WO2015077278A1 - Compositions and methods comprising chromobacterium for controlling plant nematode pests and plant insect pests - Google Patents

Abstract

Disclosed herein are pest controlling compositions comprising a novel strain of Chromobacterium spp. Further disclosed are methods of using such compositions for controlling plant pests and other invasive pests.

Description

COMPOSITIONS AND METHODS COMPRISING CHROMOBACTERIUM FOR CONTROLLING PLANT NEMATODE PESTS AND PLANT INSECT PESTS

FIELD OF THE INVENTION

Disclosed herein are compositions comprising a novel strain of Chromobacterium for controlling plant pests and other invasive pests. Further disclosed are methods of using such compositions for controlling crop damaging plant nematode pests and insect pests in agricultural environments.

BACKGROUND OF THE INVENTION

Pests, such as nematodes, insects, and acari (mites and ticks) are a major problem for the agriculture industry, limiting productivity, often significantly. Although chemical solutions have been used to control pests, excessive use of chemicals leaves residues in soil, water, and air, and has additional adverse effects on non-target organisms and the ecological balance. In addition, pests can develop resistance to chemical pesticides, limiting their effectiveness and application. Public concern over potential health hazards of chemical pesticides and the increase in cost of chemical pesticides has also led to the exploration of more eco-friendly pest management tactics.

Microbial pesticides have been developed for use as an alternative, or in some cases as a supplement, to chemical pesticides. Microbial pesticides are living organisms (e.g., bacteria and fungi) that intervene in the life cycle of pests (by killing or disabling the pest). Examples of microbial pesticides include microbial nematicides, and in particular, nematophagous bacteria including strains of Chromobacterium spp.

Tian, B.; Yang, J.; Zhang, K. (2007). Bacteria used in the biological control of plant- parasitic nematodes: populations, mechanisms of action, and future prospects. FEMS Microbiol Ecol 61 : 197-213 discusses a group of important natural enemies of nematode pests, nematophagous bacteria.

Soby, S.D.; et al, (2013). Chromobacterium vaccinii sp. nov., isolated from native and cultivated cranberry (Vaccinium macrocarpon Ait.) bogs and irrigation ponds. International Journal of Systematic and Evolutionary Microbiology 63: 1840-1846 discusses a number of Gram-negative, motile, mesophilic, violacein-producing bacteria isolated from the soils and roots of Vaccinium macrocarpon Ait. and Kalmia angustifolia L. plants and from irrigation ponds associated with wild and cultivated cranberry bogs in Massachusetts, USA. Phylogenetic analyses of 16S rRNA gene sequences placed these isolates in a clade with Chromobacterium species, but the specialized environment from which they were isolated, indicate that the cranberry and Kalmia isolates comprise a separate species of Chromobacterium, for which the name Chromobacterium vaccinii sp. nov. is proposed. Martin, P.A.W; Gundersen-Rindal, D.; Blackburn, M.; Buyer, J. (2007). Chrombacterium subtsugae sp. nov., a betaproteobacterium toxic to Colorado potato beetle and other insect pests. International Journal of Systematic and Evolutionary Microbiology 57: 993-999 discusses that Chromobacterium subtsugae sp. nov., a motile, Gram-negative, violet-pigmented bacterium isolated from Maryland forest soil, was found to be orally toxic to Colorado potato beetle larvae and other insects.

Martin, P.A.W. ; Hirose, E.; Aldrich, J.R. (2007). Toxicity of Chromobacterium subtsugae to Southern Green Stink Bug (Heteroptera: Pentatomidae) and Corn Rootworm (Coleoptera: Chrvsomelidae). J. Econ. Entomol. 100 (3): 680-684 discusses a new species of Chromobacterium, Chromobacterium subtsugae, found to be toxic to Colorado potato beetle, Leptinotarsa decemlineata, and describes the toxicity of these bacteria in laboratory assays to other pest insects: two corn rootworm species and southern green stink bug.

U.S. Patent No.: 7,244,607 discloses compositions for controlling an insect population including an insect food stuff and an insecticidally-effective amount of at least one Gram negative bacteria, viable, dead or alive, and/or an extract thereof, where the composition is applied to an area accessible to the insects and results in insect death. The compositions are ideally suited for the control of fire ants, cockroaches, carpenter ants and termites.

U.S. Patent No.: 7,037,494 discloses Chromobacterium subtsuga sp. nov., a new species of the genus Chromobacterium which possesses insecticidal activity, is described. The invention also relates to insecticidally-active metabolites obtained from the strain and to insecticidal compositions comprising cultures of the strain and/or supernatants, filtrates, and extracts obtained from the strain, and use thereof to control insect pests.

U.S. Patent Application Publication No.: 2012/0100236 discloses bioactive compounds and metabolites derived from Chromobacterium species culture responsible for controlling pests, compositions containing these compounds, methods for obtaining these compounds and methods of using these compounds and compositions for controlling pests.

U.S. Patent Application Publication No.: 2013/0074735 discloses a process for producing purple-blue natural pigment containing violacein and its derivative (deoxyviolacein) using Chromobacterium sp. NIIST-CKK-01 (MTCC 5522, NCIM 5341 ; Genbank Accession No. FJ982784). The method comprises the steps of maintaining and growing the bacterium in a specific medium under defined conditions of pH, temperature and agitation. At the end of incubation, pigment and biomass is separated from the culture broth, pigment is recovered from the biomass through solvent extraction and finally pigment is concentrated by drying.

Published PCT Patent Application No.: WO 2012/140212 discloses combinations suitable for agricultural use can include (I) a nematode-antagonistic biocontrol agent and (II) one or more agents selected, independently of each other, from any one of (A) to (H): (A) at least one fungicide; (B) at least one insecticide; (C) at least one synthetic nematicide; (D) bacterium of the genus Bacillus; (E) Harpin; (F) Isoflavones; (G) Plant growth regulators; and/or (H) Plant activators.

Published PCT Patent Application No.: WO 2012/037352 discloses compositions for reducing pathogens in soil, and to methods of using such compositions to treat soils. The composition comprises intracellular components of lysed, beneficial, crop and non-crop rhizosphere-inhabiting yeast cells and whole or lysed, beneficial, crop and non-crop rhizosphere-inhabiting bacteria cells. The composition can also include a fertilizer, and a carrier suitable for delivering the composition to the soil.

EP Patent Application Publication No.: EP 0171381 discloses bacteria which are capable of proliferating in an environment which is infested with nematodes (such as Pseudomonads which colonize the surfaces of plant roots) under conditions of microbial competition, and which inhibit nematodes. This invention also relates to a method of using such bacteria to inhibit nematodes This invention also involves the transformation of bacteria with foreign genes which express glycosidase enzymes (such as β-galactosidase) and/or chitinase, which can inhibit one or more types of nematodes. Methods are disclosed herein for (1 ) selecting suitable bacteria which can colonize a target environment, such as the root surfaces of a particular type of plant; (2) transforming a selected bacteria with foreign genes which encode chitinase and/or glycosidase enzymes, preferably under the control of a strong promoter; and (3) determining whether the bacteria effectively inhibit a selected type of nematodes in the target environment.

KR Patent No.: 100802494 discloses a pest controlling agent comprising violacein is provided to inhibit the growth of plant pathogenic fungi and plant parasitic nematode, thereby being usefully used for effective controlling of anthracnose, pythium blight, stem rot, and bean sprouts damping-off. The violacein is derived from Chromobacterium violaceum, Alteromonas luteoviolacea or Janthinobacterium lividum. The method for preparing violacein comprises the steps of: (a) culturing Chromobacterium violaceum; (b) centrifuging the culture solution obtained from the step(a); (c) extracting the supernatant obtained from the step(b) with an organic solvent; and (d) concentrating the extracted solution obtained from the step(c) under reduced pressure.

As natural agents, microbial pesticides offer more eco-friendly solutions for controlling pests and/or for use in combination with chemical pesticide. A need exists for a superior microbial pesticide that can control nematode pests and insect pests.

SUMMARY OF THE INVENTION

Disclosed herein is a novel strain of Chomobacterium spp., Chromobacterium vaccinii strain NRRL B-50880. Further disclosed are compositions and methods which offer an improved microbial pesticide for controlling damage caused to crops by pest populations. The composition will comprise an agriculturally suitable carrier, and a bacterial strain of Chromobacterium vaccinii having the deposit accession number NRRL B-50880. In an embodiment, additional optional ingredients (e.g., beneficial microbes, signal molecules, pesticides, fungicides, nematicides, and combinations thereof) may also be used in combination with the compositions described herein, including as part of the same composition or applied as a separate treatment.

Disclosed herein are also methods for controlling pests. In an embodiment, the method comprises contacting a soil with a strain of Chromobacterium vaccinii having the deposit accession number NRRL B-50880 or compositions described herein which comprise a strain of Chromobacterium vaccinii having the deposit accession number NRRL B-50880. In aother embodiment, the method comprises contacting a plant or plant part with a strain of Chromobacterium vaccinii having the deposit accession number NRRL B-50880 or compositions described herein which comprise a strain of Chromobacterium vaccinii having the deposit accession number NRRL B-50880. In still yet another embodiment, the method comprises contacting a pest with a strain of Chromobacterium vaccinii having the deposit accession number NRRL B-50880 or compositions described herein which comprise a strain of Chromobacterium vaccinii having the deposit accession number NRRL B-50880.

Further disclosed are seeds coated with a strain of Chromobacterium vaccinii having the deposit accession number NRRL B-50880 or compositions described herein which comprise a strain of Chromobacterium vaccinii having the deposit accession number NRRL B- 50880.

BRIEF DESCRIPTION OF THE DRAWINGS

Figure 1 illustrates an example phylogenetic tree.

Figure 2 illustrates an example phylogenetic tree.

Figure 3 illustrates example bioactivity of strain NRRL B-50880, Escherichia coli and control (water) against Caenorhabditis elegans after 8 hours (A) and 3 days (B).

DETAILED DESCRIPTION OF THE INVENTION

The disclosed embodiments relate to compositions and methods for controlling infestations of arthropod pests, particularly infestations of bed bugs in human dwellings. Definitions:

As used herein, the singular forms "a", "an" and "the" are intended to include the plural forms as well, unless the context clearly indicates otherwise.

As used herein, the term "agriculturally beneficial ingredient(s)" is intended to mean any agent or combination of agents capable of causing or providing a beneficial and/or useful effect in agriculture other than a biologically active ingredient. As used herein, "biologically active ingredient(s)" is intended to mean any biological organism or chemical element, molecule, or compound, or mixture thereof, which has a biological activity in a seed, a plant, or a plant part (e.g., plant signal molecules, other microorganisms, gluconolactones, glutathiones, etc.). Non-limiting examples of "biological activity" include N₂ fixation, phosphate solubilization, plant growth-enhancement, bio- pesticidal activity, etc.

As used herein, the terms "enhanced plant growth", "increased plant growth", "plant growth-enhancement", or "plant growth-enhancing", which may all be used interchangeably, are intended to refer to increased plant yield (e.g., increased biomass, increased fruit number, or a combination thereof as measured by bushels per acre), increased root number, increased root mass, increased root volume, increased leaf area, increased plant stand, increased plant vigor, faster seedling emergence (i.e., enhanced emergence), faster germination, (i.e., enhanced germination), or combinations thereof.

As used herein the terms "signal molecule(s)" or "plant signal molecule(s)", which may be used interchangeably with "plant growth-enhancing agent(s)," broadly refers to any agent, both naturally occurring in plants or microbes, and synthetic (and which may be non-naturally occurring) that directly or indirectly activates or inactivates a plant biochemical pathway, resulting in increased or enhanced plant growth, compared to untreated plants or plant parts (e.g., seeds and plants harvested from untreated seeds). Non-limiting examples of signal molecules include LCOs, COs, chitinous compounds, flavonoids, jasmonic acid or derivatives thereof, linoleic acid or derivatives thereof, linolenic acid or derivatives thereof, kerrikins, etc.

As used herein, the term "pesticidal" means any agent or combination of agents that is pathogenic to at least one target pest (e.g., a nematode, an insect, an acari, a fungal pest, a bacterial pest, a viral pest, etc.).

As used herein, the terms "microbial pesticide" or "biopesticide", which can be used interchangeably, means any microorganism, whether in a vegetative state, a dormant state (e.g., spore) or a whole broth culture, any substance derived from a microorganism (e.g., metabolites), or any fermentation product (e.g., supernatants, filtrates, extracts, etc.) that are pathogenic to a pest (e.g., capable of attacking, infecting, killing, disabling, causing disease, and/or causing injury to a pest), and is thus able to be used in the control of a pest by adversely affecting the viability or growth of the target pest. Non-limiting examples of "microbial pesticides" include microbial nematocides, microbial insecticides, microbial fungicides, microbial bactericides, and microbial viricides).

As used herein, "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. In the event that the "source" is an organism, "derived from" means that it may be isolated or obtained from the organism itself or medium used to culture or grow said organism.

As used herein, "whole broth culture" 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.

As used herein, the term "supernatant" refers to the liquid remaining when cells 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.

As defined herein, "filtrate" refers to liquid from a whole broth culture that has passed through a membrane.

As defined herein, "extract" refers to liquid substance removed from cells by a solvent (water, detergent, buffer) and separated from the cells by centrifugation, filtration or other method.

As used herein, "metabolite" refers to a compound, substance or byproduct of a fermentation of a microorganism, or supernatant, filtrate, or extract obtained from a microorganism that has pesticidal and particularly, insecticidal activity. As defined herein, an "isolated compound" is essentially free of other compounds or substances, e.g., at least about 20% pure, preferably at least about 40% pure, more preferably about 60% pure, even more preferably about 80% pure, most preferably about 90% pure, and even most preferably about 95% pure, as determined by analytical methods, including but not limited to chromatographic methods, electrophoretic methods

As used herein, the terms "spore", "microbial spore", etc., has its normal meaning which is well known and understood by those of skill in the art (i.e., a microorganism in its dormant, protected state).

As used herein, the term "herbicide(s)" is intended to refer to any agent or combination of agents capable of killing weeds and/or inhibiting the growth of weeds (the inhibition being reversible under certain conditions).

As used herein, the term "fungicide(s)" is intended to refer to any agent or combination of agents capable of killing fungi and/or inhibiting fungal growth.

As used herein, the term "nematicide" or "nematicidal" is intended to refer to any agent or combination of agents capable of killing one or more nematodes and/or inhibiting the growth of one or more nematodes.

As used herein, the term "insecticide" or "insecticidal" is intended to refer to any agent or combination of agents capable of killing one or more insects and/or inhibiting the growth of one or more insects. As used herein, the term "acaricide" or "acaricidal" is intended to refer to any agent or combination of agents capable of killing one or more acarids and/or inhibiting the growth of one or more acarids.

As used herein in, a "cuticle degrading enzyme" is an enzyme that is able to at least partially degrade a cuticle of a pest, such as, the epicuticle and/or the procuticle. The exogenously applied cuticle degrading enzyme can increase the efficacy of the fungal pesticide by increasing the ability of the fungal pesticide to colonize and/or or bore through the pest's cuticle to reach the pest's body cavity.

The term "pest" refers to any animal of the scientific classification (phylum) Nematoda (e.g., root-knot nematode, soybean cyst nematode, etc.), Arthropoda including Insecta, (e.g., white flies, thrips, weevils, etc.) and/or Arachnida, (e.g., mites, ticks, spiders, etc.).

As used herein, "exogenously applied" means that the cuticle degrading enzyme is applied independently (that is, as a separate ingredient) from the compositions disclosed herein and any enzyme produced by fungal pesticide.

The "exogenously applied" cuticle degrading enzyme is in the form of an "isolated" enzyme composition.

The term "isolated" means the enzyme is in a form or environment which does not occur in nature, that is, the enzyme is at least partially removed from one or more or all of the naturally occurring constituents with which it is associated in nature. Thus, although enzymes produced endogenously by the fungal pesticide will impact efficacy, an isolated enzyme does not encompass an enzyme endogenously produced by the fungal pesticide during treatment of a pest in the processes of the present invention. An isolated enzyme may be present in the form of a purified enzyme composition or a fermentation broth sample that contains the enzyme.

As used herein, the term "control" or "controlling" as in e.g., the phrase: the "control" of pests or pest populations, or "controlling" pests or pest populations, or as in the phrase: "controlling" pests, refers to preventing, reducing, killing, inhibiting the growth of, or elimination of a pest or population of pests as defined herein. Indeed, "control" or "controlling" as used herein refers to any indicia of success in prevention, killing, inhibition, elimination, reduction or amelioration of a pest or pest population.

As used herein, the terms "effective amount", "effective concentration", or "effective dosage" are defined as the amount, concentration, or dosage of the bacterial strain (i.e., the Chromobacterium strain, and variants thereof described herein) sufficient to cause infection in the pest which will then lead to the controlling of pests. The actual effective dosage in absolute value depends on factors including, but not limited to, the mortality rate of the pest or pests relative to the rate at which the bacterials strain is applied, synergistic or antagonistic interactions between the other active or inert ingredients which may increase or reduce the activity of the bacterial strain, the inherent susceptibility of the life stage and species of pest, and the stability of the bacterial strain in compositions. The "effective amount", "effective concentration", or "effective dosage" of the bacterial strain may be determined, e.g., by a routine dose response experiment.

As used herein, the term "nitrogen fixing organism(s)" is intended to refer to any organism capable of converting atmospheric nitrogen (N₂) into ammonia (NH₃).

As used herein, the term "phosphate solubilizing organism" is intended to refer to any organism capable of converting insoluble phosphate into a soluble phosphate form.

As used herein, the term "inoculum" is intended to mean any form of microbial cells, or spores, which is capable of propagating on or in a substrate (e.g., a soil) when the conditions of temperature, moisture, etc., are favorable for microbial growth.

As used herein, the term "isomer(s)" is intended to include all stereoisomers of the compounds and/or molecules referred to herein (e.g., flavonoids, LCOs, COs, chitinous compounds, jasmonic acid or derivatives thereof, linoleic acid or derivatives thereof, linolenic acid or derivatives thereof, kerrikins, gluconolactones, glutathiones, etc.), including enantiomers, diastereomers, as well as all conformers, roatmers, and tautomers, unless otherwise indicated. The compounds and/or molecules disclosed herein include all enantiomers in either substantially pure levorotatory or dextrorotatory form, or in a racemic mixture, or in any ratio of enantiomers. Where embodiments disclose a (D)-enantiomer, that embodiment also includes the (L)-enantiomer; where embodiments disclose a (L)- enantiomer, that embodiment also includes the (D)-enantiomer. Where embodiments disclose a (+)-enantiomer, that embodiment also includes the (-)-enantiomer; where embodiments disclose a (-)-enantiomer, that embodiment also includes the (+)-enantiomer. Where embodiments disclose a (S)-enantiomer, that embodiment also includes the (R)- enantiomer; where embodiments disclose a (R)-enantiomer, that embodiment also includes the (S)-enantiomer. Embodiments are intended to include any diastereomers of the compounds and/or molecules referred to herein in diastereomerically pure form and in the form of mixtures in all ratios. Unless stereochemistry is explicitly indicated in a chemical structure or chemical name, the chemical structure or chemical name is intended to embrace all possible stereoisomers, conformers, rotamers, and tautomers of compounds and/or molecules depicted.

As used herein, the term "carrier" is intended to refer to an "agronomically acceptable carrier." An "agronomically acceptable carrier" is intended to refer to any material which can be used to deliver the actives (e.g. , the bacterial strains, such as Chromobacterium strain, and variants thereof described herein) to a plant or plant part (e.g., foliage or seed). As used herein, the term "soil-compatible carrier" is intended to refer to any material which can be added to a soil without causing/having an adverse effect on plant growth, soil structure, soil drainage, or the like.

As used herein, the term "seed-compatible carrier" is intended to refer to any material which can be added to a seed without causing/having an adverse effect on the seed, the plant that grows from the seed, seed germination, or the like.

As used herein, the term "foliar-compatible carrier" is intended to refer to any material which can be added to a plant or plant part without causing/having an adverse effect on the plant, plant part, plant growth, plant health, or the like.

As used herein, the terms "plant(s)" and "plant part(s)" are intended to refer to all plants and plant populations such as desired and undesired wild plants or crop plants (including naturally occurring crop plants). Crop plants can be plants, which can be obtained by conventional plant breeding and optimization methods or by biotechnological and genetic engineering methods or by combinations of these methods, including the transgenic plants and including the plant cultivars protectable or not protectable by plant breeders' rights. Plant parts are to be understood as meaning all parts and organs of plants above and below the ground, such as shoot, leaf, flower and root, examples which may be mentioned being leaves, needles, stalks, stems, flowers, fruit bodies, fruits, seeds, roots, tubers and rhizomes. The plant parts also include harvested material and vegetative and generative propagation material (e.g., cuttings, tubers, rhizomes, off-shoots and seeds, etc.).

As used herein, the term "foliage" is intended to mean all parts and organs of plants above the ground. Non-limiting examples include leaves, needles, stalks, stems, flowers, fruit bodies, fruits, etc. As used herein, the term "foliar application", "foliarly applied", and variations thereof, is intended to include application of an active ingredient to the foliage or above ground portions of the plant, (e.g., the leaves of the plant). Application may be effected by any means known in the art (e.g., spraying the active ingredient).

As used herein, the term "phosphorus source" is intended to mean a compound of that element which, at least in the soil conditions under consideration, does not make the element fully available for plant uptake.

As used herein, the term "nutrient(s)" is intended to refer to any nutrient (e.g., vitamins, macrominerals, micronutrients, trace minerals, organic acids, etc.) which are needed for plant growth, plant health, and/or plant development.

As used herein, the term "biostimulant(s)" is intended to refer to any agent or combination of agents capable of enhancing metabolic or physiological processes within plants and soils. BACTERIAL STRAINS/CULTURES

In one embodiment, the strain described herein is a microbial pesticide. In another embodiment, the strain is a microbial nematicide. In a particular embodiment, the strain is a nematophagous bacteria. In a more particular embodiment, the strain is a strain of Chromobacterium spp. In an even more particular embodiment, the strain is a strain of Chromobacterium vaccinii. In still an even more particular embodiment, the strain is a strain of Chromobacterium vaccinii sp. nov.

In yet an even more particular embodiment, the strain is a strain of Chromobacterium vaccinii having the deposit accession number NRRL B-50880 (deposited with the American Type Culture Collection). Cultures of the deposited strain may consist of viable bacteria, including whole broth cultures. In another embodiment, the deposited strain(s) is a biologically pure culture (e.g., cultures having a purity of at least 60%, of at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, up to 100% pure). In another embodiment, the strain is a variant of Chromobacterium vaccinii having the deposit accession number NRRL B-50880. As used herein, the term variant shall mean a microbe which is (i) a progeny (unmodified descendents) of the strain of Chromobacterium vaccinii having the deposit accession number NRRL B-50880 and (ii) modified descendents of the strain of Chromobacterium vaccinii having the deposit accession number NRRL B-50880.

The Chromobacterium vaccinii described herein, and in particular, the strain having deposit accession number NRRL B-50880 and variants thereof, are cultivated in nutrient medium using methods known in the art. Suitable media are available may be available from commercial sources or prepared according to published compositions. Non-limiting examples of acceptable growth media include blood agar plates, King's medium B (KMB) agar (Soby, S.D.; et al, (2013)), tryptic soy medium, yeast extract mannitol medium (YEM), glycerol yeast extract (GYEA), yeast extract-peptone-glycerol (YPG), peptone yeast extract glucose (PYG) MacConkey agar, or malt extract agar, or in flasks containing suitable liquid media such as tryptic soy broth, YEM broth, KMB broth, GYEA broth. YPG broth, PYG broth, etc.

The organisms may be cultivated by shake flask cultivation, small scale or large scale fermentation (including but not limited to continuous, batch, fed-batch, or solid state fermentations) in laboratory or industrial fermentors performed in suitable medium and under conditions allowing cell growth. The cultivation may take place in suitable nutrient medium comprising carbon and nitrogen sources and inorganic salts, using procedures known in the art. These culture methods may be used in the preparation of an inoculum of Chromobacterium spp. for coating seeds and/or application to carrier to be applied to plants, plant parts, or soil. The above mentioned deposited strain was isolated from the Pawnee Grasslands in Colorado State and deposited on November 6, 2013, under terms of the Budapest Treaty on the International Recognition of the Deposit of Microorganisms for the Purposes of Patent Procedure at the Agricultural Research Service Culture Collection, 1815 North University Street, Peoria, Illinois 61604, U.S.A.

COMPOSITIONS:

The the compositions used in the embodiments disclosed herein comprise an agriculturally suitable carrier and the bacterial strain disclosed herein (i.e., Chromobacterium vaccinni strain having deposit accession number NRRL B-50880, as well as variants thereof). In a particular embodiment the compositions comprise a pesticidially effective amount of the Chromobacterium vaccinni strain having deposit accession number NRRL B-50880, as well as variants thereof, described herein.

In a particular embodiment, the compositions will comprise an agriculturally suitable carrier and whole broth cultures, liquid cultures, or suspensions of a strain from a Chromobacterium vaccinii, e.g., a strain having the identifying characteristics of Chromobacterium vaccinii and more particularly, having the identifying characteristics of NRRL B-50880, as well as supernatants, filtrates or extracts obtained from a strain of a Chromobacterium vaccinni., e.g., a strain having the identifying characteristics of Chromobacterium vaccinii and more particularly, having the identifying characteristics of NRRL B-50880, or the supernatant, filtrate and/or extract or one or more metabolites or isolated compounds derived from a strain of a Chromobacterium vaccinii or combinations of the foregoing which in particular have nematocidal activity and or insecticidal activity.

The compositions described herein will have the benefit of controlling pests, and in particular plant nematode pests such as root-knot, cyst, lesion, sting, and ring nematodes, including: Meloidogyne spp., Heterodera spp., Globodera spp., Pratylenchus spp., Belonolaimus spp., and Criconemella spp., more particularly, Meloidogyne incognita (i.e, root knot nematode), Heterodera glycine (i.e., soybean cyst nematode), and Belonolaimus longicaudatus (i.e., sting nematode). The compositions will further have the benefit of controlling insect pests, and in particular, agriculturally damaging insect pests e.g., white flies, thrips, mites, weevils, ticks, chinch bugs, etc.).

The compositions described herein can be of any form so long as the composition is able to support the desired activity (effective amount) of the bacterial strain disclosed here in (i.e., the Chromobacterium vaccinni strain having deposit accession number NRRL B-50880), regardless of form, and the composition can be applied to control a target pest. The carrier may be used to provide an environment to support the viability of the bacterial strain including by providing the proper environmental conditions and protecting the strain from harmful environmental conditions (e.g., excess oxygen, moisture and/or ultraviolet radiation, etc.). Unless the compositions are generated immediately prior to use, the carrier may be used to maintain the activity of the bacterial strain during storage (e.g., in a container for the entire shelf-life of the formulated product). The carrier may also be used to maintain the activity of the bacterial strain after the compositions described throughout have been applied to the application surface (e.g., a plant, plant part, and/or a soil). In particular embodiments, the carrier provides an environment such that the bacterial strain will not have more than a 1-log loss of the original viable content (prior to including in a carrier) over at least a one year period.

In certain embodiments, the compositions described herein may be in the form of a gel, a foam, a solid (such as a powder, granule, particle, etc.), or a liquid. In a particular embodiment, the composition is in the form of a liquid.

Carrier(s):

The carrier will have the correct values (and range of values) for rheological measurements (e.g. , viscosity, yield value, storage modulus, and loss modulus) to allow the bacterial strain to remain efficacious (e.g., capable of pesticidal activity) and viable once formulated. Non-limiting examples of carriers described herein include liquids, gels, slurries, or solids (including wettable powders or dry powders). The selection of the carrier material will depend on the intended application. In a particular embodiment, the carrier is an agronomically acceptable carrier. In another embodiment, the carrier is a foliar-compatible carrier. In still another particular embodiment, the carrier is a soil-compatible carrier. In still yet another particular embodiment, the carrier is a seed-compatible carrier. In yet still another particular embodiment, the carrier is an agonomically acceptable carrier, a foliarly-acceptable carrier, a soil-acceptable carrier, and a seed-compatible carrier.

In a particular embodiment, the carrier may be an aqueous or non-aqueous liquid carrier. Non-limiting examples of liquids useful as carriers for the compositions disclosed herein include water, an aqueous solution (e.g., sugar water), or a non-aqueous solution. In one embodiment, the carrier is water. In another embodiment the carrier is an aqueous solution. In another embodiment, the carrier is a non-aqueous liquid.

In a particular embodiment, the carrier is a non-aqueous liquid (e.g., an oil, etc.). The non-aqueous liquid may be a biodegradable non-aqueous liquid. The non-aqueous liquid may be a "Low Vapor Pressure Volatile Organic Compounds (LVP-VOC)," which is a chemical "compound" or "mixture of compounds" containing (1 ) a vapor pressure less than 0.1 mm Hg at 20 °C, (2) composed of chemical compounds with more than 12 carbon atoms and/or (3) a boiling point greater than 216 °C. See the definition of LVP-VOC provided by the California Air Resources Board (CARB). The non-aqueous liquid may be a biodegradable LVP-VOC non-aqueous liquid. Non-limiting examples of non-aqueous liquids suitable as a carrier for the compositions described herein include silicone oils, paraffinic/parrafin oils, mineral oils, hexylene glycol, glycerol, linoleic acid, oleic acid, and any combination thereof. Non-limiting examples of a commercial mineral/paraffinic oils include BRITOL 50 (available from Sonneborn, Inc., Mahwah, NJ), Ultra-Fine Spray oil (available from Sunoco, Petronas Lubricants, Belgium NV), SunSpray 6N oil (available from Sunoco, Petronas Lubricants, Belgium NV), SunSpray 7E Range oil (available from Sunoco, Petronas Lubricants, Belgium NV), SunSpray 7N oil, (available from Sunoco, Petronas Lubricants, Belgium NV), SunSpray 1 1 E Range oil (available from Sunoco, Petronas Lubricants, Belgium NV), SunSpray 1 1 N oil (available from Sunoco, Petronas Lubricants, Belgium NV), Banana Spray oil (available from Sunoco, Petronas Lubricants, Belgium NV), and BioSpray oil (available from Sunoco, Petronas Lubricants, Belgium NV). An example of a silicone oil is DM Fluid 100 CS (available from Shin-Etsu Chemical Co., LtD., Tokyo, Japan).

If a liquid carrier is used, the liquid carrier may further include growth media to culture one or more microbial strains which could be useful to the compositions described. Non- limiting examples of suitable growth media for microbial strains include KMB, tryptic soy broth, YEM broth, KMB broth, GYEA broth. YPG broth, PYG broth, etc, Czapek-Dox medium, potato dextrose broth, or any media known to those skilled in the art to be compatible with, and/or provide growth nutrients to microbial strain which may be included to the compositions described herein.

In an embodiment, composition, may be formed of 1.00 wt. % to 99.99 wt. % of carrier. There may be minor variances when measuring the weight percentage of the carrier and the composition may be formed of about 1 .00 wt. % to about 99.99 wt. % of carrier. In still another embodiment, the composition may be formed of 50.00 wt. % to 99.99 wt. % of carrier. Again, there may be minor variances when measuring the weight percentage of the carrier and the composition may be formed of about 50.00 wt. % to about 99.99 wt. % of carrier. In still yet another embodiment, the composition is formed of 50.00 wt. % to 80.00 wt. % of carrier. Yet again, there may be minor variances when measuring the weight percentage of the carrier and the composition may be formed of about 50.00 wt. % to about 80.00 wt. % of carrier.

Optional Ingredients:

The compositions described herein may further comprise one or more optional ingredients that are physically and/or chemically compatible with the composition embodied herein. Non-limiting optional ingredients include biologically active ingredients (e.g., beneficial plant signal molecules, beneficial microorganisms, enzymes, gluconolactones, glutathiones, etc.), agriculturally beneficial ingredients (micronutrients, biostimulants, preservatives, polymers, wetting agents, surfactants, herbicides, fungicides, insecticides, acaricides, nematicides, anti-freezing agents, insect growth regulators, preservatives etc.), and combinations thereof. Such ingredients are known to those skilled in the art.

Biologically Active Ingredient(s):

The compositions described herein may further comprise one or more biologically active ingredients other than the Chromobacterium vaccinii strain described herein, in particular the Chromobacterium vaccinii strain having the deposit accession number NRRL B-50880 and variants thereof.

Non-limiting examples of biologically active ingredients include plant signal molecules (e.g., lipo-chitooligosaccharides (LCO), chitooligosaccharides (CO), chitinous compounds, flavonoids, jasmonic acid or derivatives thereof, linoleic acid or derivatives thereof, linolenic acid or derivatives thereof, karrikins, etc.), beneficial microorganisms (e.g., Rhizobium spp., Bradyrhizobium spp., Sinorhizobium spp., Azorhizobium spp., Glomus spp., Gigaspora spp. , Hymenoscyphous spp., Oidiodendron spp., Laccaria spp., Pisolithus spp., Rhizopogon spp., Scleroderma spp., Rhizoctonia spp., Acinetobacter spp., Arthrobacter spp,, Arthrobotrys spp., Aspergillus spp., Azospirillum spp., Bacillus spp, Burkholderia spp., Candida spp., Chryseomonas spp., Chromobacterim spp., Enterobacter spp., Eupenicillium spp., Exiguobacterium spp., Klebsiella spp., Kluyvera spp., Microbacterium spp., Mucor spp., Paecilomyces spp., Paenibacillus spp., Penicillium spp., Pseudomonas spp., Serratia spp., Stenotrophomonas spp., Streptomyces spp., Streptosporangium spp., Swaminathania spp., Thiobacillus spp., Torulospora spp., Vibrio spp., Xanthobacter spp., Xanthomonas spp., etc.), and enzymes.

Plant Signal Molecule(s):

In at least one embodiment, the compositions described herein may comprise one or more plant signal molecules. Alternatively, the one or more plant signal molecules may be applied either simultaneously or applied sequentially, with the compositions disclosed herein. In an embodiment, the compositions described herein include one or more plant signal molecules. In one embodiment, the one or more plant signal molecules are one or more LCOs. In another embodiment, the one or more plant signal molecules are one or more COs. In still another embodiment, the one or more plant signal molecules are one or more chitinous compounds. In yet another embodiment, the one or more plant signal molecules are one or more flavonoids or derivatives thereof. In still yet another embodiment, the one or more plant signal molecules are one or more non-flavonoid nod gene inducers (e.g., jasmonic acid, linoleic acid, linolenic acid, and derivatives thereof). In still yet another embodiment, the one or more plant signal molecules are one or more karrikins or derivatives thereof. In still another embodiment, the one or more plant signal molecules are one or more LCOs, one or more COs, one or more chitinous compounds, one or more flavonoids and derivatives thereof, one or more non-flavonoid nod gene inducers and derivatives thereof, one or more karrikins and derivatives thereof, or any signal molecule combination thereof.

LCOs:

Lipo-chitooligosaccharide compounds (LCOs), also known in the art as symbiotic Nod signals or Nod factors, consist of an oligosaccharide backbone of 3-l,4-linked /V-acetyl-D-glucosamine ("GlcNAc") residues with an N-linked fatty acyl chain condensed at the non-reducing end. LCO's differ in the number of GlcNAc residues in the backbone, in the length and degree of saturation of the fatty acyl chain, and in the substitutions of reducing and non-reducing sugar residues. LCOs are intended to include all LCOs as well as isomers, salts, and solvates thereof. An example of an LCO is presented below as formula I:

in which:

G is a hexosamine which can be substituted, for example, by an acetyl group on the nitrogen, a sulfate group, an acetyl group and/or an ether group on an oxygen,

R-i , R₂, R₃, R₅, R₆ and R₇, which may be identical or different, represent H, CH₃ CO-, C_x H_y CO- where x is an integer between 0 and 17, and y is an integer between 1 and 35, or any other acyl group such as for example a carbamyl,

R₄ represents a mono-, di-, tri- and tetraunsaturated aliphatic chain containing at least 12 carbon atoms, and n is an integer between 1 and 4.

LCOs may be obtained (isolated and/or purified) from bacteria such as Rhizobia, e.g., Rhizobium spp., Bradyrhizobium spp., Sinorhizobium spp. and Azorhizobium spp. LCO structure is characteristic for each such bacterial species, and each strain may produce multiple LCO's with different structures. For example, specific LCOs from S. meliloti have also been described in U.S. Patent 5,549,718 as having the formula II:

in which R represents H or CH₃ CO- and n is equal to 2 or 3.

Even more specific LCOs include NodRM, NodRM-1 , NodRM-3. When acetylated (the R=CH₃ CO-), they become AcNodRM-1 , and AcNodRM-3, respectively (U.S. Patent 5,545,718).

LCOs from Bradyrhizobium japonicum are described in U.S. Patents 5,175,149 and 5,321 ,01 1. Broadly, they are pentasaccharide phytohormones comprising methylfucose. A number of these B. japonicum-denved LCOs are described: BjNod-V (C_18:i); BjNod-V (A_c, Ci8:i), BjNod-V (Ci6:i); and BjNod-V (A_c, Ci_6:o), with "V" indicating the presence of five N-acetylglucosamines; "Ac" an acetylation; the number following the "C" indicating the number of carbons in the fatty acid side chain; and the number following the ":" the number of double bonds.

LCOs used in compositions of the invention may be obtained (i.e., isolated and/or purified) from bacterial strains that produce LCO's, such as strains of Azorhizobium, Bradyrhizobium (including B. japonicum), Mesorhizobium, Rhizobium (including R. leguminosarum), Sinorhizobium (including S. meliloti), and bacterial strains genetically engineered to produce LCO's.

Also encompassed by the present invention are compositions using LCOs obtained (i.e., isolated and/or purified) from a mycorrhizal fungus, such as fungi of the group Glomerocycota, e.g., Glomus intraradicus. The structures of representative LCOs obtained from these fungi are described in WO 2010/049751 and WO 2010/049751 (the LCOs described therein also referred to as "Myc factors").

Further encompassed by compositions of the present invention is use of synthetic LCO compounds, such as those described in WO 2005/063784, and recombinant LCO's produced through genetic engineering. The basic, naturally occurring LCO structure may contain modifications or substitutions found in naturally occurring LCO's, such as those described in Spaink, Crit. Rev. Plant Sci. 54:257-288 (2000) and D'Haeze, et al. , Glycobiology 72:79R-105R (2002). Precursor oligosaccharide molecules (COs, which as described below, are also useful as plant signal molecules in the present invention) for the construction of LCOs may also be synthesized by genetically engineered organisms, e.g., as in Samain, et al., Carb. Res. 302:35-42 (1997); Samain, et al., J. Biotechnol. 72:33-47 (1999).

LCO's may be utilized in various forms of purity and may be used alone or in the form of a culture of LCO-producing bacteria or fungi. Methods to provide substantially pure LCO's include simply removing the microbial cells from a mixture of LCOs and the microbe, or continuing to isolate and purify the LCO molecules through LCO solvent phase separation followed by HPLC chromatography as described, for example, in U.S. Patent 5,549,718. Purification can be enhanced by repeated HPLC, and the purified LCO molecules can be freeze-dried for long-term storage.

COs:

Chitooligosaccharides (COs) are known in the art as β-1-4 linked N-actylglucosamine structures identified as chitin oligomers, also as N-acetylchitooligosaccharides. CO's have unique and different side chain decorations which make them different from chitin molecules [(C₈H₁₃N0₅)n, CAS No. 1398-61-4], and chitosan molecules [(C₅HnN0₄)n, CAS No. 9012-76-4]. Representative literature describing the structure and production of COs is as follows: Van der Hoist, et al., Current Opinion in Structural Biology, 77:608-616 (2001 ); Robina, et al., Tetrahedron 58:521-530 (2002); Hanel, et al. , Planta 232:787-806 (2010); Rouge, et al. Chapter 27, "The Molecular Immunology of Complex Carbohydrates" in Advances in Experimental Medicine and Biology, Springer Science; Wan, et al., Plant Cell 27:1053-69 (2009); PCT/F100/00803 (9/21/2000); and Demont-Caulet, et al., Plant Physiol. 120(1 ):83-92 (1999). The COs may be synthetic or recombinant. Methods for preparation of recombinant COs are known in the art. See, e.g., Samain, et al. (supra.); Cottaz, et al., Meth. Eng. 7(4j:31 1-7 (2005) and Samain, et al., J. Biotechnol. 72:33-47 (1999). COs are intended to include isomers, salts, and solvates thereof. Chitinous Compounds:

Chitins and chitosans, which are major components of the cell walls of fungi and the exoskeletons of insects and crustaceans, are also composed of GlcNAc residues. Chitinous compounds include chitin, (lUPAC: N-[5-[[3-acetylamino-4,5-dihydroxy-6- (hydroxymethyl)oxan-2yl]methoxymethyl]-2-[[5-acetylamino-4,6-dihydroxy-2- (hydroxymethyl)oxan-3-yl]methoxymethyl]-4-hydroxy-6-(hydroxymethyl)oxan-3- ys]ethanamide), chitosan, (lUPAC: 5-amino-6-[5-amino-6-[5-amino-4,6-dihydroxy- 2(hydroxymethyl)oxan-3-yl]oxy-4-hydroxy-2-(hydroxymethyl)oxan-3-yl]oxy- 2(hydroxymethyl)oxane-3,4-diol), and isomers, salts, and solvates thereof.

These compounds may be obtained commercially, e.g., from Sigma-Aldrich, or prepared from insects, crustacean shells, or fungal cell walls. Methods for the preparation of chitin and chitosan are known in the art, and have been described, for example, in U.S. Patent 4,536,207 (preparation from crustacean shells), Pochanavanich, et al., Lett. Appl. Microbiol. 35:17-21 (2002) (preparation from fungal cell walls), and U.S. Patent 5,965,545 (preparation from crab shells and hydrolysis of commercial chitosan). Deacetylated chitins and chitosans may be obtained that range from less than 35% to greater than 90% deacetylation, and cover a broad spectrum of molecular weights, e.g., low molecular weight chitosan oligomers of less than 15kD and chitin oligomers of 0.5 to 2kD; "practical grade" chitosan with a molecular weight of about 15kD; and high molecular weight chitosan of up to 70kD. Chitin and chitosan compositions formulated for seed treatment are also commercially available. Commercial products include, for example, ELEXA® (Plant Defense Boosters, Inc.) and BEYOND™ (Agrihouse, Inc.).

Flavonoids:

Flavonoids are phenolic compounds having the general structure of two aromatic rings connected by a three-carbon bridge. Flavonoids are produced by plants and have many functions, e.g., as beneficial signaling molecules, and as protection against insects, animals, fungi and bacteria. Classes of flavonoids include are known in the art. See, Jain, et al., J. Plant Biochem. & Biotechnol. 77:1 -10 (2002); Shaw, et al., Environmental Microbiol. 77:1867-80 (2006). Flavonoid compounds are commercially available, e.g. , from Novozymes BioAg, Saskatoon, Canada; Natland International Corp., Research Triangle Park, NC; MP Biomedicals, Irvine, CA; LC Laboratories, Woburn MA. Flavonoid compounds may be isolated from plants or seeds, e.g. , as described in U.S. Patents 5,702,752; 5,990,291 ; and 6,146,668. Flavonoid compounds may also be produced by genetically engineered organisms, such as yeast, as described in Ralston, et al., Plant Physiology 737:1375-88 (2005). Flavonoid compounds are intended to include all flavonoid compounds as well as isomers, salts, and solvates thereof. The one or more flavonoids may be a natural flavonoid (i.e., not synthetically produced), a synthetic flavonoid (e.g., a chemically synthesized flavonoid) or a combination thereof. In a particular embodiment, the compositions described herein comprise a flavanol, a flavone, an anthocyanidin, an isoflavonoid, a neoflavonoid and combinations thereof, including all isomer, solvate, hydrate, polymorphic, crystalline form, non-crystalline form, and salt variations thereof.

In an embodiment, the compositions described herein may comprise one or more flavanols. In still another embodiment, the compositions described herein may comprise one or more flavanols selected from the group consisting of flavan-3-ols (e.g., catechin (C), gallocatechin (GC), catechin 3-gallate (Cg), gallcatechin 3-gallate (GCg), epicatechins (EC), epigallocatechin (EGC) epicatechin 3-gallate (ECg), epigallcatechin 3-gallate (EGCg), etc.), flavan-4-ols, flavan-3,4-diols (e.g., leucoanthocyanidin), proanthocyanidins (e.g., includes dimers, trimer, oligomers, or polymers of flavanols), and combinations thereof. In still yet another embodiment, the compositions described herein may comprise one or more flavanols selected from the group consisting of catechin (C), gallocatechin (GC), catechin 3- gallate (Cg), gallcatechin 3-gallate (GCg), epicatechins (EC), epigallocatechin (EGC) epicatechin 3-gallate (ECg), epigallcatechin 3-gallate (EGCg), flavan-4-ol, leucoanthocyanidin, and dimers, trimers, olilgomers or polymers thereof.

In another embodiment, the compositions described herein may comprise one or more flavones. In still another embodiment, the compositions described herein may comprise one or more flavones selected from the group consisting of flavones (e.g., luteolin, apigenin, tangeritin, etc.), flavonols (e.g., quercetin, quercitrin, rutin, kaempferol, kaempferitrin, astragalin, sophoraflavonoloside, myricetin, fisetin, isorhamnetin, pachypodol, rhamnazin, etc.), flavanones (e.g. hesperetin, hesperidin, naringenin, eriodictyol, homoeriodictyol, etc.), and flavanonols (e.g., dihydroquercetin, dihydrokaempferol, etc.). In still yet another embodiment, the compositions described herein may comprise one or more flavones selected from the group consisting of luteolin, apigenin, tangeritin, quercetin, quercitrin, rutin, kaempferol, kaempferitrin, astragalin, sophoraflavonoloside, myricetin, fisetin, isorhamnetin, pachypodol, rhamnazin, hesperetin, hesperidin, naringenin, eriodictyol, homoeriodictyol, dihydroquercetin, dihydrokaempferol, and combinations thereof.

In still another embodiment, the compositions described herein may comprise one or more anthocyanidins. In yet another embodiment, the compositions described herein may comprise one or more anthocyanidins selected from the group selected from the group consisting of cyanidins, delphinidins, malvidins, pelargonidins, peonidins, petunidins, and combinations thereof.

In another embodiment, the compositions described herein may comprise one or more isoflavonoids. In still yet another embodiment, the compositions described herein comprise one or more isoflavonoids selected from the group consisting of phytoestrogens, isoflavones (e.g., genistein, daidzein, glycitein, etc.), and isoflavanes (e.g., equol, lonchocarpane, laxiflorane, etc.), and combinations thereof. In yet another embodiment the compositions described herein may comprise one or more isoflavonoids selected from the group consisting of genistein, daidzein, glycitein, equol, lonchocarpane, laxiflorane, and combinations thereof.

In another embodiment, the compositions described herein may comprise one or more neoflavonoids. In yet another embodiment, the compositions described herein may comprise one or more neoflavonoids selected from the group consisting of neoflavones (e.g., calophyllolide), neoflavenes (e.g., dalbergichromene), coutareagenins, dalbergins, nivetins, and combinations thereof. In still yet another embodiment, the compositions described herein may comprise one or more neoflavonoids selected from the group consisting of calophyllolide, dalbergichromene, coutareagenin, dalbergin, nivetin, and combinations thereof.

In another embodiment, the compositions described herein may comprise one or flavonoids selected from the group consisting of catechin (C), gallocatechin (GC), catechin 3- gallate (Cg), gallcatechin 3-gallate (GCg), epicatechins (EC), epigallocatechin (EGC) epicatechin 3-gallate (ECg), epigallcatechin 3-gallate (EGCg), flavan-4-ol, leucoanthocyanidin, proanthocyanidins, luteolin, apigenin, tangeritin, quercetin, quercitrin, rutin, kaempferol, kaempferitrin, astragalin, sophoraflavonoloside, myricetin, fisetin, isorhamnetin, pachypodol, rhamnazin, hesperetin, hesperidin, naringenin, eriodictyol, homoeriodictyol, dihydroquercetin, dihydrokaempferol, cyanidins, delphinidins, malvidins, pelargonidins, peonidins, petunidins, genistein, daidzein, glycitein, equol, lonchocarpane, laxiflorane, calophyllolide, dalbergichromene, coutareagenin, dalbergin, nivetin, and combinations thereof. In still another embodiment, the compositions described herein may comprise one or more flavonoids selected from the group consisting of hesperetin, hesperidin, naringenin, genistein, daidzein, and combinations thereof. In a particular embodiment, the composition described herein may comprise the flavonoid hesperetin. In another particular embodiment, the composition described herein may comprise the flavonoid hesperidin. In still another particular embodiment, the composition described herein may comprise the flavonoid naringenin. In still yet another particular embodiment, the composition described herein may comprise the flavonoid genistein. In yet still another particular embodiment, the composition described herein may comprise the flavonoid daidzein.

Non-Flavonoid Nod-Gene Inducer(s):

Jasmonic acid (JA, [1 R-[1a,23(Z)]]-3-oxo-2-(pentenyl)cyclopentaneacetic acid) and its derivatives, linoleic acid ((Z,Z)-9,12-Octadecadienoic acid) and its derivatives, and linolenic acid ((Z,Z,Z)-9,12,15-octadecatrienoic acid) and its derivatives, may also be used in the compositions described herein. Non-flavonoid nod-gene inducers are intended to include not only the non-flavonoid nod-gene inducers described herein, but isomers, salts, and solvates thereof.

Jasmonic acid and its methyl ester, methyl jasmonate (MeJA), collectively known as jasmonates, are octadecanoid-based compounds that occur naturally in plants. Jasmonic acid is produced by the roots of wheat seedlings, and by fungal microorganisms such as Botryodiplodia theobromae and Gibbrella fujikuroi, yeast (Saccharomyces cerevisiae), and pathogenic and non-pathogenic strains of Escherichia coli. Linoleic acid and linolenic acid are produced in the course of the biosynthesis of jasmonic acid. Jasmonates, linoleic acid and linoleic acid (and their derivatives) are reported to be inducers of nod gene expression or LCO production by rhizobacteria. See, e.g. , Mabood, Fazli, Jasmonates induce the expression of nod genes in Bradyrhizobium japonicum, May 17, 2001 ; and Mabood, Fazli, "Linoleic and linolenic acid induce the expression of nod genes in Bradyrhizobium japonicum," USDA 3, May 17, 2001 .

Useful derivatives of linoleic acid, linolenic acid, and jasmonic acid that may be useful in compositions of the present invention include esters, amides, glycosides and salts. Representative esters are compounds in which the carboxyl group of linoleic acid, linolenic acid, or jasmonic acid has been replaced with a -COR group, where R is an -OR¹ group, in which R¹ is: an alkyl group, such as a Ci-C₈ unbranched or branched alkyl group, e.g., a methyl, ethyl or propyl group; an alkenyl group, such as a C₂-C₈ unbranched or branched alkenyl group; an alkynyl group, such as a C₂-C₈ unbranched or branched alkynyl group; an aryl group having, for example, 6 to 10 carbon atoms; or a heteroaryl group having, for example, 4 to 9 carbon atoms, wherein the heteroatoms in the heteroaryl group can be, for example, N, O, P, or S. Representative amides are compounds in which the carboxyl group of linoleic acid, linolenic acid, or jasmonic acid has been replaced with a -COR group, where R is an NR²R³ group, in which R² and R³ are independently: hydrogen; an alkyl group, such as a C C₈ unbranched or branched alkyl group, e.g. , a methyl, ethyl or propyl group; an alkenyl group, such as a C₂-C₈ unbranched or branched alkenyl group; an alkynyl group, such as a C₂-C₈ unbranched or branched alkynyl group; an aryl group having, for example, 6 to 10 carbon atoms; or a heteroaryl group having, for example, 4 to 9 carbon atoms, wherein the heteroatoms in the heteroaryl group can be, for example, N, O, P, or S. Esters may be prepared by known methods, such as acid-catalyzed nucleophilic addition, wherein the carboxylic acid is reacted with an alcohol in the presence of a catalytic amount of a mineral acid. Amides may also be prepared by known methods, such as by reacting the carboxylic acid with the appropriate amine in the presence of a coupling agent such as dicyclohexyl carbodiimide (DCC), under neutral conditions. Suitable salts of linoleic acid, linolenic acid, and jasmonic acid include e.g., base addition salts. The bases that may be used as reagents to prepare metabolically acceptable base salts of these compounds include those derived from cations such as alkali metal cations (e.g., potassium and sodium) and alkaline earth metal cations (e.g., calcium and magnesium). These salts may be readily prepared by mixing together a solution of linoleic acid, linolenic acid, or jasmonic acid with a solution of the base. The salt may be precipitated from solution and be collected by filtration or may be recovered by other means such as by evaporation of the solvent.

Karrikin(s):

Karrikins are vinylogous 4H-pyrones e.g., 2H-furo[2,3-c]pyran-2-ones including derivatives and analogues thereof. It is intended that the karrikins include isomers, salts, and solvates thereof. Examples of the ented by the following structure:

wherein; Z is O, S or NR₅; R-i, R₂, R3, and R₄ are each independently H, alkyl, alkenyl, alkynyl, phenyl, benzyl, hydroxy, hydroxyalkyl, alkoxy, phenyloxy, benzyloxy, CN, COR₆,

COOR=, halogen, NR₆R₇, or N0₂; and R₅, R₆, and R₇ are each independently H, alkyl or alkenyl, or a biologically acceptable salt thereof. Examples of biologically acceptable salts of these compounds may include acid addition salts formed with biologically acceptable acids, examples of which include hydrochloride, hydrobromide, sulphate or bisulphate, phosphate or hydrogen phosphate, acetate, benzoate, succinate, fumarate, maleate, lactate, citrate, tartrate, gluconate; methanesulphonate, benzenesulphonate and p-toluenesulphonic acid.

Additional biologically acceptable metal salts may include alkali metal salts, with bases, examples of which include the sodium and potassium salts. Examples of compounds embraced by the structure and which may be suitable for use in the present invention include the following: 3-methyl-2H-furo[2,3-c]pyran-2-one (where

R₂, R₃, R₄=H), 2H- furo[2,3-c]pyran-2-one (where Ri, R₂, R3, R4=H), 7-methyl-2H-furo[2,3-c]pyran-2-one (where

Ri, R₂, R₄=H, R₃=CH₃), 5-methyl-2H-furo[2,3-c]pyran-2-one (where R-,, R₂, R₃=H, R₄=CH₃),

3,7-dimethyl-2H-furo[2,3-c]pyran-2-one (where R-,, R₃=CH₃, R₂, R₄=H), 3,5-dimethyl-2H- furo[2,3-c]pyran-2-one (where Ri, R₄=CH₃, R₂, R₃=H), 3,5,7-trimethyl-2H-furo[2,3-c]pyran-2- one (where R^ R₃, R₄=CH₃, R₂=H), 5-methoxymethyl-3-methyl-2H-furo[2,3-c]pyran-2-one

(where R₂, R₃=H, R₄=CH₂OCH₃), 4-bromo-3,7-dimethyl-2H-furo[2,3-c]pyran-2-one

(where R-,, R₃=CH₃, R₂=Br, R₄=H), 3-methylfuro[2,3-c]pyridin-2(3H)-one (where Z=NH, R₂, R₃, R₄=H), 3,6-dimethylfuro[2,3-c]pyridin-2(6H)-one (where Z=N-CH₃, Ri=CH₃, R₂, R₃, R₄=H). See, U.S. Patent 7,576,213. These molecules are also known as karrikins. See, Halford, "Smoke Signals," in Chem. Eng. News (April 12, 2010), at pages 37-38 (reporting that karrikins or butenolides which are contained in smoke act as growth stimulants and spur seed germination after a forest fire, and can invigorate seeds such as corn, tomatoes, lettuce and onions that had been stored). These molecules are the subject of U.S. Patent 7,576,213.

Gluconolactones:

In at least one embodiment, the compositions disclosed herein may comprise one or more gluconolactones. Alternatively, the one or more gluconolactones may be applied either simultaneously or applied sequentially, with the compositions disclosed herein. The one or more gluconolactones may be a natural gluconolactones (i.e., not synthetically produced), a synthetic glutathione (e.g., a chemically synthesized gluconolactones) or a combination thereof. The one or more gluconolactones may also be in any form (e.g., oxidized, reduced, or a combination of oxidized and reduced species).

In one embodiment, the one or more gluconolactones have the molecular formula C₆H₁₀O₆ and a molar mass of about 178.14 g mol^"1. In another embodiment, the one or more gluconolactones may include gluconolactones having the structure:

and isomers, salts, and solvates thereof.

Glutathiones:

In at least one embodiment, the compositions disclosed herein may comprise one or more glutathiones. Alternatively, the one or more glutathiones may be applied either simultaneously or applied sequentially, with the compositions disclosed herein. The one or more glutathiones may be a natural glutathione (i.e., not synthetically produced), a synthetic glutathione (e.g., a chemically synthesized glutathione) or a combination thereof. The one or more glutathiones may also be in any form (e.g., oxidized, reduced, or a combination of oxidized and reduced species).

In one embodiment, the one or more glutathiones have the molecular formula CioH₁₇N₃0₆S and a molar mass of about 307.32 g mol^"1. In another embodiment, the one or more glutathiones may include glutathiones having the structure:

and isomers, salts, and solvates thereof.

Beneficial Microorqanism(s):

In an embodiment, the compositions described herein may comprise one or more beneficial microorganisms in addition to the Chombacterium vaccinii strain having the deposit accession number NRRL B-50880 and variants thereof. Alternatively, the one or more additional beneficial microorganisms may be applied either simultaneously or applied sequentially, with the compositions disclosed herein.

The one or more beneficial microorganisms may include any number of microorganisms having one or more beneficial properties (e.g., produce one or more of the plant signal molecules described herein, enhance nutrient and water uptake, promote and/or enhance nitrogen fixation, enhance growth, enhance seed germination, enhance seedling emergence, break the dormancy or quiescence of a plant, produce or express toxins which supplement and/or enhance the activity of the fungal pesticide (e.g. δ-endotoxin, a- exotoxin, β-exotoxin, etc. produced by Bacillus thuringiensis), provide anti-fungal activity, etc.). The one or more beneficial microorganisms may be in a spore form, a vegetative form, or a combination thereof.

In a more particular embodiment, the one or more bacteria are spore forming bacterial strains. Methods for producing stabilized microorganisms, and bacteria specifically, are known in the art. See Donnellan, J. E., Nags, E. H., and Levinson, H. S. (1964). "Chemically defined, synthetic media for sporulation and for germination and growth of Bacillus subtilis." Journal of Bacteriology 87(2):332-336; and Chen, Z., Li, Q., Liu, H. Yu, N., Xie, T., Yang, M., Shen, P., Chen, X. (2010). "Greater enhancement of Bacillus subtilis spore yields in submerged cultures by optimization of medium composition through statistical experimental designs." Appl. Microbiol. Biotechnol. 85: 1353-1360.

Non-limiting examples of spore forming bacterial strains include strains from the genera Acetonema, Alkalibacillus, Ammoniphilus, Amphibacillus, Anaerobacter, Anaerospora, Aneurinibacillus, Anoxybacillus, Bacillus, Brevibacillus, Caldanaerobacter, Caloramator, Caminicella, Cerasibacillus, Clostridium, Clostridiisalibacter, Cohnella, Dendrosporobacter, Desulfotomaculum, Desulfosporomusa, Desulfosporosinus, Desulfovirgula, Desulfunispora, Desulfurispora, Filifactor, Filobacillus, Gelria, Geobacillus, Geosporobacter, Gracilibacillus, Halonatronum, Heliobacterium, Heliophilum, Laceyella, Lentibacillus, Lysinibacillus, Mahella, Metabacterium, Moorella, Natroniella, Oceanobacillus, Orenia, Ornithinibacillus, Oxalophagus, Oxobacter, Paenibacillus, Paraliobacillus, Pelospora, Pelotomaculum, Piscibacillus, Planifilum, Pontibacillus, Propionispora, Salinibacillus, Salsuginibacillus, Seinonella, Shimazuella, Sporacetigenium, Sporoanaerobacter, Sporobacter, Sporobacterium, Sporohalobacter, Sporolactobacillus, Sporomusa, Sporosarcina, Sporotalea, Sporotomaculum, Syntrophomonas, Syntrophospora, Tenuibacillus, Tepidibacter, Terribacillus, Thalassobacillus, Thermoacetogenium, Thermoactinomyces, Thermoalkalibacillus, Thermoanaerobacter, Thermoanaeromonas, Thermobacillus, Thermoflavimicrobium, Thermovenabulum, Tuberibacillus, Virgibacillus, and/ or Vulcanobacillus.

In a particular embodiment, the one or more spore forming bacteria is a bacteria selected from the genera consisting of Acetonema, Alkalibacillus, Ammoniphilus, Amphibacillus, Anaerobacter, Anaerospora, Aneurinibacillus, Anoxybacillus, Bacillus, Brevibacillus, Caldanaerobacter, Caloramator, Caminicella, Cerasibacillus, Clostridium, Clostridiisalibacter, Cohnella, Dendrosporobacter, Desulfotomaculum, Desulfosporomusa, Desulfosporosinus, Desulfovirgula, Desulfunispora, Desulfurispora, Filifactor, Filobacillus, Gelria, Geobacillus, Geosporobacter, Gracilibacillus, Halonatronum, Heliobacterium, Heliophilum, Laceyella, Lentibacillus, Lysinibacillus, Mahella, Metabacterium, Moorella, Natroniella, Oceanobacillus, Orenia, Ornithinibacillus, Oxalophagus, Oxobacter, Paenibacillus, Paraliobacillus, Pelospora, Pelotomaculum, Piscibacillus, Planifilum, Pontibacillus, Propionispora, Salinibacillus, Salsuginibacillus, Seinonella, Shimazuella, Sporacetigenium, Sporoanaerobacter, Sporobacter, Sporobacterium, Sporohalobacter, Sporolactobacillus, Sporomusa, Sporosarcina, Sporotalea, Sporotomaculum, Syntrophomonas, Syntrophospora, Tenuibacillus, Tepidibacter, Terribacillus, Thalassobacillus, Thermoacetogenium, Thermoactinomyces, Thermoalkalibacillus, Thermoanaerobacter, Thermoanaeromonas, Thermobacillus, Thermoflavimicrobium, Thermovenabulum, Tuberibacillus, Virgibacillus, Vulcanobacillus, and combinations thereof.

In another embodiment, the one or more bacterial strains is a strain of Bacillus spp. , e.g., Bacillus alcalophilus, Bacillus alvei, Bacillus aminovorans, Bacillus amyloliquefaciens, Bacillus aneurinolyticus, Bacillus aquaemaris, Bacillus atrophaeus, Bacillus boroniphilius, Bacillus brevis, Bacillus caldolyticus, Bacillus centrosporus, Bacillus cereus, Bacillus circulans, Bacillus coagulans, Bacillus firmus, Bacillus flavothermus, Bacillus fusiformis, Bacillus globigii, Bacillus infernus, Bacillus larvae, Bacillus laterosporus, Bacillus lentus, Bacillus licheniformis, Bacillus megaterium, Bacillus, mesentericus, Bacillus mucilaginosus, Bacillus mycoides, Bacillus natto, Bacillus pantothenticus, Bacillus polymyxa, Bacillus pseudoanthracis, Bacillus pumilus, Bacillus schlegelii, Bacillus sphaericus, Bacillus sporothermodurans, Bacillus stearothermophillus, Bacillus subtilis, Bacillus thermoglucosidasius, Bacillus thuringiensis, Bacillus vulgatis, Bacillus weihenstephanensis, and combinations thereof.

In another embodiment, the one or more bacterial strains is a strain of Brevibacillus spp., e.g., Brevibacillus brevis; Brevibacillus formosus; Brevibacillus laterosporus; or Brevibacillus parabrevis, and combinations thereof.

In another embodiment, the one or more bacterial strains is a strain of Paenibacillus spp., e.g., Paenibacillus alvei; Paenibacillus amylolyticus; Paenibacillus azotofixans; Paenibacillus cookii; Paenibacillus macerans; Paenibacillus polymyxa; or Paenibacillus validus, and combinations thereof.

In one embodiment, the one or more beneficial microorganisms are diazotrophs (i.e., bacteria which are symbiotic nitrogen-fixing bacteria). In still another embodiment, the one or more diazotrophs are selected from the genera Rhizobium spp., Bradyrhizobium spp., Azorhizobium spp., Sinorhizobium spp., Mesorhizobium spp. , Azospirillum spp., and combinations thereof. In still another embodiment, the one or more beneficial microorganisms are bacteria selected from the group consisting of Rhizobium cellulosilyticum, Rhizobium daejeonense, Rhizobium etli, Rhizobium galegae, Rhizobium gallicum, Rhizobium giardinii, Rhizobium hainanense, Rhizobium huautlense, Rhizobium indigoferae, Rhizobium leguminosarum, Rhizobium loessense, Rhizobium lupini, Rhizobium lusitanum, Rhizobium meliloti, Rhizobium mongolense, Rhizobium miluonense, Rhizobium sullae, Rhizobium tropici, Rhizobium undicola, Rhizobium yanglingense, Bradyrhizobium bete, Bradyrhizobium canariense, Bradyrhizobium elkanii, Bradyrhizobium iriomotense, Bradyrhizobium japonicum, Bradyrhizobium jicamae, Bradyrhizobium liaoningense, Bradyrhizobium pachyrhizi, Bradyrhizobium yuanmingense, Azorhizobium caulinodans, Azorhizobium doebereinerae, Sinorhizobium abri, Sinorhizobium adhaerens, Sinorhizobium americanum, Sinorhizobium aborts Sinorhizobium fredii, Sinorhizobium indiaense, Sinorhizobium kostiense, Sinorhizobium kummerowiae, Sinorhizobium medicae, Sinorhizobium meliloti, Sinorhizobium mexicanus, Sinorhizobium morelense, Sinorhizobium saheli, Sinorhizobium terangae, Sinorhizobium xinjiangense, Mesorhizobium albiziae, Mesorhizobium amorphae, Mesorhizobium chacoense, Mesorhizobium ciceri, Mesorhizobium huakuii, Mesorhizobium loti, Mesorhizobium mediterraneum, Mesorhizobium pluifarium, Mesorhizobium septentrionale, Mesorhizobium temperatum, Mesorhizobium tianshanense, Azospirillum amazonense, Azospirillum brasilense, Azospirillum canadense, Azospirillum doebereinerae, Azospirillum formosense, Azospirillum halopraeferans, Azospirillum irakense, Azospirillum largimobile, Azospirillum lipoferum, Azospirillum melinis, Azospirillum oryzae, Azospirillum picis, Azospirillum rugosum, Azospirillum thiophilum, Azospirillum zeae, and combinations thereof. In a particular embodiment, the one or more diazotrophs are selected from the group consisting of Bradyrhizobium japonicum, Rhizobium leguminosarum, Rhizobium meliloti, Sinorhizobium meliloti, Azospirillum brasilense, and combinations thereof. In another embodiment, the beneficial microorganism is Bradyrhizobium japonicum. In another embodiment, the beneficial microorganism is Rhizobium leguminosarum. In another embodiment, the beneficial microorganism is Rhizobium meliloti. In another embodiment, the beneficial microorganism is Sinorhizobium meliloti. In another embodiment, the beneficial microorganism is Azospirillum brasilense.

In a particular embodiment, the one or more diazotrophs comprises one or more strains of Rhizobium leguminosarum. In another particular embodiment, the strain of R. leguminosarum comprises the strain S012A-2-(IDAC 080305-01 ). In another particular embodiment, the one or more diazotrophs comprises a strain of Bradyrhizobium japonicum. In still another particular embodiment, the strain of Bradyrhizobium japonicum comprises the strain B. japonicum USDA 532C, B. japonicum USDA 1 10, B. japonicum USDA 123, B. japonicum USDA 127, B. japonicum USDA 129, B. japonicum NRRL B-50608, B. japonicum NRRL B-50609, B. japonicum NRRL B-50610, B. japonicum NRRL B-5061 1 , B. japonicum NRRL B-50612, B. japonicum NRRL B-50592 (deposited also as NRRL B- 59571 ), B. japonicum NRRL B-50593 (deposited also as NRRL B-59572), B. japonicum NRRL B-50586 (deposited also as NRRL B-59565), B. japonicum NRRL B-50588 (deposited also as NRRL B-59567), B. japonicum NRRL B-50587 (deposited also as NRRL B-59566), B. japonicum NRRL B-50589 (deposited also as NRRL B-59568), B. japonicum NRRL B- 50591 (deposited also as NRRL B-59570), B. japonicum NRRL B-50590 (deposited also as NRRL B-59569), NRRL B-50594 (deposited also as NRRL B-50493), B. japonicum NRRL B- 50726, B. japonicum NRRL B-50727, B. japonicum NRRL B-50728, B. japonicum NRRL B- 50729, B. japonicum NRRL B-50730, and combinations thereof.

In still yet a more particular embodiment, the one or more diazotrophs comprises one or more strains of R. leguminosarum comprises the strain S012A-2-(IDAC 080305-01 ), B. japonicum USDA 532C, B. japonicum USDA 1 10, B. japonicum USDA 123, B. japonicum USDA 127, B. japonicum USDA 129, B. japonicum NRRL B-50608, B. japonicum NRRL B- 50609, B. japonicum NRRL B-50610, B. japonicum NRRL B-5061 1 , B. japonicum NRRL B-50612, B. japonicum NRRL B-50592 (deposited also as NRRL B-59571 ), B. japonicum NRRL B-50593 (deposited also as NRRL B-59572), B. japonicum NRRL B-50586 (deposited also as NRRL B-59565), B. japonicum NRRL B-50588 (deposited also as NRRL B-59567), B. japonicum NRRL B-50587 (deposited also as NRRL B-59566), B. japonicum NRRL B- 50589 (deposited also as NRRL B-59568), B. japonicum NRRL B-50591 (deposited also as NRRL B-59570), B. japonicum NRRL B-50590 (deposited also as NRRL B-59569), NRRL B- 50594 (deposited also as NRRL B-50493), B. japonicum NRRL B-50726, B. japonicum NRRL B-50727, B. japonicum NRRL B-50728, B. japonicum NRRL B-50729, B. japonicum NRRL B-50730, and combinations thereof.

In another embodiment, the one or more beneficial microorganisms comprise one or more phosphate solubilizing microorganisms. Phosphate solubilizing microorganisms include fungal and bacterial strains. In an embodiment, the phosphate solubilizing microorganism are microorganisms selected from the genera consisting of Acinetobacter spp., Arthrobacter spp, Arthrobotrys spp., Aspergillus spp. , Azospirillum spp. , Bacillus spp., Burkholderia spp., Candida spp., Chryseomonas spp., Enterobacter spp. , Eupenicillium spp. , Exiguobacterium spp., Klebsiella spp. , Kluyvera spp., Microbacterium spp., Mucor spp., Paecilomyces spp. , Paenibacillus spp. , Penicillium spp., Pseudomonas spp. , Serratia spp., Stenotrophomonas spp., Streptomyces spp., Streptosporangium spp. , Swaminathania spp. , Thiobacillus spp. , Torulospora spp., Vibrio spp., Xanthobacter spp., Xanthomonas spp., and combinations thereof. In still yet another embodiment, the phosphate solubilizing microorganism is a microorganism selected from the group consisting of Acinetobacter calcoaceticus, Arthrobotrys oligospora, Aspergillus niger, Azospirillum amazonense, Azospirillum brasilense, Azospirillum canadense, Azospirillum doebereinerae, Azospirillum formosense, Azospirillum halopraeferans, Azospirillum irakense, Azospirillum largimobile, Azospirillum lipoferum, Azospirillum melinis, Azospirillum oryzae, Azospirillum picis, Azospirillum rugosum, Azospirillum thiophilum, Azospirillum zeae, Bacillus amyloliquefaciens, Bacillus atrophaeus, Bacillus circulans, Bacillus licheniformis, Bacillus subtilis, Burkholderia cepacia, Burkholderia vietnamiensis, Candida krissii, Chryseomonas luteola, Enterobacter aerogenes, Enterobacter asburiae, Enterobacter taylorae, Eupenicillium parvum, Kluyvera cryocrescens, Mucor ramosissimus, Paecilomyces hepialid, Paecilomyces marquandii, Paenibacillus macerans, Paenibacillus mucilaginosus, Penicillium bilaiae (formerly known as Penicillium bilaii), Penicillium albidum, Penicillium aurantiogriseum, Penicillium chrysogenum, Penicillium citreonigrum, Penicillium citrinum, Penicillium digitatum, Penicillium frequentas, Penicillium fuscum, Penicillium gaestrivorus, Penicillium glabrum, Penicillium griseofulvum, Penicillium implicatum, Penicillium janthinellum, Penicillium lilacinum, Penicillium minioluteum, Penicillium montanense, Penicillium nigricans, Penicillium oxalicum, Penicillium pinetorum, Penicillium pinophilum, Penicillium purpurogenum, Penicillium radicans, Penicillium radicum, Penicillium raistrickii, Penicillium rugulosum, Penicillium simplicissimum, Penicillium solitum, Penicillium variabile, Penicillium velutinum, Penicillium viridicatum, Penicillium glaucum, Penicillium fussiporus, and Penicillium expansum, Pseudomonas corrugate, Pseudomonas fluorescens, Pseudomonas lutea, Pseudomonas poae, Pseudomonas putida, Pseudomonas stutzeri, Pseudomonas trivialis, Serratia marcescens, Stenotrophomonas maltophilia, Swaminathania salitolerans, Thiobacillus ferrooxidans, Torulospora globosa, Vibrio proteolyticus, Xanthobacter agilis, Xanthomonas campestris, and combinations thereof.

In a particular embodiment, the one or more phosphate solubilizing microorganisms is a strain of the fungus Penicillium. In another embodiment, the one or more Penicillium species is P. bilaiae, P. gaestrivorus, or combinations thereof. In a particular embodiment, the strain of Penicillium comprises P. bilaiae NRRL 50169, P. bilaiae ATCC 20851 , P. bilaiae ATCC 22348, P. bilaiae ATCC 18309, P. bilaiae NRRL 50162 and combinations thereof. In another particular embodiment, the strain of Penicillium comprises strain P. gaestrivorus NRRL 50170. In still yet another particular embodiment, the strain of Penicillium comprises P. bilaiae NRRL 50169, P. bilaiae ATCC 20851 , P. bilaiae ATCC 22348, P. bilaiae ATCC 18309, P. bilaiae NRRL 50162, P. gaestrivorus NRRL 50170, and combinations thereof.

In another embodiment the beneficial microorganism is one or more mycorrhiza. In particular, the one or more mycorrhiza is an endomycorrhiza (also called vesicular arbuscular mycorrhizas, VAMs, arbuscular mycorrhizas, or AMs), an ectomycorrhiza, or a combination thereof.

In one embodiment, the one or more mycorrhiza is an endomycorrhiza of the phylum Glomeromycota and genera Glomus and Gigaspora. In still a further embodiment, the endomycorrhiza is a strain of Glomus aggregatum, Glomus brasilianum, Glomus clarum, Glomus deserticola, Glomus etunicatum, Glomus fasciculatum, Glomus intraradices, Glomus monosporum, or Glomus mosseae, Gigaspora margarita, or a combination thereof.

In another embodiment, the one or more mycorrhiza is an ectomycorrhiza of the phylum Basidiomycota, Ascomycota, and Zygomycota. In still yet another embodiment, the ectomycorrhiza is a strain of Laccaria bicolor, Laccaria laccata, Pisolithus tinctorius, Rhizopogon amylopogon, Rhizopogon fulvigleba, Rhizopogon luteolus, Rhizopogon villosuli, Scleroderma cepa, Scleroderma citrinum, or a combination thereof.

In still another embodiment, the one or more mycorrhiza is an ericoid mycorrhiza, an arbutoid mycorrhiza, or a monotropoid mycorrhiza. Arbuscular and ectomycorrhizas form ericoid mycorrhiza with many plants belonging to the order Ericales, while some Ericales form arbutoid and monotropoid mycorrhizas. All orchids are mycoheterotrophic at some stage during their lifecycle and form orchid mycorrhizas with a range of basidiomycete fungi. In one embodiment, the mycorrhiza may be an ericoid mycorrhiza, preferably of the phylum Ascomycota, such as Hymenoscyphous ericae or Oidiodendron sp. In another embodiment, the mycorrhiza also may be an arbutoid mycorrhiza, preferably of the phylum Basidiomycota. In yet another embodiment, the mycorrhiza may be a monotripoid mycorrhiza, preferably of the phylum Basidiomycota. In still yet another embodiment, the mycorrhiza may be an orchid mycorrhiza, preferably of the genus Rhizoctonia. In still another embodiment, the one or more beneficial microorganisms are microorganisms capable of exhibiting pesticidal activity, (e.g., fungicidal activity, i.e. , biofungicides). Non-limiting examples of biofungicides include, Ampelomyces quisqualis (e.g., AQ 10® from Intrachem Bio GmbH & Co. KG, Germany), Aspergillus flavus {e.g., AFLAGUARD® from Syngenta, CH), Aureobasidium pullulans {e.g. , BOTECTOR® from bio-ferm GmbH, Germany), Bacillus amyloliquefaciens FZB24 (e.g., isolates NRRL B-50304 and NRRL B-50349 TAEGRO® from Novozymes Biologicals, Inc., USA), Bacillus subtilis {e.g., isolate NRRL B-21661 in RHAPSODY®, SERENADE® MAX and SERENADE® ASO from Bayer CropScience, Gustafson), Bacillus pumilus {e.g., isolate NRRL B-50349 from Bayer CropScience, Gustafson), Bacillus amyloliquefaciens TrigoCor (also known as "TrigoCor 1448"; e.g., isolate Embrapa Trigo Accession No. 144/88.4Lev, Cornell Accession No. Pma007BR-97, and ATCC Accession No. 202152, from Cornell University, USA), Candida oleophila I-82 (e.g., ASPIRE® from Ecogen Inc., USA), Candida saitoana {e.g., BIOCURE® (in mixture with lysozyme) and BIOCOAT® from Micro Flo Company, USA (BASF SE) and Arysta), Chitosan (e.g., ARMOUR-ZEN from BotriZen Ltd., NZ), Chromobacterium subtsugae {e.g., isolate NRRL B-30655 from United States Department of Agriculture, USA), Clonostachys rosea f. catenulata, also named Gliocladium catenulatum {e.g., isolate J1446: PRESTOP® from Verdera, Finland), Coniothyrium minitans {e.g., CONTANS® from Prophyta, Germany), Cryphonectria parasitica {e.g., Endothia parasitica from CNICM, France), Cryptococcus albidus {e.g. , YIELD PLUS® from Anchor Bio-Technologies, South Africa), Fusarium oxysporum {e.g., BIOFOX® from S.I.A.P.A., Italy, FUSACLEAN® from Natural Plant Protection, France), Metschnikowia fructicola {e.g., SHEMER® from Agrogreen, Israel), Microdochium dimerum {e.g., ANTIBOT® from Agrauxine, France), Paecilomyces fumosoroseus FE991 (in NOFLY® from FuturEco Bioscience S.L., Barcelona, Spain), Phlebiopsis gigantea {e.g., ROTSOP® from Verdera, Finland), Pseudozyma flocculosa {e.g., SPORODEX® from Plant Products Co. Ltd., Canada), Pythium oligandrum DV74 (e.g., POLYVERSUM® from Remeslo SSRO, Biopreparaty, Czech Rep.), Reynoutria sachlinensis {e.g., REGALIA® from Marrone Biolnnovations, USA), Talaromyces flavus V1 17b (e.g., PROTUS® from Prophyta, Germany), Trichoderma asperellum SKT-1 (e.g., ECO-HOPE® from Kumiai Chemical Industry Co., Ltd., Japan), T. atroviride LC52 (e.g., SENTINEL® from Agrimm Technologies Ltd, NZ), T. harzianum T-22 (e.g., PLANTSHIELD® der Firma BioWorks Inc., USA), T. harzianum TH 35 (e.g., ROOT PRO® from Mycontrol Ltd., Israel), T. harzianum T-39 (e.g., TRICHODEX® and TRICHODERMA 2000® from Mycontrol Ltd., Israel and Makhteshim Ltd., Israel), T. harzianum and T. viride {e.g., TRICHOPEL from Agrimm Technologies Ltd, NZ), T. harzianum ICC012 and T. viride ICC080 (e.g., REMEDIER® WP from Isagro Ricerca, Italy), T. polysporum and T. harzianum {e.g. , BINAB® from BINAB Bio-Innovation AB, Sweden), T. stromaticum (e.g., TRICOVAB® from C.E.P.L.A.C., Brazil), T. virens GL-21 (e.g., SOILGARD® from Certis LLC, USA), T. viride (e.g., TRIECO® from Ecosense Labs. (India) Pvt. Ltd., Indien, BIO-CURE® F from T. Stanes & Co. Ltd., Indien), T. viride TV1 (e.g., T. viride TV1 from Agribiotec srl, Italy), Streptomyces lydicus WYEC 108 (e.g., isolate ATCC 55445 in ACTI NOVATE®, ACTI NOVATE AG®, ACTI NOVATE STP®, ACTI NO-IRON®, ACTI NOVATE L&G®, and ACTINOGROW® from Idaho Reseach Foundation, USA), Streptomyces violaceusniger WYEC 108 (e.g., isolate ATCC 55660 in DE-THATCH-9®, DECOMP-9®, and THATCH CONTROL® from Idaho Reseach Foundation, USA), Streptomyces WYE 53 (e.g., isolate ATCC 55750 in DE-THATCH-9®, DECOMP-9®, and THATCH CONTROL® from Idaho Research Foundation, USA), and Ulocladium oudemansii HRU3 (e.g., BOTRY-ZEN® from Botry-Zen Ltd, NZ).

Enzymes:

In at least one embodiment, the compositions described herein may optionally comprise one or more enzymes. Alternatively, the one or more enzymes may be applied either simultaneously or applied sequentially, with the compositions disclosed herein.

The compositions described herein may comprise at least one cuticle degrading enzymes. Cuticle degrading enzymes are well known in the art, and include both naturally occurring (wild-type) enzymes and variant (modified by humans) enzymes. Non-limiting examples of cuticle degrading enzymes include proteases, peptidases, chitinases, chitosanase, cutinases, and lipases. In an embodiment, the compositions optionally comprises at least one cuticle degrading enzyme selected from the group consisting of protease, peptidase, chitinase, chitosanase, lipase, cutinase, and any combination thereof. In another embodiment the at least one cuticle degrading enzyme is a protease. In another embodiment the at least one cuticle degrading enzyme is a chitinase. In yet another embodiment the at least one cuticle degrading enzyme is a lipase. In still another embodiment the at least one cuticle degrading enzyme is a cutinase.

In at least one embodiment the compositions described herein comprise a combination of at least two cuticle degrading enzymes (e.g., two cuticle degrading enzymes, three cuticle degrading enzymes, four cuticle degrading enzymes, five cuticle degrading enzymes, etc.). In one embodiment, the compositions described herein comprise a combination of at least two different types of enzymes (e.g., a protease and chitinase). In yet another embodiment, the compositions described herein comprise a combination of at least two of the same type of enzyme (e.g., at least two different proteases, etc.). In still another embodiment, the compositions described herein comprise a combination of at least three cuticle degrading enzymes (e.g., a protease, a chitinase, a lipase, etc.).

Enzymes described herein may possess one or more cuticle degrading activities. The cuticle degrading enzyme may be obtained from any suitable source. In embodiments, the cuticle degrading enzyme may be obtained from a microorganism (e.g., a bacterial source or a fungal source). In another embodiment, the cuticle degrading enzyme is the protease described in WO 89/06279. Commercial proteases may also be used, such as, e.g. the product SAVINASE (available from Novozymes A/S).

Enzymes described herein may also be isolated from an entomopathogenic fungus or an acaripathogenic fungus.

Non-limiting examples of cuticle degrading enzymes are described in Bagga, S., et al. "Reconstructing the diversification of subtilisins in the pathogenic fungus Metarhizium anisopliae." Gene 324 (2004): 159-69; Bidochka, M. J. and M. J. Melzer. "Genetic polymorphisms in three subtilisin-like protease isoforms (Pr1A, Pr1 B, and Pr1 C) from Metarhizium strains." Canadian Journal of Microbiology 46.12 (2000): 1 138-44; Braga, G. U. L, R. Vencovsky, and C. L. Messias. "Estimates of genetic parameters related to chitinase production by the entomopathogenic fungus Metarhizium anisopliae." Genetics and Molecular Biology 21 .2 (1998): 171 -77; Clarkson, J. M. "Molecular biology of fungi for the control of insects." (1996): 123-35; Cole, S. C. J., A. K. Charnley, and R. M. Cooper. "Purification and partial characterization of a novel trypsin- like cysteine protease from Metarhizium-anisopliae." FEMS Microbiology Letters 1 13.2 (1993): 189-96; Da Silva, M. V., et al. "Cuticle-induced endo/exoacting chitinase CHIT30 from Metarhizium anisopliae is encoded by an ortholog of the chi3 gene." Research in Microbiology 156.3 (2005): 382-92; Dhar & Kaur, "Production of cuticle-degrading proteases by Beauveria bassiana and their induction in different media," African Journal of Biochemistry Research, Vol. 4(3), 65-72 (2010); Fang, W. G., et al. "Expressing a fusion protein with protease and chitinase activities increases the virulence of the insect pathogen Beauveria bassiana." Journal of Invertebrate Pathology 102.2 (2009): 155-59; Freimoser, F. M., et al. "Expressed sequence tag (EST) analysis of two subspecies of Metarhizium anisopliae reveals a plethora of secreted proteins with potential activity in insect hosts." Microbiology-Sgm 149 (2003): 239-47; Gimenez-Pecci, MdIP, et al. "Characterization of mycoviruses and analyses of chitinase secretion in the biocontrol fungus Metarhizium anisopliae." Current Microbiology 45.5 (2002): 334-39; Hu, G. and R. J. S. Leger. "A phylogenomic approach to reconstructing the diversification of serine proteases in fungi." Journal of Evolutionary Biology 17.6 (2004): 1204-14; Hutwimmer, S., et al. "Algorithm-based design of synthetic growth media stimulating virulence properties of Metarhizium anisopliae conidia." Journal of Applied Microbiology 105.6 (2008): 2026-34; Joshi, L, R. S. S. Leger, and D. W. Roberts. "Isolation of a cDNA encoding a novel subtilisin- like protease (Pr1 B) from the entomopathogenic fungus, Metarhizium anisopliae using differential display-RT-PCR." Gene (Amsterdam) 197.1-2 (1997): 1-8; Kim, H. K., et al. "Gene structure and expression of the gene from Beauveria bassiana encoding bassiasin I, an insect cuticle-degrading serine protease." Biotechnology Letters 21.9 (1999): 777-83; Kim, J. S. "A novel biopesticide production: Attagel-mediated precipitation of chitinase from Beauveria bassiana SFB-205 supernatant for thermotolerance." Applied Microbiology and Biotechnology 87.5 (2010): 1639-48; "Relation of aphicidal activity with cuticular degradation by Beauveria bassiana SFB-205 supernatant incorporated with polyoxyethylene-(3)- isotridecyl ether." Journal of Microbiology and Biotechnology 20.3 (2010): 506-09; Kim, J. S., et al. "Influence of two FPLC fractions from Beauveria bassiana SFB-205 supernatant on the insecticidal activity against cotton aphid." Biocontrol Science and Technology 20.1 (2010): 77-81 ; Kim, J. S., et al. "Correlation of the aphicidal activity of Beauveria bassiana SFB-205 supernatant with enzymes." Fungal Biology 1 14.1 (2010): 120-28; Ko, H. J., et al. Optimal production of protease from entomopathogenic fungus Beauveria bassiana." Agricultural Chemistry and Biotechnology 39.6 (1996): 449-54; Ko, H. J., et al. "Purification and characterization of protease from entomopathogenic fungus Beauveria bassiana." Agricultural Chemistry and Biotechnology 40.5 (1997): 388-94; Leal, S. C. M., et al. "Amplification and restriction endonuclease digestion of the Pr1 gene for the detection and characterization of Metarhizium strains." Mycological Research 101 .3 (1997): 257-65; Liang et al., "The crystal structures of two cuticle-degrading proteases from nematophagous fungi and their contribution to infection against nematodes," The FASEB Journal, Vol. 24, 1391 - 1400, May 2010; Manalil, N. S., et al. "Comparative analysis of the Metarhizium anisopliae secretome in response to exposure to the greyback cane grub and grub cuticles." Fungal Biology 1 14.8 (2010): 637-45; Mohanty, S. S., K. Raghavendra, and A. P. Dash. "Induction of chymoelastase (Pr1 ) of Metarhizium anisopliae and its role in causing mortality to mosquito larvae." World Journal of Microbiology and Biotechnology 24.10 (2008): 2283-88; Mustafa, U. and G. Kaur. "Extracellular Enzyme Production in Metarhizium anisopliae Isolates." Folia Microbiologica 54.6 (2009): 499-504; Nahar, P., V. Ghormade, and M. V. Deshpande. "The extracellular constitutive production of chitin deacetylase in Metarhizium anisopliae: possible edge to entomopathogenic fungi in the biological control of insect pests." Journal of Invertebrate Pathology 85.2 (2004): 80-88; Ortiz-Urquiza, A., et al. "Effects of cultural conditions on fungal biomass, blastospore yields and toxicity of fungal secreted proteins in batch cultures of Metarhizium anisopliae (Ascomycota: Hypocreales)." Pest Management Science 66.7 (2010): 725-35; Paterson, I. C, et al. "Regulation of production of a trypsin-like protease by the insect pathogenic fungus Metarhizium-anisopliae." FEMS Microbiology Letters 109.2-3 (1993): 323-27; "Specific induction of a cuticle-degrading protease of the insect pathogenic fungus Metarhizium-anisopliae." Microbiology-Uk 140. Part 1 (1994): 185- 89; "Partial characterization of specific inducers of a cuticle- degrading protease from the insect pathogenic fungus Metarhizium-anisopliae." Microbiology-Uk 140. Part 1 1 (1994): 3153-59; Pinto, F. G., et al. "Genetic variation in the cuticle-degrading protease activity of the entomopathogen Metarhizium flavoviride." Genetics and Molecular Biology 25.2 (2002): 231- 34; Qazi, S. S. and G. G. Khachatourians. "Hydrated conidia of Metarhizium anisopliae release a family of metalloproteases." Journal of Invertebrate Pathology 95.1 (2007): 48-59; Rangel, D. E. N., D. G. Alston, and D. W. Roberts. "Effects of physical and nutritional stress conditions during mycelial growth on conidial germination speed, adhesion to host cuticle, and virulence of Metarhizium anisopliae, an entomopathogenic fungus." Mycological Research 1 12 (2008): 1355-61 ; Rodriguez, C. ML and B. CE Gongora. "Transformation of Beauveria bassiana Bb9205 with pr1A, prU, and stel genes of Metarhizium anisopliae and evaluation of the pathogenicity on the coffee berry borer." REVISTA COLOMBIANA DE ENTOMOLOGIA 31.1 (2005): 51 -58; Santi, L, et al. "Differential immunoproteomics enables identification of Metarhizium anisopliae proteins related to Rhipicephalus microplus infection." Research in Microbiology 160.10 (2009): 824-28; Santi, L, et al. "Metarhizium anisopliae host-pathogen interaction: differential immunoproteomics reveals proteins involved in the infection process of arthropods." Fungal Biology 1 14.4 (2010): 312-19; Sasaki, S. D., et al. "BmSI-7, a novel subtilisin inhibitor from Boophilus microplus, with activity toward Pr1 proteases from the fungus Metarhizium anisopliae." Experimental Parasitology 1 18.2 (2008): 214-20; Screen, S. E., G. Hu, and R. J. Leger. "Transformants of Metarhizium anisopliae sf. anisopliae overexpressing chitinase from Metarhizium anisopliae sf. acridum show early induction of native chitinase but are not altered in pathogenicity to Manduca sexta." Journal of Invertebrate Pathology 78.4 (2001 ): 260-66; Segers, R., et al. "The subtilisins of the invertebrate mycopathogens Verticillium chlamydosporium and Metarhizium anisopliae are serologically and functionally related." FEMS Microbiology Letters 126.3 (1995): 227-31 ; Shah, F. A., C. S. Wang, and T. M. Butt. "Nutrition influences growth and virulence of the insect- pathogenic fungus Metarhizium anisopliae." FEMS Microbiology Letters 251.2 (2005): 259-66; Small, C. L. and M. J. Bidochka. "Up-regulation of Pr1 , a subtilisin-like protease, during conidiation in the insect pathogen Metarhizium anisopliae." Mycological Research 109 (2005): 307-13; Smithson, S. L., et al. "Cloning and characterization of a gene encoding a cuticle-degrading protease from the insect pathogenic fungus Metarhizium anisopliae." Gene (Amsterdam) 166.1 (1995): 161 -65; St Leger, R. J. "The role of cuticle-degrading proteases in fungal pathogenesis of insects." Canadian Journal of Botany 73.SUPPL. 1 SECT. E-H (1995): S1 1 19-S1 125; St Leger, R. J., M. J. Bidochka, and D. W. Roberts. "Characterization of a novel carboxypeptidase produced by the entomopathogenic fungus Metarhizium anisopliae." Archives of biochemistry and biophysics 314.2 (1994): 392-98; "Germination triggers of Metarhizium anisopliae conidia are related to host species." Microbiology (Reading) 140.7 (1994): 1651 -60; St Leger, R. J., R. M. Cooper, and A. K. Charnley. "Distribution of chymoelastases and trypsin-like enzymes in five species of entomopathogenic deuteromycetes." Archives of biochemistry and biophysics 258.1 (1987): 123-31 ; St Leger, R. J., L. Joshi, and D. W. Roberts. "Adaptation of proteases and carbohydrates of saprophytic, phytopathogenic and entomopathogenic fungi to the requirements of their ecological niches." Microbiology (Reading, England) 143 ( Pt 6) (1997): 1983-92; St Leger, R. J., J. O. Nelson, and S. E. Screen. "The entomopathogenic fungus Metarhizium anisopliae alters ambient pH, allowing extracellular protease production and activity." Microbiology-Uk 145 (1999): 2691-99; St Leger, R. J. and D. W. Roberts. "Engineering improved mycoinsecticides." Trends in Biotechnology 15.3 (1997): 83-85; St Leger, R. J., M. J. Bidochka, and D. W. Roberts. "Isoforms of the cuticle-degrading pr1 proteinase and production of a metalloproteinase by Metarhizium-anisopliae." Archives of biochemistry and biophysics 313.1 (1994): 1 -7; St Leger, R. J., R. M. Cooper, and A. K. Charnley. "Analysis of aminopeptidase and dipeptidylpeptidase iv from the entomopathogenic fungus Metarhizium-anisopliae." Journal of General Microbiology 139. Part 2 (1993): 237-43; St Leger, R. J., et al. "Characterization and ultrastructural-localization of chitinases from Metarhizium-anisopliae, m-flavoviride, and Beauveria-bassiana during fungal invasion of host (manduca- sexta) cuticle." Applied and Environmental Microbiology 62.3 (1996): 907-12; St Leger, R. J., L. Joshi, and D. Roberts. "Ambient pH is a major determinant in the expression of cuticle-degrading enzymes and hydrophobin by Metarhizium- anisopliae." Applied and Environmental Microbiology 64.2 (1998): 709-13; St Leger, R. J., R. C. Staples, and D. W. Roberts. "Entomopathogenic isolates of Metarhizium-anisopliae, Beauveria-bassiana, and Aspergillus-flavus produce multiple extracellular chitinase isozymes." Journal of Invertebrate Pathology 61.1 (1993): 81 -84; St. Leger et al., "Production of Cuticle-degrading Enzymes by the Entomopathogen Metarhizium anisopliae during Infection of Cuticles from Calliphora vomitoria and Manduca sexta," Journal of General Microbiology, 133, 1371-1382 (1987); St. Leger et al., "Cuticle-degrading Enzyme of Entomopathogenic Fungi: Regulation of Production of Chitonolytic Enzymes," General Microbiology, 132, 1509-1517 (1987); St. Leger et al., "Cuticle-Degrading Enzymes of Entomopathogenic Fungi," Synthesis in Culture on Cuticle, Journal of Invertebrate Pathology, 48, 85-95 (1986); Todorova, S. I., et al. "Heterogeneity of two Beauveria bassiana strains revealed by biochemical tests, protein profiles and bio-assays on Leptinotarsa decemlineata (Col.: Chrysomelidae) and Coleomegilla maculata lengi (Col.: Coccinellidae) larvae." Entomophaga 39.2 (1994): 159-69; Valadares, M. C. C. and J. L. Azevedo. "Production of amylases and proteases by wild-type and mutant strains of Metarhizium anisopliae var. anisopliae." Revista de Microbiologia 27.4 (1996): 237-41 ; Valadares-lnglis, M. C. and J. L. Azevedo. "Amylase and protease secretion in recombinant strains of Metarhizium anisopliae var. anisopliae following parasexual crosses." Brazilian Journal of Genetics 20.2 (1997): 171 - 75; Valadares-lnglis, M. C. and J. F. Peberdy. "Location of chitinolytic enzymes in protoplasts and whole cells of the entomopathogenic fungus Metarhizium anisopliae." Mycological Research 101 .1 1 (1997): 1393-96; Wang, C. S., M. A. Typas, and T. M. Butt. "Detection and characterisation of pr1 virulent gene deficiencies in the insect pathogenic fungus Metarhizium anisopliae." FEMS Microbiology Letters 213.2 (2002): 251-55; Wei, Z., Y. Q. Cao, and Y. X. Xia. "Cloning of the subtilisin Pr1A gene from a strain of locust specific fungus, Metarhizium anisopliae, and functional expression of the protein in Pichia pastoris." World Journal of Microbiology and Biotechnology 24.1 1 (2008): 2481 -88; U.S. Patent No. 5,962,765; WO/2008/06301 1.

Agriculturally Beneficial Ingredients:

The compositions disclosed herein may comprise one or more agriculturally beneficial ingredients. Non-limiting examples of agriculturally beneficial ingredients include one or more micronutrients, biostimulants, preservatives, polymers, wetting agents, surfactants, herbicides, fungicides, insecticides, acaricides, nematicides, anti-freezing agents, preservatives or combinations thereof.

Micronutrient(s):

In an embodiment, the compositions described herein may comprise one or more micronutrients. Alternatively, the one or more micronutrients may be applied either simultaneously or applied sequentially, with the compositions disclosed herein.

Non-limiting examples of micronutrients for use in the compositions described herein include vitamins, (e.g., vitamin A, vitamin B complex (i.e., vitamin B-i , vitamin B₂, vitamin B₃, vitamin B₅, vitamin B₆, vitamin B₇, vitamin B₈, vitamin B₉, vitamin B₁₂, choline) vitamin C, vitamin D, vitamin E, vitamin K, carotenoids (ocarotene, β-carotene, cryptoxanthin, lutein, lycopene, zeaxanthin, etc.), macrominerals (e.g., phosphorus, calcium, magnesium, potassium, sodium, iron, etc.), trace minerals (e.g., boron, cobalt, chloride, chromium, copper, fluoride, iodine, iron, manganese, molybdenum, selenium, zinc, etc.), organic acids (e.g., acetic acid, citric acid, lactic acid, malic aclid, taurine, etc.), and combinations thereof. In a particular embodiment, the compositions may comprise phosphorus, boron, chlorine, copper, iron, manganese, molybdenum, zinc or combinations thereof.

In certain embodiments, where the compositions described herein may comprise phosphorus, it is envisioned that any suitable phosphorus source may be provided. In one embodiment, the phosphorus may be derived from a phosphorus source. In another embodiment, suitable phosphorus sources include phosphorus sources capable of solubilization by one or more microorganisms (e.g., Penicillium bilaiae, etc.).

In one embodiment, the phosphorus source may be a rock phosphate source. In another embodiment the phosphorus source may be one or more fertilizers comprising one or more phosphorus sources. Commercially available manufactured phosphate fertilizers are of many types. Some common ones are those containing rock phosphate, monoammonium phosphate, diammonium phosphate, monocalcium phosphate, super phosphate, triple super phosphate, and/or ammonium polyphosphate. All of these fertilizers are produced by chemical processing of insoluble natural rock phosphates in large scale fertilizer- manufacturing facilities and the product is expensive. By means of the present invention it is possible to reduce the amount of these fertilizers applied to the soil while still maintaining the same amount of phosphorus uptake from the soil.

In still another embodiment, the phosphorus source may be an organic phosphorus source. In a further particular embodiment, the phosphorus source may include an organic fertilizer. An organic fertilizer refers to a soil amendment derived from natural sources that guarantees, at least, the minimum percentages of nitrogen, phosphate, and potash. Non- limiting examples of organic fertilizers include plant and animal by-products, rock powders, seaweed, inoculants, and conditioners. These are often available at garden centers and through horticultural supply companies. In particular the organic phosphorus source is from bone meal, meat meal, animal manure, compost, sewage sludge, or guano, or combinations thereof.

In still another embodiment, the phosphorus source be derived from a combination of phosphorus sources including, but not limited to, rock phosphate, fertilizers comprising one or more phosphorus sources (e.g., monoammonium phosphate, diammonium phosphate, monocalcium phosphate, super phosphate, triple super phosphate, ammonium polyphosphate, etc.) one or more organic phosphorus sources, and combinations thereof.

Biostimulant(s):

In an embodiment, the compositions described herein may comprise one or more biostimulants. Alternatively, the one or more biostimulants may be applied either simultaneously or applied sequentially, with the compositions disclosed herein.

Biostimulants may enhance metabolic or physiological processes such as respiration, photosynthesis, nucleic acid uptake, ion uptake, nutrient delivery, or a combination thereof. Non-limiting examples of biostimulants include seaweed extracts (e.g., ascophyllum nodosum), humic acids (e.g., potassium humate), fulvic acids, myo-inositol, glycine, and combinations thereof. In another embodiment, the compositions comprise seaweed extracts, humic acids, fulvic acids, myo-inositol, glycine, and combinations thereof.

Polymer(s):

In an embodiment, the compositions described herein may comprise one or more polymers. Alternatively, the one or more polymers may be applied either simultaneously or applied sequentially, with the compositions disclosed herein.

Non-limiting uses of polymers in the agricultural industry include agrochemical delivery (e.g., use as an aqueous dispersant), heavy metal removal, water retention and/or water delivery, and combinations thereof. Pouci, et al., Am. J. Agri. & Biol. Sci., 3(7):299-314 (2008). In one embodiment, the one or more polymers is a natural polymer (e.g., agar, starch, alginate, pectin, cellulose, etc.), a synthetic polymer, a biodegradable polymer (e.g., polycaprolactone, polylactide, poly (vinyl alcohol), etc.), or a combination thereof.

For a non-limiting list of polymers useful for the compositions described herein, see Pouci, et al., Am. J. Agri. & Biol. Sci., 3(7):299-314 (2008). In one embodiment, the compositions described herein comprise cellulose, cellulose derivatives, methylcellulose, methylcellulose derivatives, starch, agar, alginate, pectin, polyvinylpyrrolidone, polymeric surfactants, and combinations thereof.

In a particular embodiment, the compositions may comprise one or more polymeric surfactants. Polymeric surfactants that may be suitable for the compositions described herein may include one or more nonionic polymeric surfactants, anionic polymeric surfactants, amphoteric polymeric surfactants, cationic polymeric surfactants, and combinations thereof. Particularly useful polymeric surfactants to the compositions described herein are polymeric surfactants that are capable of functioning as an aqueous dispersant.

Nonionic Polymeric Surfactants:

Non-limiting examples of nonionic polymeric surfactants include polyalkylene oxide block copolymers, butyl block copolymers, nonionic block copolymers, acrylic copolymer solutions, nonionic random polymeric polymers, polyoxyethylene polyarl phenols, and nonionic polymeric dispersants. Commercially available nonionic polymeric surfactants include, but are not limited to, Atlas® G-5000, Atlas® G-5002L, Atlox® 4894, Atlox® 4912, Atlox® 4912-SF, Atlox® 4913, Atlox® 4914, Cresplus® DP, Hypermer® B206, Hypermer® B210, Hypermer® B246SF, Zyphrym® PD2206, Zyphrym® PD3315, and Zyphrym® PD7000.

Anionic Polymeric Surfactants:

Non-limiting examples of anionic polymeric surfactants include styrene acrylic polymers, modified styrene acrylic polymers, and anionic polymeric dispersants. Commercially available anionic polymeric surfactants include, but are not limited to, Atlox® Metasperse 100L, Atlox® Metasperse 500L, Atlox® Metasperse 550S, and Atlox® LP-1.

Polymeric Amphoteric Surfactants:

Polymeric amphoteric surfactants suitable for the compositions described herein include, but are not limited to, polymeric amphoteric dispersants. A commercially available polymeric amphoteric dispersant includes, but is not limited to, Atlox® 4915.

Cationic Polymeric Surfactants:

Cationic polymeric surfactants suitable for the compositions described herein include, but are not limited to, polyester/polyamine condensation polymers. A commercially available cationic polymeric surfactant includes Hypermer® KD-1. Surfactant(s):

In one embodiment, the compositions described herein may further comprise one or more surfactants. Surfactants may be useful as a component in a seed coating and/or processes for coating seeds. Surfactants suitable for the compositions described herein may be non-ionic surfactants (e.g., semi-polar and/or anionic and/or cationic and/or zwitterionic). The surfactants can wet and emulsify soil(s) and/or dirt(s). It is envisioned that the surfactants used in described composition have low toxicity for any microorganisms contained within the formulation. It is further envisioned that the surfactants used in the described composition have a low phytotoxicity (i.e., the degree of toxicity a substance or combination of substances has on a plant). A single surfactant or a blend of several surfactants can be used.

Anionic surfactants

Anionic surfactants or mixtures of anionic and nonionic surfactants may also be used in the compositions. Anionic surfactants are surfactants having a hydrophilic moiety in an anionic or negatively charged state in aqueous solution. The compositions described herein may comprise one or more anionic surfactants. The anionic surfactant(s) may be either water soluble anionic surfactants, water insoluble anionic surfactants, or a combination of water soluble anionic surfactants and water insoluble anionic surfactants.

Non-limiting examples of water soluble anionic surfactants include alkyl sulfates, alkyl ether sulfates, alkyl amido ether sulfates, alkyl aryl polyether sulfates, alkyl aryl sulfates, alkyl aryl sulfonates, monoglyceride sulfates, alkyl sulfonates, alkyl amide sulfonates, alkyl aryl sulfonates, benzene sulfonates, toluene sulfonates, xylene sulfonates, cumene sulfonates, alkyl benzene sulfonates, alkyl diphenyloxide sulfonate, alpha-olefin sulfonates, alkyl naphthalene sulfonates, paraffin sulfonates, lignin sulfonates, alkyl sulfosuccinates, ethoxylated sulfosuccinates, alkyl ether sulfosuccinates, alkylamide sulfosuccinates, alkyl sulfosuccinamate, alkyl sulfoacetates, alkyl phosphates, phosphate ester, alkyl ether phosphates, acyl sarconsinates, acyl isethionates, N-acyl taurates, N-acyl-N-alkyltaurates, alkyl carboxylates, or a combination thereof.

Commercially available anionic surfactants suitable for the compositions described herein include Ninate 60E. In an embodiment, the composition comprises Ninate 60E.

Nonionic surfactants

Nonionic surfactants are surfactants having no electrical charge when dissolved or dispersed in an aqueous medium. In at least one embodiment of the composition described herein, one or more nonionic surfactants are used as they provide the desired wetting and emulsification actions and do not significantly inhibit spore stability and activity. The nonionic surfactant(s) may be either water soluble nonionic surfactants, water insoluble nonionic surfactants, or a combination of water soluble nonionic surfactants and water insoluble nonionic surfactants.

Water insoluble nonionic surfactants

Non-limiting examples of water insoluble nonionic surfactants include alkyl and aryl: glycerol ethers, glycol ethers, ethanolamides, sulfoanylamides, alcohols, amides, alcohol ethoxylates, glycerol esters, glycol esters, ethoxylates of glycerol ester and glycol esters, sugar-based alkyl polyglycosides, polyoxyethylenated fatty acids, alkanolamine condensates, alkanolamides, tertiary acetylenic glycols, polyoxyethylenated mercaptans, carboxylic acid esters, polyoxyethylenated polyoxyproylene glycols, sorbitan fatty acid esters, sorbitol ethoxylate esters, or combinations thereof. Also included are EO/PO block copolymers (EO is ethylene oxide, PO is propylene oxide), EO polymers and copolymers, polyamines, and polyvinylpyrrolidones.

Commercially available water insoluble nonionic surfactants that may be suitable for the compositions described herein include Tomadol® 91-2.5, Tomadol® 23-1 , Tomadol® 23-3, Span™ 20, Span™ 40, Span™ 60, Span™ 65, Span™ 80, Span™ 85, Arlatone® TV, Atlas® G-1086, Atlas® G-1096, Atlox® 1045A, Cirrasol® G-1086, Cirrasol® G-1096, and combinations thereof.

Water soluble nonionic surfactants

Non-limiting examples of water soluble nonionic surfactants include sorbitan fatty acid alcohol ethoxylates and sorbitan fatty acid ester ethoxylates. In one embodiment, the composition comprises at least one water soluble nonionic surfactant that is a linear primary, or secondary or branched alcohol ethoxylate having the formula: RO(CH₂CH₂0)nH, wherein R is the hydrocarbon chain length and n is the average number of moles of ethylene oxide. In an embodiment, R can be a linear primary, or secondary, or branched alcohol ethoxylates having a hydrocarbon chain length in the range from C9 to C16 and n ranges from 6 to 13. In another embodiment the composition comprises at least one alcohol ethoxylate where R is linear C9-C1 1 hydrocarbon chain length, and n is 6. In still another embodiment, when the composition described herein comprise more than one water soluble surfactant, the water soluble surfactants are of substantially the same carbon chain length.

Commercially available water soluble nonionic surfactants that may be suitable for the compositions described herein include Tomadol® 9-1 1 , Tomadol® 23-7, Tomadol® 91-6, Tween® 20, Tween® 21 , Tween® 40, Tween® 60, Tween® 80, Surfonic L24-4, and combinations thereof.

Combination of nonionic surfactants

In one embodiment, the compositions described herein comprise at least one or more nonionic surfactants. In one embodiment, the compositions comprise at least one water insoluble nonionic surfactant and at least one water soluble nonionic surfactant. In still another embodiment, the compositions comprise a combination of nonionic surfactants having hydrocarbon chains of substantially the same length.

Other Surfactants

In another embodiment, the compositions described herein may also comprise organosilicone surfactants, silicone-based antifoams used as surfactants in silicone-based and mineral-oil based antifoams. In yet another embodiment, the compositions described herein may also comprise alkali metal salts of fatty acids (e.g., water soluble alkali metal salts of fatty acids and/or water insoluble alkali metal salts of fatty acids).

Herbicide(s):

In an embodiment, the compositions described herein may comprise one or more herbicides. Alternatively, the one or more herbicides may be applied either simultaneously or applied sequentially, with the compositions disclosed herein.

In one embodiment, the compositions described herein may further comprise one or more herbicides. In a particular embodiment, the herbicide may be a pre-emergent herbicide, a post-emergent herbicide, or a combination thereof.

Suitable herbicides include chemical herbicides, natural herbicides (e.g., bioherbicides, organic herbicides, etc.), or combinations thereof. Non-limiting examples of suitable herbicides include, thaxtomin (e.g., thaxtomin and analogues thereof, thaxtomin A, thaxtomin A ortho isomer, thaxtomin B, and C-14 deoxythaxtomin B (thaxtomin D), and combinations of thaxtomins and derivatives and analogues thereof), bentazon, acifluorfen, chlorimuron, lactofen, clomazone, fluazifop, glufosinate, glyphosate, sethoxydim, imazethapyr, imazamox, fomesafe, flumiclorac, imazaquin, and clethodim. Commercial products containing each of these compounds are readily available. Herbicide concentration in the composition will generally correspond to the labeled use rate for a particular herbicide. Funqicide(s):

In an embodiment, the compositions described herein may comprise one or more fungicides. Alternatively, the one or more fungicides may be applied either simultaneously or applied sequentially, with the compositions disclosed herein.

Fungicides useful to the compositions described herein may be biological fungicides, chemical fungicides, or combinations thereof. Fungicides may be selected so as to be provide effective control against a broad spectrum of phytopathogenic fungi, including soil- borne fungi, which derive especially from the classes of the Plasmodiophoromycetes, Peronosporomycetes (syn. Oomycetes), Chytridiomycetes, Zygomycetes, Ascomycetes, Basidiomycetes and Deuteromycetes (syn. Fungi imperfecti). More common fungal pathogens that may be effectively targeted include Pytophthora, Rhizoctonia, Fusarium, Pythium, Phomopsis or Selerotinia and Phakopsora and combinations thereof. Non-limiting examples of biological fungicides that may be suitable for use with the compositions disclosed herein include Ampelomyces quisqualis (e.g., AQ 10® from Intrachem Bio GmbH & Co. KG, Germany), Aspergillus flavus {e.g., AFLAGUARD® from Syngenta, CH), Aureobasidium pullulans {e.g., BOTECTOR® from bio-ferm GmbH, Germany), Bacillus amyloliquefaciens FZB24 (e.g., isolates NRRL B-50304 and NRRL B- 50349 TAEGRO® from Novozymes Biologicals, Inc., USA), Bacillus subtilis {e.g., isolate NRRL B-21661 in RHAPSODY®, SERENADE® MAX and SERENADE® ASO from Bayer CropScience, Gustafson), Bacillus pumilus {e.g., isolate NRRL B-50349 from Bayer CropScience, Gustafson), Bacillus amyloliquefaciens TrigoCor (also known as "TrigoCor 1448"; e.g., isolate Embrapa Trigo Accession No. 144/88.4Lev, Cornell Accession No. Pma007BR-97, and ATCC Accession No. 202152, from Cornell University, USA), Candida oleophila I-82 (e.g., ASPIRE® from Ecogen Inc., USA), Candida saitoana {e.g., BIOCURE® (in mixture with lysozyme) and BIOCOAT® from Micro Flo Company, USA (BASF SE) and Arysta), Chitosan (e.g., ARMOUR-ZEN from BotriZen Ltd., NZ), Chromobacterium subtsugae {e.g., isolate NRRL B-30655 from United States Department of Agriculture, USA), Clonostachys rosea f. catenulata, also named Gliocladium catenulatum {e.g., isolate J 1446: PRESTOP® from Verdera, Finland), Coniothyrium minitans {e.g., CONTANS® from Prophyta, Germany), Cryphonectria parasitica {e.g., Endothia parasitica from CNICM, France), Cryptococcus albidus {e.g., YIELD PLUS® from Anchor Bio-Technologies, South Africa), Fusarium oxysporum {e.g., BIOFOX® from S.I.A.P.A., Italy, FUSACLEAN® from Natural Plant Protection, France), Metschnikowia fructicola {e.g., SHEMER® from Agrogreen, Israel), Microdochium dimerum {e.g., ANTIBOT® from Agrauxine, France), Paecilomyces fumosoroseus FE991 (in NOFLY® from FuturEco Bioscience S.L., Barcelona, Spain), Phlebiopsis gigantea {e.g., ROTSOP® from Verdera, Finland), Pseudozyma flocculosa {e.g., SPO ROD EX® from Plant Products Co. Ltd., Canada), Pythium oligandrum DV74 (e.g., POLYVERSUM® from Remeslo SSRO, Biopreparaty, Czech Rep.), Reynoutria sachlinensis {e.g., REGALIA® from Marrone Biolnnovations, USA), Talaromyces flavus V1 17b (e.g., PROTUS® from Prophyta, Germany), Trichoderma asperellum SKT-1 (e.g., ECO-HOPE® from Kumiai Chemical Industry Co., Ltd., Japan), T. atroviride LC52 (e.g., SENTINEL® from Agrimm Technologies Ltd, NZ), T. harzianum T-22 (e.g., PLANTSHIELD® der Firma BioWorks Inc., USA), T. harzianum TH 35 (e.g., ROOT PRO® from Mycontrol Ltd., Israel), T. harzianum T-39 (e.g., TRICHODEX® and TRICHODERMA 2000® from Mycontrol Ltd., Israel and Makhteshim Ltd., Israel), T. harzianum and T. viride {e.g., TRICHOPEL from Agrimm Technologies Ltd, NZ), T. harzianum ICC012 and T. viride ICC080 (e.g., REMEDIER® WP from Isagro Ricerca, Italy), T. polysporum and T. harzianum {e.g., BINAB® from BINAB Bio-Innovation AB, Sweden), T. stromaticum {e.g., TRICOVAB® from C.E.P.L.A.C., Brazil), T. virens GL-21 (e.g., SOILGARD® from Certis LLC, USA), T. viride (e.g., TRIECO® from Ecosense Labs. (India) Pvt. Ltd., Indien, BIO-CURE® F from T. Stanes & Co. Ltd., Indien), T. viride TV1 (e.g. , T. viride TV1 from Agribiotec srl, Italy), Streptomyces lydicus WYEC 108 (e.g., isolate ATCC 55445 in ACTI NOVATE®, ACTI NOVATE AG®, ACTI NOVATE STP®, ACTI NO-I RON®, ACTI NOVATE L&G®, and ACTINOGROW® from Idaho Reseach Foundation, USA), Streptomyces violaceusniger WYEC 108 (e.g., isolate ATCC 55660 in DE-THATCH-9®, DECOMP-9®, and THATCH CONTROL® from Idaho Reseach Foundation, USA), Streptomyces WYE 53 (e.g., isolate ATCC 55750 in DE-THATCH-9®, DECOMP-9®, and THATCH CONTROL® from Idaho Research Foundation, USA), and Ulocladium oudemansii HRU3 (e.g., BOTRY-ZEN® from Botry-Zen Ltd, NZ).

Representative examples of chemical fungicides that may be suitable for use in the present invention include

A) strobilurins:

azoxystrobin, coumethoxystrobin, coumoxystrobin, dimoxystrobin, enestroburin, fluoxastrobin, kresoxim-methyl, metominostrobin, orysastrobin, picoxystrobin, pyraclostrobin, pyrametostrobin, pyraoxystrobin, pyribencarb, trifloxystrobin, 2-[2-(2,5-dimethyl- phenoxymethyl)-phenyl]-3-methoxy-acrylic acid methyl ester and 2-(2-(3-(2,6- dichlorophenyl)-1-methyl-allylideneaminooxymethyl)- -phenyl)-2-methoxyimino-N-methyl- acetamide;

B) carboxamides:

carboxanilides: benalaxyl, benalaxyl-M, benodanil, bixafen, boscalid, carboxin, fenfuram, fenhexamid, flutolanil, fluxapyroxad, furametpyr, isopyrazam, isotianil, kiralaxyl, mepronil, metalaxyl, metalaxyl-M (mefenoxam), ofurace, oxadixyl, oxycarboxin, penflufen, penthiopyrad, sedaxane, tecloftalam, thifluzamide, tiadinil, 2-amino-4-methyl-thiazole-5- carboxanilide, N-(4'-trifluoromethylthiobiphenyl-2-yl)-3-difluoromethyl-1 -methyl-1 H-pyra- zole- 4-carboxamide and N-(2-(1 ,3,3-trimethylbutyl)-phenyl)-1 ,3-dimethyl-5-fluoro-1 H-pyrazole-4- carboxamide;

carboxylic morpholides: dimethomorph, flumorph, pyrimorph;

benzoic acid amides: flumetover, fluopicolide, fluopyram, zoxamide;

other carboxamides: carpropamid, dicyclomet, mandiproamid, oxytetracyclin, silthiofam and N-(6-methoxy-pyridin-3-yl) cyclopropanecarboxylic acid amide;

C) azoles:

triazoles: azaconazole, bitertanol, bromuconazole, cyproconazole, difenoconazole, diniconazole, diniconazole-M, epoxiconazole, fenbuconazole, fluquinconazole, flusilazole, flutriafol, hexaconazole, imibenconazole, ipconazole, metconazole, myclobutanil, oxpoconazole, paclobutrazole, penconazole, propiconazole, prothioconazole, simeconazole, tebuconazole, tetraconazole, triadimefon, triadimenol, triticonazole, uniconazole; imidazoles: cyazofamid, imazalil, pefurazoate, prochloraz, triflumizol;

D) heterocyclic compounds:

pyridines: fluazinam, pyrifenox, 3-[5-(4-chloro-phenyl)-2,3-dimethyl-isoxazolidin-3-yl]- pyridine, 3-[5-(4-methyl-phenyl)-2,3-dimethyl-isoxazolidin-3-yl]-pyridine;

pyrimidines: bupirimate, cyprodinil, diflumetorim, fenarimol, ferimzone, mepanipyrim, nitrapyrin, nuarimol, pyrimethanil;

piperazines: triforine;

pyrroles: fenpiclonil, fludioxonil;

morpholines: aldimorph, dodemorph, dodemorph-acetate, fenpropimorph, tridemorph; piperidines: fenpropidin;

dicarboximides: fluoroimid, iprodione, procymidone, vinclozolin;

non-aromatic 5-membered heterocycles: famoxadone, fenamidone, flutianil, octhilinone, probenazole, 5-amino-2-isopropyl-3-oxo-4-ortho-tolyl-2,3-dihydro-pyrazole-1 - carbothioic acid S-allyl ester;

others: acibenzolar-S-methyl, ametoctradin, amisulbrom, anilazin, blasticidin-S, captafol, captan, chinomethionat, dazomet, debacarb, diclomezine, difenzoquat, difenzoquat- methylsulfate, fenoxanil, Folpet, oxolinic acid, piperalin, proquinazid, pyroquilon, quinoxyfen, triazoxide, tricyclazole, 2-butoxy-6-iodo-3-propylchromen-4-one, 5-chloro-1 -(4,6-dimethoxy- pyrimidin-2-yl)-2-methyl-1 H-benzoimidazole and 5-chloro-7-(4-methylpiperidin-1 -yl)-6-(2,4,6- trifluorophenyl)-[1 ,2,4]triazolo-[1 ,5-a]pyrimidine;

E) other active substances:

guanidines: guanidine, dodine, dodine free base, guazatine, guazatine-acetate, iminoctadine, iminoctadine-triacetate, iminoctadine-tris(albesilate);

antibiotics: kasugamycin, kasugamycin hydrochloride-hydrate, streptomycin, polyoxine, validamycin A;

nitrophenyl derivates: binapacryl, dicloran, dinobuton, dinocap, nitrothal-isopropyl, tecnazen,

organometal compounds: fentin salts, such as fentin-acetate, fentin chloride or fentin hydroxide;

sulfur-containing heterocyclyl compounds: dithianon, isoprothiolane;

organophosphorus compounds: edifenphos, fosetyl, fosetyl-aluminum, iprobenfos, phosphorus acid and its salts, pyrazophos, tolclofos-methyl;

organochlorine compounds: chlorothalonil, dichlofluanid, dichlorophen, flusulfamide, hexachlorobenzene, pencycuron, pentachlorphenole and its salts, phthalide, quintozene, thiophanate-methyl, tolylfluanid, N-(4-chloro-2-nitro-phenyl)-N-ethyl-4-methyl- benzenesulfonamide; inorganic active substances: Bordeaux mixture, copper acetate, copper hydroxide, copper oxychloride, basic copper sulfate and sulfur.

Commercial fungicides are most suitably used in accordance with the manufacturer's instructions at the recommended concentrations.

Insecticide(s), Acaricide(s) Nematicide(s):

In at least one embodiment, the compositions described herein may optionally comprise one or more insecticides, acaricides, nematicides, or combinations thereof. Alternatively, the one or more insecticides, acaricides, nematicides may be applied either simultaneously or applied sequentially, with the compositions disclosed herein. Insecticides useful to the compositions described herein will suitably exhibit activity against a broad range of insects including, but not limited to, wireworms, cutworms, grubs, corn rootworm, seed corn maggots, flea beetles, chinch bugs, aphids, leaf beetles, stink bugs, and combinations thereof. The insecticides, acaricides, and nematicides described herein may be chemical or natural (e.g., biological solutions, such as fungal pesticides, etc.).

Non-limiting examples of insecticides, acaricides and nematicides that may be useful to the compositions disclosed herein include acrinathrin, alpha-cypermethrin, betacyfluthrin, cyhalothrin, cypermethrin, deltamethrin csfenvalcrate, etofenprox, fenpropathrin, fenvalerate, flucythrinat, lambda-cyhalothrin, gamma-cyhalothrin, permethrin, tau-fluvalinate, transfluthrin, zeta-cypermethrin, cyfluthrin, bifenthrin, tefluthrin, eflusilanat, fubfenprox, pyrethrin, resmethrin, imidacloprid, acetamiprid, thiamethoxam, nitenpyram, thiacloprid, dinotefuran, clothianidin, imidaclothiz, chlorfluazuron, diflubenzuron, lufenuron, teflubenzuron, triflumuron, novaluron, flufenoxuron, hexaflumuron, bistrifluoron, noviflumuron, buprofezin, cyromazine, methoxyfenozide, tebufenozide, halofenozide, chromafenozide, endosulfan, fipronil, ethiprole, pyrafluprole, pyriprole, flubendiamide, chlorantraniliprole {Rynaxypyr), Cyazypyr, emamectin, emamectin benzoate, abamectin, ivermectin, milbemectin, lepimectin, tebufenpyrad, fenpyroximate, pyridaben, fenazaquin, pyrimidifen, tolfenpyrad, dicofol, cyenopyrafen, cyflumetofen, acequinocyl, fluacrypyrin, bifenazate, diafenthiuron, etoxazole, clofentezine, spinosad, triarathen, tetradifon, propargite, hexythiazox, bromopropylate, chinomethionat, amitraz, pyrifluquinazon, pymetrozine, flonicamid, pyriproxyfen, diofenolan, chlorfenapyr, metaflumizone, indoxacarb, chlorpyrifos, spirodiclofen, spiromesifen, spirotetramat, pyridalyl, spinctoram, acephate, triazophos, profenofos, fenamiphos, 4-{[(6- chloropyrid-3-yl)methyl](2,2-difluoroethyl)amino}furan-2(5H)-one, cadusaphos, carbaryl, carbofuran, ethoprophos, thiodicarb, aldicarb, metamidophos, methiocarb, sulfoxaflor and also products based on Bacillus firmus (1-1582, BioNeem, Votivo), and combinations thereof.

In a particular embodiment, the inseciticde is a microbial insecticide. In a more particular embodiment, the microbial insecticide is a fungal insecticide. Non-limiting examples of fungal insecticides that may be used in the compositions disclosed herein are described in McCoy, C. W., Samson, R. A., and Coucias, D. G. "Entomogenous fungi. In "CRC Handbook of Natural Pesticides. Microbial Pesticides, Part A. Entomogenous Protozoa and Fungi." (C. M. Inoffo, ed.), (1988): Vol. 5, 151 -236; Samson, R. A., Evans, H.C., and Latge^', J. P. "Atlas of Entomopathogenic Fungi." (Springer-Verlag, Berlin) (1988); and deFaria, M. R. and Wraight, S. P. "Mycoinsecticides and Mycoacaricides: A comprehensive list with worldwide coverage and international classification of formulation types." Biol. Control (2007), doi: 10.1016/j.biocontrol.2007.08.001.

In one embodiment, non-limiting examples fungal insecticides that may be used in the compositions disclosed herein include species of Coelomycidium, Myiophagus, Coelemomyces, Lagenidium, Leptolegnia, Couchia, Sporodiniella, Conidiobolus, Entomophaga, Entomophthora, Erynia, Massospora, Meristacrum, Neozygites, Pandora, Zoophthora, Blastodendrion, Metschnikowia, Mycoderma, Ascophaera, Cordyceps, Torrubiella, Nectria, Hypocrella, Calonectria, Filariomyces, Hesperomyces, Trenomyces, Myriangium, Podonectria, Akanthomyces, Aschersonia, Aspergillus, Beauveria, Culicinomyces, Engyodontium, Fusarium, Gibellula, Hirsutella, Hymenostilbe, Isaria, Metarhizium, Nomuraea, Paecilomyces, Paraisaria, Pleurodesmospora, Polycephalomyces, Pseudogibellula, Sorosporella, Stillbella, Tetranacrium, Tilachlidium, Tolypocladium, Verticillium, Aegerita, Filobasidiella, Septobasidium, Uredinella, and combinations thereof.

Non-limiting examples of particular species that may be useful as a fungal insecticide in the compositions described herein include Trichoderma hamatum, Trichoderma hazarium, Alternaria cassiae, Fusarium lateritum, Fusarium solani, Lecanicillium lecanii, Aspergillus parasiticus, Verticillium lecanii, Metarhizium anisopliae, and Beauveria bassiana. In an embodiment, the compositions disclosed herein may include any of the fungal insecticides provided above, including any combination thereof.

In one embodiment, the composition comprises at least one fungal insecticide from the genus Metarhizium spp., such as, Metarhizium anisopliae (also may be referred to in the art as Metarrhizium anisopliae, Metarhizium brunneum, or "green muscadine"). In at least one embodiment, the fungal insecticide comprises the strain Metarhizium anisopliae. In another embodiment, the composition comprises spores of the strain Metarhizium anisopliae.

In a particular embodiment, the composition comprises at least one fungal pesticide comprising Metarhizium anisopliae strain F52 (also known as Metarhizium anisopliae strain 52, Metarhizium anisopliae strain 7, Metarhizium anisopliae strain 43, Metarhizium anisopliae BIO-1020,TAE-001 and deposited as DSM 3884, DSM 3885, ATCC 90448, SD 170, and ARSEF 771 1 ) (available from Novozymes Biologicals, Inc., USA). In still another particular embodiment, the composition comprises at least one fungal insecticide comprising spores of Metarhizium anisopliae strain F52. In yet another embodiment the composition may further comprise at least one fungal insecticide from the genus Beauveria spp. , such as, for example, Beauveria bassiana. In at least one embodiment, the fungal insecticide further comprises the strain Beauveria bassiana. In another embodiment, the composition further comprises spores of the strain Beauveria bassiana.

In a particular embodiment, the composition further comprises at least one fungal insecticide comprising Beauveria bassiana strain ATCC-74040. In another embodiment, the composition further comprises at least one fungal insecticide comprising spores of Beauveria bassiana strain ATCC-74040. In another particular embodiment, the composition further comprises at least one fungal insecticide comprising Beauveria bassiana strain ATCC- 74250. In still another particular embodiment, the composition further comprises at least one fungal insecticide comprising spores of Beauveria bassiana strain ATCC-74250. In yet another particular embodiment, the composition further comprises at least one fungal insecticide comprising a mixture of Beauveria bassiana strain ATCC-74040 and Beauveria bassiana strain ATCC-74250. In still another embodiment, the composition further comprises at least one fungal insecticide comprising a mixture of spores of Beauveria bassiana strain ATCC-74040 and Beauveria bassiana strain ATCC-74250.

In still yet another particular embodiment, the composition described herein may comprise a combination of fungi. In one embodiment, the composition may comprise two or more fungal insecticides that are different strains of the same species. In another embodiment, the composition comprises at least two different fungal insecticides that are strains of different species. In an embodiment, the composition comprises at least one fungal insecticide from the genus Metarhizium spp. and at least one fungal insecticide from the genus Beauveria spp.. In another embodiment, the composition comprise spores of Metarhizium spp. and Beauveria spp.

In a particular embodiment, the composition comprises at least one fungal insecticide, wherein at least one fungal insecticide is a strain of Metarhizium anisopliae and at least one fungal insecticide is a strain of Beauveria bassiana. In another embodiment, the composition comprises at least one fungal insecticide wherein the fungal insecticide comprises spores of Metarhizium anisopliae and Beauveria bassiana.

In a more particular embodiment, the composition comprises at least one fungal insecticide, wherein at least one fungal insecticide is a strain of Metarhizium anisopliae F52 and at least one fungal insecticide is a strain of the strain Beauveria bassiana ATCC-74040. In yet another embodiment, the composition comprises at least one fungal insecticide wherein the fungal insecticide comprises spores of the strain Metarhizium anisopliae F52 and the strain Beauveria bassiana ATCC-74040. In still another particular embodiment, the composition comprises at least one fungal insecticide, wherein at least one fungal insecticide is a strain of Metarhizium anisopliae F52 and at least one fungal insecticide is a strain of the strain Beauveria bassiana ATCC-74250. In yet another embodiment, the composition comprises at least one fungal insecticide wherein the fungal insecticide comprises spores of the strain Metarhizium anisopliae F52 and the strain Beauveria bassiana ATCC-74250.

In still yet another particular embodiment, the composition comprises at least one fungal insecticide, wherein at least one fungal insecticide is a strain of Metarhizium anisopliae F52, at least one fungal insecticide is a strain of the strain Beauveria bassiana ATCC-74040, and at least one fungal insecticide is a strain of the strain Beauveria bassiana ATCC-74250. In yet another embodiment, the composition comprises at least one fungal insecticide wherein the fungal insecticide comprises spores of the strain Metarhizium anisopliae F52, the strain Beauveria bassiana ATCC-74040, and the strain Beauveria bassiana ATCC-74250.

In another embodiment, the compositions disclosed herein comprise a nematicide. In a more particular embodiment, the nematicide is a microbial nematicide, more preferably a nematophagous fungus and/or nematophagous bacteria. In a particular embodiment, the microbial nematicide is a nematophagous fungus selected from the group consisting of Arthrobotrys spp., Dactylaria spp., Harposporium spp., Hirsutella spp., Monacrosporium spp., Nematoctonus spp., Meristacrum spp., Myrothecium spp., Paecilomyces spp., Pasteuria spp., Pochonia spp., Trichoderma spp., Verticillium spp., and combinations thereof. In still a more particular embodiment, the nematophagous fungus is selected from the group consisting of Arthrobotrys dactyloides, Arthrobotrys oligospora, Arthrobotrys superb, Arthrobotrys dactyloides, Dactylaria Candida, Harposporium anguillulae, Hirsutella rhossiliensis, Hirsutella minnesotensis, Monacrosporium cionopagum, Nematoctonus geogenius, Nematoctonus leiosporus, Meristacrum asterospermum, Myrothecium verrucaria, Paecilomyces lilacinus, Paecilomyces fumosoroseus, Pasteuria penetrans, Pasteuria usgae, Pochonia chlamydopora, Trichoderma harzianum, Verticillium chlamydosporum, and combinations thereof.

In a more particular embodiment, the microbial nematicide is a nematophagous bacteria selected from the group consisting of Actinomycetes spp., Agrobacterium spp., Arthrobacter spp., Alcaligenes spp., Aureobacterium spp., Azobacter spp., Beijerinckia spp., Burkholderia spp., Chromobacterium spp., Clavibacter spp., Clostridium spp., Comomonas spp., Corynebacterium spp., Curtobacterium spp., Desulforibtio spp., Enterobacter spp., Flavobacterium spp., Gluconobacter spp., Hydrogenophage spp., Klebsiella spp., Methylobacterium spp., Phyllobacterium spp., Phingobacterium spp., Photorhabdus spp., Serratia spp. Stenotrotrophomonas spp., Xenorhadbus spp. Variovorax spp., Streptomyces spp., Pseudomonas spp., Paenibacillus spp., and combinations thereof. In still a more particular embodiment, the microbial nematicide is a nematophagous bacteria selected from the group consisting of Chromobacterium subtsugae, Chromobacterium violaceum, Streptomyces lydicus, Streptomyces violaceusniger, and combinations thereof. In a particular embodiment, the strain of Chromobacterium subtsugae is a strain of Chromobacterium subtsugae sp. nov., more particularly, the strain of Chromobacterium subtsugae sp. nov. has the deposit accession number NRRL B-30655. In still another particular embodiment, the strain of Streptomyces is a strain of Streptomyces lydicus WYEC 108, a strain of Streptomyces violaceusniger Y ^'CED 9, or a combination thereof.

Insect Growth Regulators

In at least one embodiment, the compositions described herein may optionally comprise one or more insect growth regulators. Alternatively, the one or more insect growth regulators may be applied either simultaneously or applied sequentially, with the compositions disclosed herein. Non-limiting examples of insect growth regulators include pyripoxyfen, ethofenprox, cold-pressed neem oil, S-hydroprene, chitin synthesis inhibitors, juvenile hormone analogs (e.g. methoprene) and combinations thereof.

Anti-freezing Aqent(s):

In at least one embodiment, the compositions described herein may optionally comprise one or more anti-freezing agents. Alternatively, the one or more anti-freezing agents may be applied either simultaneously or applied sequentially, with the compositions disclosed herein. In one embodiment, the compositions described herein may further comprise one or more anti-freezing agents. Non-limiting examples of anti-freezing agents include ethylene glycol, propylene glycol, urea, glycerin, and combinations thereof.

Preservatives

In at least one embodiment, the compositions described herein may optionally comprise one or more preservatives. Alternatively, the one or more preservatives may be applied either simultaneously or applied sequentially, with the compositions disclosed herein. As used herein, the term "preservative" includes a biocide (i.e., a bacteriostats or a bactericides). Non-limiting examples of biocides include the following:

Bactericides:

As used herein, a bactericide is an agent that kills bacteria. A bactericide may be a disinfectant, antiseptic or antibiotic.

Non-limiting examples of a bactericidal disinfectant may be:

active chlorine (i.e., hypochlorites, chloramines, dichloroisocyanurate and trichloroisocyanurate, wet chlorine, chlorine dioxide, etc.),

active oxygen (peroxides, such as peracetic acid, potassium persulfate, sodium perborate, sodium percarbonate and urea perhydrate), iodine (iodpovidone (povidone-iodine, Betadine), Lugol's solution, iodine tincture, iodinated nonionic surfactants),

concentrated alcohols (mainly ethanol, 1 -propanol, called also n-propanol and 2- propanol, called isopropanol and mixtures thereof; further, 2-phenoxyethanol and 1- and 2- phenoxypropanols),

phenolic substances (such as phenol (also called "carbolic acid"), cresols (called "Lysole" in combination with liquid potassium soaps), halogenated (chlorinated, brominated) phenols, such as hexachlorophene, triclosan, trichlorophenol, tribromophenol, pentachlorophenol, Dibromol and salts thereof),

cationic surfactants, such as some quaternary ammonium cations (such as benzalkonium chloride, cetyl trimethylammonium bromide or chloride, didecyldimethylammonium chloride, cetylpyridinium chloride, benzethonium chloride) and others, non-quarternary compounds, such as chlorhexidine, glucoprotamine, octenidine dihydrochloride, etc.),

strong oxidizers, such as ozone and permanganate solutions;

heavy metals and their salts, such as colloidal silver, silver nitrate, mercury chloride, phenylmercury salts, copper sulfate, copper oxide-chloride, etc. Heavy metals and their salts are the most toxic, and environment-hazardous bactericides and therefore, their use is strongly oppressed or eliminated; further, also

properly concentrated strong acids (phosphoric, nitric, sulfuric, amidosulfuric, toluenesulfonic acids) and

alkalis (sodium, potassium, calcium hydroxides), such as of pH <1 or >13, particularly under elevated temperature (above 60°C), kills bacteria.

Non-limiting examples of a bactericidal antiseptic may be:

properly diluted chlorine preparations (e.g., Daquin's solution, 0.5% sodium or potassium hypochlorite solution, pH-adjusted to pH 7-8, or 0.5-1 % solution of sodium benzenesulfochloramide (chloramine B)),

some iodine preparations, such as iodopovidone in various galenics (ointment, solutions, wound plasters), in the past also Lugol's solution,

peroxides as urea perhydrate solutions and pH-buffered 0.1-0.25% peracetic acid solutions,

alcohols with or without antiseptic additives, used mainly for skin antisepsis, weak organic acids such as sorbic acid, benzoic acid, lactic acid and salicylic acid, some phenolic compounds, such as hexachlorophene, triclosan and Dibromol, and cation-active compounds, such as 0.05-0.5% benzalkonium, 0.5-4% chlorhexidine, 0.1-2% octenidine solutions. Non-limiting examples of a bactericidal antibiotic may be penicillin, cephalosporins, and aminoglycosidic antibiotics.

Other bactericidal antibiotics include the fluoroquinolones, nitrofurans, vancomycin, monobactams, co-trimoxazole, and metronidazole.

Preferred bactericides are:

Halogen containing compounds such as:

Bronopol - active 2-bromo-2-nitro-1 ,3-propanadiol

Dowicil 75 - active 1-(3-chloroallyl)-3,5,7-triaza-1 -azoniaadamantane chloride

DBNPA - active dibromonitrilopropionamide

OrganoSulfurs - includes Isothaizolones such as:

Proxel (Nipacide) - active 1 ,2-benzisothiazolin-3-one

Kathon - active 5-chloro-2-methyl-4-isosthiazolin-3-one, 2-methyl-4-isosthiazolin-3- one

Nitrogen containing compounds such as:

Germall II (Diazolidinyl urea)

Tris nitro (tris(hydroxymethyl)nitromethane)

Phenolics such as:

Dowicide (sodium o-phenylphenate)

Preventol D2® (benzyl-hemiformal)

Inorganics such as:

copper arsenates

cuprous oxide

Organometallics such as:

compounds of arsenic, copper, mercury

Quaternary ammonium compounds.

Bacteriostats:

As used herein, a bacteriostat is an agent, usually chemical, that prevents the growth of bacteria but that does not necessarily kill them or their spores. Upon removal of the bacteriostat, the bacteria usually start to grow again.

Non-limiting examples of bacteriostats include sodium azide and thimerosol.

Non-Naturallv Occurring Components

Example compositions may contain one or more non-naturally occurring components. Non-naturally occurring components generally are not products of nature or are different than products of nature. Non-naturally occurring components may differ in structure from components that are naturally occurring. Non-naturally occurring components may differ in function from components that are naturally occurring, or may cause components, substances or compositions to which they are mixed or added to have a function different than would the component/substance/composition if the non-naturally occurring component had not been mixed or added. Mixing or adding of non-naturally occurring components with or to substances or compositions may cause the initial substances/compositions to have a function different than would the substance/composition if a naturally-occurring component had been mixed or added. In one example, addition of one or more non-naturally occurring components to an initial composition that contains naturally occurring components or exclusively naturally occurring components, makes the composition, as a whole, markedly or significantly different than the initial composition. The markedly or significantly different composition, as a whole, may not be found in nature and/or may function differently than a product of nature. At least some of the substances/components/ingredients that are disclosed in this application may be non-naturally occurring components.

METHODS

In another aspect, methods of using the deposited strain, including variants thereof, and compositions described herein are disclosed.

In one embodiment a method of controlling one or more plant pests is described. The method comprises introducing into the soil one or more bacterial strains described herein. In an embodiment, the method comprises introducing into the soil one or more microbial pesticides described herein. In another embodiment, the method comprises introducing into the soil one or more microbial nematicides. In a particular embodiment, the method comprises introducing into the soil one or more nematophagous bacteria. In a more particular embodiment, the one or more nematophagous bacteria are one or more strains of Chromobacterium spp. In an even more particular embodiment, the one or more nematophagous bacteria are one or more strains of Chromobacterium vaccinii. In still an even more particular embodiment, the one or more nematophagous bacteria is one or more strains of Chromobacterium vaccinii sp. nov. In yet an even more particular embodiment, the one or more nematophagous bacteria is the strain of Chromobacterium vaccinii having the deposit accession number NRRL B-50880, variants of the strain of Chromobacterium vaccinii having the deposit accession number NRRL B-50880, or combinations thereof.

In particular aspect of the embodiment, the method comprises introducing into the soil an inoculum of one or more bacterial strains described herein. In an embodiment, the method comprises introducing into the soil an inoculum of one or more microbial pesticides described herein. In another embodiment, the method comprises introducing into the soil an inoculum of one or more microbial nematicides. In a particular embodiment, the method comprises introducing into the soil an inoculum of one or more nematophagous bacteria. In a more particular embodiment, the inoculum of one or more nematophagous bacteria is one or more strains of Chromobacterium spp. In an even more particular embodiment, the inoculum of one or more nematophagous bacteria is one or more strains of Chromobactehum vaccinii. In still an even more particular embodiment, the inoculum of one or more nematophagous bacteria is one or more strains of Chromobactehum vaccinii sp. nov. In yet an even more particular embodiment, the inoculum of one or more nematophagous bacteria is the strain of Chromobactehum vaccinii having the deposit accession number NRRL B-50880, variants of the strain of Chromobactehum vaccinii having the deposit accession number NRRL B-50880, or combinations thereof.

In yet another particular aspect of the embodiment, the method comprises introducing into the soil one or more of the compositions described herein.

The bacterial strain(s), inoculum(s) or composition(s) may be introduced into the soil according to methods known to those skilled in the art. Non-limiting examples of introducing the one or more bacterial strain(s), inoculum(s) or composition(s) into the soil include in- furrow introduction, spraying, coating seeds, foliar introduction, etc. In a particular embodiment, the introducing step comprises in-furrow introduction of one or more the bacterial strain(s), inoculum(s), or compositions described herein. In another particular embodiment, the introducing step comprises foliar introduction of one or more the bacterial strain(s), inoculum(s), or compositions described herein. In still another particular embodiment, the introducing step comprises spraying the soil with one or more of the bacterial strain(s), inoculum(s), or compositions described herein. In still yet another particular embodiment, the introducing step comprises coating seeds with one or more of the bacterial strain(s), inoculum(s), or compositions described herein.

In some embodiments, the step of introducing into the soil one or more the bacterial strain(s), inoculum(s), or compositions described herein comprises introducing an effective amount of the one or more the bacterial strain(s), inoculum(s), or compositions described herein. In certain embodiments, the step of introducing into the soil one or more the bacterial strain(s), inoculum(s), or compositions described herein comprises introducing the one or more the bacterial strain(s), inoculum(s), or compositions described herein in an amount of 1 x 10¹ - 1 x 10⁸, more preferably 1 x 10⁶ - 1 x 10¹² colony forming units per hectare. In other certain embodiments, the step of introducing into the soil one or more the bacterial strain(s), inoculum(s), or compositions described herein comprises introducing the one or more the bacterial strain(s), inoculum(s), or compositions described herein as a seed coated with 1 x 10¹— 1 x 10⁸, more preferably 1 x 10² - 1 x 10⁶ colony forming units per seed.

In another aspect, a method of controlling one or more pests comprises contacting one or more plants or plant parts with one or more bacterial strains described herein. In an embodiment, the method contacting one or more plants or plant parts with one or more microbial pesticides described herein. In another embodiment, the method comprises contacting one or more plants or plant parts with one or more microbial nematicides. In a particular embodiment, the method comprises contacting one or more plants or plant parts with one or more nematophagous bacteria. In a more particular embodiment, the one or more nematophagous bacteria are one or more strains of Chromobacterium spp. In an even more particular embodiment, the one or more nematophagous bacteria are one or more strains of Chromobacterium vaccinii. In still an even more particular embodiment, the one or more nematophagous bacteria is one or more strains of Chromobacterium vaccinii sp. nov. In yet an even more particular embodiment, the one or more nematophagous bacteria is the strain of Chromobacterium vaccinii having the deposit accession number NRRL B-50880, variants of the strain of Chromobacterium vaccinii having the deposit accession number NRRL B-50880, or combinations thereof.

In particular aspect of the embodiment, the method comprises contacting one or more plants or plant parts with an inoculum of one or more bacterial strains described herein. In an embodiment, the method comprises contacting one or more plants or plant parts with an inoculum of one or more microbial pesticides described herein. In another embodiment, the method comprises contacting one or more plants or plant parts with an inoculum of one or more microbial nematicides. In a particular embodiment, the method comprises contacting one or more plants or plant parts with an inoculum of one or more nematophagous bacteria. In a more particular embodiment, the inoculum of one or more nematophagous bacteria is one or more strains of Chromobacterium spp. In an even more particular embodiment, the inoculum of one or more nematophagous bacteria is one or more strains of Chromobacterium vaccinii. In still an even more particular embodiment, the inoculum of one or more nematophagous bacteria is one or more strains of Chromobacterium vaccinii sp. nov. In yet an even more particular embodiment, the inoculum of one or more nematophagous bacteria is the strain of Chromobacterium vaccinii having the deposit accession number NRRL B-50880, variants of the strain of Chromobacterium vaccinii having the deposit accession number NRRL B-50880, or combinations thereof.

In yet another particular aspect of the embodiment, the method comprises contacting one or more plants or plant parts with one or more of the compositions described herein.

Contacting one or more plants or plant parts with one or more of the bacterial strain(s), inoculum(s) or composition(s) described herein may be performed according to methods known to those skilled in the art. Non-limiting examples of contacting one or more plants or plant parts with one or more of the bacterial strain(s), inoculum(s) or composition(s) described herein include in-furrow contact, coating seeds, foliar contact (e.g., spraying, or dusting the bacterial strain(s), inoculum(s) or composition(s) described herein onto one or more plants or plant parts. In a particular embodiment, the contacting step comprises foliarly applying the one or more plants or plant parts with one or more of the bacterial strain(s), inoculum(s), or compositions described herein. In another particular embodiment, the contacting step comprises spraying the one or more plants or plant parts with one or more of the bacterial strain(s), inoculum(s), or compositions described herein. In still another particular embodiment, the contacting step comprises dusting the one or more plants or plant parts with one or more of the bacterial strain(s), inoculum(s), or compositions described herein. In still yet another particular embodiment, the contacting step comprises in-furrow contact with one or more plants or plant parts with one or more of the bacterial strain(s), inoculum(s), or compositions described herein.

In yet another aspect, a method of controlling one or more pests comprises contacting one or more pests with one or more bacterial strains described herein. In an embodiment, the method contacting one or more pests with one or more microbial pesticides described herein. In another embodiment, the method comprises contacting one or more pests with one or more microbial nematicides. In a particular embodiment, the method comprises contacting one or more pests with one or more nematophagous bacteria. In a more particular embodiment, the one or more nematophagous bacteria are one or more strains of Chromobacterium spp. In an even more particular embodiment, the one or more nematophagous bacteria are one or more strains of Chromobacterium vaccinii. In still an even more particular embodiment, the one or more nematophagous bacteria is one or more strains of Chromobacterium vaccinii sp. nov. In yet an even more particular embodiment, the one or more nematophagous bacteria is the strain of Chromobacterium vaccinii having the deposit accession number NRRL B-50880, variants of the strain of Chromobacterium vaccinii having the deposit accession number NRRL B-50880, or combinations thereof.

In particular aspect of the embodiment, the method comprises contacting one or more pests with an inoculum of one or more bacterial strains described herein. In an embodiment, the method comprises contacting one or more pests with an inoculum of one or more microbial pesticides described herein. In another embodiment, the method comprises contacting one or more pests with an inoculum of one or more microbial nematicides. In a particular embodiment, the method comprises contacting one or more pests with an inoculum of one or more nematophagous bacteria. In a more particular embodiment, the inoculum of one or more nematophagous bacteria is one or more strains of Chromobacterium spp. In an even more particular embodiment, the inoculum of one or more nematophagous bacteria is one or more strains of Chromobacterium vaccinii. In still an even more particular embodiment, the inoculum of one or more nematophagous bacteria is one or more strains of Chromobacterium vaccinii sp. nov. In yet an even more particular embodiment, the inoculum of one or more nematophagous bacteria is the strain of Chromobacterium vaccinii having the deposit accession number NRRL B-50880, variants of the strain of Chromobacterium vaccinii having the deposit accession number NRRL B-50880, or combinations thereof.

In yet another particular aspect of the embodiment, the method comprises contacting one or more pests with one or more of the compositions described herein. Contacting one or more pests with one or more of the bacterial strain(s), inoculum(s) or composition(s) described herein may be performed according to methods known to those skilled in the art. Non-limiting examples of contacting one or more pests with one or more of the bacterial strain(s), inoculum(s) or composition(s) described herein include foliar applications such as spraying, or dusting the bacterial strain(s), inoculum(s) or composition(s) described herein onto one or more pests. In a particular embodiment, the contacting step comprises foliarly applying the one or more pests with one or more of the bacterial strain(s), inoculum(s), or compositions described herein. In another particular embodiment, the contacting step comprises spraying the one or more pests with one or more of the bacterial strain(s), inoculum(s), or compositions described herein. In still another particular embodiment, the contacting step comprises dusting the one or more pests with one or more of the bacterial strain(s), inoculum(s), or compositions described herein.

Plant Pests:

The methods described herein are potentially useful for controlling pests (e.g., plant nematode pests, plant insect pests, plant acarai pests, etc.) and generally improving the growth conditions and/or yield for any type of plant.

Plant Nematode Pests:

In a particular embodiment, the one or more bacterial strain(s), inoculum(s) or composition(s) described herein are useful to control plant nematode pests. Non-limiting examples of plant nematode pests include root-knot, cyst, lesion, sting, and ring nematodes, including: Meloidogyne spp., Heterodera spp., Globodera spp., Pratylenchus spp., Belonolaimus spp., and Criconemella spp.

One or more of the bacterial strain(s), inoculum(s) or composition(s) described herein are also useful to control Tylenchulus semipenetrans, Trichodorus spp., Longidorus spp., Rotylenchulus spp., Xiphinema spp., Belonolaimus spp. (such as B. longicaudatus), Criconemoides spp., Tylenchorhynchus spp., Hoplolaimus spp., Rotylenchus spp., Helicotylenchus spp., Radopholus spp. (such as R. citrophilis and R. similis), Ditylenchus spp. and other plant nematode pests.

In particular embodiments the targets are cyst nematodes, such as Heterodera glycines (soybean cyst nematodes), Heterodera schachtii (beet cyst nematode), Heterodera avenae (Cereal cyst nematode), Meloidogyne incognita (Cotton (or southern) root knot nematode), Globodera rostochiensis and Globodera pallida (potato cyst nematodes), and Belonolaimus longicaudatus and Belonolaimus gracilis (sting nematodes). In still other particular embodiments, the targets are root knot nematodes, such as M. incognita (cotton root knot nematode), M. javanica (Javanese root knot nematode), M. hapia (Northern root knot nematode), and M. arenaria (peanut root knot nematode).

Plant Insect Pests:

In a particular embodiment, the one or more bacterial strain(s), inoculum(s) or composition(s) described herein are useful to control plant insect pests. Non-limiting examples of plant insect pests include:

Hemiptera harmful insects:

Planthoppers {Delphacidae) such as small brown planthopper (Laodelphax striatellus), brown rice planthopper (Nilaparvata lugens), white-backed rice planthopper (Sogatella furcifera) and the like; leafhoppers (Deltocephalidae) such as green rice leafhopper (Nephotettix cincticeps), green rice leafhopper (Nephotettix virescens) and the like; aphids {Aphididae) such as cotton aphid {Aphis gossypii), green peach aphid {Myzus persicae), cabbage aphid (Brevicoryne brassicae), potato aphid (Macrosiphum euphorbiae), foxglove aphid (Aulacorthum solani), oat bird-cherry aphid (Rhopalosiphum padi), tropical citrus aphid (Toxoptera citricidus) and the like; stink bugs (Pentatomidae) such as green stink bug (Nezara antennata), bean bug {Riptortus clavetus), rice bug {Leptocorisa chinensis), white spotted spined bug (Eysarcoris parvus), stink bug (Halyomorpha mista), tarnished plant bug (Lyus lineolarxs) and the like; whiteflies (Aleyrodidae) such as greenhouse whitefly (Trialeurodes vaporariorum), sweetpotato whitefly (Bemisia tabaci), silverleaf whitefly (Bemisia argentifolii) and the like; scales (Coccidae) such as Calfornia red scale (Aonidiella aurantii), San Jose scale {Comstockaspis perniciosa) , citrus north scale {Unaspis citri), red wax scale (Ceroplastes rubens), cottonycushion scale (Icerya purchasi) and the like; lace bugs (Tingidae); psyllids (Psyllidae); etc.

Lepidoptera harmful insects:

Pyralid moths (Pyralidae) such as rice stem borer (Chilo suppressalis), yellow rice borer (Tryporyza incertulas), rice leafroller (Cnaphalocrocis medinalis), cotton leafroller (Notarcha derogata), Indian meal moth (Plodia interpunctella), oriental corn borer (Ostrinia furnacalis), European corn borer (Ostrinianubilaris), cabbage webworm (Hellula undalis), bluegrass webworm (Pediasia teterrellus) and the like; owlet moths (Noctuidae) such as common cutworm (Spodoptera litura), beet armyworm (Spodoptera exigua), armyworm (Pseudaletia separata), cabbage armyworm (Mamestra brassicae), black cutworm (Agrotis ipsilon), beet semi-looper (Plusia nigrisigna), Thoricoplusia spp., Heliothis spp., Helicoverpa spp. and the like; white butterflies (Pieridae) such as common white (Pieris rapae) and the like; tortricid moths (Tortricidae) such as Adoxophyes spp., oriental fruit moth (Grapholita molesta), soybean pod borer (Leguminivora glycinivorella), azuki bean podworm (Matsumuraeses azukivora), summer fruit tortrix (Adoxophyes orana fasciata), smaller tea tortrix (Adoxophyes spp.), oriental tea tortrix (Homona magnanima), apple tortrix (Archips fuscocupreanus), codling moth (Cydia pomonella) and the like; leafblotch miners (Gracillariidae) such as tea leafroller (Caloptilia theivora), apple leafminer (Phyllonorycter ringoneella) and the like; Carposinidae such as peach fruit moth (Carposina niponensis) and the like; lyonetiid moths (Lyonetiidae) such as Lyonetia spp. and the like; tussock moths (Lymantriidae) such as Lymantria spp., Euproctis spp. and the like; yponomeutid moths (Yponomeutidae) such as diamondback (Plutella xylostella) and the like; gelechiid moths (Gelechiidae) such as pink bollworm (Pectinophora gossypiella), potato tubeworm (Phthorimaea operculella) and the like; tiger moths and allies (Arctiidae) such as fall webworm (Hyphantria cunea) and the like; tineid moths (Tineidae) such as casemaking clothes moth (Tinea translucens) , webbing clothes moth (Tineola bisselliella) and the like; etc.

Thysanoptera harmful insects:

Thrips (Thripidae) such as yellow citrus thrips (Frankliniella occidentalis), melon thrips (Thrips palmi), yellow tea thrips (Scirtothrips dorsalis), onion thrips (Thrips tabaci), flower thrips (Frankliniella intonsa), tobacco thrips (Frankliniella fusca) and the like, etc.

Diptera harmful insects:

House flies (Musca domestica) , common house mosquito (Culex popiens pallens), horsefly (Tabanus trigonus), onion fly (Hylemya antiqua), seedcorn maggot (Hylemya platura), asian tiger mosquito (Anopheles sinensis); leafminer flies (Agromyzidae) such as rice leafminer (Agromyza oryzae), little rice leafminer (Hydrellia griseola), rice stemmaggot (Chlorops oryzae), legume leafminer (Liriomyza trifolii) and the like; melon fly (Dacus cucurbitae), Meditteranean fruit fly (Ceratitis capitata), etc.;

Coleoptera harmful insects:

Twenty-eight-spotted ladybird (Epilachna vigintioctopunctata), cucurbit leaf beetle (Aulacophora femoralis), striped flea beetle (Phyllotreta striolata) , rice leaf beetle (Oulema oryzae), rice curculio (Echinocnemus squameus), rice water weevil (Lissorhoptrus oryzophilus), boll weevil (Anthonomus grandis), azuki bean weevil (Callosobruchus chinensis), hunting billbug (Sphenophorus venatus), Japanese beetle (Popxllia japonica), cupreous chafer (Anomala cuprea), Corn root worms (Diabrotica spp.), Colorado potato beetle (Leptinotarsa decemlineata), click beetles (Agriotes spp.), cigarette beetle (Lasioderma serricorne), varied carper beetle (Anthrenus verbasci), red flour beetle (Tribolium castaneum), powder-post beetle (Lyctus brunneus), white-spotted longicorn beetle (Anoplophora malasiaca), pine shoot beetle (Tomicus piniperda), etc.;

Orthoptera harmful insects:

Asiatic locust (Locusta migratoria), African mole cricket (Gryllotalpa africana), rice grasshopper (Oxya yezoensis), rice grasshopper (Oxya japonica), etc.; Hvmenoptera harmful insects:

Cabbage sawfly (Athalia rosae), leaf-cutting ant (Acromyrmex spp.), fire ant {Solenopsis spp.), etc.;

Blattodea harmful insects:

German cockroach (Blattella germanica), smokybrown cockroach (Periplaneta fuliginosa), American cockroach (Periplaneta americana), Periplaneta brunnea, oriental cockroach (Blatta orientalis), etc.

Particular examples of the above-described harmful arthropods include aphids (Aphididae), Thrips (Thripidae), leafminer flies (Agromyzidae), horsehair worms (Paragordius tricuspidatus), Colorado potato beetle (Leptinotarsa decemlineata), Japanese beetle (Popillia japonica), cupreous chafer (Anomala cuprea), boll weevil (Anthonomus grandis), rice water weevil (Lissorhoptrus oryzophilus), tobacco thrips (Frankliniella fusca) , Corn root worms (Diabrotica spp.), diamondback (Plutella xylostella), cabbageworms, soybean pod borer (Leguminivora glycinivorella), and the like.

The methods of the present invention are applicable to both leguminous and non- leguminous plants or plant parts. In a particular embodiment the plants or plant parts are selected from the group consisting of alfalfa, rice, wheat, barley, rye, oat, cotton, canola, sunflower, peanut, corn, potato, sweet potato, bean, pea, chickpeas, lentil, chicory, lettuce, endive, cabbage, brussel sprout, beet, parsnip, turnip, cauliflower, broccoli, turnip, radish, spinach, onion, garlic, eggplant, pepper, celery, carrot, squash, pumpkin, zucchini, cucumber, apple, pear, melon, citrus, strawberry, grape, raspberry, pineapple, soybean, tobacco, tomato, sorghum, and sugarcane. The methods described herein are also used to treat perennial plants, including plantation crops such as banana and coffee and those present in forests, parks or landscaping.

In one particular embodiment the plant is selected from the group consisting of non- legumes, legumes, Brassica spp., cereals, fruits, vegetables, nuts, flowers, and turf. Particularly the cereals are wheat, corn, rice, oat, rye, barley. Particularly legumes are lentil, chickpeas, beans, soybeans, peas, and alfalfa.

In another particular embodiment the plants are selected from the group consisting of alfalfa, rice, wheat, barley, rye, oat, cotton, sunflower, peanut, corn, potato, sweet potato, bean, pea, chickpeas, lentil, chicory, lettuce, endive, cabbage, brussel sprout, beet, parsnip, turnip, cauliflower, broccoli, turnip, radish, spinach, onion, garlic, eggplant, pepper, celery, carrot, squash, pumpkin, zucchini, cucumber, apple, pear, melon, citrus, strawberry, grape, raspberry, pineapple, soybean, tobacco, tomato, sorghum, and sugarcane.

SEED COATINGS

In another aspect, seeds are coated with one or more microbial pesticides described herein. In another embodiment, seeds are coated with one or more pests with one or more microbial nematicides. In a particular embodiment, the seeds are coated with one or more nematophagous bacteria. In a more particular embodiment, seeds are coated with one or more strains of Chromobacterium spp. In an even more particular embodiment, seeds are coated with one or more strains of Chromobacterium vaccinii. In still an even more particular embodiment, seeds are coated with one or more strains of Chromobacterium vaccinii sp. nov. In yet an even more particular embodiment, seeds are coated with the strain of Chromobacterium vaccinii having the deposit accession number NRRL B-50880, variants of the strain of Chromobacterium vaccinii having the deposit accession number NRRL B-50880, or combinations thereof.

In one embodiment, seeds may be treated with composition(s) described herein in several ways but preferably via spraying or dripping. Spray and drip treatment may be conducted by formulating compositions described herein and spraying or dripping the composition(s) onto a seed(s) via a continuous treating system (which is calibrated to apply treatment at a predefined rate in proportion to the continuous flow of seed), such as a drum- type of treater. Batch systems, in which a predetermined batch size of seed and composition(s) as described herein are delivered into a mixer, may also be employed. Systems and apparati for performing these processes are commercially available from numerous suppliers, e.g., Bayer CropScience (Gustafson).

In another embodiment, the treatment entails coating seeds. One such process involves coating the inside wall of a round container with the composition(s) described herein, adding seeds, then rotating the container to cause the seeds to contact the wall and the composition(s), a process known in the art as "container coating". Seeds can be coated by combinations of coating methods. Soaking typically entails using liquid forms of the compositions described. For example, seeds can be soaked for about 1 minute to about 24 hours (e.g., for at least 1 min, 5 min, 10 min, 20 min, 40 min, 80 min, 3 hr, 6 hr, 12 hr, 24 hr). In certain embodiments, a seed(s) coated with one or more of the compositions described herein will comprise 1 x 10¹ - 1 x 10⁸, more preferably 1 x 10² - 1 x 10⁶ colony forming units of one or more of the microbial pesticides strains per seed, in particular, the strain of Chromobacterium vaccinii having the deposit accession number NRRL B-50880, variants of the strain of Chromobacterium vaccinii having the deposit accession number NRRL B-50880, or combinations thereof.

EXAMPLES

The following examples are provided for illustrative purposes and are not intended to limit the scope of the invention as claimed herein. Any variations in the exemplified examples which occur to the skilled artisan are intended to fall within the scope of the present invention. Example 1. Isolation of, and phylogenetic studies on, Chromobacterium spp. isolate PS1

(NRRL B-50880)

Two thousand bacterial isolates were obtained from soil samples in Virginia and Colorado and were used to screen for activity against the nematode Caenorhabditis elegans. The soil samples were suspended in phosphate buffer bottles and diluted to plate for single isolates onto SMA (Standard Method Agar) plates. Single isolates were re-isolated onto SMA plates and the strains were used to screen against C. elegans. A 10 μΙ inoculating loop was used to harvest cells from the plates and suspended in 1 ml of phosphate buffer. One hundred μΙ of the suspension was plated onto SMA plates. After the plates were dried, a square piece of agar containing C. elegans was transferred to SMA plates containing individual isolates. Observations of the health of the C. elegans were made on days 1 , 2, 3, 4, and 7. After several screening rounds, three isolates were found to inhibit or completely kill C. elegans. Those isolates were sent for 16S rDNA sequence analysis. Based on 500 base pair 16S rDNA sequence analysis, the three isolates were identified as Brevibacillus laterosporus, Bacillus thuringenesis, and Chromobacterium spp. Preliminary studies indicated the Chromobacterium isolate, designated PS1 (also as NRRL B-50880), was toxic to C. elegans, and additional studies were performed on this isolate.

To better understand the identity of the PS1 Chromobacterium isolate, full length (1500 bp) 16S rDNA sequences were obtained for PS1 ). The sequence alignment program, ClustalW, was used to compare the PS1 sequence to full length sequences of three different strains of Chromobacterium vaccinium (MWU300, MWU328, MWU205), along with Chromobacterium subtsugae. After completion of the alignment, a phylogenetic tree was generated and the PS1 (NRRL B-50880) isolate of Chromobacterium spp. belongs to the species Chromobacterium vaccinium (Figure 1 ).

In addition, the PS1 isolate was compared to C. violaceum using the ClustalW program. Again full length 16S rDNA sequences were obtained and comparisons were made to produce the phylogenetic tree. Based on the analysis, the isolate Chromobacterium spp (PS1 ) was closer to C. vaccinium than either C. violaceum or C. subtsugae (Figure 2). Therefore, the isolate PS1 isolate was classified as C. vaccinium and deposited into the USDA culture collection as NRRL B-50880. Example 2. Bioactivitv of NRRL B-50880 toward Caenorhabditis elegans in vitro

Bacterial isolates were plated onto SMA plates from -80°C stock cultures and incubated at 30°C overnight. In addition, these isolates were compared to Chromobacterium subtsugae and Escherichia coli (K12) as controls. A 10 μΙ loop of cells was inoculated into fresh 5 ml select medium in disposable culture tubes for all bacterial isolates except E. coli. which was grown in 5 ml Luria broth medium. All culture tubes were incubated overnight at 35°C with shaking. One-half ml of each overnight culture was centrifuged on a benchtop centrifuge at 13,000 rpm for 1 minute in a microcentrifuge tube. Four tenths of a ml of the supernatant was disposed while the cells were resuspended in the remaining 100 μΙ of supernatant. The concentrated cells were plated onto 1.2% noble agar plates to cover the entire plate. Each isolate was plated onto three individual plates. Once the plates were dried, square pieces of agar (-0.5 square inches) containing juvenile stage C. elegans were transferred to the isolate plates. The health of the C. elegans was observed at 8 hours, 1 day, 3 days, and 4 days under microscopy. At 8 hours, and days 1 and 2, C. elegans were still healthy and motile for all isolates tested. After 3 days of exposure, no C. elegans could be observed for NRRL B-50880. Pictures were taken for observations made at 8 hours and 3 days (Figure 3). The results also showed that the Chromobacterium vaccinium was very effective in killing or lysing C. elegans nematodes within 3 days. At the same time, C. elegans nematodes were alive in control plates (water and E. coli K12 controls).

Example 3. Bioactivitv of NRRL B-50880 toward pathogenic root-knot nematodes (Meloidogyne incognita)

An experiment was performed to determine the nematicidal effect of Brevibacillus laterosporus, Bacillus thuringenesis, and Chromobacterium spp. on southern root-knot nematodes (Meloidogyne incognita). The in vitro bioactivities of those three bacterial species were compared to the effects of Escherichia coli K12, Streptomyces lydicus (WYEC108), Dursban (chemical standard), media control, and water, on the nematodes.

Each bacterium was grown in two different media and the cultures were mixed 50:50 before the in vitro testing was conducted. The media were select medium (10 g dextrose, 1 g tryptone, 1 g yeast extract, 1 g beef extract, 0.01 g iron sulfate, pH 7.2) and potato dextrose medium. In the experiment, fifty second-stage M. incongnita juveniles were suspended in each treatment and each treatment was repeated five times. Mortality of the nematodes was counted after one hour and one day of exposure to each treatment. Efficacy of each treatment was evaluated based on the corrected percent mortality. The equation used to calculate corrected percent mortality was Abbott's Formula (Corrected percent mortality = (Percent alive in control - Percent alive in treated) / (Percent alive in control)). The results of this experiment, shown in Table 1 , indicated that the bioactivity of Chromobacterium vaccinium NRRL B-50880 on root-knot nematodes was better than Brevibacillus laterosporus and Bacillus thuringenesis, and similar to the chemical standard.

Table 1 : Corrected mortality from each treatment at different time after exposure to the treatment

'Means followed by the same letter among rows and columns do not significantly differ (p=0.05).

A similar experiment was performed, also comparing the nematicidal activity of Chromobacterium vaccinium. The results of this experiment, shown in Table 2, indicated the activity of NRRL B-50880 was similar or possibly better than that of Chromobacterium subtsugae.

Table 2: Corrected mortality from each treatment at different time after exposure to the treatment

'Means followed by the same letter among rows and columns do not significantly differ (p=0.05).

In addition to the mortality of juvenile nematodes, the inhibition of nematode egg hatching was tested using the same bacteria. The results of this experiment, shown in Table 3, demonstrated that Chromobacterium vaccinium NRRL B50880 and B. thuringenesis exhibited egg hatching inhibition.

Table 3: Corrected e hatchin inhibition after ex osure to the treatment % )

'Means followed by the same letter among rows and columns do not significantly differ (p=0.05).

Claims

CLAIMS:

1. An isolated bacterial strain of Chromobacterium vaccinii having a deposit accession number NRRL B-50880.

2. A composition comprising a carrier and a bacterial strain of Chromobacterium vaccinii.

3. The composition of claim 2, wherein the composition includes a non- naturally occurring component.

4. The composition of claim 2, wherein the carrier is a non-naturally occurring component.

5. The composition of claim 2, wherein the bacterial strain is the strain of Chromobacterium vaccinii having a deposit accession number NRRL B-50880.

6. The composition of claim 2, wherein the composition further comprises one or more beneficial microorganisms.

7. The composition of claim 6, wherein the one or more beneficial microorganisms is a nitrogen fixing microorganism, a phosphate solubilizing microorganism, or a combination thereof.

8. The composition of claim 2, wherein the composition further comprises one or more fungicides.

9. The composition of claim 8, wherein the fungicide is a biofungicide.

10. The composition of claim 2, wherein the composition further comprises one or more nematicides.

1 1. The composition of claim 10, wherein the one or more nematicides is a chemical nematicide, a natural nematicide, a bionematicide, or combinations thereof.

12. The composition of claim 1 1 , wherein the bionematicide is a nematophagous fungus and/or nematophagous bacteria.

13. The composition of claim 2, wherein the composition further comprises one or more plant signal molecules.

14. The composition of claim 13, wherein the one or more plant signal molecules is a lipo-chitooligosaccharide (LCO).

15. The composition of claim 13, wherein the plant signal molecule is a chitinous compound.

16. The composition of claim 15, wherein the chitinous compound is a chito-oligomer (CO).

17. The composition of claim 13, wherein the plant signal molecule is a flavonoid.

18. The composition of claim 2, further comprising one or more insecticides.

19. A method of controlling one or more plant pests, comprising introducing into soil a strain of Chromobacterium vaccinii having a deposit accession number NRRL B-50880.

20. The method of claim 19, wherein the step of introducing into soil the Chromobacterium vaccinii comprises introducing an inoculum of the Chromobacterium vaccinii strain into soil in an amount of 1 x 10¹ - 1 x 10¹², more preferably 1 x 10⁶ - 1 x 10¹² colony forming units per hectare.

21 . A method of controlling one or more plant pests, comprising introducing into soil the composition of any of claims 2-18.

22. The method of claim 21 , wherein the plant pest is a nematode.

23. The method of claim 21 wherein the plant pest is an insect.

24. A seed coated with a bacterial strain of Chromobacterium vaccinii having a deposit accession number NRRL B-50880.

25. A seed coated with a composition of any of claims 2-18.