Agricultural Utilization Research Agricultural Research Service, U S. Department of Agriculture, 1815 North University Street, Peoria, Illinois 61604 U.S.A. on June 4, 2019 and assigned NRRL No.B-67780.
AIP035573 was deposited with the Patent Depository of the National Center for
Agricultural Utilization Research Agricultural Research Service, U S. Department of Agriculture, 1815 North University Street, Peoria, Illinois 61604 U.S.A. on June 14, 2019 and assigned NRRL No. _ .
AIP071234 was deposited with the Patent Depository of the National Center for
Agricultural Utilization Research Agricultural Research Service, U S. Department of Agriculture, 1815 North University Street, Peoria, Illinois 61604 U.S.A. on June 4, 2019 and assigned NRRL No.B-67793.
AIP080021 was deposited with the Patent Depository of the National Center for
Agricultural Utilization Research Agricultural Research Service, U S. Department of Agriculture, 1815 North University Street, Peoria, Illinois 61604 U.S.A. on June 4, 2019 and assigned NRRL No.B-67803.
AIP001237 was deposited with the Patent Depository of the National Center for
Agricultural Utilization Research Agricultural Research Service, U S. Department of Agriculture, 1815 North University Street, Peoria, Illinois 61604 U.S.A. on June 4, 2019 and assigned NRRL No.B-67794.
AIP050674 was deposited with the Patent Depository of the National Center for
Agricultural Utilization Research Agricultural Research Service, U S. Department of Agriculture, 1815 North University Street, Peoria, Illinois 61604 U.S.A. on June 4, 2019 and assigned NRRL No.B-67795.
AIP071546 was deposited with the Patent Depository of the National Center for
Agricultural Utilization Research Agricultural Research Service, U S. Department of Agriculture, 1815 North University Street, Peoria, Illinois 61604 U.S.A. on June 4, 2019 and assigned NRRL No.B-67796.
AIP049805 was deposited with the Patent Depository of the National Center for
Agricultural Utilization Research Agricultural Research Service, U S. Department of Agriculture, 1815 North University Street, Peoria, Illinois 61604 U.S.A. on June 14, 2019 and assigned NRRL No. AIPO 16229 was deposited with the Patent Depository of the National Center for
Agricultural Utilization Research Agricultural Research Service, U S. Department of Agriculture, 1815 North University Street, Peoria, Illinois 61604 U.S.A. on June 4, 2019 and assigned NRRL No.B-67798.
AIP081435 was deposited with the Patent Depository of the National Center for
Agricultural Utilization Research Agricultural Research Service, U S. Department of Agriculture, 1815 North University Street, Peoria, Illinois 61604 U.S.A. on June 4, 2019 and assigned NRRL No.B-67805.
AIP082140 was deposited with the Patent Depository of the National Center for
Agricultural Utilization Research Agricultural Research Service, U S. Department of Agriculture, 1815 North University Street, Peoria, Illinois 61604 U.S.A. on June 4, 2019 and assigned NRRL No.B-67779.
AIP000817 was deposited with the Patent Depository of the National Center for
Agricultural Utilization Research Agricultural Research Service, U S. Department of Agriculture, 1815 North University Street, Peoria, Illinois 61604 U.S.A. on June 4, 2019 and assigned NRRL No.B-67789.
AIP060333 was deposited with the Patent Depository of the National Center for
Agricultural Utilization Research Agricultural Research Service, U S. Department of Agriculture, 1815 North University Street, Peoria, Illinois 61604 U.S.A. on June 4, 2019 and assigned NRRL No.B-67801.
AIP070494 was deposited with the Patent Depository of the National Center for
Agricultural Utilization Research Agricultural Research Service, U S. Department of Agriculture, 1815 North University Street, Peoria, Illinois 61604 U.S.A. on June 4, 2019 and assigned NRRL No.B-67790.
AIPO 15104 was deposited with the Patent Depository of the National Center for
Agricultural Utilization Research Agricultural Research Service, U S. Department of Agriculture, 1815 North University Street, Peoria, Illinois 61604 U.S.A. on June 4, 2019 and assigned NRRL No.B-67802.
AIPO 11586 was deposited with the Patent Depository of the National Center for
Agricultural Utilization Research Agricultural Research Service, U S. Department of Agriculture, 1815 North University Street, Peoria, Illinois 61604 U.S.A. on June 4, 2019 and assigned NRRL No.B-67778.
AIPO 10884 was deposited with the Patent Depository of the National Center for
Agricultural Utilization Research Agricultural Research Service, U S. Department of Agriculture, 1815 North University Street, Peoria, Illinois 61604 U.S.A. on June 4, 2019 and assigned NRRL No.B-67777.
AIP082287 was deposited with the Patent Depository of the National Center for
Agricultural Utilization Research Agricultural Research Service, U S. Department of Agriculture, 1815 North University Street, Peoria, Illinois 61604 U.S.A. on June 4, 2019 and assigned NRRL No.B-67804.
AIP088026 was deposited with the Patent Depository of the National Center for
Agricultural Utilization Research Agricultural Research Service, U S. Department of Agriculture, 1815 North University Street, Peoria, Illinois 61604 U.S.A. on June 4, 2019 and assigned NRRL No.B-67791.
AIP065969 was deposited with the Patent Depository of the National Center for
Agricultural Utilization Research Agricultural Research Service, U S. Department of Agriculture, 1815 North University Street, Peoria, Illinois 61604 U.S.A. on June 4, 2019 and assigned NRRL No.B-67785.
AIP018514 was deposited with the Patent Depository of the National Center for
Agricultural Utilization Research Agricultural Research Service, U S. Department of Agriculture, 1815 North University Street, Peoria, Illinois 61604 U.S.A. on June 4, 2019 and assigned NRRL No.B-67782.
AIP033041 was deposited with the Patent Depository of the National Center for
Agricultural Utilization Research Agricultural Research Service, U S. Department of Agriculture, 1815 North University Street, Peoria, Illinois 61604 U.S.A. on June 4, 2019 and assigned NRRL No.B-67806.
AIP092281 was deposited with the Patent Depository of the National Center for
Agricultural Utilization Research Agricultural Research Service, U S. Department of Agriculture, 1815 North University Street, Peoria, Illinois 61604 U.S.A. on June 4, 2019 and assigned NRRL No.B-67788.
AIP081114 was deposited with the Patent Depository of the National Center for
Agricultural Utilization Research Agricultural Research Service, U S. Department of Agriculture, 1815 North University Street, Peoria, Illinois 61604 U.S.A. on June 4, 2019 and assigned NRRL No.B-67786.
AIP046403 was deposited with the Patent Depository of the National Center for
Agricultural Utilization Research Agricultural Research Service, U S. Department of Agriculture, 1815 North University Street, Peoria, Illinois 61604 U.S.A. on June 4, 2019 and assigned NRRL No.B-67787. AIP022635 was deposited with the Patent Depository of the National Center for
Agricultural Utilization Research Agricultural Research Service, U S. Department of Agriculture, 1815 North University Street, Peoria, Illinois 61604 U.S.A. on June 4, 2019 and assigned NRRL No.B-67792.
AIP070925 was deposited with the Patent Depository of the National Center for
Agricultural Utilization Research Agricultural Research Service, U S. Department of Agriculture, 1815 North University Street, Peoria, Illinois 61604 U.S.A. on June 4, 2019 and assigned NRRL No.B-67800.
AIP039063 was deposited with the Patent Depository of the National Center for
Agricultural Utilization Research Agricultural Research Service, U S. Department of Agriculture, 1815 North University Street, Peoria, Illinois 61604 U.S.A. on June 4, 2019 and assigned NRRL No.B-67783.
Each of the deposits identified above will be maintained under the terms of the Budapest Treaty on the International Recognition of the Deposit of Microorganisms for the Purposes of Patent Procedure. Each deposit was made merely as a convenience for those of skill in the art and is not an admission that a deposit is required under 35 U.S.C. §112.
The term "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.
As used herein, 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. In some embodiments, 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. In specific embodiments, a culture of bacteria contains no other bacterial species in quantities to be detected by normal bacteriological techniques.
By "population" is intended a group or collection that comprises two or more individuals (i.e., 10, 100, 1,000, 10,000, lxlO6, lxlO7, or lxlO8 or greater) of a given bacterial strain. Various compositions are provided herein that comprise a population of at least one bacterial strain or a mixed population of individuals from more than one bacterial strain. In specific embodiments, the population of at least one of a bacterial strain (i.e., cells of AIP031898, AIP023234, AIP024552, AIP035573, AIP071234, AIP080021, AIP001237, AIP050674, AIP071546, AIP049805,
AIR016229, AIP081435, AIP082140, AIP000817, AIP060333, AIP070494, AIP015104,
AIP011586, AIR010884, AIP082287, AIP088026, AIP065969, AIP018514, AIP033041,
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,
AIP082140, AIP000817, AIP060333, AIP070494, AIP015104, AIP011586, AIP010884,
AIP082287, AIP088026, AIP065969, AIP018514, AIP033041, AIP092281, AIP081114,
AIP046403, AIP022635, AIP070925, and AIP039063, or an active variant of any thereof) comprises a concentration of at least about 105 CFU/ml to about 10u CFU/ml, about 105 CFU/ml to about 1010 CFU/ml, about 105 CFU/ml to about 1012 CFU/ml, about 105 CFU/ml to about 106 CFU/ml, about 106 CFU/ml to about 107 CFU/ml, about 107 CFU/ml to about 108 CFU/ml, about 108 CFU/ml to about 109 CFU/ml, about 109 CFU/ml to about 1010 CFU/ml, about 1010 CFU/ml to about 1011 CFU/ml, about 1011 CFU/ml to about 1012 CFU/ml. In other embodiments, the concentration of the bacterial strain provided herein or active variant thereof comprises at least about 105 CFU/ml, at least about 106 CFU/ml, at least about 107 CFU/ml, at least about 108 CFU/ml, at least about 109 CFU/ml, at least about 1010 CFU/ml, at least about 1011 CFU/ml, or at least about 1012 CFU/ml.
A“spore” refers to at least one dormant (at application) but viable reproductive unit of a bacterial species. Non-limiting methods by which spores are formed from each of AIP031898, AIP023234, AIP024552, AIP035573, AIP071234, AIP080021, AIP001237, AIP050674,
AIP071546, AIP049805, AIP016229, AIP081435, AIP082140, AIP000817, AIP060333,
AIP070494, AIP015104, AIP011586, AIP010884, AIP082287, AIP088026, AIP065969,
AIP018514, AIP033041, AIP092281, AIP081114, AIP046403, AIP022635, AIP070925, and AIP039063 (or variants of any thereof) 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.
The compositions comprising a cell of a bacterial strain (i.e., at least one of AGR031898, AIP023234, AIP024552, AIP035573, AIP071234, AIP080021, AIP001237, AIP050674,
AIP071546, AIP049805, AIP016229, AIP081435, AIP082140, AIP000817, AIP060333,
AIP070494, AIP015104, AIP011586, AIP010884, AIP082287, AIP088026, AIP065969,
AIP018514, AIP033041, AIP092281, AIP081114, AIP046403, AIP022635, 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 AGR031898, AIP023234, AIP024552, AIP035573, AIP071234, AIP080021, AIP001237, AIP050674, AIP071546, AIP049805, AIP016229,
AIP081435, AIP082140, AIP000817, AIP060333, AIP070494, AIP015104, AIP011586,
AIP010884, AIP082287, AIP088026, AIP065969, AIP018514, AIP033041, AIP092281,
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.
In one embodiment, carriers include polar or non-polar liquid carriers such as water, mineral oils and vegetable oils. Additional carriers are disclosed elsewhere herein.
A. Active Variants of a Bacterial Strain
Further provided are active variants of AIP031898, AIP023234, AIP024552, AIP035573, AIP071234, AIP080021, AIP001237, AIP050674, AIP071546, AIP049805, AIP016229,
AIP081435, AIP082140, AIP000817, AIP060333, AIP070494, AIP015104, AIP011586,
AIP010884, AIP082287, AIP088026, AIP065969, AIP018514, AIP033041, AIP092281,
ATP081 1 14, AIP046403, AGR022635, AIP070925, and ATP039063 Such variants will retain the ability to control one or more plant pests or improve one or more agronomic traits of interest in a plant. Thus, in some embodiments, the active variants of the bacterial strains provided herein will retain pesticidal activity against a plant pest. As used herein,“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. In some embodiments, variants will retain the ability to control one or more insect pests or nematode pests. In particular embodiments, 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.
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,
AIP000817, AIP060333, AIP070494, AIP015104, AIP011586, AIP010884, AIP082287,
AIP088026, AIP065969, AIP018514, AIP033041, AIP092281, AIP081114, AIP046403,
AIP022635, AIP070925, and AIP039063.
In specific embodiments, 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. In instances where the bacterial strain is not compatible with a biocide of interest, if desired, 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.
By“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. In some cases the modified bacterial strain comprises any one of AIP031898, AIP023234, AIP024552,
AIP035573, AIP071234, AIP080021, AIP001237, AIP050674, AIP071546, AIP049805,
AIPO 16229, AIP081435, AIP082140, AIP000817, AIP060333, AIP070494, AIP015104,
AIP011586, AIPO 10884, AIP082287, AIP088026, AIP065969, AIP018514, AIP033041,
AIP092281, AIP081114, AIP046403, AIP022635, AIP070925, and AIP039063, or an active variant of any thereof. In specific embodiments, 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 (e.g., a herbicide, insecticide, fungicide, pesticide, or other crop protection chemical resistance) 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. In specific embodiments, the increase in resistance to a biocide is demonstrated in the presence of an agriculturally effective amount of the biocide.
In such instances, the modified bacterial strain having resistance to one or more biocides is useful for enhancing the competitiveness of bacterial strains particularly over other microbial agents which are not resistant to herbicides, insecticides, fungicides, pesticides, or other crop protection chemicals. Therefore, 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.
Thus, 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. (1983) Studies on transformation of Escherichia coli with plasmids J Mol. Biol. 166, 557-77; Seidman, C.E. (1994) In: Current Protocols in Molecular Biology, Ausubel, F.M. et al. eds., John Wiley and Sons, NY; Choi et al. (2006) J. Microbiol. Methods 64:391-397; Wang et al. 2010. J. Chem. Technol.
Biotechnol. 85:775-778. Transformation may occur by natural uptake of naked DNA by competent cells from their environment in the laboratory. Alternatively, 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. 6,758,044; 7,250,561; 7,935,869; and 8, 124,846), or other such genes known in the art. The disclosures of WO 96/38576, U.S. Patent No. 5,792,931, U.S. Patent No. 6,758,044; U.S. Patent No. 7,250,561; U.S. Patent No. 7,935,869; and U.S. Patent No. 8,124,846 are herein incorporated by reference. The bar gene encodes resistance to the herbicide basta, the nptll gene encodes resistance to the antibiotics kanamycin and geneticin, and the ALS-gene mutants encode resistance to the sulfonylurea herbicides including chlorsulfuron, metsulfuron, sulfometuron, nicosulfuron, rimsulfuron, flazasulfuron, sulfosulfuron, and triasulfuron, and the imadizolinone herbicides including imazethapyr, imazaquin, imazapyr, and imazamethabenz.
To identify and produce a modified population of bacterial strains through selection, 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.
Further active variants of the various bacteria provided herein can be identified employing, for example, methods that determine the sequence identity relatedness between the 16S ribosomal RNA, methods to identify groups of derived and functionally identical or nearly identical strains include Multi-locus sequence typing (MLST), concatenated shared genes trees, Whole Genome Alignment (WGA), Average Nucleotide Identity, and MinHash (Mash) distance metric.
In one aspect, the active variants of the bacterial strain AIP031898, AGR023234,
AIP024552, AIP035573, AIP071234, AIP080021, AIP001237, AIP050674, AIP071546,
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
7(1)1741-7007-7-3. Thus, in specific embodiments, 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).
In another aspect, 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 (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) and 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. Thus, in specific embodiments, an active variant of bacterial strain
AIP031898, AIP023234, AIP024552, AIP035573, AIP071234, AIP080021, AIP001237,
AIP050674, AIP071546, AIP049805, AIP016229, AIP081435, AIP082140, AIP000817,
AIP060333, AIP070494, AIP015104, AIP011586, AIP010884, AIP082287, AIP088026,
AIP065969, AIP018514, AIP033041, AIP092281, AIP081114, AIP046403, AIP022635,
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).
In another aspect, 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,
AIP060333, AIP070494, AIP015104, AIP011586, AIP010884, AIP082287, AIP088026,
AIP065969, AIP018514, AIP033041, AIP092281, AIP081114, AIP046403, AIP022635,
AIP070925, and AIP039063) on the basis of 16S rDNA sequence identity. See Stackebrandt E, el al,‘"Report of the ad hoc committee for the re-evaluation of the species definition in
bacteriology,” Int J Syst Evol Microbiol. 52(3): 1043-7 (2002) regarding use of 16S rDNA sequence identity for determining relatedness in bacteria. In an embodiment, 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).
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.
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,
AIP060333, AIP070494, AIP015104, AIP011586, AIP010884, AIP082287, AIP088026,
AIP065969, AIP018514, AIP033041, AIP092281, AIP081114, AIP046403, AIP022635,
AIP070925, and AIP039063 on the basis of the Minhash (Mash) distance between complete genome DNA sequences. Thus, in specific embodiments, 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. In other embodiments, 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. In other embodiments, 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.
As used herein,“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,
Dupont et al. (2012) ISMKJ 6: 1625- 1628, and the CheckM framework (Parks et al. (2015) Genome Research 25: 1043-1055); each of these references is herein incorporated in their entirety.
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.
Exemplary steps to obtain a distance estimate between the genomes in question are as follows: (1) 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. (2) 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. (3) Confirm that the Mash distance between the two genomes is less than 0.005, 0.010, 0.015, 0.020, 0.025, or 0.030. 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).
III. Formulations
The bacterial strains provided herein (i.e., cells of AIP031898, AIP023234, AGR024552, AIP035573, AIP071234, AIP080021, AIP001237, AIP050674, AIP071546, AIP049805,
AIR016229, AIP081435, AIP082140, AIP000817, AIP060333, AIP070494, AIP015104,
AIP011586, AIR010884, AIP082287, AIP088026, AIP065969, AIP018514, AIP033041,
AIP092281, AIP081114, AIP046403, AIP022635, AIP070925, and AIP039063, or active variants of any thereof, or a spore or a forespore or a combination of cells, forespores and/or spores, from any 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, AIP033041, AIP092281, AIP081114, AIP046403, AIP022635,
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. In some embodiments, 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. In particular embodiments, 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. An adjuvant in this context is a component which enhances the biological effect of the formulation, without the component itself having a biological effect. Examples of 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). If the extender used is water, it is also possible to employ, for example, organic solvents as auxiliary solvents. Essentially, non-limiting liquid 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.
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. 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. Non-limiting examples are those 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. Examples of 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.
Further 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. 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.
Furthermore, the 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. Further possible auxiliaries include mineral and vegetable oils.
There may possibly be further auxiliaries present in the formulations and the application forms derived from them. Examples of such additives include fragrances, protective colloids, binders, adhesives, thickeners, thixotropic substances, penetrants, retention promoters, stabilizers, sequestrants, complexing agents, humectants and spreaders. Generally speaking, 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.
The various 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,
AIP082140, AIP000817, AIP060333, AIP070494, AIP015104, AIP011586, AIP010884,
AIP082287, AIP088026, AIP065969, AIP018514, AIP033041, AIP092281, AIP081114,
AIP046403, AIP022635, AIP070925, and AIP039063, or 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, AIP024552, AIP035573, AIP071234, AIP080021, AIP001237, AIP050674, AIP071546, AIP049805, AIP016229, AIP081435, AIP082140, AIP000817, AIP060333, AIP070494, AIP015104, AIP011586, AIP010884, AIP082287, AIP088026, AIP065969, AIP018514, AIP033041, AIP092281, AIP081114, AIP046403, AIP022635, AIP070925, and AIP039063, or an active variant of any thereof. Such an amount can comprise a concentration of the bacterial strain of at least about 104 to about 1011, at least about 105 CFU/gram to about 1011 CFU/gram, about 105 CFU/gram to about 1010 CFU/gram, about 105 CFU/gram to about 1012 CFU/gram, about 105 CFU/gram to about 106 CFU/gram, about 106 CFU/gram to about 107 CFU/gram, about 107 CFU/gram to about 108 CFU/gram, about 108 CFU/gram to about 109 CFU/gram, about 109 CFU/gram to about 1010 CFU/gram, about 1010 CFU/gram to about 1011 CFU/gram, or about 10u CFU/gram to about 1012 CFU/gram. In other embodiments, the concentration of the bacterial strain comprises at least about 104 CFU/gram, at least about 105 CFU/gram, at least about 106 CFU/gram, at least about 107 CFU/gram, at least about 108
CFU/gram, at least about 109 CFU/gram, at least about 1010 CFU/gram, at least about 1011 CFU/gram, at least about 1012 CFU/gram. Such 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.
In some embodiments, 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,
AIP023234, AIP024552, AIP035573, AIP071234, AIP080021, AIP001237, AIP050674, AIP071546, AIP049805, AIP016229, AIP081435, AIP082140, AIP000817, AIP060333, AIP070494, AIP015104, AIP011586, AIP010884, AIP082287, AIP088026, AIP065969, AIP018514, AIP033041, AIP092281, AIP081114, AIP046403, AIP022635, AIP070925, and AIP039063, or 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, AIP024552, AIP035573, AIP071234, AIP080021, AIP001237, AIP050674, AIP071546, AIP049805, AIP016229, AIP081435, AIP082140, AIP000817, AIP060333, AIP070494, AIP015104, AIP011586, AIP010884, AIP082287, AIP088026, AIP065969, AIP018514, AIP033041, AIP092281,
AIP081114, AIP046403, AIP022635, AIP070925, and AIP039063, or an active variant of any thereof. In liquid forumations, the amount of bacterial strain, or active variant thereof, disclosed herein can comprise a concentration of at least about 104 to about 1011 CFU/mL, at least about 105 CFU/mL to about 1011 CFU/ mL, about 105 CFU/ mL to about 1010 CFU/ mL, about 105 CFU/ mL to about 1012 CFU/ mL, about 105 CFU/ mL to about 106 CFU/ mL, about 106 CFU/ mL to about 107 CFU/ mL, about 107 CFU/ mL to about 108 CFU/ mL, about 108 CFU/ mL to about 109 CFU/ mL, about 109 CFU/ mL to about 1010 CFU/ mL, about 1010 CFU/ mL to about 1011 CFU/ mL, or about 1011 CFU/ mL to about 1012 CFU/ mL or at least about 104 CFU/ mL, at least about 105 CFU/ mL, at least about 106 CFU/ mL, at least about 107 CFU/ mL, at least about 108 CFU/ mL, at least about 109 CFU/ mL, at least about 1010 CFU/ mL, at least about 1011 CFU/ mL, at least about 1012 CFU/ mL.
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,
AIP081435, AIP082140, AIP000817, AIP060333, AIP070494, AIP015104, AIP011586,
AIP010884, AIP082287, AIP088026, AIP065969, AIP018514, AIP033041, AIP092281,
AIP081114, AIP046403, AIP022635, AIP070925, and AIP039063, or 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, AIP024552, AIP035573, AIP071234, AIP080021, AIP001237,
AIP050674, AIP071546, AIP049805, AIP016229, AIP081435, AIP082140, AIP000817,
AIP060333, AIP070494, AIP015104, AIP011586, AIP010884, AIP082287, AIP088026,
AIP065969, AIP018514, AIP033041, AIP092281, AIP081114, AIP046403, AIP022635,
AIP070925, and AIP039063, or an active variant of any thereof. The amount of the bacterial strain in the dry formulation (e.g., cell pastes, wettable powders, and/or spray dried formulations) can comprise a concentration of the bacterial strain of at least about 105 CFU/gram to about 1011 CFU/gram, about 107 CFU/gram to about 1010 CFU/gram, about 107 CFU/gram to about 1011 CFU/gram, about 106 CFU/gram to about 1010 CFU/gram, about 106 CFU/gram to about 1011 CFU/gram, about 1011 CFU/gram to about 1012 CFU/gram, about 105 CFU/gram to about 1010 CFU/gram, about 105 CFU/gram to about 1012 CFU/gram, about 105 CFU/gram to about 106 CFU/gram, about 106 CFU/gram to about 107 CFU/gram, about 107 CFU/gram to about 108 CFU/gram, about 108 CFU/gram to about 109 CFU/gram, about 109 CFU/gram to about 1010 CFU/gram, about 1010 CFU/gram to about 1011 CFU/gram, or about 1011 CFU/gram to about 1012 CFU/gram. In some embodiments, the concentration of the bacterial strain comprises at least about 105 CFU/gram, at least about 106 CFU/gram, at least about 107 CFU/gram, at least about 108 CFU/gram, at least about 109 CFU/gram, at least about 1010 CFU/gram, at least about 1011
CFU/gram, at least about 1012 CFU/gram, or at least about 1013 CFU/gram.
As used herein, a“cell paste” comprises a population of cells that has been centrifuged and/or filtered or otherwise concentrated.
Further provided is 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,
AIP033041, AIP092281, AIP081114, AIP046403, AIP022635, 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, AIP024552, AIP035573, AIP071234,
AIP080021, AIP001237, AIP050674, AIP071546, AIP049805, AIP016229, AIP081435,
AIP082140, AIP000817, AIP060333, AIP070494, AIP015104, AIP011586, AIP010884,
AIP082287, AIP088026, AIP065969, AIP018514, AIP033041, AIP092281, AIP081114,
AIP046403, AIP022635, AIP070925, and AIP039063, or an active variant of any thereof, wherein said bacterial strain or the active variant thereof is present on the seed at about 105 CFU/seed to about 107 CFU/seed, at about 104 CFU/seed to about 108 CFU/seed, at about 104 CFU/seed to about 105 CFU/seed, at about 105 CFU/seed to about 106 CFU/seed, at about 106 CFU/seed to about 107 CFU/seed, or at about 107 CFU/seed to about 108 CFU/seed. Various plants of interest are disclosed elsewhere herein.
In particular embodiments, 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,
AIP050674, AIP071546, AIP049805, AIP016229, AIP081435, AIP082140, AIP000817,
AIP060333, AIP070494, AIP015104, AIP011586, AIP010884, AIP082287, AIP088026,
AIP065969, AIP018514, AIP033041, AIP092281, AIP081114, AIP046403, AIP022635,
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,
AIP024552, AIP035573, AIP071234, AIP080021, AIP001237, AIP050674, AIP071546,
AIP049805, AIP016229, AIP081435, AIP082140, AIP000817, AIP060333, AIP070494,
AIP015104, AIP011586, AIP010884, AIP082287, AIP088026, AIP065969, AIP018514,
AIP033041, AIP092281, AIP081114, AIP046403, AIP022635, AIP070925, and AIP039063, or an active variant of any thereof, wherein said bacterial strain or the active variant thereof is present on the seed at about 105 CFU/seed to about 107 CFU/seed, at about 104 CFU/seed to about 108 CFU/seed, at about 104 CFU/seed to about 105 CFU/seed, at about 105 CFU/seed to about 106 CFU/seed, at about 106 CFU/seed to about 107 CFU/seed, or at about 107 CFU/seed to about 108 CFU/seed. As used herein,“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. In particular embodiments,“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. For example, 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.
The various formulations disclosed herein can be stable for at least 30, 40, 50, 60, 70, 80,
90, 100, 125, 150, 200, 225, 250, 275, 300, 325, 350 days, 1 .5 years, 2 years or longer. By stable is intended that the formulation retains viable bacteria and/or retains an effective amount of a biologically active bacterial population. Biological activity as used herein refers to the ability of the formulation to improve an agronomic trait of interest or control a plant pest. In one
embodiment, 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. In another embodiment, 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. In another embodiment, 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. In still another embodiment, 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. In particular embodiemnts,“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. For example, 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.
The various formulations disclosed herein can be stable for at least 30, 40, 50, 60, 70, 80,
90, 100, 125, 150, 200, 225, 250, 275, 300, 325, 350 days, 1.5 years, 2 years or longer. By stable is intended that the formulation retains viable bacteria and/or retains an effective amount of a biologically active bacterial population. Biological activity as used herein refers to the ability of the formulation to improve an agronomic trait of interest or control a plant pest. In one
embodiment, 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. In another embodiment, 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. In another embodiment, 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. In still another embodiment, 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.
Moreover, 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. In some cases, 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. In other embodiments, the biocontrol agents described herein can be mixed with other biocontrol agents.
In specific embodiments, the bacterial strain or active variant thereof is compatible with agricultural chemicals used to improve performance of biocides. Such 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. In instances where the bacterial strain or active variant is not compatible with an agricultural chemical of interest, if desired, 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. In one embodiment, 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; [Lactic acid, n-butyl ester, (S)]; Butyl lactate [Lactic acid, n-butyl ester]; Butyl stearate [Octadecanoic acid, butyl ester]; Calcareous shale; Calcite (Ca(Co3)); Calcium acetate; Calcium acetate monohydrate [Acetic acid, calcium salt,
monohydrate]; Calcium benzoate [Benzoic acid, calcium salt]; Calcium carbonate; Calcium citrate [Citric acid, calcium salt]; Calcium octanoate; Calcium oxide silicate (Ca30(Si04)); Calcium silicate [Silicic acid, calcium salt]; Calcium stearate [Octadecanoic acid, calcium salt]; Calcium sulfate; Calcium sulfate dehydrate; Calcium sulfate hemihydrate; Canary seed; Carbon; Carbon dioxide; Carboxymethyl cellulose [Cellulose, carboxymethyl ether]; Cardboard; Carnauba wax; Carob gum [Locust bean gum]; Carrageenan; Caseins; Castor oil; Castor oil, hydrogenated; Cat food; Cellulose; Cellulose acetate; Cellulose, mixture with cellulose carboxymethyl ether, sodium salt; Cellulose, pulp; Cellulose, regenerated; Cheese; Chlorophyll a; Chlorophyll b; Citrus meal; Citric acid; Citric acid, monohydrate; Citrus pectin; Citrus pulp; Clam shells; Cocoa; Cocoa shell flour; Cocoa shells; Cod-liver oil; Coffee grounds; Cookies; Cork; Corn cobs; Cotton; Cottonseed meal; Cracked wheat; Decanoic acid, monoester with l,2,3-propanetriol; Dextrins; Diglyceryl monooleate [9-Octadecenoic acid, ester with l,2,3-propanetriol]; Diglyceryl monostearate [9- Octadecanoic acid, monoester with xybis(propanediol)]; Dilaurin [Dodecanoic acid, diester with l,2,3-propanetriol]; Dipalmitin [Hexadecanoic acid, diester with l,2,3-propanetriol]; Dipotassium citrate [Citric acid, dipotassium salt]; Disodium citrate [Citric acid, disodium salt]; Disodium sulfate decahydrate ; Diatomaceous earth (less than 1% crystalline silica); Dodecanoic acid, monoester with l,2,3-propanetriol; Dolomite; Douglas fir bark; Egg shells; Eggs; (+)-Ethyl lactate [Lactic acid, ethyl ester, (S)]; Ethyl lactate [Lactic acid, ethyl ester]; Feldspar; Fish meal; Fish oil (not conforming to 40 CFR 180.950) ; Fuller's earth; Fumaric acid; gamma-Cyclodextrin; Gelatins; Gellan gum; Glue (as depolymd. animal collagen); 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, ester with 1,2,3-propanetriol]; Granite; Graphite; Guar gum; Gum Arabic; Gum tragacanth; Gypsum; Hematite (Fe203); Humic acid; Hydrogenated cottonseed oil;
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 (Fe2Mg04);
Iron oxide (Fe20 ); Iron oxide (Fe20 ); Iron oxide (Fe304); Iron oxide (FeO); Isopropyl alcohol [2- Propanol]; Isopropyl myristate; Kaolin; Lactose; Lactose monohydrate; Lanolin; Latex rubber; Laurie acid; Lecithins; Licorice extract; Lime (chemical) dolomitic; Limestone; Linseed oil;
Magnesium carbonate [Carbonic acid, magnesium salt (1 : 1); Magnesium benzoate; Magnesium oxide; Magnesium oxide silicate (Mg30(Si205)2), monohydrate; Magnesium silicate; Magnesium silicate hydrate; Magnesium silicon oxide (Mg2Si308); 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-dihydroxypropyl ester]; Monomyristin [Decanoic acid, diester with 1,2,3-propanetriol]; Monopalmitin [Hexadecanoic acid, monoester with 1,2,3- propanetriol]; Monopotassium citrate [Citric acid, monopotassium salt; Monosodium citrate [Citric acid, monosodium salt]; Montmorillonite; Myristic acid; Nepheline syenite; Nitrogen; Nutria meat; Nylon; Octanoic acid, potassium salt; Octanoic acid, sodium salt; Oils, almond; Oils, wheat; Oleic acid; Oyster shells; Palm oil; Palm oil, hydrogenated; Palmitic acid [Hexadecanoic acid]; Paraffin wax; Peanut butter; Peanut shells; Peanuts; Peat moss; Pectin; Perlite; Perlite, expanded; Plaster of parts; Polyethylene; Polyglyceryl oleate; Polyglyceryl stearate; Potassium acetate [Acetic acid, potassium salt]; Potassium aluminum silicate, anhydrous; Potassium benzoate [Benzoic acid, potassium salt]; Potassium bicarbonate [Carbonic acid, monopotassium salt]; Potassium chloride; Potassium citrate [Citric acid, potassium salt]; Potassium humate [Humic acids, potassium salts]; Potassium myristate [Tetradecanoic acid, potassium salt]; Potassium oleate [9-Octadecenoic acid (9Z )-, potassium salt; Potassium ricinoleate [9-Octadecenoic acid, 12-hydroxy-, monopotassium salt,(9Z,l2R)-]; Potassium sorbate [Sorbic acid, potassium salt ]; Potassium stearate [Octadecanoic acid, potassium salt]; Potassium sulfate; Potassium sulfate [Sulfuric acid, monopotassium salt]; 1 ,2- Propylene carbonate [l,3-Dioxolan-2-one, 4-methyl-]; Pumice; Red cabbage color (expressed from edible red cabbage heads via a pressing process using only acidified water); Red cedar chips; Red dog flour; Rubber; Sawdust; Shale; Silica, amorphous, fumed (crystalline free); Silica, amorphous, precipated and gel; Silica (crystalline free); Silica gel; Silica gel, precipitated, crystalline-free; Silica, hydrate; Silica, vitreous; Silicic acid (H2Si03), magnesium salt (1 : 1); Soap (The water soluble sodium or potassium salts of fatty acids produced by either the saponification of fats and oils, or the neutralization of fatty acid); Soapbark [Quillaja saponin]; Soapstone; Sodium acetate [Acetic acid, sodium salt]; Sodium alginate; Sodium benzoate [Benzoic acid, sodium salt]; Sodium bicarbonate; Sodium carboxym ethyl cellulose [Cellulose, carboxymethyl ether, sodium salt];
Sodium chloride; Sodium citrate; Sodium humate [Humic acids, sodium salts]; Sodium oleate; Sodium ricinoleate [9-Octadecenoic acid, 12-hydroxy-, monosodium salt, (9Z,l2R)-]; Sodium stearate [Octadecanoic acid, sodium salt]; Sodium sulfate; Sorbitol [D-glucitol]; Soy protein; Soya lecithins [Lecithins, soya]; Soybean hulls; Soybean meal; Soybean, flour; Stearic acid
[Octadecanoic acid]; Sulfur; Syrups, hydrolyzed starch, hydrogenated; Tetragylceryl monooleate [9-Octadecenoic acid (9Z)-, monoester with tetraglycerol]; Tri calcium citrate [Citric acid, calcium salt (2:3)]; Triethyl citrate [Citric acid, triethyl ester; Tripotassium citrate [Citric acid, tripotassium salt]; Tripotassium citrate monohydrate [Citric acid, tripotassium salt, monohydrate]; Trisodium citrate [Citric acid, trisodium salt]; Trisodium citrate dehydrate [Citric acid, trisodium salt, dehydrate]; Trisodium citrate pentahydrate [Citric acid, trisodium salt, pentahydrate]; Ultramarine blue [C.I. 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]
IV. Methods of Use
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. In other instances, 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 per plant, a detectable modulation in the level of a metabolite, a detectable modulation in the level of a transcript, or a detectable modulation in the proteome relative to a reference plant.
In one non-limiting embodiment, 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. By "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. By“a plant susceptible to a pest” is meant that a pest is able to infect or damage the plant. For example, a plant susceptible to a pest can be susceptible to damage caused by a fungal, insect, or nematode pest as disclosed elsewhere herein.
Examples of 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 hypogaea), cotton ( Gossypium barbadense, Gossypium hirsutum ), sweet potato ( Ipomoea batatus ), cassava ( Manihot esculenta), coffee ( Coffea spp.), coconut ( Cocos nucifera), pineapple {Ananas comosus), citrus trees {Citrus spp.), cocoa {Theobroma cacao), tea {Camellia sinensis), banana {Musa spp.), avocado {Persea americand), fig {Ficus casica), guava {Psidium guajava ), mango ( Mangifera indica ), olive {Olea europaea ), papaya {Carica papaya), grape {Vitus spp.), strawberry {Fragaria x ananassa), cherry {Prunus spp.), apple {Malus domestica), orange {Citrus x .v/// /?.v/.v)=cashew {Anacardium occidentale), macadamia (Macadamia integrifolia ), almond ( Prunus amygdalus), sugar beets ( Beta vulgaris ), sugarcane {Saccharum spp.), oats, barley, vegetables, ornamentals, and conifers.
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). 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). In specific embodiments, plants of the present invention are crop plants (for example, com, alfalfa, sunflower, Brassica, soybean, cotton, safflower, peanut, sorghum, wheat, millet, tobacco, etc.). In other embodiments, com and soybean plants are optimal, and in yet other embodiments com plants are optimal.
Other 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.
A. Non-limitinz Plant Pests
In specific embodiments, 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.In specific embodiments, the bacterial strains provided herein are those that target one or more insect or insect pests. The term "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 spp., Tenebrio spp., Tribolium spp. and Trogoderma spp. In specific embodiments, 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 Scaribaeidae (e.g., Japanese beetle {Popillia japonica Newman) and European chafer {Rhizotrogus majalis Razoumowsky));
wireworms from the family Elateridae and bark beetles from the family Scolytidae.
As disclosed herein, insect pests include Coleoptera pests of the corn rootworm complex: Western corn rootworm, Diabrotica virgifera virgifera ; northern com rootworm, I) barberi ;
Southern com rootworm or spotted cucumber beetle, Diabrotica undecimpunctata howardi; and the Mexican com rootworm, I) virgifera zeae. In specific embodiments, 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,
Pseudatomoscelis seriatus , Adelphocoris rapidus , Poecilocapsus lineatus , Blissus leucopterus , Nysius ericae , Nysius raphanus , Euschistus servus , Nezara viridula , Eurygaster, Coreidae, Pyrrhocoridae, Tinidae, Blostomatidae, Reduviidae, Cimicidae; Aleurocanthus woglumi, citrus blackfly; Aleyrodes proletella , cabbage whitefly; Bemisia argentifolii , silverleaf whitefly;
Trialeurodes vaporariorum , greenhouse whitefly, and Bemisia tabaci , sweet potato whitefly.
Insect pests of interest also include Araecerus fasciculatus, coffee bean weevil;
Acanthoscelides obtectus, bean weevil; Bruchus rufmanus , broadbean weevil; Bruchus pisorum , pea weevil; Zabrotes subfasciatus , Mexican bean weevil; Diabrotica balteata , banded cucumber beetle; Cerotoma trifurcata, bean leaf beetle; Diabrotica virgifera, Mexican corn rootworm; Epitrix cucumeris, potato flea beetle; Chaetocnema confinis, sweet potato flea beetle; Hypera postica , alfalfa weevil; Anthonomus quadrigibbus , apple curculio; Sternechus paludatus , bean stalk weevil; Hypera brunnipennis, Egyptian alfalfa weevil; Sitophilus granaries, granary weevil; Craponius inaequalis, grape curculio; Sitophilus zeamais , maize weevil; Conotrachelus nenuphar, plum curculio; Euscepes postfaciatus , West Indian sweet potato weevil; Maladera castanea , Asiatic garden beetle; Rhizotrogus majalis , European chafer; Macrodactylus subspinosus , rose chafer; Tribolium confusum , confused flour beetle; Tenebrio obscurus, dark mealworm; Tribolium castaneum , red flour beetle; Tenebrio molitor , yellow mealworm and the family Drosophilidae including Drosophila suzukii , spotted wing drosophila.
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,
Anticar sia gemmatalis, Archips sp., Argyrotaenia sp., Athetis mindara, Bombyx mori, Bucculatrix thurberiella , Cadra cautella, Choristoneura sp., Cochylls hospes, Colias eurytheme , Corcyra cephalonica, Cydia latiferreanus, Cydia pomonella, Datana integerrima, Dendrolimus sibericus, Desmiafeneralis, Diaphania hyalinata , Diaphania nitidalis , Diatraea grandiose lla, Diatraea saccharalis, Ennomos subsignaria, Eoreuma loflini, Esphestia elutella, Erannis tilaria, Estigmene acrea, Eulia salubricola, Eupocoellia ambiguella, Eupoecilia ambiguella, Euproctis chrysorrhoea, Euxoa messoria, Galleria meUoneUa, Grapholita molesta, Harrisina americana, Helicoverpa subflexa , Helicoverpa zea, Heliothis virescens, Hemilenca oliviae, Homoeosoma electellum, Hyphantia cunea, Keiferia lycopersicella, Lambdina fiscellaria fiscellaria, Lambdina fiscellaria lugubrosa , Leucoma salicis , Lobesia botrana, Loxostege such calls, Lymantria dispar , Macalla thyrisalis, Malacosoma sp., Mamestra brassicae , Marne stra configurata , Manduca
quinquemaculata, Manduca sexta, Maruca testulalis, Melanchra picta, Operophtera brumata, Orgyia sp., Ostrinia nubilalis, Paleacrita vernal a, Papilio cresphontes , Pectinophora gossypiella, Phryganidia californica , Phyllonorycter blancardella , Pieris napi, Pieris rapae, Plathypena scabra, Platynota flouendana, Platynota stultana, Platyptilia carduidactyla, Plodia interpunctella, Plutella xylostella , Pontia protodice , Pseudaletia unipuncta , Pseudoplasia includens , Sabulodes aegrotata , Schizura concinna , Sitotroga cerealella, Spilonta ocellana, Spodoptera sp.,
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,
Mallophaga, Homoptera, Hemiptera, Orthroptera, Thysanoptera, Dermaptera, Isoptera, Anoplura, Siphonaptera, and Trichoptera. Insect pests of the present invention can further include those of the order Acari including, but not limited to, mites and ticks. In specific embodiments, 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 grub); Cyclocephala immaculata , southern masked chafer (white grub); Popillia japonica, Japanese beetle; Chaetocnema pulicaria, corn flea beetle;
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; Elasmopalpus lignosellus, lesser cornstalk borer; Feltia subterranea, granulate cutworm; Phyllophaga crinita, white grub; Eleodes, Conoderus, and Aeolus spp., wireworms; Oulema melanopus, cereal leaf beetle; Chaetocnema pulicaria, corn flea beetle; Sphenophorus maidis , maize billbug;
Rhopalosiphum maidis ; corn leaf aphid; Sipha flava, yellow sugarcane aphid; chinch bug, e.g., Blissus leucopterus leucopterus, Contarinia sorghicola, sorghum midge; Tetranychus
cinnabarinus, carmine spider mite; Tetranychus urticae, two-spotted spider mite; Wheat:
Pseudaletia unipunctata , armyworm; Spodoptera frugiperda, fall armyworm; Elasmopalpus lignosellus, lesser cornstalk borer; Agrotis orthogonia, pale western cutworm; Elasmopalpus lignosellus, lesser cornstalk borer; Oulema melanopus, cereal leaf beetle; Hypera punctata, clover leaf weevil; southern corn rootworm, e.g., Diabrotica undecimpunctata howardi ; Russian wheat aphid; Schizaphis graminum, greenbug; Macrosiphum avenae, English grain aphid; Melanoplus femurrubrum, redlegged grasshopper; Melanoplus differ entialis, differential grasshopper;
Melanoplus sanguinipes, migratory grasshopper; Mayetiola destructor, Hessian fly; Sitodiplosis mosellana, wheat midge; Meromyza americana, wheat stem maggot; Hylemya coarctata, wheat bulb fly; Frankliniella fusca, tobacco thrips; Cephus cinctus, wheat stem sawfly; Aceria tulipae, wheat curl mite; Sunflower: Cylindrocupturus adspersus, sunflower stem weevil; Smicronyx fulus, red sunflower seed weevil; Smicronyx sordidus, gray sunflower seed weevil; Suleima helianthana, sunflower bud moth; Homoeosoma electellum , sunflower moth; Zygogramma exclamationis, sunflower beetle; Bothyrus gibbosus , carrot beetle; Neolasioptera murtfeldtiana , sunflower seed midge; Cotton: Heliothis virescens, tobacco budworm; Helicoverpa zea , cotton bollworm;
Spodoptera exigua, beet armyworm; Pectinophora gossypiella, pink bollworm; boll weevil, e.g., Anthonomus grandis ; Aphis gossypii , cotton aphid; Pseudatomoscelis ser talus. cotton fleahopper; Trialeurodes abutilonea, banded winged whitefly; Lygus lineolaris, tarnished plant bug;
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;
Sitophilus oryzae, rice weevil; Nephotettix nigropictus , rice leafhopper; chinch bug, e.g., Blissus leucopterus leucopterus, Acrosternum hilare, green stink bug; Soybean: Pseudoplusia includens, soybean looper; Anticar sia gemmatalis , velvetbean caterpillar; Plathypena scabra , green cl overworm; Ostrinia nubilalis , European com borer; Agrotis ip si Ion , black cutworm; Spodoptera exigua, beet armyworm; Heliothis virescens, tobacco budworm; Helicoverpa zea, cotton bollworm; Epilachna varivestis, Mexican bean beetle; Myzus persicae, green peach aphid; Empoasca fabae, potato leafhopper; Acrosternum hilare, green stink bug; Melanoplus femurrubrum, redlegged grasshopper; Melanoplus differ entialis, differential grasshopper; Hylemya platura, seedcorn maggot; Sericothrips variabilis, soybean thrips; Thrips tabaci, onion thrips; Tetranychus turkestani, strawberry spider mite; Tetranychus urticae, two-spotted spider mite; Barley: Ostrinia nubilalis, European com borer; Agrotis ipsilon, black cutworm; Schizaphis graminum, greenbug; chinch bug, e.g., Blissus leucopterus leucopterus ; Acrosternum hilare, green stink bug; Euschistus servus, brown stink bug; Jylemya platura, seedcorn maggot; Mayetiola destructor, Hessian fly; Petrobia latens, brown wheat mite; Oil Seed Rape: Vrevicoryne brassicae, cabbage aphid; Phyllotreta cruciferae, crucifer flea beetle; Phyllotreta striolata, striped flea beetle; Phyllotreta nemorum, striped turnip flea beetle; Meligethes aeneus, rapeseed beetle; and the pollen beetles Meligethes rufimanus, Meligethes nigrescens, Meligethes canadianus, and Meligethes viridescens; Potato: Leptinotarsa decemlineata , Colorado potato beetle; Sweet potato: Spartocera batatas, giant sweet potato bug; Charidotella (=Metriona) bicolor, golden tortoise beetle; Cylas formicarius, sweet potato weevil; Cylas puncticollis, sweet potato weevil; Cylas brunneus, sweet potato weevil Naupactus (=Graphognathus) spp., whitefringed beetles; Conoderus rudis, wireworm; Conoderus scissus, peanut wireworm; Blosyrus spp., rough sweet potato weevil; Acraea acerata, sweet potato butterfly; Agrius convolvuli, sweet potato hornworm; Spodoptera exigua, armyworm; Spodoptera eridania, armyworm; Synanthedon spp., clearwing moth; Hairiness and eriophyid mites; Euscepes postfasciatus, West Indian sweetpotato weevil; Peloropus batatae, Peloropus weevil; Omphisia anastomasalis, sweet potato stemborer, and white grubs- larvae of various species of scarabid beetles.
The term "insect" 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.
In some embodiments, the compositions and methods provided herein control nematode plant pests. 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),
Chrysanthemum Root-Lesion Nematode (Pratylenchus fallax ), Pratylenchus cojfeae , Pratylenchus loosi and Walnut Root-Lesion Nematode (Pratylenchus vulnus)] 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 Aphelenchus such as Aphelenchus avenae ; Nematodes of the species Radopholus, such as the Burrowing-Nematode (Radopholus similis)] Nematodes of the species Tylenchulus such as Tylenchulus semipenetrans, Nematodes of the species Rotylenchulus such as Rotylenchulus re n [for mis: Nematodes living in trees such as Bursaphelenchus xylophilus and the Red Ring Nematode (Bursaphelenchus cocophilus) etc. and Globodera spp.; particularly members of the cyst nematodes, including, but not limited to
Globodera rostochiensis and Globodera pailida (potato cyst nematodes); Spiral
(Helicotylenchus spp) Burrowing (Radopholus similis ); Bulb and stem (Ditylenchus dipsaci ); Reniform (Rotylenchulus reniformis), Dagger (Xiphinema spp)] Bud and leaf
(Aphelenchoides spp), and Pine Wilt Disease (Bursaphelenchus xylophilus). Lesion nematodes include Pratylenchus spp. The term "nematode" encompasses eggs, larvae, juvenile and mature forms of nematodes.
In specific embodiments, a cell of the bacterial strain AIP031898, AIP023234, AIP024552, AIP035573, AIP071234, AIP080021, AIP001237, AIP050674, AIP071546, AIP049805,
AIPO 16229, AIP081435, AIP082140, AIP000817, AIP060333, AIP070494, AIP015104,
AIP011586, AIPO 10884, AIP082287, AIP088026, AIP065969, AIP018514, AIP033041,
AIP092281, AIP081114, AIP046403, AIP022635, 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 control an insect or nematode pest. Thus, in some embodiments, the plant pest disclosed herein is an insect pest from the order Coleoptera. For example, a cell of the bacterial strain AIP031898, AIP023234, AIP024552, AIP035573, AIP071234, AIP080021, AIP001237, AIP050674,
AIP071546, AIP049805, AIP016229, AIP081435, AIP082140, AIP000817, AIP060333,
AIP070494, AIP015104, AIP011586, AIP010884, AIP082287, AIP088026, AIP065969,
AIP018514, AIP033041, AIP092281, AIP081114, AIP046403, AIP022635, 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 can control com rootworm, Colorado potato beetle, and weevils. In specific embodiments, a cell of the bacterial strain AIP031898, AIP023234, AGR024552,
AIP035573, AIP071234, AIP080021, AIP001237, AIP050674, AIP071546, AIP049805,
AIPO 16229, AIP081435, AIP082140, AIP000817, AIP060333, AIP070494, AIP015104,
AIP011586, AIPO 10884, AIP082287, AIP088026, AIP065969, AIP018514, AIP033041,
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. In particular embodiments, AIP031898, AIP023234, AIP024552, AIP035573, AIP071234, AIP080021, AIP001237, AIP050674, AIP071546, AIP049805,
AIPO 16229, AIP081435, AIP082140, AIP000817, AIP060333, AIP070494, AIP015104,
AIP011586, AIPO 10884, AIP082287, AIP088026, AIP065969, AIP018514, AIP033041,
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.
The methods and compositions disclosed herein can be used to control one or more fungal pests. 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
sylvaticum, Pythium myriotylum, Pythium ultimum, Phytophthora spp., Phytophthora capsici, Phytophthora nicotianae, Phytophthora infestans, Phytophthora tropicalis, Phytophthora sojae, Fusarium spp., Fusarium graminearum, Fusarium solani, Fusarium oxysporum, Fusarium graminicola, Gibber ella zeae, Colletotrichum graminicola, Penicillium spp., Phakopsora sp., Phakopsora meibomiae, Phakopsora pachyrizi, Puccinia triticina, Puccinia recondita, Puccinia striiformis, Puccinia graminis, Puccinia spp.,Sclerotium spp., Sclerotinia ssp., Venturia inaequalis, Verticillium spp, Erwinia amylovora, Monilinia spp., Monilinia fructicola, Monilinia lax, and Monilinia fructigena.
In some embodiments, 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
aphanidermatum, Pythium sylvaticum, Pythium myriotylum, Pythium ultimum, Phytophthora nicotianae, Phytophthora infestans, Phytophthora tropicalis, Phytophthora sojae, Fusarium graminearum, Fusarium solani, Phakopsora pachyrizi, and Venturia inaequalis
In further embodiments, 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.
B. Methods of Controlling. Plant Pests and Treatins or Preventing Plant Disease
Provided herein are methods 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. Also provided herein are 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. Provided herein are methods of treating or preventing 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. In particular embodiments, the plant disease is a disease caused by an insect pest, such as a coleopteran pest. In certain embodiments, 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,
AIP081435, AIP082140, AIP000817, AIP060333, AIP070494, AIP015104, AIP011586,
AIP010884, AIP082287, AIP088026, AIP065969, AIP018514, AIP033041, AIP092281,
AIP081114, AIP046403, AIP022635, 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, AIP024552, AIP035573, AIP071234, AIP080021, AIP001237, AIP050674, AIP071546, AIP049805, AIP016229, AIP081435, AIP082140, AIP000817,
AIP060333, AIP070494, AIP015104, AIP011586, AIP010884, AIP082287, AIP088026,
AIP065969, AIP018514, AIP033041, AIP092281, AIP081114, AIP046403, AIP022635,
AIP070925, and AIP039063, or an active variant of any thereof. In some embodiments, the effective amount of the bacterial strain or active variant thereof comprises at least about 1012 to 1016 CFU per hectare or least about 104 to 1016 CFU per hectare, or least about 105 to 10u 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. In some methods, the bacterial strain controls one, two, three, four, five or more insect pests, such as Coleoptera pests. In some embodiments, 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. Therefore, 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. In yet another example, Table 1 shows that Colletotrichum cereal, Apiognomonia errabunda, Apiognomonia veneta , Colletotrichum gloeosporiodes , Discula fraxinea cause Anthracnose leaf spot. Therefore, 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.
Table 1
Figure imgf000039_0001
Figure imgf000040_0001
In specific embodiments, the bacterial strain provided herein or active variants thereof controls one or more nematode pests. For example, 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.
The term“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.
The terms“controlling” a plant pest 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. As such, 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. In particular embodiments,“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. As such, 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.
The term“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. In this instance, the composition is applied before exposure to the plant pests.
The term“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. In general, the term refers to the improvement in a damaged or diseased plant’s physiological state.
The term "inhibit" 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. The term "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.
The terms "delay", "retard" and all variations thereof are intended to encompass 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. In other instances, 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%,
11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%,
28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%.
45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%,
62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%,
79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%,
96%, 97%, 98%, 99%, or about 100% greater when compared to non-treated control plants.
Methods for assessing plant damage or disease severity are known, and 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. In other instances, 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%,
24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%,
75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or about 100% greater when compared to non-treated control plants. Methods for measuring the number of plant pests are known, and 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)
J. of Economic Entomology 78:290-293 and U.S. Pat. No. 5,743,477, all of which are herein incorporated by reference in their entirety.
In some embodiments, the bacterial strains and active variants thereof provided herein have pesticidal activity against an insect pest (i.e., insecticidal activity). In some of these embodiments, the insecticidal activity is activity against a Coleopteran species. In one embodiment, the insecticidal activity is against a Lepidoptera insect. In one embodiment, the insecticidal activity is against a Hemiptera species. In some embodiments, the 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.
In specific embodiments, 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. Hence, 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. For example, a plant expressing a pesticidal polypeptide or exposed to a pesticidal composition is challenged with a pest of interest. Over time, 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.
Furthermore, 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.
Chem. 267: 2228-2233, both of which are herein incorporated by reference).
C. Methods of Inducing Pest and/or Disease Resistance in Plants and/or for Improving an Agronomic Trait of Interest
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. Provided herein are 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. In certain embodiments, 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,
AIR016229, AIP081435, AIP082140, AIP000817, AIP060333, AIP070494, AIP015104,
AIP011586, AIR010884, AIP082287, AIP088026, AIP065969, AIP018514, AIP033041,
AIP092281, AIP081114, AIP046403, AIP022635, 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, AIP024552, AIP035573, AIP071234, AIP080021, AIP001237, AIP050674, AIP071546, AIP049805, AIP016229, AIP081435, AIP082140,
AIP000817, AIP060333, AIP070494, AIP015104, AIP011586, AIP010884, AIP082287, AIP088026, AIP065969, AIP018514, AIP033041, AIP092281, AIP081114, AIP046403,
AIP022635, AIP070925, and AIP039063, or an active variant of any thereof. In certain
embodiments, the bacterial strain provided herein or active variant thereof promotes a defensive response to the pest that causes the plant disease or damage. In some embodiments, the effective amount of the bacterial strain provided herein or active variant thereof comprises at least about 1012 to 1016 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.
In some methods, 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.
By "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. By "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. Further provided are methods of improving plant health and/or improving an agronomic trait of interest comprising applying to a plant an effective amount of at least one bacterial strain provided herein or an active variant thereof or an active derivative thereof. In certain embodiments, 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, AIP081435, AIP082140, AIP000817,
AIP060333, AIP070494, AIP015104, AIP011586, AIP010884, AIP082287, AIP088026,
AIP065969, AIP018514, AIP033041, AIP092281, AIP081114, AIP046403, AIP022635,
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,
AIP024552, AIP035573, AIP071234, AIP080021, AIP001237, AIP050674, AIP071546,
AIP049805, AIP016229, AIP081435, AIP082140, AIP000817, AIP060333, AIP070494,
AIP015104, AIP011586, AIP010884, AIP082287, AIP088026, AIP065969, AIP018514,
AIP033041, AIP092281, AIP081114, AIP046403, AIP022635, AIP070925, and AIP039063, or an active variant of any thereof. In some embodiments, the effective amount of the bacterial strain provided herein or active variant thereof comprises at least about 1012 to 1016 CFU per hectare.
In particular embodiments, the agronomic trait of interest that is improved by the bacterial strains or active variants thereof described herein is improved plant health. By“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. Non-limiting examples of improved agronomic traits are disclosed elsewhere herein. In specific embodiments, 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.
D. Methods of Application to a Plant or Plant Part
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. In specific embodiments, 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. In other embodiments, 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.
Generally, the rate of bacterial strain provided herein or active variant thereof is 107 to 1016 colony forming units (CFU) per hectare. In other embodiments, for a field inoculation, the rate of bacterial strain provided herein or active variant thereof application is 3 x 107 to 1 x 1011 colony forming units (CFU) per hectare. (This corresponds to about 1 Kg to 10kg of formulated material per hectare). In other embodiments, for a field inoculation, the rate of bacterial strain provided herein or active variant thereof application is 3 x 107 to 1 x 1016 colony forming units (CFU) per hectare; about lxlO12 to about lxlO13 colony forming units (CFU) per hectare, about lxlO13 to about lxlO14 colony forming units (CFU) per hectare, about lxlO14 to about lxlO15 colony forming units (CFU) per hectare, about lxlO15 to about lxlO16 colony forming units (CFU) per hectare, about lxlO16 to about lxlO17 colony forming units (CFU) per hectare; about lxlO4 to about lxlO14 colony forming units (CFU) per hectare; about lxlO5 to about lxlO13 colony forming units (CFU) per hectare; about lxlO6 to about lxlO12 colony forming units (CFU) per hectare; about lxlO9 to about lxlO11 colony forming units (CFU) per hectare; or about lxlO9 to about lxlO11 colony forming units (CFU) per hectare. In other embodiments, for a field inoculation, the rate of bacterial strain provided herein or active variant thereof application is at least about lxlO4, about lxlO5, about lxlO6, about lxlO7, about lxlO8, about lxlO9, about lxlO10, about lxlO11, about
1c10121c1013, about lxlO14, lxlO15, about lxlO16, or about lxlO17 colony forming units (CFU) per hectare. In other embodiments, for a field inoculation, the rate of bacterial strain provided herein or active variant thereof application is at least lxlO7 to at least about lxlO12 CFU/hectare. In specific embodiments, 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,
AIP000817, AIP060333, AIP070494, AIP015104, AIP011586, AIP010884, AIP082287,
AIP088026, AIP065969, AIP018514, AIP033041, AIP092281, AIP081114, AIP046403,
AIP022635, 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, AIP024552, AIP035573, AIP071234, AIP080021, AIP001237, AIP050674,
AIP071546, AIP049805, AIP016229, AIP081435, AIP082140, AIP000817, AIP060333,
AIP070494, AIP015104, AIP011586, AIP010884, AIP082287, AIP088026, AIP065969,
AIP018514, AIP033041, AIP092281, AIP081114, AIP046403, AIP022635, AIP070925, and AIP039063, or an active derivative of any thereof. 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. Moreover, 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.
Further provided is a method for controlling or inhibiting the growth of a plant pest, such as those that cause plant disease, by applying a composition comprising a bacterial strain provided herein or active variant thereof provided herein (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, AIP033041, AIP092281, AIP081114, AIP046403, AIP022635, 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, AIP024552, AIP035573, AIP071234, AIP080021, AIP001237, AIP050674, AIP071546, AIP049805, AIP016229,
AIP081435, AIP082140, AIP000817, AIP060333, AIP070494, AIP015104, AIP011586,
AIP010884, AIP082287, AIP088026, AIP065969, AIP018514, AIP033041, AIP092281,
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.
Furthermore, 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). In a specific embodiment, 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.
In some embodiments, 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.
In some embodiments, 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. For example, 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. In other embodiments, 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. In specific embodiments, 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
(suppressing growth, feeding, fecundity, or viability), reducing (reducing the pest infestation, reducing the pest feeding activities on a particular crop) or killing (causing the morbidity, mortality, or reduced fecundity of) a plant pest (e.g., an insect pest, such as a Coleoptera pest).
In other embodiments, 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. For example, 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). In specific embodiments, 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). In some embodiments, 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.
In other embodiments, 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.
In some embodiments, 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
( Meloidogyne incognita), Javanese Root-Knot nematode (Meloidogyne javanica ), Northern Root- Knot Nematode (Meloidogyne hapla) and Peanut Root-Knot Nematode.
As used herein, 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.
In specific embodiments, 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,
AIP081435, AIP082140, AIP000817, AIP060333, AIP070494, AIP015104, AIP011586,
AIP010884, AIP082287, AIP088026, AIP065969, AIP018514, AIP033041, AIP092281,
AIP081114, AIP046403, AIP022635, 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, AIP024552, AIP035573, AIP071234, AIP080021, AIP001237, AIP050674, AIP071546, AIP049805, AIP016229, AIP081435, AIP082140, AIP000817,
AIP060333, AIP070494, AIP015104, AIP011586, AIP010884, AIP082287, AIP088026,
AIP065969, AIP018514, AIP033041, AIP092281, AIP081114, AIP046403, AIP022635,
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 aboutlO5 CFU/gram to about 1011 CFU/gram, about 107 CFU/gram to about 1010 CFU/gram, about 107 CFU/gram to about 1011 CFU/gram, about 106 CFU/gram to about 1010 CFU/gram, about 106 CFU/gram to about 1011 CFU/gram, about 1011 CFU/gram to about 1012 CFU/gram, about 105 CFU/gram to about 1010 CFU/gram, about 105 CFU/gram to about 1012 CFU/gram, about 105 CFU/gram to about 106 CFU/gram, about 106 CFU/gram to about 107 CFU/gram, about 107 CFU/gram to about 108 CFU/gram, about 108 CFU/gram to about 109 CFU/gram, about 109 CFU/gram to about 1010 CFU/gram, about 1010 CFU/gram to about 1011 CFU/gram, or about 1011 CFU/gram to about 1012 CFU/gram. In some embodiments, the concentration of the bacterial strain comprises at least about 105 CFU/gram, at least about 106 CFU/gram, at least about 107 CFU/gram, at least about 108 CFU/gram, at least about 109 CFU/gram, at least about 1010 CFU/gram, at least about 1011 CFU/gram, at least about 1012 CFU/gram, or at least about 1013 CFU/gram. In specific embodiments, 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.
In specific embodiments, 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,
AIP081435, AIP082140, AIP000817, AIP060333, AIP070494, AIP015104, AIP011586,
AIP010884, AIP082287, AIP088026, AIP065969, AIP018514, AIP033041, AIP092281,
AIP081114, AIP046403, AIP022635, 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, AIP024552, AIP035573, AIP071234, AIP080021, AIP001237, AIP050674, AIP071546, AIP049805, AIP016229, AIP081435, AIP082140, AIP000817,
AIP060333, AIP070494, AIP015104, AIP011586, AIP010884, AIP082287, AIP088026,
AIP065969, AIP018514, AIP033041, AIP092281, AIP081114, AIP046403, AIP022635,
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.
Various methods are provided for controlling 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,
AIP060333, AIP070494, AIP015104, AIP011586, AIP010884, AIP082287, AIP088026,
AIP065969, AIP018514, AIP033041, AIP092281, AIP081114, AIP046403, AIP022635,
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,
AIP024552, AIP035573, AIP071234, AIP080021, AIP001237, AIP050674, AIP071546,
AIP049805, AIP016229, AIP081435, AIP082140, AIP000817, AIP060333, AIP070494,
AIP015104, AIP011586, AIP010884, AIP082287, AIP088026, AIP065969, AIP018514,
AIP033041, AIP092281, AIP081114, AIP046403, AIP022635, AIP070925, and AIP039063, or an active variant of any thereof), wherein the effective amount of the bacterial strain provided herein or active variant thereof controls the plant pest without significantly affecting the plant. In specific embodiments, the effective amount comprises at least about 1012 to 1016 colony forming units (CFU) per hectare. Further provided is a method for growing a plant susceptible to a plant pest or a plant disease caused by a plant pest. The method 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. In certain embodiments, 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,
AIP081435, AIP082140, AIP000817, AIP060333, AIP070494, AIP015104, AIP011586,
AIP010884, AIP082287, AIP088026, AIP065969, AIP018514, AIP033041, AIP092281,
AIP081114, AIP046403, AIP022635, 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, AIP024552, AIP035573, AIP071234, AIP080021, AIP001237, AIP050674, AIP071546, AIP049805, AIP016229, AIP081435, AIP082140, AIP000817,
AIP060333, AIP070494, AIP015104, AIP011586, AIP010884, AIP082287, AIP088026,
AIP065969, AIP018514, AIP033041, AIP092281, AIP081114, AIP046403, AIP022635,
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 1012 to 1016 colony forming units (CFU) per hectare.
Methods for increasing plant yield are provided. The "yield" of the plant refers to the quality and/or quantity of biomass produced by the plant. By "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 is also provided and 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,
AIP082140, AIP000817, AIP060333, AIP070494, AIP015104, AIP011586, AIP010884,
AIP082287, AIP088026, AIP065969, AIP018514, AIP033041, AIP092281, AIP081114,
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,
AIP071546, AIP049805, AIP016229, AIP081435, AIP082140, AIP000817, AIP060333,
AIP070494, AIP015104, AIP011586, AIP010884, AIP082287, AIP088026, AIP065969,
AIP018514, AIP033041, AIP092281, AIP081114, AIP046403, AIP022635, AIP070925, and AIP039063, or an active variant of any thereof, wherein said effective amount comprises at least about 1012 to 1016 colony forming units (CFU) per hectare, and wherein said composition controls a plant pest, thereby increasing yield.
As used herein, 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.
In other embodiments, a plant of interest (i.e., plant susceptible to a plant pest or plant disease caused by a plant pest) and/or 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. 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. Furthermore, 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). Moreover, 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
(sequential), so long as the desired effect is achieved.
In one non-limiting embodiment, the active variant comprises a bacterial strain provided herein that is resistant to one or more biocide. In specific embodiments, 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,
AIP033041, AIP092281, AIP081114, AIP046403, AIP022635, 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, AIP024552, AIP035573, AIP071234, AIP080021, AIP001237, AIP050674, AIP071546, AIP049805, AIP016229, AIP081435, AIP082140,
AIP000817, AIP060333, AIP070494, AIP015104, AIP011586, AIP010884, AIP082287,
AIP088026, AIP065969, AIP018514, AIP033041, AIP092281, AIP081114, AIP046403,
AIP022635, AIP070925, and AIP039063, or an active variant of any thereof) is resistant to glyphosate. In such methods, 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.
In another non-limiting embodiment, the active variant comprises a bacterial strain provided herein that is resistant to glufosinate. In such methods, 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. In such embodiments, 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. In a further embodiment, the bacterial strain provided herein or active variant thereof comprises an effective amount of 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, AIP033041, AIP092281, AIP081114, AIP046403, AIP022635,
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,
AIP024552, AIP035573, AIP071234, AIP080021, AIP001237, AIP050674, AIP071546,
AIP049805, AIP016229, AIP081435, AIP082140, AIP000817, AIP060333, AIP070494,
AIP015104, AIP011586, AIP010884, AIP082287, AIP088026, AIP065969, AIP018514,
AIP033041, AIP092281, AIP081114, AIP046403, AIP022635, AIP070925, and AIP039063, or an active variant of any thereof.
V. Biocides for Use in Combination with the Bacterial Strain Provided Herein or Active Variants Thereof
As discussed elsewhere herein, 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). In such instances, the bacterial strain provided herein or active variant thereof is compatible with the biocide of interest.
By“herbicide, insecticide, fungicide, pesticide, insecticide or other crop protection chemical tolerance or herbicide, fungicide, pesticide, insecticide or other crop protection chemical resistance” is intended the ability of an organism (i.e., the plant and/or the bacterial strain provided herein or active variant thereof etc.) to survive and reproduce following exposure to a dose of the herbicide, insecticide, fungicide, pesticide, insecticide, or other crop protection chemical that is normally lethal to the wild type organism.
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 (GC)); Glutamine Synthesis inhibitor (Phosphinic Acid (PA)); DOXP synthase inhibitor (Isoxazolidinone (IA)); HPPD inhibitor (Pyrazole (PA)), Triketone (TE)); PPO inhibitors (Diphenyl ether (DE), N- phenylphthalimide (NP) (Ary triazinone (AT)); VLFA inhibitors (chloroacetamide (CA)), Oxyacetamide (OA)); Photosystem I inhibitor (Bipyridyliums (BP)); and the like.
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, and thiamethoxam, spinosyns such as spinosad, cyclodiene organochlorines such as camphechlor, chlordane, endosulfan, gamma-HCH, HCH, heptachlor, lindane, methoxychlor, fiproles such as acetoprole, ethiprole, fipronil, vaniliprole, chloride-channel, 6.1 mectins such as avermectin, emamectin, emamectin-benzoate, ivermectin, and milbemycin, juvenile-hormone mimics such as diofenolan, epofenonane, fenoxycarb, hydroprene, kinoprene, methoprene, pyriproxyfen, and triprene, ecdysone agoni sts/di sruptors, diacylhydrazine, chromafenozide, halofenozide, methoxyfenozide, tebufenozide, chitin biosynthesis inhibitors, benzoylureas such as bistrifluron, chlorfluazuron, diflubenzuron, fluazuron, fluey cl oxuron, flufenoxuron, hexaflumuron, lufenuron, novaluron, noviflumuron, penfluron, teflubenzuron, triflumuron, buprofezin, cyromazine, oxidative phosphorylation inhibitors, ATP disruptors, diafenthiuron, organotins such as azocyclotin, cyhexatin, fenbutatin-oxide, pyrroles such as chlorfenapyr, dinitrophenols such as binapacryl, dinobuton, dinocap, DNOC, site-I electron transport inhibitors, METI's such as fenazaquin, fenpyroximate, pyrimidifen, pyridaben, tebufenpyrad, tolfenpyrad, hydramethyinon, dicofol, rotenone, acequinocyl, fluacrypyrim, spirodiclofen, spiromesifen, tetramic acids, carboxamides such as flonicamid, octopaminergic agonists such as amitraz, magnesium-stimulated ATPase inhibitors such as propargite, BDCA's such as N2-[l,l-dimethyl-2-(methylsulfonyl)ethyl]-3-iodo- Nl-[2-methyl-4-[l,2,2,2— tetrafluoro-l-(trifluoromethyl)ethyl]phenyl]-l, 2-benzene, nereistoxin analogues such as thiocyclam hydrogen oxalate, and thiosultap sodium. Preferably the insecticide is one or more of chlorpyrifos and tefluthrin.
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, isamidofos, mecarphon, phorate, thionazin, triazophos, diamidafos, fosthietan, phosphamidon), and certain fungicides, such as captan, thiophanate-methyl and thiabendazole.
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, pefurazoate, valifenalate); anilide fungicides (benalaxyl, benalaxyl-M, bixafen, boscalid, carboxin, fenhexamid, fluxapyroxad, isotianil, metalaxyl, metalaxyl-M, metsulfovax, ofurace, oxadixyl, oxycarboxin, penflufen, pyracarbolid, sedaxane, thifluzamide, tiadinil, vanguard); benzanilide fungicides (benodanil, flutolanil, mebenil, mepronil, salicylanilide, tecloftalam); furanilide fungicides (fenfuram, furalaxyl, furcarbanil, methfuroxam); sulfonanilide fungicides (flusulfamide); benzamide fungicides (benzohydroxamic acid, fluopicolide, fluopyram, tioxymid, trichlamide, zarilamid, zoxamide); furamide fungicides (cyclafuramid, furmecyclox); phenylsulfamide fungicides (dichlofluanid, tolylfluanid); sulfonamide fungicides (amisulbrom, cyazofamid); valinamide fungicides (benthiavalicarb, iprovalicarb);
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
pentachlorophenoxide, tecnazene, trichlorotrinitrobenzenes); arsenical fungicides (asomate, urbacide); aryl phenyl ketone fungicides (metrafenone, pyriofenone); benzimidazole fungicides (albendazole, benomyl, carbendazim, chlorfenazole, cypendazole, debacarb, fuberidazole, mecarbinzid, rabenzazole, thiabendazole); benzimidazole precursor fungicides (furophanate, thiophanate, thiophanate-methyl); benzothiazole fungicides (bentaluron, benthiavalicarb, benthiazole, chlobenthiazone, probenazole); botanical fungicides (allicin, berberine, carvacrol, carvone, osthol, sanguinarine, santonin); bridged diphenyl fungicides (bithionol, dichlorophen, diphenylamine, hexachlorophene, parinol); carbamate fungicides (benthiavalicarb, furophanate, iodocarb, iprovalicarb, picarbutrazox, propamocarb, pyribencarb, thiophanate, thiophanate-methyl, tolprocarb); benzimidazolylcarbamate fungicides (albendazole, benomyl, carbendazim, cypendazole, debacarb, mecarbinzid); carbanilate fungicides (diethofencarb, pyraclostrobin, pyrametostrobin, triclopyricarb); conazole fungicides, conazole fungicides (imidazoles)
(climbazole, clotrimazole, imazalil, oxpoconazole, prochloraz, triflumizole); conazole fungicides (triazoles) (azaconazole, bromuconazole, cyproconazole, diclobutrazol, difenoconazole, diniconazole, diniconazole-M, epoxiconazole, etaconazole, fenbuconazole, fluquinconazole, flusilazole, flutriafol, furconazole, furconazole-cis, hexaconazole, imibenconazole, ipconazole, metconazole, myclobutanil, penconazole, propi conazole, prothioconazole, quinconazole, simeconazole, tebuconazole, tetraconazole, triadimefon, triadimenol, triticonazole, uniconazole, uniconazole-P); copper fungicides (acypetacs-copper, Bordeaux mixture, Burgundy mixture, Cheshunt mixture, copper acetate, copper carbonate, basic, copper hydroxide, copper naphthenate, copper oleate, copper oxychloride, copper silicate, copper sulfate, copper sulfate, basic, copper zinc chromate, cufraneb, cuprobam, cuprous oxide, mancopper, oxine-copper, saisentong, thiodiazole- copper); cyanoacrylate fungicides (benzamacril, phenamacril); dicarboximide fungicides
(famoxadone, fluoroimide); dichlorophenyl dicarboximide fungicides (chlozolinate, dichlozoline, iprodione, isovaledione, myclozolin, procymidone, vinclozolin); phthalimide fungicides (captafol, captan, ditalimfos, folpet, thiochlorfenphim); dinitrophenol fungicides (binapacryl, dinobuton, dinocap, dinocap-4, dinocap-6, meptyldinocap, dinocton, dinopenton, dinosulfon, dinoterbon, DNOC); dithiocarbamate fungicides (amobam, asomate, azithiram, carbamorph, cufraneb, cuprobam, disulfiram, ferbam, metam, nabam, tecoram, thiram, urbacide, ziram); cyclic dithiocarbamate fungicides (dazomet, etem, milneb); polymeric dithiocarbamate fungicides (mancopper, mancozeb, maneb, metiram, polycarbamate, propineb, zineb); dithiolane fungicides (isoprothiolane, saijunmao); fumigant fungicides (carbon disulfide, cyanogen, dithioether, methyl bromide, methyl iodide, sodium tetrathiocarbonate); hydrazide fungicides (benquinox, saijunmao); imidazole fungicides (cyazofamid, fenamidone, fenapanil, glyodin, iprodione, isovaledione, pefurazoate, triazoxide); conazole fungicides (imidazoles) (climbazole, clotrimazole, imazalil, oxpoconazole, prochloraz, triflumizole); inorganic fungicides (potassium azide, potassium thiocyanate, sodium azide, sulfur, see also copper fungicides, see also inorganic mercury fungicides); mercury fungicides; inorganic mercury fungicides (mercuric chloride, mercuric oxide, mercurous chloride); organomercury fungicides ((3-ethoxypropyl)mercury bromide, ethylmercury acetate, ethylmercury bromide, ethylmercury chloride, ethylmercury 2,3-dihydroxypropyl mercaptide, ethylmercury phosphate, Ar-(ethylmercury)-p-toluenesulphonanilide, hydrargaphen, 2- methoxyethylmercury chloride, methylmercury benzoate, methylmercury dicyandiamide, methylmercury pentachlorophenoxide, 8-phenylmercurioxyquinoline, phenylmercuriurea, phenylmercury acetate, phenylmercury chloride, phenylmercury derivative of pyrocatechol, phenylmercury nitrate, phenylmercury salicylate, thiomersal, tolylmercury acetate); morpholine fungicides (aldimorph, benzamorf, carbamorph, dimethomorph, dodemorph, fenpropimorph, flumorph, tridemorph); organophosphorus fungicides (ampropylfos, ditalimfos, EBP, edifenphos, fosetyl, hexylthiofos, inezin, iprobenfos, izopamfos, kejunlin, phosdiphen, pyrazophos, tolclofos- methyl, triamiphos); organotin fungicides (decafentin, fentin, tributyltin oxide); oxathiin fungicides (carboxin, oxycarboxin); oxazole fungicides (chlozolinate, dichlozoline, drazoxolon, famoxadone, hymexazol, metazoxolon, myclozolin, oxadixyl, oxathiapiprolin, pyrisoxazole, vinclozolin);
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 fungicides (bupirimate, diflumetorim, dimethirimol, ethirimol, fenarimol, ferimzone, nuarimol, triarimol);
anilinopyrimidine fungicides (cyprodinil, mepanipyrim, pyrimethanil); pyrrole 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); thiadiazole fungicides
(etridiazole, saisentong, thiodiazole-copper, zinc thiazole); thiazole fungicides (ethaboxam, isotianil, metsulfovax, octhilinone, oxathiapiprolin, thiabendazole, thifluzamide); thiazolidine fungicides (flutianil, thiadifluor); thiocarbamate fungicides (methasulfocarb, prothiocarb);
thiophene fungicides (ethaboxam, isofetamid, silthiofam); triazine fungicides (anilazine); triazole fungicides (amisulbrom, bitertanol, fluotrimazole, triazbutil); conazole fungicides (triazoles) (azaconazole, bromuconazole, cyproconazole, diclobutrazol, difenoconazole, diniconazole, diniconazole-M, epoxiconazole, etaconazole, fenbuconazole, fluquinconazole, flusilazole, flutriafol, furconazole, furconazole-cis, hexaconazole, huanjunzuo, imibenconazole, ipconazole, metconazole, myclobutanil, penconazole, propi conazole, prothioconazole, quinconazole, simeconazole, tebuconazole, tetraconazole, triadimefon, triadimenol, triticonazole, uniconazole, uniconazole-P); triazolopyrimidine fungicides (ametoctradin); urea fungicides (bentaluron, pencycuron, quinazamid); zinc fungicides (acypetacs-zinc, copper zinc chromate, cufraneb, mancozeb, metiram, polycarbamate, polyoxorim-zinc, propineb, zinc naphthenate, zinc thiazole, zinc trichlorophenoxide, zineb, ziram); unclassified fungicides (acibenzolar, acypetacs, allyl alcohol, benzalkonium chloride, bethoxazin, bromothalonil, chitosan, chloropicrin, DBCP, dehydroacetic acid, diclomezine, diethyl pyrocarbonate, ethylicin, fenaminosulf, fenitropan, fenpropidin, formaldehyde, furfural, hexachlorobutadiene, methyl isothiocyanate, nitrostyrene, nitrothal-isopropyl, OCH, pentachlorophenyl laurate, 2-phenylphenol, phthalide, piperalin, propamidine, proquinazid, pyroquilon, sodium orthophenylphenoxide, spiroxamine, sultropen, thicyofen, tricyclazole), mefenoxam, or acetemiprid.
Non-limiting embodiments of the invention include:
1. 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,
AIP035573, AIP071234, AIP080021, AIP001237, AIP050674, AIP071546, AIP049805,
AIPO 16229, AIP081435, AIP082140, AIP000817, AIP060333, AIP070494, AIP015104,
AIP011586, AIPO 10884, AIP082287, AIP088026, AIP065969, AIP018514, AIP033041,
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 cell, spore, forespore, or combination of cells, spores and/or forespores or the active variant of any thereof is present at about 105 CFU/gram to about 1012 CFU/gram or at about 105 CFU/ml to about 1012 CFU/ml, and wherein an effective amount of said composition controls a plant pest or improves an agronomic trait of interest of a plant.
2. The composition of embodiment 1, wherein said plant pest comprises an insect pest.
3. The composition of embodiment 2, wherein said insect pest comprises a Coleoptera insect or Hemiptera insect.
4. The composition of embodiment 3, wherein said Coleoptera insect is a corn rootworm, Colorado potato beetle, or a weevil.
5. The composition of embodiment 3, wherein said Coleoptera insect is a Western com rootworm, Colorado potato beetle, or sweetpotato weevil.
6. The 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 105 CFU/gram to about 1010 CFU/gram or at about 105 CFU/ml to about 1010 CFU/ml. 7. The composition of any of embodiments 1-6, wherein said composition comprises a cell paste.
8. The composition of any one of embodiments 1-7, wherein said composition comprises a wettable powder.
9. 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,
AIP071546, AIP049805, AIP016229, AIP081435, AIP082140, AIP000817, AIP060333,
AIP070494, AIP015104, AIP011586, AIP010884, AIP082287, AIP088026, AIP065969,
AIP018514, AIP033041, 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 an effective amount of said composition controls a plant pest or improves an agronomic trait of interest of a plant.
10. The composition of embodiment 9, wherein said plant pest comprises an insect pest.
11. The composition of embodiment 10, wherein said insect pest comprises a Coleoptera insect or Hemiptera insect.
12. The composition of embodiment 11, wherein said Coleoptera insect is a corn rootworm, Colorado potato beetle, or a weevil.
13. The composition of embodiment 11, wherein said Coleoptera insect is a Western com rootworm, Colorado potato beetle, or sweetpotato weevil.
14. 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,
AIP071546, AIP049805, AIP016229, AIP081435, AIP082140, AIP000817, AIP060333,
AIP070494, AIP015104, AIP011586, AIP010884, AIP082287, AIP088026, AIP065969,
AIP018514, AIP033041, 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 an effective amount of said composition controls a plant pest or improves an agronomic trait of interest of a plant. 15. The composition of embodiment 14, wherein said plant pest comprises at least one insect pest.
16. The composition of embodiment 15, wherein said insect pest comprises a Coleoptera insect or Hemiptera insect.
17. The composition of embodiment 16, wherein said Coleoptera insect is a corn rootworm, Colorado potato beetle, or a weevil.
18. The composition of embodiment 16, wherein said Coleoptera insect is a Western com rootworm, Colorado potato beetle, or sweetpotato weevil.
19. The composition of any one of embodiments 14-18, wherein said active variant is resistant to at least one herbicide, nematicide, fungicide, pesticide, insecticide or other crop protection chemical.
20. The 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.
21. The composition of any one of embodiments 14-20, wherein said active variant has been transformed with a herbicide resistance gene rendering the cell, spore, forespore, or combination of cells, forespores and/or spores herbicide resistant. .
22. The 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).
23. 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,
AIP024552, AIP035573, AIP071234, AIP080021, AIP001237, AIP050674, AIP071546,
AIP049805, AIP016229, AIP081435, AIP082140, AIP000817, AIP060333, AIP070494,
AIP015104, AIP011586, AIP010884, AIP082287, AIP088026, AIP065969, AIP018514,
AIP033041, AIP092281, AIP081114, AIP046403, 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;
wherein 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.
24. The isolated biologically pure culture of embodiment 23, wherein said bacterial strain is resistant to a biocide selected from a herbicide, a fungicide, a pesticide, insecticide, or a crop protection chemical, wherein said culture is produced by growing in the presence of said biocide. 25. The isolated biologically pure culture of embodiment 24, wherein said biologically pure culture is able to grow in the presence of glyphosate.
26. The isolated biologically pure culture of any one of embodiments 23-25, wherein said plant pest comprises an insect pest.
27. The isolated biologically pure culture of embodiment 26, wherein said insect pest comprises a Coleoptera insect or Hemiptera insect.
28. The isolated biologically pure culture of embodiment 27, wherein said Coleoptera insect is a com rootworm, Colorado potato beetle, or a weevil.
29. The isolated biologically pure culture of embodiment 27, wherein said Coleoptera insect is a Western corn rootworm, Colorado potato beetle, or sweetpotato weevil.
30. A bacterial culture grown from a cell, a spore, a forespore, or a combination of cells, spores and/or forespores of AIP031898, AIP023234, AIP024552, AIP035573, AIP071234, AIP080021, AIP001237, AIP050674, AIP071546, AIP049805, AIP016229, AIP081435, AIP082140,
AIP000817, AIP060333, AIP070494, AIP015104, AIP011586, AIP010884, AIP082287,
AIP088026, AIP065969, AIP018514, AIP033041, 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 bacterial culture is able to grow in the presence of glufosinate; and
wherein an effective amount of said bacterial culture controls a plant pest or improves an agronomic trait of interest of a plant.
31. The bacterial culture of embodiment 30, wherein said plant pest comprises an insect pest.
32. The bacterial culture of embodiment 31, wherein said insect pest comprises a Coleoptera insect or Hemiptera insect.
33. The bacterial culture of embodiment 32, wherein said Coleoptera insect is a corn rootworm, Colorado potato beetle, or a weevil.
34. The bacterial culture of embodiment 32, wherein said Coleoptera insect is a Western corn rootworm, Colorado potato beetle, or sweetpotato weevil.
35. 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.
36. 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,
AIP082140, AIP000817, AIP060333, AIP070494, AIP015104, AIP011586, AIP010884,
AIP082287, AIP088026, AIP065969, AIP018514, AIP033041, 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 effective amount comprises at least about 1012 to 1016 colony forming units (CFU) per hectare; and
wherein said effective amount controls said plant pest or improves the agronomic trait of interest.
37. The method of embodiment 36, wherein said method increases yield of the plant susceptible to the plant pest.
38. The method of embodiment 36 or 37, wherein said plant is a soybean.
39. The method of any one of embodiments 36-38, wherein said plant pest comprises an insect pest.
40. The method of embodiment 39, wherein said insect pest comprises a Coleoptera insect or Hemiptera insect.
41. The method of embodiment 40, wherein said Coleoptera insect is a com rootworm,
Colorado potato beetle, or a weevil.
42. The method of embodiment 40, wherein said Coleoptera insect is a Western com rootworm, Colorado potato beetle, or sweetpotato weevil.
43. A method of controlling a plant pest in an area of cultivation comprising:
(a) planting the area of cultivation with seeds or plants susceptible to the plant pest; and
(b) applying to the seed or plant susceptible to the plant pest an effective amount of a cell, a spore, a forespore, or a combination of cells, spores and/or forespores of at least one bacterial strain comprising AIP031898, AIP023234, AIP024552, AIP035573, AIP071234, AIP080021, AIP001237, AIP050674, AIP071546, AIP049805, AIP016229, AIP081435, AIP082140,
AIP000817, AIP060333, AIP070494, AIP015104, AIP011586, AIP010884, AIP082287,
AIP088026, AIP065969, AIP018514, AIP033041, AIP092281, AIP081114, AIP046403,
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 1012 to 1016 colony forming units (CFU) per hectare.
44. The method of embodiment 43, where said seed or plant is a soybean.
45. The method of embodiment 43 or 44, wherein said plant pest comprises an insect pest.
46. The method of embodiment 45, wherein said insect pest comprises a Coleoptera insect or
Hemiptera insect.
47. The method of embodiment 46, wherein the Coleoptera insect is a corn rootworm, Colorado potato beetle, or a weevil.
48. The method of embodiment 46, wherein the Coleoptera insect is a Western corn rootworm, Colorado potato beetle, or sweetpotato weevil.
49. The method of any one of embodiments 43-48, wherein said method further comprises applying an effective amount of a biocide, wherein said effective amount of the biocide selectively controls an organism of interest while not significantly damaging said seed or plant.
50. The method of embodiment 49, wherein said cell, spore, forespore, or combination of cells, spores and/or forespores or active variant of any thereof and the biocide are applied simultaneously.
51. The method of embodiment 49, wherein said cell, spore, forespore, or combination of cells, spores and/or forespores or active variant of any thereof and the biocide are applied sequentially.
52. The method of any one of embodiments 49-51 where the biocide is a nematicide.
53. A method of making a modified bacterial strain comprising:
(a) providing a population of a cell, a spore, a forespore, or a combination of cells, spores and/or forespores of at least one bacterial strain comprising AIP031898, AIP023234, AIP024552, AIP035573, AIP071234, AIP080021, AIP001237, AIP050674, AIP071546, AIP049805,
AIR016229, AIP081435, AIP082140, AIP000817, AIP060333, AIP070494, AIP015104,
AIP011586, AIR010884, AIP082287, AIP088026, AIP065969, AIP018514, AIP033041,
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;
(b) culturing said population in the presence of a biocide of interest; and,
(c) selecting a modified bacterial strain having an increased resistance to said biocide of interest.
54. The method of embodiment 53, where said culturing comprises increasing the concentration of the biocide over time.
55. The method of embodiment 53 or 54, where said biocide is glyphosate or glufosinate. 56. 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,
AIP000817, AIP060333, AIP070494, AIP015104, AIP011586, AIP010884, AIP082287,
AIP088026, AIP065969, AIP018514, AIP033041, AIP092281, AIP081114, AIP046403,
AIP022635, AIP070925, and AIP039063, or an active variant of any thereof, wherein the active variant comprises a bacterial strain having a genome within a Mash distance of about 0.015;
wherein said effective amount comprises at least about 1012 to 1016 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.
57. The method of embodiment 56, wherein said plant is a soybean.
58. The method of embodiment 56 or 57, wherein said bacterial strain or active variant thereof treats or prevents one or more plant diseases.
59. The method of embodiment 58, wherein said one or more plant diseases are caused by an insect pest.
60. The method of embodiment 59, wherein said insect pest is a Coleoptera insect or a
Hemiptera insect.
61. The method of embodiment 60, wherein said Coleoptera insect is a com rootworm,
Colorado potato beetle, or a weevil.
62. The method of embodiment 60, wherein said Coleoptera insect is a Western com rootworm, Colorado potato beetle, or sweetpotato weevil.
The following examples are offered by way of illustration and not by way of limitation.
EXAMPLES
Example 1. Microbial strains and Methods of Culturing
Table 2. A bacterial strain selected for evaluation of inhibition of pest activity.
Figure imgf000066_0001
Figure imgf000067_0001
Figure imgf000068_0001
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 107 CFU/ml, at least 108 CFU/ml, at least 109 CFU/ml, at least 1010 CFU/ml, at least 1011 CFU/ml, or at least 1012 CFU/ml.
Table 3. Culture conditions
Figure imgf000068_0002
Figure imgf000069_0001
Example 2. Evaluation of Bacterial Strains Against Asian Soybean Rust
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 mitioΐ 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.
Strain evaluation:
Evaluation on detached-leaf. Briefly, leaf disks (3.5-cm diameter each) were sprayed with 120 pl of each bacterial strain of interest (1 x 108 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). 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. Boxes with 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 104 urediniospores/mL of sterile distilled water) using an atomizer attached to an air compressor (Twizeyimana and Hartman, 2010). After inoculation, 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.
Table 4. Effects on Asian Soybean Rust
Figure imgf000069_0002
Figure imgf000070_0001
Figure imgf000071_0001
Example 3. Rhizoctonia Damping-Off Assay— Soybean Mock Seed Treatment/In-Furrow
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. Once treatments are applied, 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.
Data Collection and Results: After 10-12 days, the assay is evaluated to determine the number of seeds that germinated. Data is reported as the % of seeds that germinated out of a total of 6 seeds per treatment.
Example 4 Insecticidal Activity
Microbe growth:
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.
Twenty-five mΐ from each well of the starter culture was added to the assay culture blocks. 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.
Lepidopteran Diet Overlay Assay:
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. 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. For all strains, 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.
Colorado Potato Beetle fCPB) assay:
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 wells were scored for leaf damage 48 h after treatment and introduction of larvae. The positive controls were AFS092623, AFS046061, bacteria that consistently shows activity against CPB and the negative control was AFS094370, a bacterium that consistently is inactive against CPB. Bacteria were considered active if leaf damage was <20%. Results are provided in Table 5. Western corn rootworm Assay:
The bacterial cultures were evaluated in an insect diet overlay bioassay in wells of 24-well plates (Cellstar, 24-well, Greiner Bio One. 60 mΐ 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 mΐ 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, and the positive control was AFS037424, a bacterium that consistently shows activity against WCR. The results are set forth in Table 5.
Stink Bug feedine assay:
Southern Green stink bugs (SGSB) were reared at the AgBiome facility in RTP, NC. Only healthy second instar SGSB were used in the assay. One ml of a 50:50 whole culture
microbe: 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.
Mortality was assessed for each sample. A microbe is considered active for SGSB when mortality is greater than 50% in three or more independent repetitions. The results are set forth in Table 5.
Table 5.
Figure imgf000073_0001
Figure imgf000074_0001
Example 5. Field Trials for the Various Bacterial Strains or Active Variants Thereof
The various bacterial strain recited in Table 2 are applied to soybeans in the field.
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.
Treatments:
1. Untreated Check
2. Inoculated Check
3. Quadri s at 6.2oz/acre
4. Quadri s at 2.1 oz/acre
5. AIP031898 at 7.5g/L
6. AIP023234 at 7.5g/L
7. AIP024552 at 7.5g/L
8. AIP035573 at 7.5g/L
9. AIP071234 at 7.5g/L
10. AIP080021 at 7.5g/L
11. AIP00l237 at 7.5g/L
12. AIP050674 at 7.5g/L 13. AIP071546 at 7.5g/L
14. AIP049805 at 7.5g/L
15. AIPO 16229 at 7.5g/L
16. AIP081435 at 7.5g/L
17. AIP082140 at 7.5g/L
18. AIP000817 at 7.5g/L
19. AIP060333 at 7.5g/L
20. AIP070494 at 7.5g/L
21. AIP015104 at 7.5g/L
22. AIPO 11586 at 7.5g/L
23. AIPO 10884 at 7.5g/L
24. AIP082287 at 7.5g/L
25. AIP088026 at 7.5g/L
26. AIP065969 at 7.5g/L
27. AIPO 18514 at 7.5g/L
28. AIP033041 at 7.5g/L
29. AIP092281 at 7.5g/L
30. AIP081114 at 7.5g/L
31. AIP046403 at 7.5g/L
32. AIP022635 at 7.5g/L
33. AIP070925 at 7.5g/L
34. AIP039063 at 7.5g/L
Example 6. Field Trials Against Various Fungal Pathogens for the Various Bacterial Strains
The various bacterial strains recited in Table 2 are applied to the crops listed in Table 6 in the field under the current agronomic practices as listed in Table 6 to achieve uniform plant coverage and follow proper agronomic practices. Treatments are applied preventatively and/or curatively at the appropriate timings per disease.
Table 6
Figure imgf000075_0001
Figure imgf000076_0001
Figure imgf000077_0001
The specific treatments are outlined below:
Foliar Pathogen Treatment List: Early Blight
6-10 treatments
Treatment Volume: 100 gallons/acre
Treatment List:
1. Non-Inoculated, untreated Check
2. Inoculated Check
3. Chemical control chosen by cooperator applied at label instructions
4. Biological control Serenade applied at label instructions
5. Experimental Biological Foliar treatment(s) at 5g/L plus Capsil at 3oz/l00 gallons
Example 7. Field Trials Against Various Fungal Pathogens for the Various Bacterial Strains or
Active Variants Thereof employing Seed Treatments
The various bacterial strains recited in Table 2 are applied to the crops listed in Table 7 as seed treatments prior to being planted into the field. Bacterial strain treatments are applied for preventative control of the diseases and at the application rates in Table 8. The specific treatments are outlined below.
Table 7
Figure imgf000077_0002
Seed Treatment Trial Treatment List:
1. Non-inoculated Check
2. Inoculated Check
3. Disease appropriate Seed Treatment Chemical Check chosen and applied by cooperator
5. Biological Experimental Seed Treatment(s)
Table 8
Figure imgf000078_0001
Example 8. Field Trials Against Various Fungal Pathogens for the Various Bacterial Strains or Active Variants Thereof Employing In-Furrow Treatments
The various bacterial strains or active variants thereof recited in Table 2 are applied to the crops listed in Table 9 as in-furrow treatments at time of planting as preventative control for the diseases and at the treatment rates listed in Table 9. The specific treatments are outlined below: In-Furrow Trial Treatment List:
1. Non-inoculated Check
2. Inoculated Check
3. In-Furrow Biological Treatment(s) 5g/L + Capsil at 6oz/l00 Gallons at 15 Gallons/Acre
4. Disease appropriate In-Furrow Chemical Check as chosen and applied by cooperator.
Table 9
Figure imgf000078_0002
Figure imgf000079_0001
Example 9. Biological Control Strain Seed Treatment Protocol
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.
Example 10. Wettable Powder Formulations
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.
Example 11. Pythium Field Trials.
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:
Treatments:
1. Untreated Check
2. Inoculated Check
3. Quadris at 0.4 fluid ounces/ Acre
4. AGR0 1898 seed treatment
5. AIP023234 seed treatment
6. AIP024552 seed treatment
7. AIP035573 seed treatment
8. AIP071234 seed treatment
9. AIP080021 seed treatment
10. AIP001237 seed treatment
11. AIP050674 seed treatment
12. AIP071546 seed treatment
13. AIP049805 seed treatment
14. AIPO 16229 seed treatment
15. AIP081435 seed treatment
16. AIP082140 seed treatment
17. AIP000817 seed treatment
18. AIP060333 seed treatment
19. AIP070494 seed treatment
20. AIPO 15104 seed treatment
21. AIPO 11586 seed treatment 22. AIPO 10884 seed treatment
23. AIP082287 seed treatment
24. AIP088026 seed treatment
25. AIP065969 seed treatment
26. AIPO 18514 seed treatment
27. AIP033041 seed treatment
28. AIP092281 seed treatment
29. AIP081114 seed treatment
30. AIP046403 seed treatment
31. AIP022635 seed treatment
32. AIP070925 seed treatment
33. AIP039063 seed treatment
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.
After growth, 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.
The supernatant is then poured off so that only the pellet remained. Pellets are then re-suspended in 35 ml of nematode water for a 10X concentration or 116 ml of nematode water for a 3X concentration. These concentrated microbe solutions are then used in the on-plant assay. All microbial preparations are applied within 12 h of preparation.
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. At one week, the cucumber plants received a drench application with 10X microbial concentration or 3X microbial concentration, prepared as described above. Alternatively, 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. Four hours after the drench application, plants are infected with 1500 stage 2 RKN juveniles per plant. 48 hrs after the plants are infected, watering and fertilizing for plant care resumed as needed. Plants are then allowed to grow in a growth chamber under standard conditions for 28 days. At 28 days, plants are assessed for galling and egg counts per test condition. Galling of the test microbes is compared to the positive and negative controls. The indication of activity is stated as having reduced galling or no reduction in galling as compared to the negative control. Egg counts are also completed and reported as total egg count for each test condition. Results are provided in Table 10.
Table 10.
Figure imgf000083_0001

Claims

What is claimed is:
1. 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 AGR031898, AIP023234, AIP024552, AIP035573, AIP071234, AIP080021, AIP001237, AIP050674, AIP071546, AIP049805,
AIPO 16229, AIP081435, AIP082140, AIP000817, AIP060333, AIP070494, AIP015104,
AIP011586, AIPO 10884, AIP082287, AIP088026, AIP065969, AIP018514, AIP033041,
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 cell, spore, forespore, or combination of cells, spores and/or forespores or the active variant of any thereof is present at about 105 CFU/gram to about 1012 CFU/gram or at about 105 CFU/ml to about 1012 CFU/ml, and wherein an effective amount of said composition controls a plant pest or improves an agronomic trait of interest of a plant.
2. The composition of claim 1, wherein said composition comprises a cell paste or a wettable powder.
3. 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, AIP024552, AIP035573, AIP071234, AIP080021, AIP001237, AIP050674, AIP071546,
AIP049805, AIP016229, AIP081435, AIP082140, AIP000817, AIP060333, AIP070494,
AIP015104, AIP011586, AIP010884, AIP082287, AIP088026, AIP065969, AIP018514,
AIP033041, AIP092281, AIP081114, AIP046403, 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;
wherein 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.
4. The composition of claim 1 or 2, or the isolated biologically pure culture of claim 3, wherein said plant pest comprises an insect pest.
5. The composition of claim 1 or 2, or the isolated biologically pure culture of claim 3, wherein said plant pest comprises a Coleoptera insect or Hemiptera insect.
6. The composition of claim 1 or 2, or the isolated biologically pure culture of claim 3, wherein said Coleoptera insect is a corn rootworm, Colorado potato beetle, or a sweet potato weevil.
7. The composition of claim 1 or 2, or the isolated biologically pure culture of claim 3, wherein said bacterial strain or active variant thereof is resistant to a biocide selected from a herbicide, a fungicide, a pesticide, insecticide, or a crop protection chemical.
8. The composition or the isolated biologically pure culture of claim 7, wherein said bacterial strain or active variant thereof 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.
9. The composition or the isolated biologically pure culture of claim 7, wherein said bacterial strain or active variant has been transformed with a herbicide resistance gene rendering the cell, spore, forespore, or combination of cells, spores, and/or forespores or active variant thereof herbicide resistant.
10. A method for controlling a plant pest population comprising contacting said population with an effective amount of the composition of any one of claims 1, 2, or 4-9 or the isolated biologically pure culture of any one of claims 3-9, wherein said bacterial strain controls said plant pest.
11. 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,
AIP082140, AIP000817, AIP060333, AIP070494, AIP015104, AIP011586, AIP010884,
AIP082287, AIP088026, AIP065969, AIP018514, AIP033041, 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 effective amount controls said plant pest or improves the agronomic trait of interest.
12. A method of controlling a plant pest in an area of cultivation comprising:
(a) planting the area of cultivation with seeds or plants susceptible to the plant pest; and
(b) applying to the seed or plant susceptible to the plant pest an effective amount of a cell, a spore, a forespore, or a combination of cells, spores and/or forespores of at least one bacterial strain comprising AIP031898, AIP023234, AIP024552, AIP035573, AIP071234, AIP080021, AIP001237, AIP050674, AIP071546, AIP049805, AIP016229, AIP081435, AIP082140,
AIP000817, AIP060333, AIP070494, AIP015104, AIP011586, AIP010884, AIP082287,
AIP088026, AIP065969, AIP018514, AIP033041, AIP092281, AIP081114, AIP046403,
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 1012 to 1016 colony forming units (CFU) per hectare.
13. 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,
AIP000817, AIP060333, AIP070494, AIP015104, AIP011586, AIP010884, AIP082287,
AIP088026, AIP065969, AIP018514, AIP033041, AIP092281, AIP081114, AIP046403,
AIP022635, AIP070925, and AIP039063, or an active variant of any thereof, wherein the active variant comprises a bacterial strain having a genome within a Mash distance of about 0.015;
wherein said effective amount comprises at least about 1012 to 1016 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.
14. The method of any one of claims 11-13, where said seed or plant is a soybean.
15. The method of any one of claims 11-14, wherein said plant pest comprises an insect pest.
16. The method of claim 15, wherein said insect pest comprises a Coleoptera insect or Hemiptera insect.
17. The method of claim 16, wherein the Coleoptera insect is a corn rootworm,
Colorado potato beetle, or a sweet potato weevil.
18. The method of any one of claims 11-17, wherein said method further comprises applying an effective amount of a biocide, wherein said effective amount of the biocide selectively controls an organism of interest while not significantly damaging said seed or plant.
19. A method of making a modified bacterial strain comprising:
(a) providing a population of a cell, a spore, a forespore, or a combination of cells, spores and/or forespores of at least one bacterial strain comprising AIP031898, AIP023234, AIP024552, AIP035573, AIP071234, AIP080021, AIP001237, AIP050674, AIP071546, AIP049805,
AIPO 16229, AIP081435, AIP082140, AIP000817, AIP060333, AIP070494, AIP015104,
AIP011586, AIPO 10884, AIP082287, AIP088026, AIP065969, AIP018514, AIP033041,
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;
(b) culturing said population in the presence of a biocide of interest; and,
(c) selecting a modified bacterial strain having an increased resistance to said biocide of interest.
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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2022225925A1 (en) 2021-04-19 2022-10-27 AgBiome, Inc. Compositions and methods for improving plant health and controlling plant disease
WO2022245786A1 (en) 2021-05-18 2022-11-24 AgBiome, Inc. Compositions and methods for improving plant health and controlling plant disease
WO2024026305A1 (en) 2022-07-26 2024-02-01 AgBiome, Inc. Compositions and methods based on pseudomonas chlororaphis for improving plant health and controlling plant disease

Citations (20)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4094097A (en) 1976-09-10 1978-06-13 Cornell Research Foundation, Inc. Method for preparing a pesticidally resistant rhizobium and agronomic composition thereof
US4306027A (en) 1976-09-10 1981-12-15 Cornell Research Foundation, Inc. Pesticidally resistant rhizobium and agronomic use thereof
WO1994010845A2 (en) 1992-11-17 1994-05-26 Ciba-Geigy Ag Synergistic microbicidal compositions
WO1995028085A1 (en) * 1994-04-18 1995-10-26 Svenska Lantmännen, riksförbund ek.för. Composition and method for controlling plant diseases
WO1996038576A1 (en) 1995-06-01 1996-12-05 Signal Pharmaceuticals, Inc. Human oligodendroglial progenitor cell line
US5614395A (en) 1988-03-08 1997-03-25 Ciba-Geigy Corporation Chemically regulatable and anti-pathogenic DNA sequences and uses thereof
US5743477A (en) 1992-08-27 1998-04-28 Dowelanco Insecticidal proteins and method for plant protection
US5792931A (en) 1994-08-12 1998-08-11 Pioneer Hi-Bred International, Inc. Fumonisin detoxification compositions and methods
US5849320A (en) 1996-06-13 1998-12-15 Novartis Corporation Insecticidal seed coating
EP1241247A1 (en) * 2001-03-13 2002-09-18 C.C.H. S.A. Isolated bacteria for the protection of plants against phytopathogenic fungi and bacteria
US6758044B2 (en) 2001-10-10 2004-07-06 Goodrich Pump & Engine Control Systems, Inc. Control system for positioning compressor inlet guide vanes
US7250561B1 (en) 1996-07-16 2007-07-31 Bayer Cropscience S.A. Chimera gene with several herbicide resistant genes, plant cell and plant resistant to several herbicides
WO2007103076A1 (en) 2006-03-01 2007-09-13 Bayer Cropscience, Lp A polymer based seed coating
US8124846B2 (en) 2000-10-30 2012-02-28 Bayer CropScience S.A.S. Herbicide-tolerant plants through bypassing metabolic pathway
CN103614312A (en) * 2013-09-22 2014-03-05 山东农业大学 Pseudomonas chloroaphis with effects of effective zea mays sheath blight control and zea mays growth promotion
US20140173979A1 (en) 2012-12-21 2014-06-26 Dow Agrosciences Llc Method of coating seed
CN104017744A (en) * 2013-11-07 2014-09-03 上海交通大学 Preparation method and application of pseudomonas chlororaphis for resisting disease and promoting growth
US20140336049A1 (en) 2009-09-03 2014-11-13 Fbsciences Holdings, Inc. Seed treatment compositions & methods
US20150033811A1 (en) 2013-08-05 2015-02-05 Specialty Fertilizer Products, Llc Micronutrient-enhanced polymeric seed coatings
RU2603281C1 (en) * 2015-12-09 2016-11-27 Федеральное бюджетное учреждение науки Государственный научный центр прикладной микробиологии и биотехнологии (ФБУН ГНЦ ПМБ) PHOSPHATE-DISSOLVING STRAIN Pseudomonas chlororaphis ssp chlororaphis Vsk-26a3, WITH FUNGICIDAL AND BACTERICIDAL ACTIVITY

Patent Citations (21)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4306027A (en) 1976-09-10 1981-12-15 Cornell Research Foundation, Inc. Pesticidally resistant rhizobium and agronomic use thereof
US4094097A (en) 1976-09-10 1978-06-13 Cornell Research Foundation, Inc. Method for preparing a pesticidally resistant rhizobium and agronomic composition thereof
US5614395A (en) 1988-03-08 1997-03-25 Ciba-Geigy Corporation Chemically regulatable and anti-pathogenic DNA sequences and uses thereof
US5743477A (en) 1992-08-27 1998-04-28 Dowelanco Insecticidal proteins and method for plant protection
WO1994010845A2 (en) 1992-11-17 1994-05-26 Ciba-Geigy Ag Synergistic microbicidal compositions
WO1995028085A1 (en) * 1994-04-18 1995-10-26 Svenska Lantmännen, riksförbund ek.för. Composition and method for controlling plant diseases
US5792931A (en) 1994-08-12 1998-08-11 Pioneer Hi-Bred International, Inc. Fumonisin detoxification compositions and methods
WO1996038576A1 (en) 1995-06-01 1996-12-05 Signal Pharmaceuticals, Inc. Human oligodendroglial progenitor cell line
US5849320A (en) 1996-06-13 1998-12-15 Novartis Corporation Insecticidal seed coating
US7935869B2 (en) 1996-07-16 2011-05-03 Bayer S.A.S. Chimeric gene with several herbicide tolerance genes, plant cell and plant resistant to several herbicides
US7250561B1 (en) 1996-07-16 2007-07-31 Bayer Cropscience S.A. Chimera gene with several herbicide resistant genes, plant cell and plant resistant to several herbicides
US8124846B2 (en) 2000-10-30 2012-02-28 Bayer CropScience S.A.S. Herbicide-tolerant plants through bypassing metabolic pathway
EP1241247A1 (en) * 2001-03-13 2002-09-18 C.C.H. S.A. Isolated bacteria for the protection of plants against phytopathogenic fungi and bacteria
US6758044B2 (en) 2001-10-10 2004-07-06 Goodrich Pump & Engine Control Systems, Inc. Control system for positioning compressor inlet guide vanes
WO2007103076A1 (en) 2006-03-01 2007-09-13 Bayer Cropscience, Lp A polymer based seed coating
US20140336049A1 (en) 2009-09-03 2014-11-13 Fbsciences Holdings, Inc. Seed treatment compositions & methods
US20140173979A1 (en) 2012-12-21 2014-06-26 Dow Agrosciences Llc Method of coating seed
US20150033811A1 (en) 2013-08-05 2015-02-05 Specialty Fertilizer Products, Llc Micronutrient-enhanced polymeric seed coatings
CN103614312A (en) * 2013-09-22 2014-03-05 山东农业大学 Pseudomonas chloroaphis with effects of effective zea mays sheath blight control and zea mays growth promotion
CN104017744A (en) * 2013-11-07 2014-09-03 上海交通大学 Preparation method and application of pseudomonas chlororaphis for resisting disease and promoting growth
RU2603281C1 (en) * 2015-12-09 2016-11-27 Федеральное бюджетное учреждение науки Государственный научный центр прикладной микробиологии и биотехнологии (ФБУН ГНЦ ПМБ) PHOSPHATE-DISSOLVING STRAIN Pseudomonas chlororaphis ssp chlororaphis Vsk-26a3, WITH FUNGICIDAL AND BACTERICIDAL ACTIVITY

Non-Patent Citations (35)

* Cited by examiner, † Cited by third party
Title
"Pesticide Specifications, Manual on development and use of FAO and WHO specifications for pesticides, FAO Plant Production and Protection Papers - 173", 2004, CROP LIFE INTERNATIONAL
ANDERSON ANNE J ET AL: "Biopesticides produced by plant-probiotic Pseudomonas chlororaphis isolates", CROP PROTECTION, vol. 105, 23 November 2017 (2017-11-23), pages 62 - 69, XP085317644, ISSN: 0261-2194, DOI: 10.1016/J.CROPRO.2017.11.009 *
ANDREWS ET AL., BIOCHEM. J., vol. 252, 1988, pages 199 - 206
BAUR ET AL., PESTICIDE SCIENCE, vol. 51, 1997, pages 131 - 152
BISHOP ET AL., BMC BIOLOGY, vol. 7, no. 1, 2009, pages 1741 - 7007,7-3
CAMMUE ET AL., J. BIOL. CHEM., vol. 267, 1992, pages 2228 - 2233
CAMPBELL ET AL., PNAS USA, vol. 110, no. 14, 2013, pages 5540 - 45
CHEMICAL ABSTRACTS, Columbus, Ohio, US; abstract no. 66733-21-9
CHOI ET AL., J. MICROBIOL. METHODS, vol. 64, 2006, pages 391 - 397
CZAPLALANG, J. ECON. ENTOMOL., vol. 83, no. 6, 1990, pages 2480 - 2485
D J HILL ET AL: "Evaluation of AtEze for suppression of fusarium wilt of chrysanthemum.", CANADIAN JOURNAL OF PLANT PATHOLOGY, vol. 21, 1 June 1999 (1999-06-01), CA, pages 193 - 197, XP055619222, ISSN: 0706-0661, DOI: 10.1080/07060669909501212 *
DATABASE WPI Week 201430, Derwent World Patents Index; AN 2014-H05173, XP002794059 *
DATABASE WPI Week 201479, Derwent World Patents Index; AN 2014-V38708, XP002794060 *
DATABASE WPI Week 201705, Derwent World Patents Index; AN 2016-77308B, XP002794061 *
DUPONT ET AL., ISMEJ, vol. 6, 2012, pages 1625 - 1628
G. BERG ET AL: "Evaluation of potential biocontrol rhizobacteria from different host plants of Verticillium dahliae Kleb", JOURNAL OF APPLIED MICROBIOLOGY., vol. 91, no. 6, 1 December 2001 (2001-12-01), GB, pages 963 - 971, XP055285946, ISSN: 1364-5072, DOI: 10.1046/j.1365-2672.2001.01462.x *
GIRIJA GANESHAN ET AL: "Pseudomonas fluorescens, a potential bacterial antagonist to control plant diseases", JOURNAL OF PLANT INTERACTIONS, vol. 1, no. 3, 1 September 2005 (2005-09-01), pages 123 - 134, XP055619133, ISSN: 1742-9145, DOI: 10.1080/17429140600907043 *
GODOY ET AL., FITOPATOL. BRAS., vol. 31, no. 1, 2006, pages 63 - 68
HANAHAN, D.: "Studies on transformation of Escherichia coli with plasmids", J. MOL. BIOL., vol. 166, 1983, pages 557 - 77
HU ET AL., PLANTMOL. BIOL., vol. 34, 1997, pages 949 - 959
JANG HOON LEE ET AL: "Nematicidal Activity of a Nonpathogenic Biocontrol Bacterium,O6", CURRENT MICROBIOLOGY, SPRINGER-VERLAG, NE, vol. 62, no. 3, 21 October 2010 (2010-10-21), pages 746 - 751, XP019886191, ISSN: 1432-0991, DOI: 10.1007/S00284-010-9779-Y *
JEFFREY D. PALUMBO ET AL: "Inhibition of Aspergillus flavus in Soil by Antagonistic Pseudomonas Strains Reduces the Potential for Airborne Spore Dispersal", PHYTOPATHOLOGY, vol. 100, no. 6, 1 June 2010 (2010-06-01), US, pages 532 - 538, XP055265142, ISSN: 0031-949X, DOI: 10.1094/PHYTO-100-6-0532 *
KONSTANTINIDIS, K.T. ET AL., PNAS USA, vol. 102, no. 7, 2005, pages 2567 - 72
LIU ET AL., PLANT BIOLOGY, vol. 91, 1994, pages 1888 - 1892
MARRONE ET AL., J. OF ECONOMIC ENTOMOLOGY, vol. 78, 1985, pages 290 - 293
MARRONE ET AL., JOURNAL OF ECONOMIC ENTOMOLOGY, vol. 78, 1985, pages 290 - 293
ONDOV ET AL., GENOME BIOLOGY, vol. 17, 2016, pages 132
PARKS ET AL., GENOME RESEARCH, vol. 25, 2015, pages 1043 - 1055
RICHTER, M. ET AL., PNAS, vol. 106, no. 45, 2009, pages 19126 - 31
STACKEBRANDT E: "Report of the ad hoc committee for the re-evaluation of the species definition in bacteriology", INF J SYST EVOL MICROBIOL., vol. 52, no. 3, 2002, pages 1043 - 7
THOMMA ET AL., PLANT BIOLOGY, vol. 95, 1998, pages 15107 - 15111
TWIZEYIMANAHARTMAN, PLANT DIS, vol. 94, 2010, pages 1453 - 1460
U. SCHELLENBERG ET AL: "A selective insecticidal protein from Pseudomonas for controlling corn rootworms", SCIENCE, NIH AUTHOR MANUSCRIPT, vol. 354, no. 6312, 3 November 2016 (2016-11-03), US, pages 634 - ^637, XP055342083, ISSN: 0036-8075, DOI: 10.1126/science.aaf6056 *
VARGHESE, N.J. ET AL., NUCLEIC ACIDS RESEARCH, 6 July 2015 (2015-07-06), pages gkv657
WANG ET AL., J. CHEM. TECHNOL. BIOTECHNOL., vol. 85, 2010, pages 775 - 778

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WO2022225926A1 (en) 2021-04-19 2022-10-27 AgBiome, Inc. Compositions and methods for improving plant health and controlling plant disease
WO2022245786A1 (en) 2021-05-18 2022-11-24 AgBiome, Inc. Compositions and methods for improving plant health and controlling plant disease
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