Classifications

• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
  • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
  • C12N1/20—Bacteria; Culture media therefor
  • C12N1/205—Bacterial isolates
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
  • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
  • C12N1/20—Bacteria; Culture media therefor
• • 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
  • A01N63/22—Bacillus
  • A01N63/23—B. thuringiensis
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
  • C12R2001/00—Microorganisms ; Processes using microorganisms
  • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
  • C12R2001/07—Bacillus
  • C12R2001/075—Bacillus thuringiensis

Definitions

• the invention relates to bacterial strains and populations for controlling plant pests and disease and/or improving an agronomic trait of interest in a plant.
• Plant diseases and 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 diseases and the pathogens that cause them and to effectively control plant pests.
• compositions and methods for controlling plant diseases and plant pests and/or for improving at least one agronomic trait of interest in a plant comprise a population of biocontrol agents or bacterial strains that control one or more pathogens that cause plant disease, one or more plant pests, and/or improve at least one agronomic trait of interest.
• the biological agents or bacterial strains can be used as an inoculant for plants.
• Methods for growing a plant susceptible to plant disease and methods and compositions for controlling plant disease and plant pests are also provided. Further provided are methods and compositions of increasing disease resistance and pest resistance in plants. Methods and compositions for improving plant health and/or improving at least one agronomic trait of interest are also provided.
• compositions and methods for improving at least one agronomic trait of interest and/or improving plant health and/or for controlling one or more plant diseases and/or plant pests are provided.
• a biological agent, biocontrol agent, bacterial strain, modified bacterial strain, modified biological agent, or modified biocontrol agent or active variant thereof, and/or a composition derived therefrom are used herein to describe a microorganism that is used to control plant pests, disease-causing plant pathogens and/or improve at least one agronomic trait of interest and/or improve plant health. //.
• biocontrol agents or bacterial strains are provided which can be used to control one or more plant pest, one or more plant disease, and/or improve at least one agronomic trait of interest and/or improve plant health.
• bacterial strains include AIP000648 (a Bacillus thuringiensis strain),
• AIP097852 (a. Bacillus thuringiensis strain), and AIP051459 (a. Bacillus thuringiensis strain).
• Cell populations comprising one or more of AIP000648, AIP097852, and AIP051459 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 AIP000648, AIP097852, and AIP051459, or an active variant of any thereof.
• AIP000648 was deposited with the Patent Depository of the National Center for Agricultural Utilization Research Agricultural Research Service, U.S. Department of Agriculture, 1815 North
• AIP097852 was deposited with the Patent Depository of the National Center for Agricultural Utilization Research Agricultural Research Service, U.S. Department of Agriculture, 1815 North
• AIP051459 was deposited with the Patent Depository of the National Center for Agricultural Utilization Research Agricultural Research Service, U.S. Department of Agriculture, 1815 North
• 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
• compositions of the invention comprise substantially pure cultures of bacterial strain AIP000648, AIP097852, or AIP051459.
• the compositions of the invention also provide progeny of substantially pure cultures of bacterial strain AIP000648, AIP097852, or AIP051459, wherein the culture has all of the physiological and morphological characteristics of AIP000648, AIP097852, or AIP051459, respectively.
• population is intended a group or collection that comprises two or more individuals (i.e., 10, 100, 1,000, 10,000, lxlO 6 , lxlO 7 , or lxlO 8 or greater) of a given bacterial strain.
• compositions are provided herein that comprise a population of at least one bacterial strain or a mixed population of individual from more than one bacterial strain.
• the population of at least one of a bacterial strain comprises a concentration of 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/ml, about
• the concentration of the bacterial strain provided herein or active variant thereof, and/or a composition derived therefrom 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.
• Non-limiting methods by which spores are formed from each of AIP000648, AIP097852, and AIP051459 (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.
• derived from means directly isolated or obtained from a particular source or alternatively having identifying characteristics of a substance or organism isolated or obtained from a particular source.
• source is an organism
• derived from means that it may be isolated or obtained from the organism itself or a culture broth, suspension, or medium used to culture or grow said organism.
• a compound or composition "derived from” or “obtainable from” means that the compound or composition may be isolated from or produced by a cell culture or a whole cell broth, or a suspension, filtrate, supernatant, fraction, or extract derived from a cell culture or a whole cell broth.
• whole broth culture or whole cell broth refers to a liquid culture containing both cells and media. If bacteria are grown on a plate, the cells can be harvested in water or other liquid, whole culture.
• whole broth culture and “whole cell broth” are used interchangeably.
• the supernatant may be diluted with another composition, such as water, buffer, fresh media, and/or a formulation.
• the diluted supernatant is still considered a supernatant of the invention.
• filtrate refers to liquid from a whole broth culture that has passed through a membrane.
• the filtrate may comprise a concentrated amount of an effective compound or metabolite compared to the concentration of the effective compound or metabolite in the whole broth culture or supernatant.
• extract refers to liquid substance removed from cells by a solvent (water, detergent, buffer, and/or organic solvent, for example) and separated from the cells by centrifugation, filtration, or other method known in the art.
• the extract may comprise a concentrated amount of an effective compound or metabolite compared to the concentration of the effective compound or metabolite in the cells prior to extraction.
• the filtrate or extract may then be diluted with another composition, such as water, buffer, fresh media, and/or a formulation. Such diluted filtrates or extracts are still considered filtrates and extracts of the invention.
• metabolite refers to a compound, substance, or byproduct of fermentation of a bacterial strain (i.e., at least one of AIP000648, AIP097852, AIP051459, or an active variant of any thereof).
• An effective compound or metabolite is a compound present in the supernatant, whole cell broth, or bacterial strain which may improve any agronomic trait of interest of a plant, or which controls a plant pest or plant pathogen that causes a plant disease, when applied to a plant of interest at an effective amount.
• a composition of the invention comprises a filtrate or extract derived from fermentation of a bacterial strain, wherein said composition comprises a concentrated amount of an effective compound or metabolite compared to the amount in a whole cell broth or supernatant of said bacterial strain, wherein the bacterial is at least one of AIP000648, AIP097852, AIP051459, or an active variant of any thereof.
• a composition of the invention comprises a diluted filtrate, diluted extract, or diluted supernatant derived from the fermentation of a bacterial strain, wherein said composition comprises a diluted amount of the effective compound or metabolite compared to the amount whole cell broth or undiluted supernatant of said bacterial strain, wherein the bacterial is at least one of AIP000648, AIP097852, AIP051459, or an active variant of any thereof.
• the diluted filtrate, diluted extract, or diluted supernatant may still comprise an effective amount of the effective compound or metabolite.
• compositions and methods described herein comprise or are derived from a bacterial strain (i.e., at least one of AIP000648, AIP097852, AIP051459, or an active variant of any thereof, or a spore or a forespore or a combination of cells, forespores or/and spores, from any one of AIP000648, AIP097852, AIP051459, or an active variant of any thereof).
• Methods comprise cultivating at least one of these bacterial strains.
• at least one of these bacterial strains is cultivated and compounds and/or compositions are obtained by isolating these compounds and/or compositions from the culture of at least one of these bacterial strains.
• At least one bacterial strain is cultivated in nutrient medium using methods known in the art.
• the bacterial strain can be cultivated by shake flask cultivation or by small scale or large scale fermentation (including but not limited to continuous, batch, fed-batch, or solid state fermentation) in laboratory or industrial fermenters performed in a suitable medium and under conditions allowing for bacterial cell growth.
• the cultivation can take place in suitable nutrient medium comprising carbon and nitrogen sources and inorganic salts, using prodedures known in the art Suitable media are available from commercial sources or are prepared according to publications well-known in the art.
• compounds, metabolites, and/or compositions can be extracted from the culture broth.
• the extract can be fractionated by chromatography.
• the extract can be further purified using methods well-known in the art.
• the extract can also be diluted using methods well-known in the art.
• compositions comprising a bacterial strain i.e., at least one of AIP000648, AIP097852, and AIP051459 or an active variant of any thereof, or a spore or a forespore or a combination of cells, forespores and/or spores, and/or a composition derived from any one of AIP000648, AIP097852, and AIP051459, or an active variant of any thereof
• a bacterial strain i.e., at least one of AIP000648, AIP097852, and AIP051459 or an active variant of any thereof, or a spore or a forespore or a combination of cells, forespores and/or spores, and/or a composition derived from any one of AIP000648, AIP097852, and AIP051459, or an active variant of any thereof
• an agriculturally acceptable carrier i.e., at least one of AIP000648, AIP097852,
• 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 a plant pest or plant disease of interest (i.e., root knot nematode (RKN)), or any other pest or disease disclosed herein 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.
• Such variants will retain the ability to control one or more plant diseases (i.e., reduce disease severity and/or reduce disease development), control one or more plant pests (e.g., insect pests or nematode pests), and/or control one or more plant pathogens.
• variants will retain the ability to control one or more fungal plant diseases and/or one or more fungal pathogens.
• variants will retain the ability to control root knot nematode (RKN).
• RKN root knot nematode
• 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 corn rootworms (e.g., Western corn rootworm), Colorado potato beetle, weevils (e.g., sweet potato weevil), or hemipteran insect pests (e.g., Southern Green stink bug).
• variants will retain the ability to control nematode pests, including RKN or soybean cyst nematode (SCN).
• Active variants of the various bacterial strains provided herein include, for example, any isolate or mutant of AIP000648, AIP097852, or AIP051459.
• mutant refers to a variant of the parental stran as well as methods for obtaining a mutant or variant in which the pesticidal activity is greater than that expressed by the parental strain.
• the "parent strain” is the original strain before mutagenesis. To obtain such mutants the parental strain may be treated with a chemical such as N-methyl-N'-nitro-N-nitrosoguanidine, ethylmethanesulfone (EMS), or by irradiation using gamma, x-ray, or UV-irradiation, or by other means well known in the art.
• the active variant contains at least mutation in at least one gene, relative to the deposited strain.
• the gene(s) may have a role in, for example, biofilm formation, motility, chemotaxis, extracellular secretion, transport (for example ABC transporter proteins), stress responses, volatiles, transcription (for example alternative sigma factors and global transcription regulators), root colonization, ability to stimulate induced systemic resistance in a plant, and/or secondary metabolism including synthesis of lipopeptides, polyketides, macromolecular hydrolases (for example proteases and/or carbohydrases), and/or antimicrobial compounds including antibiotics.
• transport for example ABC transporter proteins
• stress responses for example alternative sigma factors and global transcription regulators
• transcription for example alternative sigma factors and global transcription regulators
• root colonization ability to stimulate induced systemic resistance in a plant, and/or secondary metabolism including synthesis of lipopeptides, polyketides, macromolecular hydrolases (for example proteases and/or carbohydrases), and/or antimicrobial compounds including antibiotics.
• Secondary metabolism refers to both non-ribosomal and ribosomal synthesis of antimicrobial compounds, including cyclic lipopeptides, polyketides, iturins, bacteriocins (for example plantazolicin and amylocyclicin) and dipeptides (for example bacilysin).
• An example of an active variant is a cell of bacterial strain AIP000648, AIP097852, or
• AIP051459 wherein the cell further comprises a mutation in the swrA gene that results in loss of function.
• the swrA mutation which affects biofilm formation (Kearns et al, Molecular Microbiology (2011) 52(2): 357-369) may result in an active variant of a strain of the invention which has enhanced ability to control a plant pest or improve an agronomic trait of interest of a plant.
• genes that are involved in biofilm formation such as sfp, epsC, degQ, and a plasmid gene called rapP (see for example, McLoon ei al, J of Bacteriology, (2011) 193(8): 2027-2034), may also be mutated in an active variant of a bacterial strain of the invention.
• 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; herbicides; insecticides, 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 AIP000648, AIP097852, AIP051459, or an active variant of any thereof.
• 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 (i.e., bacterial cell or spore) to survive and reproduce following exposure to a dose of the biocide (e.g, herbicide, 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, 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 disease and further are able to grow in the presence of at least one biocide.
• Recombinant bacterial strains having resistance to an herbicide, 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;
• 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, 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, 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, fungicide, pesticide, or other crop protection chemical. Thus, their identification is readily possible by comparison with characteristics of the known sensitive strain.
• the active variants of the bacterial strain(s) AIP000648, AIP097852, AIP051459 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.
• 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%.
• 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.
• an active variant of bacterial strain AIP000648, AIP097852, and AIP051459 disclosed herein includes a bacterial stain 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 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 disease severity, reducing plant disease development, and/or increasing pest resistance.
• 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 AIP000648, AIP097852, or AIP051459) on the basis of 16S rDNA sequence identity. See Stackebrandt E, et al, "Report of the ad hoc committee for the re-evaluation of the species definition in bacteriology," IntJ Syst Evol Microbiol. 52(3): 1043-7 (2002) regarding use of 16S rDNA sequence identity for determining relatedness in bacteria.
• the at least one strain 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 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 disease severity, increasing pest resistance, and/or reducing plant disease development.
• 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).
• the Mash distance estimates the mutation rate between two sequences directly from their MinHash sketches (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 Average Nucleotide Identity method
• Active variants of the bacterial strain(s) disclosed herein include strains that are closely related to any of AIP000648, AIP097852, or AIP051459 on the basis of the Minhash (Mash) distance between complete genome DNA sequences.
• 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 0.001, 0.0025, 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.
• 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.
• 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 0.001, 0.0025, 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 Illumina 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.
• 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 Illumina 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.
• a Mash distance of 0.015 between two genomes means the expected mutation rate is 0.015 mutations per homologous position.
• Active variants of the bacteria identified by such methods will retain the ability 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 disease severity, reducing plant disease development, and/or increasing pest resistance.
• the bacteria strains provided herein i.e., AIP000648, AIP097852, AIP051459, or active variant of any thereof, or a spore or a forespore or a combination of cells, forespores, and/or spores, and/or a composition derived, from any one of AIP000648, AIP097852, AIP051459, or an active variant of any thereof
• Such formulations will comprise the bacteria provided herein or an active variant thereof, and/or a composition derived therefrom 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), suspo-emulsions (SE), flowable concentrates for seed treatment (FS), oil dispersions (OD), water-dispersible granules (WG), granules (GR), capsule concentrates (CS), water-dispersible granules (WG), granules (GR), block baits (BB), water-soluble granules (SG), and mixed formulations of CS and SC (ZC).
• SL water-soluble liquids
• EC emulsifiable concentrates
• EW emulsions in water
• SC suspension concentrates
• SE suspo-emulsions
• FS flowable concentrates for seed treatment
• OD oil dispersions
• WG water-dispersible granules
• GR granules
• CS capsule concentrates
• BB block baits
• 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
• 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
• 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.
• Non-limiting solvents are, for example, aromatic hydrocarbons, such as xylene, toluene or alkylnaphthalenes, for example, chlorinated aromatic or aliphatic hydrocarbons such as chlorobenzenes, chloroethylenes or methylene chloride, aliphatic hydrocarbons such as cyclohexane or paraffin
• 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.
• aliphatic hydrocarbons such as cyclohexane
• paraffins petroleum fractions, mineral and vegetable oils
• alcohols such as methanol, ethanol, isopropanol, butanol or glycol
• ketones such as acetone, methyl ethyl ketone, methyl isobutyl ketone or cyclohexan
• 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.
• 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 naphthalene sulphonic 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, alkylsulphonates, alkyl sulphates, arylsulphonates, protein hydrophos
• 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 al., 1997, Pesticide Science 51 : 131-152).
• compositions and formulations disclosed herein can comprise an amount of a bacterial strain, such as AIP000648,
• AIP097852, AIP051459, or active variant of any thereof or a spore or a forespore or a combination of cells, forespores or/and spores, and/or can comprise an amount of a composition derived from any one of AIP000648, AIP097852, AIP051459, or an active variant of any thereof.
• 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 CFU/gram to about 10 CFU/gram, about 10 CFU/gram to about 10 CFU/gram, about 10 CFU/gram to about 10 10 CFU/gram, about 10 10 CFU/gram to about 10 11 CFU/gram, or about 10 11 CFU/gram to about 10 12 CFU/g ram.
• 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, at least about 10 4 CFU/gram.
• 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, cell paste, wettable granule, or freeze-dried formulation.
• the bacterial strain can occur in a liquid formulation.
• Liquid formulations can comprise an amount of a cell of a bacterial strain, such as AIP000648, AIP097852, AIP051459, 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 AIP000648, AIP097852, AIP051459, or an active variant of any thereof, and/or a composition derived therefrom.
• the amount of bacterial strain, or active variant thereof, and/or a composition derived therefrom, 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/ mL, or about 10 11 CFU/ mL to about 10 12 CFU/ mL or at
• Dry formulations such as cell pastes, wettable powders, and spray dried formulations can comprise a bacterial strain, such as AIP000648, AIP097852, AIP051459, or active variant of any thereof, or a spore or a forespore or a combination of cells, forespores or/and spores of any thereof, and/or can comprise a composition derived from any one of AIP000648, AIP097852, AIP051459, or an active variant of any thereof.
• the amount of the bacterial strain in the cell paste or wettable powder 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 CFU/gram to about 10 CFU/gram, about 10 CFU/gram to about 10 CFU/gram, about 10 CFU/gram to about 10 10 CFU/gram, about 10 CFU/gram to about 10 10 11 CFU/gram, or about 10 11 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.
• 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 at least one bacterial strain, such as AIP000648, AIP097852, AIP051459, or active variant of any thereof, or a spore or a forespore or a combination of cells, forespores or/and spores, and/or can comprise a composition derived from any one of AIP000648, AIP097852, AIP051459, or an active variant of any thereof, wherein said bacterial strain or the active variant thereof is present on the seed at about 10 5 CFU/seed to about 10 7 CFU/seed, at about 10 4 CFU/seed to about 10 8 CFU/seed, at about 10 4 CFU/seed to about 10 5 CFU/seed, at about 10 5 CFU/seed to about 10 6 CFU/seed, at about 10 6 CFU/seed to about 10 7 CFU/seed, or at about 10 7 CFU/seed to about 10 8
• a seed coating can further comprise at least at least one nutrient, at least one herbicide or at least one pesticide, or at least one biocide. See, for example, US App Pub. 20040336049, 20140173979, and 20150033811.
• compositions comprising a whole cell broth, supernatant, filtrate, or extract derived from at least one of bacterial strain AIP000648, AIP097852, AIP051459, or an active variant of any thereof, wherein an effective amount of the composition improves an agronomic trait of interest of a plant or controls a plant pest or plant pathogen that causes disease.
• the composition contains effective compound(s), metabolite(s), and/or protein(s) which improve an agronomic trait of interest of a plant or controls a plant pest or plant pathogen that causes disease.
• the supernatant refers to the liquid remaining when cells are grown in broth or are harvested in another liquid from an agar plate and are removed by centrifugation, filtration, sedimentation, or other means well known in the art.
• the supernatant may be further concentrated to produce a filtrate.
• the filtrate may comprise a concentrated amount of an effective compound or metabolite compared to the concentration of the effective compound or metabolite in the supernatant or whole cell broth.
• the supernatant, filtrate, or extract may be processed to a wettable powder, spray dried formulation, and/or seed coating.
• the supernatant, filtrate, or extract may be concentrated (e.g., water is removed) but remain in a liquid formulation.
• the composition described above can be applied alone or in combination with another substance, in an effective amount to control a plant pest or improve an agronomic trait of interest of a plant.
• 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.
• stable is intened that the formulation retains viable bacteria and/or retains an effective amount of a biologically active bacterial population.
• Biological activity refers to the ability of the formulation to improve an agronomic trait of interest or control a plant pest or plant pathogen that causes a plant disease.
• the stable formlation 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 variant 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 bacterial strain or active variant thereof, and/or a composition derived therefrom 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, active variant thereof, and/or a composition derived therefrom can be used to significantly improve at least one agronomic trait of interest (i.e, reduce disease caused by a plant pathogen or reduce susceptibility to plant pests, such as insect and nematode pests).
• the bacterial strain provided herein, active variant thereof, and/or a composition derived therefrom 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; [Lactic acid, n- butyl ester, (S)]; Butyl lactate [Lactic acid, n-butyl ester]; Butyl stearate [Octadecanoic acid, butyl ester]; Calcareous shale; Calcite (Ca(Co 3 )); Calcium acetate; Calcium a
• Dilaurin [Dodecanoic acid, diester with 1,2,3-propanetriol]; Dipalmitin [Hexadecanoic acid, diester with 1,2,3-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 1,2,3-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;
• 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;
• 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, active variants thereof, and/or compositions derived therefrom provided herein can be employed with any plant species to improve an agronomic trait of interest.
• Agonomic 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, 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, pathogen resistance, pest resistance, 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
• the bacterial strain, active variant thereof, and/or a composition derived therefrom provided herein can be employed to decrease or reduce the level of a plant pest.
• pests includes but is not limited to, insects, fungi, bacteria, nematodes, acarids, protozoan pathogens, animal -parasitic liver flukes, and the like.
• pest resistance is intended that the bacterial strain, active variant thereof, and/or a composition derived therefrom 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.
• the bacterial strain, active variant thereof, and/or a composition derived therefrom provided herein can be employed with any plant species susceptible to a plant disease.
• a plant susceptible to a plant disease is meant that the causative pathogen(s) of the plant disease are able to infect the plant.
• a plant susceptible to a plant disease can be susceptible to a plant disease caused by an insect or nematode pest as disclosed elsewhere herein.
• plant species of interest include, but are not limited to, corn (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 annuus), safflower (Carthamus tinctorius), wheat ⁇ Triticum aestivum), soybean ⁇ Glycine max), tobacco (Nicotiana tabacum), potato ⁇ Solarium tuberosum), peanuts ⁇ A
• 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 Locopersicon esculentum
• lettuce e.g., Lactuca sativa
• green beans Phaseolus vulgaris
• lima beans 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 (Chamaecyparis nootkatensis).
• pines such as loblolly pine (Pinus taeda), slash pine (P
• plants of the present invention are crop plants (for example, corn, 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 com, 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 nematode or nematode pests.
• nematode encompasses eggs, larvae, juvenile, and mature forms of nematodes.
• Bacterial strains or active variants thereof can be tested for pesticidal activity on eggs, larvae, juvenile, and mature forms of nematodes.
• Nematodes include plant parasitic nematodes such as root-knot, cyst, and lesion nematodes, including species of Me loidogyne 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); nematode species of Ditylenchus such as Ditylenchus destructor and Ditylenchus dipsaci; species of Pratylenchus such as the Cob Root-Lesion Nematode ⁇ Pratylenchus penetrans), Chrysanthemum Root-Lesion Nematode ⁇ Pratylenchus fallax), Pratylenchus coffeae, Pratylenchus loosi and Walnut
• Heterodera avenae (cereal cyst nematode); species of Aphelenchoides such as the Rice White-tip Nematode ⁇ Aphelenchoides besseyi), Aphelenchoides ritzemabosi and Aphelenchoides fragariae; species of Aphelenchus such as Aphelenchus avenae; species of Radopholus , such as the Burrowing-Nematode ⁇ Radopholus similis); species of Tylenchulus such as Tylenchulus semipenetrans; species of
• Rotylenchulus such as Rotylenchulus reniformis
• nematodes living in trees such as Bursaphelenchus xylophilus and the Red Ring Nematode ⁇ Bursaphelenchus cocophilus
• Globodera spp. particularly members of the cyst nematodes, including but not limited to Globodera rostochiensis, Globodera pallida (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 the causal nematode of Pine Wilt Disease ⁇ Bursaphelenchus xylophilus).
• the bacterial strains or active variants thereof i.e., AIP000648,
• the bacterial strain or modified biological agents disclosed herein i.e., AIP000648, AIP097852, AIP051459, 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 AIP000648, AIP097852, AIP051459, or an active variant of any thereof
• a composition derived from any one of AIP000648, AIP097852, AIP051459, or an active variant of any thereof control at least one, at least two, or all Meloidogyne spp., Heterodera spp., Ditylenchus spp., Aphelencoides spp., Tylenchulus spp., Rotylenchulus spp., Globodera spp.,
• Pratylenchus spp. Helicotylenchus spp., Radopholus spp., Ditylenchus spp., Bursaphelenchus spp., and Xiphinema spp.
• the bacterial strain or modified biological agents disclosed herein i.e., AIP000648, AIP097852, AIP051459, 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 AIP000648, AIP097852, AIP051459, or an active variant of any thereof
• a composition derived from any one of AIP000648, AIP097852, AIP051459, or an active variant of any thereof control at least one, at least two, or all Meloidogyne spp.
• the bacterial strains or modified bacterial strains disclosed herein i.e., AIP000648, AIP097852, AIP051459, 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 AIP000648, AIP097852, AIP051459, or an active variant of any thereof
• AIP000648, AIP097852, AIP051459, or an active variant of any thereof control Meloidogyne incognita, Meloidogyne hapla, Meloidogyne javanica, and/ 'or Meloidogyne enterolobii.
• 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.
• 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.
• the term “instar” is used to denote the developmental stage of the larval or nymphal forms of insects.
• Insect pests include insects selected from the orders Coleoptera, Lepidoptera, Hemiptera, Diptera, Hymenoptera, Lepidoptera, Mallophaga, Homoptera, Hemiptera, Orthroptera, Thysanoptera, Trombidiformes, Dermaptera, Isoptera, Anoplura, Siphonaptera, Trichoptera, etc.
• Insect pests of the order Coleoptera include, but are 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.
• 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 corn rootworm ⁇ Diabrotica virgifera virgifera LeConte)); chafers and other beetles from the family Scaribaeidae (e.
• insect pests include Coleoptera pests of the corn rootworm complex:
• the insect pest is Western corn rootworm, Diabrotica virgifera virgifera; northern corn rootworm, D. barberi; Southern corn rootworm or spotted cucumber beetle, Diabrotica undecimpunctata howardi; and the Mexican corn rootworm, D. virgifera zeae.
• the insect pest is Western corn rootworm,
• Diabrotica virgifera virgifera virgifera.
• Insect pests that can be controlled with the compositions and methods disclosed herein further include, but are not limited to, insects of the order Lepidoptera, e.g. Achoroia grisella, Acleris gloverana, Acleris variana, Adoxophyes orana, Agrotis ipsilon, Alabama argillacea, Alsophila pometaria, Amyelois transitella, Anagasta kuehniella, Anarsia lineatella, Anisota senatoria, Antheraea pernyi, Anticarsia gemmatalis, Archips spp ., Argyrotaenia spp., Athetis mindara, Bombyx mori, Bucculatrix thurberiella, Cadra cautella, Choristoneura spp., Cochylls hospes, Colias eurytheme, Corcyra cephalonica, Cydia latiferreanus , Cydia pomonella
• Thaurnstopoea pityocampa Tinsola bisselliella, Trichoplusia hi, Tuta absoluta, Udea rubigalis, Xylomyges curtails, and Yponomeuta padella.
• the methods and compositions provided herein can also be used against insect pests of the order Hemiptera including, but not limited to, Lygus spp. including Lygus hesperus, Lygus lineolaris, Lygus pratensis, Lygus rugulipennis, and Lygus pabulinus, Calocoris norvegicus, Orthops compestris, Plesiocoris rugicollis, Cyrtopeltis modestus, Cyrtopeltis notatus, Spanagonicus albofasciatus,
• Lygus spp. including Lygus hesperus, Lygus lineolaris, Lygus pratensis, Lygus rugulipennis, and Lygus pabulinus
• Calocoris norvegicus Orthops compestris, Plesiocoris rugicollis, Cyrtopeltis modestus, Cyrtopeltis notatus, Spanagonicus albofasciat
• Thysenoptera can also be used against insect pests of the order Thysenoptera including, but not limited to, thrips species, including Frankliniella spp., for example Western Flower thrips ⁇ Frankliniella occidentalis (Pergande)); Thrips spp., for example Thrips tabaci; Scirtothrips spp., for example Scirtothrips dorsalis; Klambothrips spp., for example Klambothrips myopori; Echinothrips spp., for example Echinothrips americanus; and Megalurothrips spp., for example Megalurothrips usitatus.
• thrips species including Frankliniella spp., for example Western Flower thrips ⁇ Frankliniella occidentalis (Pergande)
• Thrips spp. for example Thrips tabaci
• Scirtothrips spp. for example Scirtothrips dor
• the methods and compositions provided herein can also be used against insect pests of the order Trombidiformes including, but are not limited to, plant feeding mites, including six-spooted spider mite ⁇ Eutetranychus sexmaculatus), Texas citrus mite ⁇ Eutetranychus banksi), Citrus red mite (Panonychus citri), European red mite ⁇ Panonychus ulmi), McDaniel mite ⁇ Tetranychus mcdanieli), Pacific spider mite ⁇ Tetranychus pacificus), Strawberry spider mite ⁇ Tetranychus urticae), Spruce spider mite ⁇ Oligonychus ununguis), Sugi spider mite ⁇ Oligonychus nondonensisi), and Tetranychus evansi.
• plant feeding mites including six-spooted spider mite ⁇ Eutetranychus sexmaculatus), Texas citrus mite ⁇ Eutetranychus
• 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 com rootworm; Epitrix cucumeris, potato flea beetle; Chaetocnema confinis, sweet potato flea beetle; Hypera postica, alfalfa weevil; Anthonomus
• Insect pests of the present invention can further include those of the order Acari including, but not limited to, mites and ticks.
• 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;
• western corn rootworm e.g., Diabrotica virgifera virgifera
• northern com rootworm e.g., Diabrotica longicornis barberi
• southem com rootworm
• Rhopalosiphum maidis com leaf aphid; Anuraphis maidiradicis, com root aphid; Myzus 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, seedcom 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;
• Rhopalosiphum maidis com 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
• ( 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.
• Bacterial strains or active variants thereof can be tested for pesticidal activity of compositions of the invention 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 bacterial strains AIP000648, AIP097852, AIP051459, or an active variant thereof, and/or a composition derived therefrom, or a spore, or a forespore or a combination of cells, forespores and/or spores control an insect or nematode pest.
• the plant pathogen disclosed herein is a nematode pest.
• the bacterial strains AIP000648, AIP097852, AIP051459, or an active variant thereof, and/or a composition derived therefrom, or a spore, or a forespore or a combination of cells, forespores and/or spores can control root-knot nematode, Western corn rootworm, Colorado potato beetle, sweet potato weevil, and/or Southern green stink bug.
• the bacterial strains AIP000648, AIP097852, AIP051459, or an active variant thereof, and/or a composition derived therefrom can have activity against root-knot nematode, Western corn rootworm, Colorado potato beetle, sweet potato weevil, and/or Southern green stink bug.
• AIP000648, AIP097852, AIP051459, or an active variant thereof, and/or a composition derived therefrom have activity against root-knot nematodes.
• the bacterial strains provided herein are those that target one or more plant pathogens.
• Plant pathogens of the invention include, but are not limited to, viruses or viroids, bacteria, insects, nematodes, oomycetes, fungi, and the like.
• the various bacterial strains provided herein target one or more fungal or opathogens that cause plant disease.
• plant diseases which can be treated or reduced or prevented include, but are not limited to, plant diseases caused by fungi, oomycetes, viruses or viroids, bacteria, insects, nematodes, protozoa, and the like.
• fungal plant diseases 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, Corn Stalk Rot, Brown Rust, Black Rust, Yellow Rust, Wheat Rust, Rust, Apple Scab, Verticillium Wilt, Fire Blight, and Brown Rot, to name a few.
• ASR Asian Soybean Rust
• gray mold leaf spot
• Frogeye Leaf Spot Early Blight
• Damping off complex Brown Patch
• Brown Patch black scurf
• root rot belly rot
• Sheath blight Powdery Mildew
• Anthracnose leaf spot Downy Milde
• any of the bacterial strain provided herein or active variant thereof, and/or a composition derived therefrom can have antifungal activity against one, two, three, four, five, or more fungal pathogens and/or fungal diseases described herein.
• a fungal pathogen can be, but is not limited to, a fungus selected from the group consisting of Botrytis spp., Botrytis cinerea, 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 irregular e, Pythium sylvaticum, Py
• the fungal pathogen is selected from the group consisting of Botrytis cinerea, Cercospora sojina, Alternaria solani, Rhizoctonia solani, Erysiphe necator, Podosphaera xanthii, Colletotrichum cereale, 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
• the fungal pathogen is Phakopsora sp., including Phakopsora pachyrhizi and/or Phakopsora meibomiae.
• the bacterial strains or active variants thereof i.e., AIP000648,
• the bacterial strains AIP000648, AIP097852, AIP051459, or active variants thereof, or an active variant of any thereof, or a spore, or a forespore or a combination of cells, forespores and/or spores, and/or a composition derived from any one of AIP000648, AIP097852, AIP051459, or an active variant of any thereof control at least one fungal disease and/or fungal pathogen.
• the bacterial strains or active variants thereof i.e., AIP000648,
• the bacterial strain or active variant thereof, and/or a composition derived therefrom control at least one, two, three, four, five, or more fungal diseases selected from the group consisting of Asian Soybean Rust, gray mold, Frogeye Leaf Spot, Early Blight, Damping off complex, Brown Patch, Powdery Mildew, Anthracnose leaf spot, Downy Mildew, Pythium Blight, Late Blight, Fusarium Head Blight, SDS, and Apple Scab.
• the bacterial strains or active variants thereof i.e., AIP000648,
• Tritici Erysiphe necator, Podosphaera xanthii, Golovinomyces cichoracearum, Erysiphe lager stroemiae, Sphaerotheca pannosa, Colletotrichum cereale, Apiognomonia errabunda, Apiognomonia veneta, Colletotrichum gloeosporiodes, Discula fraxinea, Plasmopara viticola, P seudoperonospora cubensis, Peronospora belbahrii, Bremia lactucae, Peronospora lamii, Plasmopara obduscens, Pythium cryptoirregulare, Pythium aphanidermatum, Pythium irregulare, Pythium sylvaticum, Pythium myriotylum, Pythium ultimum, Phytophthora capsici, Phytophthora nicotianae, Phy
• the bacterial strains or active variants thereof control at least one, two, three, four, five, or more fungal pathogens selected from the group consisting of Botrytis cinerea, Cercospora sojina, Alternaria solani, Rhizoctonia solani, Erysiphe necator, Podosphaera xanthii, Colletotrichum cereale, Plasmopara viticola, Peronospora belbahrii, Pythium aphanidermatum, Pythium sylvaticum, Pythium myriotylum, Pythium
• the bacterial strains or active variants thereof i.e., AIP000648,
• the bacterial strain or modified biological agents disclosed herein i.e., AIP000648, AIP097852, AIP051459, 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 AIP000648, AIP097852, AIP051459, or an active variant of any thereof
• the bacterial strains or modified bacterial strains disclosed herein i.e., AIP000648, AIP097852, AIP051459, 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 AIP000648, AIP097852, AIP051459, or an active variant of any thereof) control Phakopsora pachyrhizi.
• a plant disease comprising applying to a plant having a plant disease or at risk of developing a plant disease an effective amount of at least one bacterial strain provided herein or an active variant thereof, and/or a composition derived therefrom wherein the bacterial strain controls a plant pathogen that causes the plant disease.
• the plant pathogen may be a fungal pathogen.
• methods of controlling a plant pest or preventing plant damage caused by a plant pest comprising applying to a plant an effective amount of at least one bacterial strain provided herein or an active variant thereof, and/or a composition derived therefrom wherein the bacterial strain controls a plant pest.
• the plant pest may be a nematode or 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 disease 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, and/or a composition derived therefrom wherein the bacterial strain controls the plant pest, such as a nematode pest or an insect pest.
• the plant damage is caused by an insect pest, such as a coleopteran pest or a hemipteran pest.
• the plant damage is caused by a nematode pest.
• the bacterial strain provided herein or active variant thereof, and/or a composition derived therefrom may comprise at least one of AIP000648, AIP097852, AIP051459, 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 AIP000648, AIP097852, AIP051459, or an active variant or any thereof.
• the effective amount of the bacterial strain or active variant thereof, and/or a composition derived therefrom 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 11 CFU per hectare.
• the composition is derived from a bacterial strain provided herein or active variant thereof which may comprise a cell of at least one of AIP000648, AIP097852, AIP051459, 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 AIP000648, AIP097852, AIP051459, or an active variant of any thereof.
• the bacterial strain provided herein or an active variant thereof, and/or a composition derived therefrom is an agent that treats or prevents one, two, three, four, five or more plant diseases, infections, or infestations by plant pests.
• the bacterial strain provided herein or an active variant thereof, and/or a composition derived therefrom is an antifungal agent that treats or prevents one, two, three, four, five or more fungal plant diseases or infections caused by insect pests or nematode pests.
• the insect pests are hemipteran or coleopteran pests.
• the nematode pests are Me loidogyne spp., Heterodera spp., Ditylenchus spp.,
• the nematode pests are RKN nematodes or SCN nematodes.
• the bacterial strain provided herein or an active variant thereof, and/or a composition derived therefrom can be employed with any plant species susceptible to a plant disease of interest and/or susceptible to a plant pest of interest.
• Examples of diseases causes by the fungal pathogens described herein are provided in Table 1.
• Table 1 shows that Bortrytis cinerea causes gray mold on all flowering crops. Therefore, a bacterial strain provided herein or active variant thereof, and/or a composition derived therefrom 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.
• Table 1 shows that Rhizoctonia solani causes Damping off complex in corn, Damping off complex in soybean, Brown Patch in turf, and Damping off complex in ornamentals.
• a bacterial strain provided herein or active variant thereof, and/or a composition derived therefrom 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 cereale, Apiognomonia errabunda,
• Apiognomonia veneta, Colletotrichum gloeosporiodes, Discula fraxinea cause Anthracnose leaf spot. Therefore, a bacterial strain provided herein or active variant thereof, and/or a composition derived therefrom that controls one or more of Colletotrichum cereale, 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.
• ASR Asian Soybean Rust
• methods of treating or preventing Asian Soybean Rust comprising applying to a plant having ASR or at risk of developing ASR an effective amount of at least one bacterial strain provided herein or an active variant thereof, and/or a composition derived therefrom comprising AIP000648, AIP097852, AIP051459, 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 AIP000648, AIP097852, AIP051459, or an active variant of any thereof.
• the effective amount of the bacterial strain provided herein or an active variant thereof, and/or a composition derived therefrom comprises at least about 10 12 to 10 16 CFU per hectare and wherein the bacterial strain provided herein or active variant thereof, and/or a composition derived therefrom controls a plant pathogen that causes ASR.
• an effective amount of at least one bacterial strain provided herein or active variant thereof, and/or a composition derived therefrom provided herein is used as a foliar application on a plant to treat or prevent ASR.
• the bacterial strain provided herein or an active variant thereof, and/or a composition derived therefrom or modified bacterial strain provided herein can be employed with any plant species susceptible to ASR.
• a plant susceptible to Asian Soybean Rust (ASR) is meant that the causative pathogen(s) of ASR are able to infect the plant.
• soybean Glycine max
• common bean Phaseolus vulgaris
• lima beans Phaseolus limensis
• butter beans Phaseolus lunatus
• cowpeas Vigna unguiculata
• pigeon peas Cajanus cajan
• yam beans such as jicama (Pachyrhizus erosus).
• a soybean plant is employed.
• the bacterial strain provided herein or an active variant thereof, and/or a composition derived therefrom controls one or more fungi that causes ASR (such as, for example, Phakopsora).
• ASR is caused by one or more fungal pathogens of the genus Phakopsora.
• the fungal pathogens that cause ASR are Phakopsora pachyrhizi or Phakopsora meibomiae.
• the ASR pathogen is well adapted for long-distance dispersal, because the spores can be readily carried by the wind, making it an ideal means for introduction to new, rust-free regions.
• the primary means of dissemination are spores, which can be carried by wind or splashed rain. These pathogens are obligate pathogens surviving and reproducing only on live hosts.
• the first symptoms are light-brown polygonal lesions of 2 to 5 mm on the adaxial leaf surface. These lesions develop into volcano-shaped lesions known as pustules that appear on the abaxial surface of the leaf, where uredospores are produced.
• the bacterial strain provided herein or an active variant thereof, and/or a composition derived therefrom controls Phakopsora pachyrhizi. In yet further embodiments, the bacterial strain provided herein or active variant thereof, and/or a composition derived therefrom controls
• Phakopsora meibomiae Phakopsora meibomiae.
• Assays to measure such activity are disclosed elsewhere herein.
• the bacterial strains provided herein or active variants thereof controls one or more nematode pests.
• the bacterial strains 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 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 and/or a composition derived therefrom 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 and/or a composition derived therefrom may show a reduced severity or reduced development of disease or damage in the presence of plant pests by a statistically significant amount.
• prevent 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 pathogens or 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 pathogen or pest growth and/or an improvement in the diseased plant height, weight, number of leaves, root system, or yield.
• the term refers to the improvement in a diseased plant 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.
• 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.
• 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, and/or a composition derived therefrom may show a reduced disease severity or reduced disease development in the presence of plant pathogens or plant pest by a statistically significant amount.
• a reduced disease severity or reduced disease development 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, and/or a composition derived therefrom may show a reduced disease severity or reduced disease development in the presence of plant pathogen 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%,
• a plant, plant part, or area of cultivation treated with the bacterial strain provided herein or an active variant thereof, and/or a composition derived therefrom 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, and/or a composition derived therefrom may show a 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%
• 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) 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.
• the insecticidal activity is against lepidopteran species. In one embodiment, the insecticidal activity is against a coleopteran species. In one embodiment, the insecticidal activity is against a hemipteran species. In some embodiments, the insecticidal activity is against one or more insect pests, such as the Western corn rootworm, the Colorado potato beetle, the sweet potato weevil, or the Southern green stink bug.
• the bacterial strains, active variants thereof, and/or a composition derived therefrom 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 : 15107-151 1 1, herein incorporated by reference.
• in vitro pesticidal assays include, for example, the addition of varying
• compositions and methods for inducing disease resistance in a plant to plant pathogens are also provided. Accordingly, the compositions and methods are also useful in protecting plants against fungal pathogens, viruses, nematodes, and insects.
• methods of inducing disease resistance against a plant pathogen comprising applying to a plant that is susceptible to a plant disease caused by the plant pathogen an effective amount of at least one bacterial strain provided herein or active variant thereof, and/or a composition derived therefrom.
• Also provided herein are methods of inducing disease resistance against a plant pest or inducing resistance to infection by a plant pest comprising applying to a plant that is susceptible to a plant disease or infection caused by the plant pest an effective amount of at least one bacterial strain provided herein or active variant thereof, and/or a composition derived therefrom.
• the bacterial strain provided herein or active variant thereof, and/or a composition derived therefrom may comprise at least one of AIP000648, AIP097852, AIP051459, 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 AIP000648, AIP097852, AIP051459, or an active variant of any thereof.
• the bacterial strain provided herein or active variant thereof, and/or a composition derived therefrom promotes a defensive response to the pathogen that causes the plant disease.
• the effective amount of the bacterial strain provided herein or active variant thereof comprises at least about 10 5 to 10 12 CFU per hectare. In some embodiments, 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, and/or a composition derived therefrom to the plant, but prior to pathogen challenge and/or after pathogen challenge of the plant treated with the bacterial strain provided herein or active variant thereof, and/or a composition derived therefrom.
• a defensive response in the plant can be triggered after applying the bacterial strain provided herein or active variant thereof, and/or a composition derived therefrom 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, and/or a composition derived therefrom.
• the bacterial strain provided herein or active variant thereof, and/or a composition derived therefrom induces resistance to one, two, three, four, five or more plant pathogens and/or pests described herein. In other methods, the bacterial strain provided herein or active variant thereof, and/or a composition derived therefrom induces resistance to one, two, three, four, five or more fungal plant pathogens, insect pests or nematode pests described herein.
• disease resistance is intended that the plants avoid the disease symptoms that result from plant-pathogen interactions. That is, pathogens are prevented from causing plant diseases and the associated disease symptoms, or alternatively, the disease symptoms caused by the pathogen are minimized or lessened as compared to a control.
• pest resistance is intended that the plants avoid the symptoms that result from infection 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, and/or a composition derived therefrom may comprise at least one of AIP000648, AIP097852, AIP051459, 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 AIP000648, AIP097852, AIP051459, or an active variant of any thereof.
• the effective amount of the bacterial strain provided herein or active variant thereof comprises at least about 10 5 to 10 12 CFU per hectare. In some embodiments, 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 composition is derived from a bacteria strain provided herein or active variant thereof which may comprise a cell of at least one of AIP000648, AIP097852, AIP051459, 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 AIP000648, AIP097852, AIP051459, or an active variant of any thereof.
• 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 pathogen (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, and/or a composition derived therefrom 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 strain provided herein or active variant thereof, and/or a composition derived therefrom are applied in an effective amount.
• An effective amount of a bacterial strain provided herein or active variant thereof, and/or a composition derived therefrom is an amount sufficient to control, treat, prevent, inhibit the pathogen or pest that causes a plant disease, and/or reduce plant disease severity or reduce plant disease development.
• the effective amount of the bacterial strain provided herein or active variant thereof, and/or a composition derived therefrom 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.
• the rate of application of the bacterial strain provided herein or active variant thereof, and/or a composition derived therefrom may vary according to the pathogen or pest being targeted, the crop to be protected, the efficacy of the bacterial strain provided herein or active variant thereof, and/or a composition derived therefrom, the severity of the disease, the climate conditions, the agronomic trait of interest to improve, and the like.
• 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 Ixl0 12 lxl0 13 , about lxlO 14 , lxlO 15 , about lxlO 16 , or about lxlO 17 colony forming units (CFU) per hectare.
• CFU colony forming units
• the rate of bacterial strain provided herein or active variant thereof application is at least lxl0 7 to at least about lxlO 12 CFU/hectare.
• the bacterial strain provided herein or active variant thereof applied comprises the strain deposited as AIP000648, AIP097852, AIP051459, 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 AIP000648, AIP097852, AIP051459, or an active derivative of any thereof.
• the applied composition is derived from a bacterial strain or active variant thereof comprising a strain deposited as AIP000648, AIP097852, AIP051459, 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 AIP000648, AIP097852, AIP051459, or an active derivative of any thereof.
• the applied composition may be a substantially pure culture, whole cell broth, supernatant, filtrate, extract, or compound derived from a bacterial strain of the invention or an active variant thereof.
• the applied composition may be applied alone or in combination with another substance, in an effective amount for controlling a plant pest or for improving an agronomic trait of interest in a plant.
• An effective amount of the applied composition is the quantity of microorganism cells, supernatant, whole cell broth, filtrate, cell fraction or extract, metabolite, and/or compound alone or in combination with another pesticidal substance that is sufficient to modulate plant pest infestation or the performance of an agronomic trait of interest in the plant.
• the amount that will be within an effective range can be determined by laboratory or field tests by one skilled in the art.
• the effective amount when the composition is applied directly to the seed, is a concentration of about 0.05-25%, or about 0.1-20%, or about 0.5-15%, or about 1-10%, or about 2- 5% of the active ingredient per 100 g of seed. In some embodiments, the effective amount is about 0.5- 1% of the active ingredient per 100 g of seed.
• the effective amount when the composition is applied to the soil by, for example, in furrow, is about 0.1-50 oz. of the active ingredient per 1000 ft row. In another embodiment, the effective amount for soil application is about 1-25 oz. of the active ingredient per 1000 ft row. In another embodiment, the effective amont is about 2-20 oz, or about 3-15 oz, or about 4-10 oz, or about 5-8 oz, of the active ingredient per 1000 ft row. In yet another embodiment, the effective amount is about 14 or 28 oz of the active ingredient per 1000 ft row.
• Any appropriate agricultural application rate for a biocide can be applied in combination with the bacterial strain provided herein or active variant thereof, and/or a composition derived therefrom disclosed herein.
• Methods to assay for the effective amount of the bacterial strain provided herein or active variant thereof, and/or a composition derived therefrom include, for example, any statistically significant increase in the control of the pathogen or pest targeted by the biocide. Methods to assay for such control are known.
• compositions comprising at least one bacterial strain provided herein or active variant thereof, and/or a composition derived therefrom provided herein (i.e., AIP000648, AIP097852, AIP051459, or an active variant or any thereof, or a spore, or a forespore or a combination of cells, forespores and/or spores from any one of AIP000648, AIP097852, AIP051459, or an active variant any of thereof).
• a composition comprising at least one bacterial strain provided herein or active variant thereof, and/or a composition derived therefrom provided herein (i.e., AIP000648, AIP097852, AIP051459, or an active variant or any thereof, or a spore, or a forespore or a combination of cells, forespores and/or spores from any one of AIP000648, AIP097852, AIP051459, or an active variant any of thereof).
• applying is intended contacting an effective amount of the bacterial strain provided herein or active variant thereof, and/or a composition derived therefrom to a plant, area of cultivation, seed and/or weed with one or more of the bacterial strain provided herein or active variant thereof, and/or a composition derived therefrom so that a desired effect is achieved.
• the application of the bacterial strain provided herein or active variant thereof, and/or a composition derived therefrom 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, and/or a composition derived therefrom is a foliar application. Therefore, a further embodiment of the invention provides a method for controlling or inhibiting the growth of a plant pathogen by applying the population of bacterial strain provided herein or active variant thereof, and/or a composition derived therefrom to an environment in which the plant pathogen and/or 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, and/or a composition derived therefrom provided herein is used as a foliar application to control or inhibit growth of one or more plant pathogens from the group consisting of Botrytis cinerea, Cercospora sojina, Alternaria solani, Rhizoctonia solani, Erysiphe necator,
• Podosphaera xanthii Colletotrichum cereale, 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.
• an effective amount of at least one bacterial strain provided herein or active variant thereof, and/or a composition derived therefrom provided herein is used as a foliar application to control or inhibit growth of one or more insect pests from the group consisting of Nezara spp. (including Nezara viridula), Lygus spp. (including Lygus hesperus), Euschistus spp. (including Euschistus servus and Euschistus heros), Dichelops spp. (including Dichelops melacantus and Dichelops furcatus), and Halyomorpha spp. (including Halyomorpha halys).
• Nezara spp. including Nezara viridula
• Lygus spp. including Lygus hesperus
• Euschistus spp. including Euschistus servus and Euschistus heros
• Dichelops spp. including Dichelops melacantus and Dichelops furcatus
• an effective amount of at least one bacterial strain provided herein or active variant thereof, and/or a composition derived therefrom provided herein is used as a foliar application to control or inhibit growth of one or more insect or insect pests.
• an effective amount of at least one bacterial strain provided herein or active variant thereof, and/or a composition derived therefrom 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 nematodes or nematode pests.
• an effective amount of at least one bacterial strain provided herein or active variant thereof, and/or a composition derived therefrom provided herein is applied to the plant propagule (i.e. seed, slip, stem cutting, corn etc.) from which the plant to be protected are growing or will grow to control or inhibit growth of one or more insect or insect pests.
• an effective amount of at least one bacterial strain provided herein or active variant thereof, and/or a composition derived therefrom 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, and/or a composition derived therefrom 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 plant 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 ⁇ Meloidogyne arenaria).
• an effective amount of at least one bacterial strain provided herein or active variant thereof, and/or a composition derived therefrom provided herein is applied to the plant after harvest to control or inhibit growth of one or more pathogens selected from the group consisting of Southern Root-Knot nematode ⁇ Meloidogyne incognita), Javanese Root-Knot nematode (Meloidogyne javanica), Northern Root-Knot Nematode ⁇ Meloidogyne hapld) and Peanut Root-Knot Nematode ⁇ Meloidogyne arenarid).
• pathogens selected from the group consisting of Southern Root-Knot nematode ⁇ Meloidogyne incognita), Javanese Root-Knot nematode (Meloidogyne javanica), Northern Root-Knot Nematode ⁇ Meloidogyne hapld) and Peanut 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, and/or a composition derived therefrom i.e., AIP000648, AIP097852, AIP051459, 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 AIP000648, AIP097852, AIP051459, or an active variant thereof, and/or a composition derived therefrom
• the timing of application can vary depending on the conditions and geographical location.
• the bacterial strain provided herein or active variant thereof, and/or a composition derived therefrom is applied at the Rl (beginning flowering stage) of soybean development or may be applied earlier depending on disease onset and the disease severity.
• the biocide to a crop, area of cultivation, or field it is intended that one or more of a particular field, plant crop, seed and/or weed is treated with one or more of the bacterial strain provided herein or active variant thereof, and/or a composition derived therefrom and one or more biocide so that a desired effect is achieved.
• Various methods are provided for controlling a plant pathogen and/or plant pest that causes a plant disease in an area of cultivation containing a plant susceptible to the plant disease.
• 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, and/or a composition derived therefrom (i.e., AIP000648, AIP097852,
• the effective amount of the bacterial strain provided herein or active variant thereof, and/or a composition derived therefrom controls the plant disease without significantly affecting the crop.
• the effective amount comprises at least about 10 5 to 10 12 colony forming units (CFU) per hectare. In other embodiments, the effective amount comprises at least about 10 to 10 colony forming units (CFU) per hectare.
• the composition is derived from a bacterial strain provided herein or active variant thereof and may comprise a cell of at least one of AIP000648, AIP097852, AIP051459, 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 AIP000648, AIP097852, AIP051459, or an active variant of any thereof.
• 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, and/or a composition derived therefrom.
• the bacterial strain provided herein or active variant thereof, and/or a composition derived therefrom may comprise at least one of AIP000648, AIP097852, AIP051459, 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 AIP000648, AIP097852, AIP051459, or an active variant of any thereof.
• the effective amount of the bacterial strain provided herein or active variant thereof comprises at least about 10 5 to 10 12 colony forming units (CFU) per hectare, or 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, and/or a composition derived therefrom.
• 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 AIP000648, AIP097852, AIP051459, 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 AIP000648, AIP097852, AIP051459, or an active variant of any thereof, wherein said effective amount comprises at least about 10 5 to 10 12 colony forming units (CFU) per hectare, or at least about 10 12 to 10 16 colony forming units (CFU) per hectare, and wherein said composition controls a plant pathogen or other plant pest, thereby increasing yield.
• a composition comprising at least one bacterial strain comprising AIP000648, AIP097852, AIP051459, or an active variant of any thereof, a spore or a forespore or a combination of cells, forespor
• a method for increasing yield in a plant comprises applying to a crop or an area of cultivation an effective amount of a composition derived from at least one bacterial strain comprising AIP000648, AIP097852, AIP051459, 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 AIP000648, AIP097852, AIP051459, or an active variant of any thereof, wherein said composition controls a plant pest, thereby increasing yield.
• 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 coated seed which comprises a seed and a coating on the seed, wherein the coating comprises at least one bacterial strain provided herein or active variant thereof, and/or a composition derived therefrom.
• the bacterial strain provided herein or active variant thereof, and/or a composition derived therefrom may comprise at least one of AIP000648, AIP097852, AIP051459, 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 AIP000648, AIP097852, AIP051459, or an active variant of any thereof.
• said bacterial strain provided herein or active variant thereof is present on the seed at about 10 5 CFU/seed to about 10 7 CFU/seed, at about 10 4 CFU/seed to about 10 8 CFU/seed, at about 10 4 CFU/seed to about 10 5 CFU/seed, at about 10 5 CFU/seed to about 10 6 CFU/seed, at about 10 6 CFU/seed to about 10 7 CFU/seed, or at about 10 7 CFU/seed to about 10 8 CFU/seed.
• the seed coating can be applied to any seed of interest (i.e., for a monocot or a dicot). Various plants of interest are disclosed elsewhere herein.
• a seed coating can further comprise at least at least one nutrient, at least one herbicide or at least one pesticide, or at least one biocide. See, for example, US App Pub. 20040336049, 20140173979, and 20150033811.
• a plant of interest i.e., plant susceptible to the plant disease
• 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/or a composition derived therefrom and an effective amount of a biocide.
• a combination of or “applying a combination of a bacterial strain provided herein or active variant thereof, and/or a composition derived therefrom and a biocide 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 strain provided herein or active variant thereof, and/or a composition derived therefrom and one or more biocide so that a desired effect is achieved.
• the application of one or both of the bacterial strain provided herein or active variant thereof, and/or a composition derived therefrom and the biocide can occur prior to the planting of the crop (for example, to the soil, or the plant).
• the application of the bacterial strain provided herein or active variant thereof, and/or a composition derived therefrom and the biocide may be simultaneous or the applications may be at different times (sequential), so long as the desired effect is achieved.
• the active variant comprises a bacterial strain provided herein that is resistance to one or more biocide.
• the bacterial strain provided herein or active variant thereof, and/or a composition derived therefrom i.e., AIP000648, AIP097852, AIP051459, 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 AIP000648, AIP097852, AIP051459, or an active variant of any thereof
• AIP000648, AIP097852, AIP051459 or an active variant of any thereof
• 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, and/or a composition derived therefrom 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, and/or a composition derived therefrom 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, and/or a composition derived therefrom 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, and/or a composition derived therefrom 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, and/or a composition derived therefrom comprises an effective amount of AIP000648, AIP097852, AIP051459, 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 AIP000648, AIP097852, AIP051459, or an active variant of any thereof.
• the bacterial strain provided herein or active variant thereof, and/or a composition derived therefrom can be used in combination with a biocide (i.e., an herbicide, fungicide, pesticide, or other crop protection chemical).
• a biocide i.e., an herbicide, fungicide, pesticide, or other crop protection chemical.
• the bacterial strain provided herein or active variant thereof, and/or a composition derived therefrom 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, and/or a composition derived therefrom 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));
• Pesticides that can be used in the various methods and compositions disclosed herein include imidacloprid clothianidin, arylpyrazole compounds (WO2007103076); 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).
• Nematicides that can be used in the various methods and compositions disclosed herein include any synthetic nematicide or biological nematicide.
• synthetic nematicides 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, terbuf
• Burkholderia spp. Burkholderia spp.; Pasteuria spp., Brevibacillus spp.; Pseudomonas spp.; Rhizobacteria; and Bacillus spp.
• 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
• 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); me
• 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
• dicarboximide fungicides (famoxadone, fluoroimide); dichlorophenyl dicarboximide fungicides
• phenylmercury salicylate thiomersal, tolylmercury acetate
• mo holine fungicides aldimorph, benzamorf, carbamo h, dimethomo h, dodemo h, ⁇ , flumo ⁇ h, tridemo ⁇ h
• 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 (
• anilinopyrimidine fungicides cyprodinil, mepanipyrim, pyrimethanil
• pyrrole fungicides dimetachlone, fenpiclonil, fludioxonil, fluoroimide
• quaternary ammonium fungicides berberine, sanguinarine
• quinoline fungicides ethoxyquin, halacrinate, 8 -hydroxy quinoline 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 metalhasulfocarb, prothiocarb
• thiophene fungicides eth
• triazolopyrimidine fungicides ametoctradin
• urea fungicides urea fungicides
• pencycuron 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), or mefenoxam.
• fungicides acibenzolar,
• kits of parts comprising a bacterial strain provided herein or active variant thereof, and/or a composition derived therefrom, and at least one biocide, in a spatially separated arrangement.
• the biocide is an herbicide, fungicide, insecticide, pesticide, or other crop protection chemical.
• composition comprising:
• bacterial strain AIP000648, AIP097852, AIP051459, or an active variant of any thereof comprises a bacterial strain having a genome within a Mash distance of about 0.015, and wherein said bacterial strain or an active variant 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;
• spore, or a forespore, or a combination of cells, forespores, and/or spores from any of AIP000648, AIP097852, AIP051459, 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, and wherein said spore, forespore, or a combination of cells, forespores, and/or spores or an active variant 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/or
• an effective amount of said composition improves an agronomic trait of interest of a plant or controls a plant pest or a plant pathogen that causes a plant disease.
• composition of embodiment 1, wherein said bacterial strain or the active variant 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 of embodiment 1 or 2 wherein said composition comprises a cell paste.
• composition comprises a wettable powder, a spray dried formulation, or a stable formulation.
• composition of embodiment 1 or 2 wherein said composition comprises a seed treatment.
• composition of any of embodiments 1-7, wherein the plant pest is a. Meloidogyne spp or a hermipteran insect.
• composition of embodiment 6 or 7, wherein said plant pest comprises one or more nematode pests selected from the group consisting of Meloidogyne incognita, Meloidogyne javanica, Meloidogyne hapla, Meloidogyne arenaria, Ditylenchus destructor, Ditylenchus dipsaci, Pratylenchus penetrans, Pratylenchus fallax, Pratylenchus coffeae, Pratylenchus loosi, Pratylenchus vulnus,
• composition of embodiment 9, wherein said plant pest comprises one or more nematode pests selected from the group consisting of Meloidogyne incognita, Meloidogyne javanica, Meloidogyne hapla, and Meloidogyne arenaria.
• composition of embodiment 6 or 7, wherein said plant pest comprises one or more hemipteran pests selected from the group consisting ⁇ Lygus spp. including Lygus hesperus, Lygus lineolaris, Lygus pratensis, Lygus rugulipennis, and Lygus pabulinus, Calocoris norvegicus, Orthops compestris, Plesiocoris rugicollis, Cyrtopeltis modestus, Cyrtopeltis notatus, Spanagonicus albofasciatus, Diaphnocoris chlorinonis, Labopidicola allii, Pseudatomoscelis seriatus, Adelphocoris rapidus,
• composition of embodiment 11, wherein said plant pest comprises one or more hemipteran pests selected from the group consisting of Nezara spp. including Nezara viridula, Euschistus spp. including Euschistus servus and Euschistus heros, Dichelops spp. including Dichelops melacantus and Dichelops furcatus, and Halyomorpha halys.
• Nezara spp. including Nezara viridula
• Euschistus spp. including Euschistus servus and Euschistus heros
• Dichelops spp. including Dichelops melacantus and Dichelops furcatus
• Halyomorpha halys selected from the group consisting of Nezara spp. including Nezara viridula, Euschistus spp. including Euschistus servus and Euschistus heros, Dichelops spp. including Dichelops melacantus and Dichelop
• composition of embodiments 13 or 14, wherein said plant pathogen comprises one or more fungal pathogens selected from the group consisting of Botrytis cinerea, Cersospora spp,
• Cercospora sojina Cercospora beticola, Alternaria solani, Rhizoctonia solani, Blumeria graminis, Erysiphe necator, Podosphaera xanthii, Golovinomyces cichoracearum, Erysiphe lager stroemiae, Sphaerotheca pannosa, Colletotrichum cereale, Apiognomonia errabunda, Apiognomonia veneta, Colletotrichum gloeosporiodes, Discula fraxinea, Plasmopara viticola, Pseudoperonospora cubensis, Peronospora belbahrii, Bremia lactucae, Peronospora lamii, Plasmopara obduscens, Pythium cryptoirregulare, Pythium aphanidermatum, Pythium irregulare, Pythium sylvaticum, Pythium
• a composition comprising a cell paste comprising:
• an effective amount of said bacterial strain composition improves an agronomic trait of interest of the plant or controls a plant pest or plant pathogen that causes a plant disease.
• the plant pest is a nematode pest or an insect pest.
• said nematode pest comprises one or more nematode pests selected from the group consisting of Meloidogyne incognita, Meloidogyne javanica, Meloidogyne hapla, Meloidogyne arenaria, Ditylenchus destructor, Ditylenchus dipsaci, Pratylenchus penetrans, Pratylenchus fallax, Pratylenchus coffeae, Pratylenchus loosi, Pratylenchus vulnus,
• Globodera rostochiensis Globodera pallida, Heterodera glycines, Heterodera schachtii, Heterodera avenae, Aphelenchoides besseyi, Aphelenchoides ritzemabosi, Aphelenchoides fragariae, Aphelenchus avenae, Radopholus similis Tylenchulus semipenetrans, Rotylenchulus reniformis, Bursaphelenchus xylophilus, Bursaphelenchus cocophilus, Helicotylenchus spp, Radopholus similis, Ditylenchus dipsaci, Rotylenchulus reniformis, Xiphinema spp., Aphelenchoides spp., Bursaphelenchus xylophilus, and Pratylenchus spp.
• composition of embodiment 18, wherein said plant pest comprises one or more nematode pests selected from the group consisting of Meloidogyne incognita, Meloidogyne javanica, Meloidogyne hapla, and Meloidogyne arenaria.
• composition of embodiment 17, wherein said insect pest comprises one or more hemipteran pests selected from the group consisting ⁇ Lygus spp. including Lygus hesperus, Lygus lineolaris, Lygus pratensis, Lygus rugulipennis, and Lygus pabulinus, Calocoris norvegicus, Orthops compestris, Plesiocoris rugicollis, Cyrtopeltis modestus, Cyrtopeltis notatus, Spanagonicus albofasciatus, Diaphnocoris chlorinonis, Labopidicola allii, Pseudatomoscelis seriatus, Adelphocoris rapidus,
• Dichelops melacantus and Dichelops furcatus including Dichelops melacantus and Dichelops furcatus, Halyomorpha halys, Lipaphis erysimi, Aphis gossypii, Macrosiphum avenae, Myzus persicae, Acyrthosiphon pisum, Aphidoidea spp, Eurygaster spp., Coreidae spp., Pyrrhocoridae spp.,
• Blostomatidae spp. Reduviidae spp., Cimicidae spp. Aleurocanthus woglumi, Aleyrodes proletella, Bemisia spp. including Bemisia argentifolii and Bemisia tabaci, and Trialeurodes vaporariorum.
• composition of embodiment 16, wherein the plant pathogen comprises at least one fungal pathogen.
• composition of embodiment 21, wherein said plant pathogen comprises one or more fungal pathogens selected from the group consisting of Botrytis cinerea, Cersospora spp, Cercospora sojina, Cercospora beticola, Alternaria solani, Rhizoctonia solani, Blumeria graminis, Erysiphe necator, Podosphaera xanthii, Golovinomyces cichoracearum, Erysiphe lager stroemiae, Sphaerotheca pannosa, Colletotrichum cereale, Apiognomonia errabunda, Apiognomonia veneta, Colletotrichum gloeosporiodes, Discula fraxinea, Plasmopara viticola, Pseudoperonospora cubensis, Peronospora belbahrii, Bremia lactucae, Peronospora lamii, Plasmopara ob
• Phytophthora tropicalis Phytophthora sojae, Fusarium graminearum, Fusarium solani, Fusarium oxysporum, Fusarium graminicola, Gibberella zeae, Colletotrichum graminicola, Phakopsora spp., Phakopsora meibomiae, Phakopsora pachyrizi, Puccinia triticina, Puccinia recondita, Puccinia striiformis, Puccinia graminis, Puccinia spp., Venturia inaequalis, Verticillium spp, Erwinia amylovora, Monilinia fructicola, Monilinia lax, and Monilinia fructigena.
• a composition comprising a wettable power comprising:
• bacterial strain AIP000648, AIP097852, AIP051459, or an active variant of any thereof comprises a bacterial strain having a genome within a Mash distance of about 0.015, and wherein said bacterial strain or an active variant 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;
• spore, or a forespore, or a combination of cells, forespores, and/or spores from any of AIP000648, AIP097852, AIP051459, 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, and wherein said spore, forespore, or a combination of cells, forespores, and/or spores or an active variant 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/or
• an effective amount of said composition improves an agronomic trait of interest of a plant or controls a plant pest or a plant pathogen that causes a plant disease.
• composition of embodiment 23, wherein the plant pest comprises at least one nematode pest or at least one insect pest.
• composition of embodiment 24, wherein the said plant pest comprises one or more nematode pests selected from the group consisting of Meloidogyne incognita, Meloidogyne javanica, Meloidogyne hapla, Meloidogyne arenaria, Ditylenchus destructor, Ditylenchus dipsaci, Pratylenchus penetrans, Pratylenchus fallax, Pratylenchus coffeae, Pratylenchus loosi, Pratylenchus vulnus,
• nematode pests selected from the group consisting of Meloidogyne incognita, Meloidogyne javanica, Meloidogyne hapla, Meloidogyne arenaria, Ditylenchus destructor, Ditylenchus dipsaci, Pratylenchus penetrans, Pratylenchus fallax, Pratylenchus coffeae, Pra
• Globodera rostochiensis Globodera pallida, Heterodera glycines, Heterodera schachtii, Heterodera avenae, Aphelenchoides besseyi, Aphelenchoides ritzemabosi , Aphelenchoides fragariae, Aphelenchus avenae, Radopholus similis Tylenchulus semipenetrans, Rotylenchulus reniformis, Bursaphelenchus xylophilus, Bursaphelenchus cocophilus, Helicotylenchus spp, Radopholus similis, Ditylenchus dipsaci, Rotylenchulus reniformis, Xiphinema spp., Aphelenchoides spp., Bursaphelenchus xylophilus, and Pratylenchus spp..
• composition of embodiment 25, wherein said plant pest comprises one or more nematode pests selected from the group consisting of Meloidogyne incognita, Meloidogyne javanica, Meloidogyne hapla, and Meloidogyne arenaria.
• composition of embodiment 24, wherein said insect pest comprises one or more hemipteran pests selected from the group consisting ⁇ Lygus spp. including Lygus hesperus, Lygus lineolaris, Lygus pratensis, Lygus rugulipennis, and Lygus pabulinus, Calocoris norvegicus, Orthops compestris, Plesiocoris rugicollis, Cyrtopeltis modestus, Cyrtopeltis notatus, Spanagonicus albofasciatus, Diaphnocoris chlorinonis, Labopidicola allii, Pseudatomoscelis seriatus, Adelphocoris rapidus,
• Dichelops melacantus and Dichelops furcatus including Dichelops melacantus and Dichelops furcatus, Halyomorpha halys, Lipaphis erysimi, Aphis gossypii, Macrosiphum avenae, Myzus persicae, Acyrthosiphon pisum, Aphidoidea spp, Eurygaster spp., Coreidae spp., Pyrrhocoridae spp.,
• Blostomatidae spp. Reduviidae spp., Cimicidae spp. Aleurocanthus woglumi, Aleyrodes proletella, Bemisia spp. including Bemisia argentifolii and Bemisia tabaci, and Trialeurodes vaporariorum.
• composition of embodiment 23, wherein the plant pathogen comprises at least one fungal pathogen.
• composition of embodiment 28, wherein said plant pathogen comprises one or more fungal pathogens selected from the group consisting of Botrytis cinerea, Cersospora spp, Cercospora sojina, Cercospora beticola, Alternaria solani, Rhizoctonia solani, Blumeria graminis, Erysiphe necator, Podosphaera xanthii, Golovinomyces cichoracearum, Erysiphe lager stroemiae, Sphaerotheca pannosa, Colletotrichum cereale, Apiognomonia errabunda, Apiognomonia veneta, Colletotrichum gloeosporiodes, Discula fraxinea, Plasmopara viticola, Pseudoperonospora cubensis, Peronospora belbahrii, Bremia lactucae, Peronospora lamii, Plasmopara ob
• Phytophthora tropicalis Phytophthora sojae, Fusarium graminearum, Fusarium solani, Fusarium oxysporum, Fusarium graminicola, Gibberella zeae, Colletotrichum graminicola, Phakopsora spp., Phakopsora meibomiae, Phakopsora pachyrizi, Puccinia triticina, Puccinia recondita, Puccinia striiformis, Puccinia graminis, Puccinia spp., Venturia inaequalis, Verticillium spp, Erwinia amylovora, Monilinia fructicola, Monilinia lax, and Monilinia fructigena.
• composition of embodiment 30, 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 32, wherein the plant pest is a nematode pest or an insect pest.
• composition of embodiment 32, wherein the plant pathogen comprises at least one fungal pathogen.
• said insect pest comprises one or more hemipteran pests selected from the group consisting of Lygus spp. including Lygus hesperus, Lygus lineolaris, Lygus pratensis, Lygus rugulipennis, and Lygus pabulinus, 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 spp.
• Lygus spp. including Lygus hesperus, Lygus lineolaris, Lygus pratensis, Lygus rugulipennis, and Lygus pa
• Nezara spp. including Nezara viridula, Acrosternum hilare, Euschistus spp. including Euschistus servus and Euschistus heros, Dichelops spp.
• Cersospora spp Cercospora sojina, Cercospora beticola, Alternaria solani, Rhizoctonia solani, Blumeria graminis, Erysiphe necator, Podosphaera xanthii, Golovinomyces cichoracearum, Erysiphe lagerstroemiae, Sphaerotheca pannosa, Colletotrichum cereale, Apiognomonia errabunda,
• Pseudoperonospora cubensis Peronospora belbahrii, Bremia lactucae, Peronospora lamii, Plasmopara obduscens, 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 graminearum, Fusarium solani, Fusarium oxysporum, Fusarium graminicola, Gibberella zeae, Colletotrichum graminicola, Phakopsora spp., Phakopsora meibomiae, Phakopsora pachyrizi, Puccinia triticina, Puccinia recondita, Puccinia striiformis, Puccinia graminis, Puccinia spp., Venturia inaequalis, Verticillium spp, Erwinia amylovora, Monilinia fructicola, Monilinia lax, and Monilinia fructigena.
• said bacterial culture has pesticidal activity against a plant pest or plant pathogen that causes a plant disease and is able to grow in the presence of glufosinate or an effective amount of said bacterial culture improves an agronomic trait of interest of the plant.
• Globodera rostochiensis Globodera pallida, Heterodera glycines, Heterodera schachtii, Heterodera avenae, Aphelenchoides besseyi, Aphelenchoides ritzemabosi, Aphelenchoides fragariae, Aphelenchus avenae, Radopholus similis Tylenchulus semipenetrans, Rotylenchulus reniformis, Bursaphelenchus xylophilus, Bursaphelenchus cocophilus, Helicotylenchus spp, Radopholus similis, Ditylenchus dipsaci, Rotylenchulus reniformis, Xiphinema spp., Aphelenchoides spp., Bursaphelenchus xylophilus, and Pratylenchus spp.
• Cercospora sojina Cercospora beticola, Alternaria solani, Rhizoctonia solani, Blumeria graminis, Erysiphe necator, Podosphaera xanthii, Golovinomyces cichoracearum, Erysiphe lager stroemiae, Sphaerotheca pannosa, Colletotrichum cereale, Apiognomonia errabunda, Apiognomonia veneta, Colletotrichum gloeosporiodes, Discula fraxinea, Plasmopara viticola, Pseudoperonospora cubensis, Peronospora belbahrii, Bremia lactucae, Peronospora lamii, Plasmopara obduscens, Pythium cryptoirregulare, Pythium aphanidermatum, Pythium irregulare, Pythium sylvaticum, Pythium
• a method for growing a plant susceptible to a plant pest or plant disease or improving an agronomic trait of interest in a plant comprising applying to the plant:
• AIP051459 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 10 12 to 10 16 colony forming units (CFU) per hectare;
• said effective amount controls a plant pest or plant pathogen that causes the plant disease or improves the agronomic trait of interest.
• said plant pest comprises one or more nematode pests selected from the group consisting of Meloidogyne incognita, Meloidogyne javanica, Meloidogyne hapla, Meloidogyne arenaria, Ditylenchus destructor, Ditylenchus dipsaci, Pratylenchus penetrans, Pratylenchus fallax, Pratylenchus coffeae, Pratylenchus loosi, Pratylenchus vulnus, Globodera rostochiensis, Globodera pallida, Heterodera glycines, Heterodera schachtii, Heterodera avenae, Aphelenchoides besseyi, Aphelenchoides ritzemabosi, Aphelenchoides fragariae, Aphelenchus avenae, Radopholus similis Tylench
• Rotylenchulus reniformis reniformis, Xiphinema spp., Aphelenchoides spp., Bursaphelenchus xylophilus, and Pratylenchus spp.
• said plant pest comprises one or more nematode pests selected from the group consisting of Meloidogyne incognita, Meloidogyne javanica, Meloidogyne hapla, Meloidogyne arenaria.
• said plant pathogen comprises one or more fungal pathogens selected from the group consisting of Botrytis cinerea, Cersospora spp, Cercospora sojina, Cercospora beticola, Alternaria solani, Rhizoctonia solani, Blumeria graminis, Erysiphe necator, Podosphaera xanthii, Golovinomyces cichoracearum, Erysiphe lager stroemiae, Sphaerotheca pannosa, Colletotrichum cereale, Apiognomonia errabunda, Apiognomonia veneta, Colletotrichum gloeosporiodes, Discula fraxinea, Plasmopara viticola, Pseudoperonospora cubensis, Peronospora belbahrii, Bremia lactucae, Peronospora lamii, Plasmopara
• Phytophthora tropicalis Phytophthora sojae, Fusarium graminearum, Fusarium solani, Fusarium oxysporum, Fusarium graminicola, Gibberella zeae, Colletotrichum graminicola, Phakopsora spp., Phakopsora meibomiae, Phakopsora pachyrizi, Puccinia triticina, Puccinia recondita, Puccinia striiformis, Puccinia graminis, Puccinia spp., Venturia inaequalis, Verticillium spp, Erwinia amylovora, Monilinia fructicola, Monilinia lax, and Monilinia fructigena.
• a method of controlling a plant pest or plant pathogen that causes a plant disease in an area of cultivation comprising:
• an effective amount of at least one bacterial strain comprising AIP000648, AIP097852, AIP051459, 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, and wherein said effective amount comprises at least about 10 12 to 10 16 colony forming units (CFU) per hectare;
• an effective amount of at least one bacterial strain comprising a spore, or a forespore, or a combination of cells, forespores and/or spores from any one of AIP000648, AIP097852, AIP051459, 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, and wherein said effective amount comprises at least about 10 12 to 10 16 colony forming units (CFU) per hectare; or
• said effective amount controls a plant pest or plant pathogen that causes a plant disease in an area of cultivation.
• 61 The method of any one of embodiments 58-60, wherein said composition controls one or more nematode pest.
• the one or more nematode pests are selected from the group consisting of Meloidogyne incognita, Meloidogyne javanica, Meloidogyne hapla, Meloidogyne arenaria, Ditylenchus destructor, Ditylenchus dipsaci, Pratylenchus penetrans,
• Pratylenchus fallax Pratylenchus coffeae, Pratylenchus loosi, Pratylenchus vulnus, Globodera rostochiensis, Globodera pallida, Heterodera glycines, Heterodera schachtii, Heterodera avenae, Aphelenchoides besseyi, Aphelenchoides ritzemabosi, Aphelenchoides fragariae, Aphelenchus avenae, Radopholus similis Tylenchulus semipenetrans, Rotylenchulus reniformis, Bursaphelenchus xylophilus, Bursaphelenchus cocophilus, Helicotylenchus spp, Radopholus similis, Ditylenchus dipsaci,
• Rotylenchulus reniformis reniformis, Xiphinema spp., Aphelenchoides spp., Bursaphelenchus xylophilus, and Pratylenchus spp.
• composition controls one or more nematode pests are selected from the group consisting of Meloidogyne incognita, Meloidogyne javanica, Meloidogyne hapla, Meloidogyne arenaria.
• the one or more fungal pathogens are selected from the group consisting of Botrytis cinerea, Cersospora spp, Cercospora sojina, Cercospora beticola, Alternaria solani, Rhizoctonia solani, Blumeria graminis, Erysiphe necator, Podosphaera xanthii, Golovinomyces dehor acearum, Erysiphe lager stroemiae, Sphaerotheca pannosa, Colletotrichum cereale, Apiognomonia errabunda, Apiognomonia veneta, Colletotrichum gloeosporiodes, Discula fraxinea, Plasmopara viticola, Pseudoperonospora cubensis, Peronospora belbahrii, Bremia lactucae,
• Peronospora lamii Plasmopara obduscens, 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 graminearum, Fusarium solani, Fusarium oxysporum, Fusarium graminicola, Gibber ella zeae, Colletotrichum graminicola, Phakopsora spp., Phakopsora meibomiae, Phakopsora pachyrizi, Puccinia triticina, Puccinia recondita, Puccinia striiformis,
• a method of making a modified bacterial strain comprising:
• the active variant comprises a bacterial strain having a genome within a Mash distance of about 0.015; 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 10 12 to 10 16 CFU per hectare; and/or (c) an effective amount of a supernatant, filtrate, or extract derived from a whole cell culture of at least one of bacterial strain AIP000648, AIP097852, AIP051459, 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
• the effective amount controls the plant pest or plant pathogen that causes the plant disease.
• nematode pests are selected from the group consisting oiMeloidogyne incognita, Meloidogyne javanica, Meloidogyne hapla, Meloidogyne arenaria, Ditylenchus destructor, Ditylenchus dipsaci, Pratylenchus penetrans, Pratylenchus fallax, Pratylenchus coffeae, Pratylenchus loosi, Pratylenchus vulnus, Globodera rostochiensis, Globodera pallida, Heterodera glycines, Heterodera schachtii, Heterodera avenae, Aphelenchoides besseyi, Aphelenchoides ritzemabosi, Aphelenchoides fragariae, Aphelenchus avenae, Rad
• Rotylenchulus reniformis reniformis, Xiphinema spp., Aphelenchoides spp., Bursaphelenchus xylophilus, and Pratylenchus spp..
• composition controls one or more nematode pests are selected from the group consisting oiMeloidogyne incognita, Meloidogyne javanica, Meloidogyne hapla, and Meloidogyne arenaria.
• Lygus spp. including Lygus hesperus, Lygus lineolaris, Lygus pratensis, Lygus rugulipennis, and Lygus pabulinus
• Calocoris norvegicus Orthops comp
• Dichelops melacantus and Dichelops furcatus including Dichelops melacantus and Dichelops furcatus, Halyomorpha halys, Lipaphis erysimi, Aphis gossypii, Macrosiphum avenae, Myzus persicae, Acyrthosiphon pisum, Aphidoidea spp, Eurygaster spp., Coreidae spp., Pyrrhocoridae spp.,
• Blostomatidae spp. Reduviidae spp., Cimicidae spp. Aleurocanthus woglumi, Aleyrodes proletella, Bemisia spp. including Bemisia argentifolii and Bemisia tabaci, and Trialeurodes vaporariorum.
• the one or more fungal pathogens are selected from the group consisting of Botrytis cinerea, Cersospora spp, Cercospora sojina, Cercospora beticola, Alternaria solani, Rhizoctonia solani, Blumeria graminis, Erysiphe necator, Podosphaera xanthii, Golovinomyces dehor acearum, Erysiphe lager stroemiae, Sphaerotheca pannosa, Colletotrichum cereale, Apiognomonia errabunda, Apiognomonia veneta, Colletotrichum gloeosporiodes, Discula fraxinea, Plasmopara viticola, Pseudoperonospora cubensis, Peronospora belbahrii, Bremia lactucae,
• Peronospora lamii Plasmopara obduscens, 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 graminearum, Fusarium solani, Fusarium oxysporum, Fusarium graminicola, Gibber ella zeae, Colletotrichum graminicola, Phakopsora spp., Phakopsora meibomiae, Phakopsora pachyrizi, Puccinia triticina, Puccinia recondita, Puccinia striiformis,
• a kit of parts comprising a biocide and:
• kits of embodiment 87, wherein the biocide is an herbicide, fungicide, insecticide, nematicide, and/or pesticide.
• Example 1 Microbial strains and methods of culturing
• Table 2 Bacterial strains selected for evaluation for inhibition of est activity.
• Bacterial strains set forth in Table 2 were cultured in LB medium.
• Table 3 summarizes the incubation time and the concentration of bacteria (CFU/ml) achieved.
• Example 2 In vitro evaluation of bacterial strains for nematicidal activity
• a starter culture was prepared by filling a 96-well block with 1-ml (per well) LB media and inoculating with a bacterial strain of Table 2. 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 -1.74 ml (per well) media. 25 ⁇ of starter culture was added to the assay culture blocks. Assay blocks 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 0.290 ml of nematode water for a 6X concentration and then used in the assay. All microbial preparations were applied within 12 h of preparation.
• Root-Knot Nematodes were assessed for motility in the presence of microbes. J2's were selected by hatching Meloidogyne incognita eggs over a nylon sieve and collecting the J2's that were motile enough to move through the nylon sieve for selection into the assay. Root-Knot Nematodes (Meloidogyne incognita) were dispensed into a 96 well assay plate with a total volume of 95 ⁇ containing approximately 100 stage 2 juveniles (J2) per well. 5 ⁇ of the microbial preparations described above were dispensed into wells containing the nematodes.
• Example 3 On-plant assays for evaluation of bacterial strains for nematicidal activity
• a starter culture was 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 was grown at 30 °C with shaking at 225 rpms for 18 h. After 18 hours 1 ml from each starter culture was passaged through 10 ml of LB media. These cultures were incubated with agitation at 30 °C with shaking at 225 rpms for 5-6 hrs. Assay cultures were prepared by filling 1 liter flasks with 350ml of media. 1.5ml of the 10ml culture was added to the assay culture flask. Flasks were grown at 30 °C for either 24, 48 or 72 h at 175 rpms.
• the flasks were 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 was then poured off so that only the pellet remained. Pellets were 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 were then used in the on-plant assay. All microbial preparations were applied within 12 h of preparation.
• Example 3.1 On-plant assay in a growth chamber
• Cucumber plants were assessed for infectivity with Root-Knot Nematodes in the presence of a bacterial strain of interest. Cucumber seeds were planted in a 4 inch plastic pot with 90% sand and 10% soil. Following germinating, plants were 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 were treated with a 1 Ox 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 were infected with 1500 stage 2 RKN juveniles per plant.
• Cucumber plants were raised from seed in propagation greenhouses, in seedling trays. These cucumber plants were then transplanted to pots containing soil infested with Root-Knot Nematodes. Shortly (1-2 hours) after the cucumber plant was planted into the infested soil, the plant received a drench application of the treatment. The treatment was either a bacterial strain of interest (prepared as described above), an application of the nematicide Ecozin ® Plus at a standard concentration, or untreated (negative control). The crop was maintained in conditions like those of commercial practice, with the appropriate use of fertilizers and maintenance pesticides. At approximately 28 days after transplanting, the plants were observed for galling caused by RKN. The experiment was repeated two times, and indicated in Table 6 as Trial 1 and Trial 2. Data shown in Table 6 is reported as a gall score, where a score of 0 is no galls and a score of 10 is severe galling with 90% of root covered in galls.
• Yellow crookneck squash are assessed for infectivity with Root-Knot Nematodes (Meloidogyne incognita) in the presence of the microbe of interest. Yellow Squash plants are raised from seed in the greenhouse and then transplanted to the field. The crop is maintained in conditions like those of commercial practice, with the appropriate use of fertilizers and maintenance pesticides. Three to six week old plants are transplanted into 1.4ft.sq. field plots with enumerated Root-Knot Nematode populations at 6 replicates per treatment in a randomized complete block design.
• Root-Knot Nematodes Malignant ne incognita
• Microbial treatments are applied at day of transplant and 14-28 days after planting at a volume of .5 to 4 pints per square foot to achieve a uniform soil drench to a depth of 2-12 inches. Treatments are applied at a concentration of 0.5 to 3 Og microbial treatment per liter of finished solution. Plants are maintained using conditions like those of commercial practice. Plants are rated for stunting on a weekly basis throughout the trial. After 50 days plants are harvested for galling compared to the positive and negative controls and egg counts are performed to compare end of trial counts from the initiation of the trial.
• Example 5 Evaluation of bacterial strains for insecticidal activity
• a starter culture was prepared by filling a 96-well block with 1-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 ⁇ 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.
• 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 corn 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). Whole culture microbial suspensions were 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 26°C at 65%RH on a 16:8 day:night 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. The results are set forth in Table 7.
• Southern Green stink bugs 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:21C, 16:8 day:night 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 7.
• Example 6 Evaulation of bacterial strains against fungal and oomvcete plant pathogens
• Example 6.1 Evaluation of bacterial strains against Podosphaera xanthii (powdery mildew on cucurbits)
• Bacterial strains AIP000648, AIP097852, and AIP021332 were stored at -80°C and then were streaked onto Luria Bertani (LB) agar (25g LB broth and 15g water agar per liter of deionized water) in petri plates. A single colony was picked and placed in 50 ml of LB liquid medium or in liquid culture (CHA medium), which consists of, per L, NaCl (5g), tryptone (10g), nutrient broth (8g), CaCl 2 (0.14mM), MgCl 2 6H 2 0 (0.2mM), and MnCl 2 4H 2 0 (0.01 mM) in a 250-ml flask.
• LB liquid medium liquid culture
• Cultures were harvested after 48 hours by pelleting cells and re-suspending to the original volume in deionized sterile water. Each culture was titered to determine CFU/mL using standard dilution plate count methods and plates were inspected for signs of contamination. Sterile distilled water was added to achieve a final concentration needed in strain evaluation.
• Leaf disks of healthy squash leaves were excised and cut into uniform leaf disks 35 mm in diameter using a large cork borer.
• An experimental unit consisted of a single leaf disk, each treated with a suspension of the selected bacterium.
• Treatments included: AgBiome strains AIP061382, AIP075655, and AIP029105 on inoculated leaf discs, an "inoculated control" which was not treated with a bacterial strain, a "non-inoculated” control which was not inoculated with the pathogen, and a control treated with the synthetic fungicide tebuconazole at 10 ppm prior to inoculation with P. xanthii.
• Bacterial strains were prepared as described above.
• Each leaf disk was sprayed with 200 of the treatment (for bacterial strains, 1 x 10 CFU/mL, suspended in magnesium chloride buffer)on the adaxial surface 24 hours before inoculation with the pathogen. After treatment, leaf disks were incubated in the dark for 24 hours at 23 °C.
• Leaf disks were inoculated by spraying a 1 x 10 6 suspension of P. xanthii conidia on the treated leaf surface. Treatments were placed into sealed clear plastic boxes and incubated for six days at 25°C with a relative humidity of 80% and a 12 hour photoperiod. This experiment was run twice (primary and secondary), with each treatment replicated two to three times. Percent disease control (%) for each experiment on each treatment is indicated in Table 8.
• Example 6.2 Evaluation of bacterial strains against Colletotrichum sublineolum (sorghum anthracnose) Sorghum cultivar, 12-GS9016-KS585, was grown in the greenhouse for a steady supply of leaf tissue for the bacterial strain evaluation. Fully expanded sorghum leaves from 4-6 weeks old plants, were excised, and cut into equal pieces, 2.5cm wide. Colletotrichum sublineolum, (obtained from the Dr. Isakeit laboratory at Texas A&M University) was grown on 20% Oatmeal agar for 14 days. The actively growing culture was flooded with sterile distilled water, dislodging the spores. The concentration of the suspension was then adjusted to lxl 0 6 spores/mL. Tween 20 was then added to the suspension at the rate of 0.05%.
• Colletotrichum sublineolum sorghum anthracnose
• Bacterial strains were prepared as described in Example 6.1. Leaf pieces were sprayed with 120 of the bacterial strains (1 ⁇ 10 8 CFU/mL, suspended in magnesium chloride buffer) using a ribbed skirt fine mist fingertip sprayer (ID- S009, Container & Packaging Supply, Eagle, ID), fitted to a 15 mL conical centrifuge tube (Fisher Scientific, Cat No. l4-59-53A). Alternatively, Pyraclostrobin was applied at 10 ppm as the fungicide control treatment. The treated leaf pieces were then plated on 1% water agar amended with 6-Benzylaminopurine (BAP) and incubated at room temperature in the dark for 24 hours.
• BAP 6-Benzylaminopurine
• the tomato cultivar Money Maker was used in the following bacterial strain evaluation.
• a detached leaf assay was used to evaluate the bacterial strains.
• Fully expanded leaflets from 4 - 5 -week- old tomato plants were excised and made into disks of equal sizes using a 3.5-cm diameter cork borer.
• To prepare the inoculum a culture of P. infestans, genotype US-23, isolated from tomatoes in North Carolina (obtained from Dr. Ristaino Laboratory at North Carolina State University) was used.
• a 2- week-old actively growing culture was flooded with sterile distilled water and mycelium scrapped off. The mycelial suspension was then ruptured for consistency.
• Bacterial strains were prepared as described in Example 6.1. Leaf disks (3.5-cm diameter each) were sprayed with 120 of the bacterial strain ( 1 ⁇ 10 8 CFU/ml, suspended in magnesium chloride buffer) using a ribbed skirt fine mist fingertip sprayer (ID- S009, Container & Packaging Supply, Eagle, ID) fitted to a 15 mL conical centrifuge tube (Fisher Scientific, Cat No.14-59-53A). Alternatively, the fungicide Mefenoxam was applied at 5 ppm for the fungicide control treatment.
• ID- S009 Container & Packaging Supply, Eagle, ID
• leaf disks 24 hours after treatment with a bacterial strain, fungicide, or neither, leaf disks were inoculated with a 30 droplet of P. infestans mycelial suspension.
• the leaf disks were placed abaxial side in contact with a saturated double layer of WhatmanTM 3MM chromatography paper, 20 ⁇ 20 cm (Fisher Scientific Cat No. 3030-861), in a plastic container (BlisterBox P5887, 20 ⁇ 20 cm, Placon, Madison, WI). Boxes with leaf disks were placed inside a double zipper gallon storage bag (ZIP IGS250-448632, AEP Industrial Inc.
• Example 7 Field Trials of bacterial strains against various fungal pests
• At least one bacterial strain recited in Table 2, or an active variant thereof is applied to the crops listed in Table 5 in the field under the current agronomic practices as listed in Table 9 to achieve uniform plant coverage and follow proper agronomic practices. Treatments are applied preventatively and/or curatively at the appropriate timings per disease.
• Table 9 A lication of a bacterial strain to control a fungal est
• Example 8 Field trials of bacterial strains against various fungal pests via seed treatments
• At least one bacterial strain recited in Table 2, or an active variant thereof, is applied to the crops listed in Table 10 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 11. The specific treatments are outlined below.
• Table 11 A lication of a bacterial strain to control a fungal est
• Example 9 Field Trials of bacterial strains against various fungal pests via in-furrow treatments
• At least one bacterial strain recited in Table 2, or an active variant thereof, is applied to the crops listed in Table 12 as in-furrow treatments at time of planting as preventative control for the diseases and at the treatment rates listed in Table 12.
• the specific treatments are outlined below:
• Table 12 Application of a bacterial strain via in-furrow treatment to control a fungal pest
• Example 10 Methods of formulation for the bacterial strains
• a wettable powder formulation may be made using a bacterial stain recited in Table 2, or an active variant thereof.
• a culture produced as described in Example 1 is centrifuged, 20 minutes, 10,000 rpm to produce a pellet. The supernatant is poured off and another volume of culture fluid is added to the previous pellet and centrifuged again.
• a wettable powder formulation is produced by adding 5% (by mass of pellet) of glycerol to the cell pellet and then mixed with a spatula. 20% (by mass) of Microcel E is transferred to a food processor and the glycerol/pellet is poured over the Microcel. This material is blended using the knife blade attachment of the food processor for not more than 10 seconds. The product is dried overnight at 40 °C to approximately an Aw of 0.3. The dried powder formulation is stored in vacuum sealed mylar pouches at 22 C. The dried powder formulation retains antifungal activity, nematicidal activity, and/or insecticidal activity.
• Example 11 Methods to form a spray dried composition
• a spray dried composition may be made using a bacterial strain recited in Table 2, or an active variant thereof.
• a culture produced as described in Example 1 is spray dried using a centrifugal atomizer (Anhydro S-l) under the following conditions: Air heater: Electrical resistance; Target inlet temperature: 160 °C; Target outlet temperature: 65 - 75 °C.
• Three 8 liter carboys are combined into a mixing tank containing the bacterial cutlure and agitated thoroughly.
• the spray dried formulations ranged from about 10 6 CFU/g to about 10 11 CFU/g.
• the spray dried formulation is then blended with up to 5% w/w with the anti -caking agent Sipernat S22.
• a seed treatment formulation may be made using a bacterial strain recited in Table 2, or an active variant thereof.
• 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.

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