Methylobacterium compositions for inhibition of plant pathogens and related methods

Abstract

Compositions comprising Methylobacterium that inhibit infection by a oomycete or fungal plant pathogen and / or reduce severity of symptoms caused by a plant pathogenic oomycete or fungus, methods of making the compositions, and methods of treating plants to inhibit and / or reduce symptoms of oomycete and / or fungal infections are provided.

Description

METHYLOBACTERIUM COMPOSITIONS FORINHIBITION OF PLANT PATHOGENS AND RELATED METHODSReference to Priority Applications

[0001] This patent application claims benefit of U.S. Patent Application No. 63 / 632,681, filed April 11, 2024.Sequence Listing Statement

[0002] A computer readable form of the Sequence Listing XML containing the file named "NLSYM195920WO Sequence Listing. xml," which is 23,993 bytes in size (as measured in MICROSOFT WINDOWS® EXPLORER) and was created on April 8, 2025, is provided herein and is herein incorporated by reference. This Sequence Listing consists of SEQ ID NOs:l-24.BACKGROUND

[0003] One-carbon organic compounds such as methane and methanol are found extensively in nature, and are utilized as carbon sources by bacteria classified as methanotrophs and methylotrophs. Methanotrophic bacteria include species in the genera Methylobacter, Methylomonas, Methylomicrobium, Methylococcus, Methylosinus, Methylocystis, Methylosphaera, Methylocaldum, and Methylocella (Lidstrom, 2006).

[0004] Methanotrophs possess the enzyme methane monooxygenase that incorporates an atom of oxygen from O2 into methane, forming methanol. All methanotrophs are obligate one-carbon utilizers that are unable to use compounds containing carbon-carbon bonds. Methylotrophs, on the other hand, can also utilize more complex organic compounds, such as organic acids, higher alcohols, sugars, and the like. Thus, methylotrophic bacteria are facultative methylotrophs. Methyl otrophic bacteria include species in the genera Methylobacterium, Methylorubrum, Hyphomicrobiiim. Methylophilus, Methylobacillus, Methylophaga, Aminobacler. Methylorhabdus, Methylopila, Methylosulfonomonas, Marinosulfonomonas, Paracoccus, Xanthobacter , Ancylobacter (also known as Microcyclus), Thiobacillus, Rhodopseudomonas, Rhodobacter, Acetobacter, Bacillus, Mycobacterium, Arthobacter, and Nocardia (Lidstrom, 2006).

[0005] Most methylotrophic bacteria of the genera Methylobacterium and Methylorubrum are pink- pigmented. They are conventionally referred to as PPFM bacteria, being pink-pigmented facultative methylotrophs. Green (2005, 2006) identified twelve validated species in the genus Methylobacterium, specifically M. aminovorans, M. chlor omethanicum, M. dichlor omethanicum, M. extorquens, M. fujisawaense, M. mesophilicum, M. organophilum, M. radiotolerans, M. rhodesianum, M. rhodinum, M. thiocyanatum, and M. zatmanii. However, M. nodulans is a nitrogen-fixing Methylobacterium that isnot a PPFM (Sy et al., 2001). Methylobacterium are ubiquitous in nature, being found in soil, dust, fresh water, sediments, and leaf surfaces, as well as in industrial and clinical environments (Green, 2006).

[0006] Biotic pressures — the combined effects of pathogens, animals, insects, and weeds — result in an annual loss of 20-40% crop productivity. Additional costs of pest control, reduced crop quality, and funding research to combat these pressures, among other factors, make it difficult to precisely quantify losses due to crop pests and diseases. Agricultural biologicals present a novel, exciting, and rapidly emerging option to combat crop pests and diseases in a manner that complements and preserves current crop germplasm resistance traits, chemical control options, and cultural practices. Important targets of such biocontrol methods include diseases caused by soilborne pathogens.

[0007] Diseases caused by soilborne pathogens such asoomycetes (heterokonts or stramenopiles in the class: Oomycota) are particularly widespread and contribute to significant crop losses. Plant pathogenic oomycetes known to cause losses in crop production include, for example, species from Pythium, Peronospora. Phytophthora, Plasmopara, Haliotidica, Saprolegnia, and Aphanomyces. Oomycetes are also known as “water molds” and are mainly dispersed as self-motile spores, called zoospores. Notable plant diseases caused by oomycetes include damping-off, seedling blights, root rots, foliar blights and downy mildews.

[0008] Many Pythium species, along with their close relatives Phytophthora, are plant pathogens of economic importance in agriculture. t / zzzzzzz-induced root rot is a common crop disease. When the organism kills newly emerged or emerging seedlings, it is known as damping off, and is a very common problem in fields and greenhouses. t / zzzzzzz-induced wilt is caused by zoospore infection of older plants, leading to biotrophic infections that become necrotrophic in response to colonization / reinfection pressures or environmental stress, which leads to wilting resulting from reduced root functioning. Pythium species tend to be very generalistic and non-specific in their range of hosts, while Phytophthora species are generally more host specific. Pythium species are also more devastating in the root rot they cause in crops because of the ability of Pythium species, which are also good saprotrophs, to survive on decaying plant matter. As a result, crop rotation alone is an ineffective approach to eradication of the pathogen.

[0009] In field crops, Pythium zoospores can spread infection in field soils to a limited amount. Zoospores can, however, spread rapidly when present in irrigation water and also travel in moist soils. In hydroponic systems in greenhouses, extensive monocultures of plants are maintained in a nutrient solution that is continuously recirculated to the crop. Zoospores can rapidly infect entire recirculating tanks in hydroponic systems and spread infection when present. Thus, in greenhouse environments, Pythium spp. can cause extensive and devastating root rot and t / zzzzzzz-induced diseases are often difficult to prevent or control.

[0010] While not all species of Pythium cause crop losses, there are many types of Pythium that effect a wide variety of crops worldwide. Pythium species have a broad host range and disease development is highly dependent on environmental conditions. Some species require, moist and cool conditions (such as Pythium irregulars and P. ultimum) while others are most severe at higher temperatures (such as Pythium aphanidermatum and P. myriotylum).[OH] Plant diseases caused by Pythium can affect plant parts in contact with the soil (roots, lower stem, seeds, tubers, and fleshy fruits), while other Pythium-m ucQ diseases affect above ground parts (leaves, young stems, and fruits). Some Pythium species infect roots of mature plants, typically causing necrotic lesions on root tips or fine feeder roots and, less commonly, on tap roots. Pythium infections are often limited to the meristematic root tips, root epidermis, cortex of roots, and fruits, but severe infections can occur when the pathogen moves deeper into the plant tissue and reaches the vascular system. If soil is infested with Pythium, pre-emergence damping-off occurs when the seeds are colonized during germination and become soft, turn brown, and disintegrate. In post-emergence damping-off, seedlings are infected right after germination.

[0012] Plant diseases caused by oomycetes are currently mitigated by chemical fungicides, although the efficacy of these chemistries is rapidly diminishing. New compositions and methods for treating oomycete related plant diseases are needed and. Microbial inoculants which can colonize plants from seed and persist on plants and in soil for the full duration of the growing season, can provide benefit alone, or in combination with existing chemistries, and offer extended windows of protection. Thus, microbial inoculants can compensate for chemical control insufficiencies, and extend the benefits of fungicide treatments through resistance management.

[0013] Fusarium species are also known to cause seedling disease on many plants hosts, including row crop plants and crops grown in hydroponic systems. Symptoms of diseases caused by Fusarium species include damping-off, root damage and vascular wilt. F. graminearum is the causal agent of Fusarium head blight (FHB) on wheat, barley, and other cereals. This pathogen is also responsible for ear and stalk rot in com. In addition to causing significant reductions in yield and grain quality, F. graminearum produces harmful mycotoxins that are a major concern in the animal feed industry. Furthermore, there is an increasing problem in farming with fungal pathogens such as F. graminearum becoming resistant to a wide range of chemical fungicides.

[0014] Sudden death syndrome of soybean is caused by the soil-borne fungus Fusarium virguliforme, previously known as Fusarium solani f. sp. glycines, and is exacerbated by conditions of high soil moisture and soil compaction. Symptoms of SDS include a mosaic-like appearance of leaf tissue in which main veins remain green while other leaf areas become chlorotic or necrotic, reddish discoloration of xylem tissue, blackening or rotting of root tissue, and significant reductions to overall plant health and yield. From 1994-2010, soybean yield losses to diseases caused by Fusarium species were estimatedat c. 36.2 million bushels / year and the majority of these losses were attributed to F. virguliforme. Thus, there exists a need in the agricultural and animal feed industries for the development of effective new approaches for inhibiting fungal or oomycete pathogens and / or reducing adverse effects of such pathogens on plants.SUMMARY OF THE INVENTION

[0015] The present invention provides methods and compositions that are shown to have the following beneficial characteristics: a) improved growth metrics, including plant size, tiller counts, plant height; b) improved yield, including panicle counts, panicle weights, and shoot biomass; c) improved methane mitigation; and d) improved colonization of beneficial microorganisms.

[0016] Moreover, the present invention provides methods to identify and use compositions useful for improved growth metrics, yield, methane mitigation, and colonization. Also provided are methods that utilize such strains for mitigating yield loss due to infection pressure, infection, or disease. For example, the present strains and methods are useful in rice cultivation environments.

[0017] The present invention provides compositions comprising a Methyl obacterium that prevents infection of plants and / or reduces severity of disease associated with one or more oomycete or fungal plant pathogens, wherein the Methyl obacterium is selected from the group consisting of: NLS0059, NLS0165, NLS0214, NLS0419, NLS0433, NLS0498, NLS0532, NLS0632, NLS0952, NLS0729, NLS1310, NLS0502, NLS0575, NLS0704, NLS0707, NLS0934, and variants thereof.

[0018] Also provided are such compositions, wherein the plant pathogen is a Pythium species or a Fusarium species.

[0019] Also provided are such compositions, wherein said plant pathogen is a species of Pythium, Peronospora, Phytophthora, Plasmopara, Haliotidica, Saprolegnia, Albugo, Pustula, Wilsoniana, Hyaloperonospora, Pseudoperonospora, Globosporangium or Aphanomyces.

[0020] Also provided are such compositions, wherein said plant pathogen is a Pythium species and said Methyl obacterium is selected from the group consisting of: NLS0059, NLS0165, NLS0214, NLS0532, NLS0632, NLS0704, and variants thereof..

[0021] Also provided are such compositions, wherein said plant pathogen is a Fusarium species and said Methyl obacterium is selected from the group consisting of: NLS0419, NLS0433, NLS0498, NLS0952, NLS0729, NLS1310, NLS0502, NLS0575, NLS0704, NLS0707, NLS0934, and variants thereof.

[0022] Also provided are such compositions, wherein the composition further comprises Methyl obacterium strain selected from the group consisting of: one or more strain in Table 1, one ormore strain in Table 1A, one or more strain in Table IB, NLS0017, NLS0064, NLS0020, NLS0021, NLS0042, NLS0066, NLS0089, NLS0109, and variants thereof.

[0023] Also provided are such compositions, wherein the composition further comprises a Methyl obacterium strain that is a variant of NLS0017 or NLS0064 has a sequence of any one of SEQ ID NOS: l-3 or 13-15.

[0024] Also provided are such compositions, wherein the fungal plant pathogen is a soilborne pathogen.

[0025] Also provided are such compositions, wherein the composition further comprises a fungicide.

[0026] Also provided are such compositions, wherein the fungicide is strobilurin, azoxystrobin, Trichoderma, metalaxyl, mefenoxam, ethaboxam, or oxathiapiprolin.

[0027] The present invention also provides methods for reducing the adverse effects on a plant caused by an oomycete or fungal plant pathogen, wherein said method comprises applying a composition comprising Methyl obacterium selected from the group consisting of: NLS0017, NLS0064, NLS0059, NLS0165, NLS0214, NLS0419, NLS0433, NLS0498, NLS0532, NLS0632, NLS0952, NLS0729, NLS1310, NLS0502, NLS0575, NLS0704, NLS0707, NLS0934, and variants thereof to a plant, a plant part, to soil or to a nutrient solution where a plant is grown, or any combination thereof in an amount that provides for i) inhibition of infection by said plant pathogen in said plant, plant part, or a plant obtained therefrom, or ii) reduction in severity of symptoms of said plant pathogen; wherein said inhibition of infection and / or reduction of severity of symptoms are relative to infection of a control plant, plant part, or plant that had not received an application of said composition or been grown in soil or nutrient solution treated with the composition.

[0028] Also provided are such methods, wherein application of said composition provides for at least about 10%, at least about 20%, at least about 25%, at least about 30%, 40%, 50%, 75%, at least 85%, or at least 95% inhibition of infection or reduction in severity of symptoms in said plant, plant part, or a plant derived therefrom relative to the control plant, plant part, or plant.

[0029] Also provided are such methods, wherein said plant part is selected from the group consisting of: a leaf, a stem, a vegetative cutting, a fruit, a flower, a root, a tuber, and a seed.

[0030] Also provided are such methods, wherein the plant or plant is selected the group consisting of: berry, cannabis, corn, floriculture / ornamental, grape, herb, hops, leafy green, lettuce, microgreen, peanut, pepper, pome fruit, rice, soybean, tomato, tree nut, turf grass, vegetable, and wheat.

[0031] Also provided are such methods, wherein said plant pathogen is a Pythium species and said Methyl obacterium is selected from the group consisting of: NLS0059, NLS0165, NLS0214, NLS0532, NLS0632, NLS0704, and variants thereof.

[0032] Also provided are such methods, wherein said plant pathogen is a Fusarium species and said Methyl obacterium is selected from the group consisting of: NLS0419, NLS0433, NLS0498, NLS0952, NLS0729, NLS1310, NLS0502, NLS0575, NLS0704, NLS0707, NLS0934, and variants thereof.

[0033] Also provided are such methods, wherein the plant pathogen is a Fusarium species and the Methyl obacterium is NLS0017 or a variant of NLS0017 comprising any one of SEQ ID NOS: 1-3, or the plant pathogen is a Pythium species and the Methyl obacterium is NLS0064, or a variant of NLS0064 comprising any one of SEQ ID NOS: 13-15.

[0034] Also provided are such methods, further comprising applying a second Methyl obacterium to a plant, a plant part, to soil where a plant is grown, or any combination thereof, wherein said second Methyl obacterium is selected from the group consisting of: one or more strain in Table 1, one or more strain in Table 1A, one or more strain in Table IB, NLS0017, NLS0064, NLS0020, NLS0021, NLS0042, NLS0066, NLS0089, NLS0109, and variants thereof

[0035] Also provided are such methods, wherein the second Methylobacterium is applied to the soil in furrow with the plant part, on a seed from which the plant is grown, to the plant foliage, as a liquid to a vegetative cutting, or in a nutrient solution in which the plant is grown.

[0036] Also provided are such methods, wherein Methylobacterium and said second Methylobacterium are applied simultaneously.

[0037] Also provided are such methods, wherein said method further comprises applying a fungicide in a composition comprising said Methylobacterium or following application of said Methylobacterium.

[0038] Also provided are such methods, wherein the fungicide is strobilurin, azoxystrobin, Trichoderma. metalaxyl, mefenoxam, ethaboxam, or oxathiapiprolin.

[0039] Also provided are such methods, wherein the fungicide is active against Pythium species.

[0040] Also provided are such methods, wherein the fungicide is active against Fusarium species.

[0041] The present invention also provides plants or plant parts that are at least partially coated with a composition herein.

[0042] Also provided are such plants or plant parts, wherein the plant or plant part is selected from the group consisting of: berry, cannabis, corn, floriculture / omamental, grape, herb, hops, leafy green, lettuce, microgreen, peanut, pepper, pome fruit, rice, soybean, seed crop, tomato, tree nut, turf grass, vegetable, and wheat.

[0043] Also provided are such plants or plant parts, wherein the plant or plant part is isolated.DESCRIPTION

[0044] Provided herein are inoculant compositions and fermentation products comprising Methylobacterium species that can be used to prevent infection of plants by oomycete and / or fungal pathogens, including plant pathogenic Pythium and Fusarium species, and / or reduce severity of diseases caused by these and other plant pathogenic species. Methods of using the compositions to suppress adverse effects of Pythium and Fusarium diseases of plants, plant parts, and plants derived therefrom, and methods of making the compositions are also provided herein. In certain embodiments, species for inhibition of plant diseases caused by soilborne pathogens can be distinguished from other Methylobacterium that are not effective for inhibiting infection of a plant by a soilborne pathogen by assaying for the ability of the Methylobacterium to inhibit infection or reduce adverse effects of such diseases in a plant or on an isolated plant part.

[0045] Comparing the genomes of Methylobacterium isolates that are effective against pathogens that cause plant diseases to Methylobacterium isolates that are not effective against the same pathogens can reveal genes and gene pathways responsible for the reduction of infection phenotype. Methylobacterium that are useful for preventing infection of a particular plant pathogen, such as a Pythium or Fusarium species, or suppressing symptoms of a disease caused by such pathogens, may also find use as biocontrol agents against other plant pathogens, including for example other soilborne oomycetes or fungal pathogens. Thus, Methylobacterium that inhibit infection and / or adverse effects caused by a Pythium species may also find use in inhibiting infection and / or reducing symptoms related to a disease caused by one or more of the following genera: Pythium, Peronospora, Phytophthora, Plasmopara, Haliotidica, Saprolegnia, Albugo, Pustula, Wilsoniana, Hyaloperonospora, Pseudoperonospora, Globosporangium and Aphanomyces .

[0046] Provided herein are compositions comprising Methylobacterium that inhibit infection caused by oomycete or fungal pathogens and / or reduce the adverse effects related to plant diseases caused by such pathogens. In some embodiments, the pathogens are species of Pythium ox Fusarium. Such compositions may comprise a single disclosed Methylobacterium isolate or may comprise a combination of different Methylobacterium isolates. Such compositions may also comprise additional components, for example, one or more agriculturally acceptable excipients and / or agriculturally acceptable adjuvants.

[0047] In certain embodiments of any of the aforementioned compositions, the pathogen is a Pythium species and the Methylobacterium is selected from the group consisting of NLS0059, NLS0165, NLS0214, NLS0532, NLS0632, NLS0704, and variants thereof. In certain embodiments, the pathogen is a Fusarium species and the Methylobacterium is selected from the group consisting of NLS0419, NLS0433, NLS0498, NLS0952, NLS0729, NLS1310, NLS0502, NLS0575, NLS0704, NLS0707, NLS0934 and variants thereof.

[0048] In certain embodiments of any of the aforementioned compositions, the composition comprises a combination of any of the aforementioned Methylobacterium species for inhibition of plant diseases caused by oomycete or fungal pathogens. In certain embodiments of any of the aforementioned compositions, the compositions further comprise a second Methylobacterium strain selected from NLS0020, NLS0021, NLS0037, NLS0042, NLS0066, NLS0089, NLS0109, or variants thereof Use of any of the aforementioned compositions for coating or partially coating a plant part (e.g., a seed) to inhibit plant diseases caused by oomycete or fungal pathogens is also provided herein.

[0049] In certain embodiments of any of the aforementioned compositions, the Methylobacterium for inhibition of infection by a Pythium or Fusarium pathogen and / or reduction of symptoms or adverse effects of a disease caused by Pythium or Fusarium pathogens is also effective in inhibiting infections by other plant pathogens. For example, Methylobacterium effective against Pythium species may also be effective against other oomycetes, such as Phytophthora species. In any of the aforementioned embodiments inhibition of the fungal pathogen may be the result of activity on spores, sporangia, hyphal swellings or hyphae stage of the pathogen, or on a combination of one or more stages of the pathogen. In any of the aforementioned embodiments, inhibition of a plant pathogenic fungus can result from an effect on an anamorphic form, teleomorphic form, or in both its anamorphic form and its teleomorphic forms. In any of the aforementioned embodiments, the compositions comprise a Methylobacterium concentration sufficient to inhibit infection by a fungal pathogen and or reduce adverse effects, ie. plant disease symptoms, caused by a fungal pathogen. In some embodiments, the fungal pathogen is a Fusarium or Pythium species. Use of any of the aforementioned compositions for coating or partially coating a plant or plant part (e.g., a seed) to inhibit a plant disease is also provided herein. In certain embodiments, a plant, plant part, or seed is coated with a Methylobacterium selected from the group consisting of NLS0059, NLS0165, NLS0214, NLS0419, NLS0433, NLS0498, NLS0532, NLS0632, NLS0952, NLS0729, NLS1310, NLS0502, NLS0575, NLS0704, NLS0707, NLS0934 and variants thereof. In certain embodiments, the fungal pathogen is a Pythium species and the Methylobacterium is selected from the group consisting of NLS0059, NLS0165, NLS0214, NLS0532, NLS0632, NLS0704, and variants thereof. In certain embodiments, the plant pathogenic fungus is a. Pythium species selected from Pythium aphanidermatum, P. dissoticum, P. myriotylum, P. ultimum, P. ultimum var. ultimum, P. catenulatum, P. arrhenomanes, P. debaryanum, P. graminicola, P. heterothallicum, P. irregulare, P. myriotylum, P. torulosum; P. lutarium, P. oligandrum, P. oopapillum, P. paddicum, P. paroecandrum, and P. sylvaticum. In certain embodiments, the fungal pathogen is a Fusarium species and the Methylobacterium is selected from the group consisting of NLS0419, NLS0433, NLS0498, NLS0952, NLS0729, NLS1310, NLS0502, NLS0575, NLS0704, NLS0707, NLS0934 and variants thereof. In certain embodiments the Fusarium species is selected from the group consisting of F. graminearum, F.verticillioides, F. oxysporum, F. virguliforme, and solani.

[0050] In certain embodiments of the aforementioned compositions the Methylobacterium is selected from the group consisting of NLS0059, NLS0165, NLS0214, NLS0419, NLS0433, NLS0498, NLS0532, NLS0632, NLS0704, NLS0952, NLS0729, NLS1310, NLS0502, NLS0575, NLS0707, NLS0934 and variants thereof, and the plant is a crop plant selected from soybean, wheat, corn, hops, cannabis, grapes, microgreens, leafy greens, lettuce, tomato, pepper and other vegetables, particularly greenhouse grown vegetables, floriculture crops and other ornamentals, and other specialty crops.

[0051] In any of the aforementioned embodiments, the composition can further comprise an antifungal compound, for example an antifungal compound having activity against Fusarium and / or Pythium species. In certain embodiments, the antifungal compound can be a metalaxyl, mefenoxam, oxathiapiprolin, ethaboxam, thiram, propamocarb, dimethomorph, or a thiophanate-methyl fungicide. In certain embodiments, the Methylobacterium compositions may further comprise fungicides active against plant pathogenic fungi such as Rhizoctonia or Fusarium species, including for example, tebuconazole, triticonazole, metconazole, ipconazole, prothioconazole, thiram, captan, penflufen, imazalil, carboxin, thiabendazole, or fluxapyroxad fungicides. Thus, compositions provided herein may comprise any of a broad number of types of chemicals for control of plant pathogenic organisms, such as Pythium species, or plant pathogenic fungi, including Rhizoctonia and / or Fusarium species. Such chemicals include azoles, acetanilides, dithiocarbamates, a strobilurins, phenylamides, thiazolecarboxamides, piperidinyl thiazole isoxazolines or benzimidazoles. Use of any of the aforementioned compositions for coating or partially coating a plant or plant part (e.g., a seed) to inhibit or reduce symptoms of diseases induced by fungal or oomycete plant pathogens is also provided herein.

[0052] Also provided are plants or plant parts that are at least partially coated with any of the aforementioned compositions comprising Methylobacterium for inhibiting disease caused by fungal pathogens such as Fusarium, oomycetes such as Pythium or other plant pathogenic oomycetes. In certain embodiments, the at least partially coated plant or plant part is selected from soybean, wheat, com, hops, cannabis, grapes, berry, leafy greens, lettuce, tomato, pepper and vegetables, particularly greenhouse grown vegetables, floriculture crops and other ornamentals. In certain embodiments, the at least partially coated plant part is a plant part from a plant selected from the group consisting of soybean, wheat, corn, hops, cannabis, grapes, leafy greens, lettuce, tomato, pepper and vegetables, particularly greenhouse grown vegetables, floriculture crops and other ornamentals. In certain embodiments, the at least partially coated plant or plant part is a dicot plant part. In certain embodiments, the dicot plant or plant part is a soybean or cannabis plant or plant part. In certain embodiments of any of the aforementioned plants or plant parts, the Methylobacterium in the composition was obtained from a plant genus, plant species, plant sub-species, or plant cultivar that is distinct from the genus, species, sub-species, or cultivar of the plant or plant part that is coated with the composition.

[0053] In certain embodiments of any of the aforementioned compositions, uses thereof, or methods, the plant or plant part comprises an amount of the Methylobacterium that is effective for inhibiting or reducing adverse effects of a disease caused by a Pythium species or Fusarium species. In certain embodiments, the amount of the Methylobacterium applied to a plant part (e.g., a seed) is about l.OxlO2, l.OxlO3, l.OxlO4, or l.OxlO5to about l.OxlO7or l.OxlO8CFUs of Methylobacterium per plant part (e.g., a seed). In certain embodiments, the Methylobacterium is heterologous to the plant or plant part. In certain embodiments of any of the aforementioned plant parts, the plant part is a leaf, a stem, a fruit, a vegetative cutting (such as used in clonal propagation, for example), a flower, a root, a seedling, a tuber, or a seed.

[0054] Also provided are methods of making any of the aforementioned compositions containing the Methylobacterium for inhibition of Pythium or / ’zz.szzz'zzz / jz-induced plant diseases or plant diseases that are induced by other plant pathogenic fungi or oomycetes. Such methods can in certain embodiments comprising the steps of combining the Methylobacterium with one or more additional components including an agriculturally acceptable excipient, an agriculturally acceptable adjuvant, a pesticide, or a second biological.

[0055] In certain embodiments, the composition used to treat the seed or plant part can contain a Methylobacterium strain and an agriculturally acceptable excipient. Agriculturally acceptable excipients include, but are not limited to, woodflours, clays, activated carbon, diatomaceous earth, fine-grain inorganic solids, calcium carbonate and the like. Clays and inorganic solids that can be used with the include, but are not limited to, calcium bentonite, kaolin, china clay, talc, perlite, mica, vermiculite, silicas, quartz powder, montmorillonite and mixtures thereof.

[0056] Agriculturally acceptable adjuvants that promote sticking to the seed that can be used include, but are not limited to, polyvinyl acetates, polyvinyl acetate copolymers, hydrolyzed polyvinyl acetates, polyvinylpyrrolidone-vinyl acetate copolymer, polyvinyl alcohols, polyvinyl alcohol copolymers, polyvinyl methyl ether, polyvinyl methyl ether-maleic anhydride copolymer, waxes, latex polymers, celluloses including ethylcelluloses and methylcelluloses, hydroxy methylcelluloses, hydroxypropylcellulose, hydroxymethylpropylcelluloses, polyvinyl pyrrolidones, alginates, dextrins, malto-dextrins, polysaccharides, fats, oils, proteins, karaya gum, jaguar gum, tragacanth gum, polysaccharide gums, mucilage, gum arabics, shellacs, vinylidene chloride polymers and copolymers, soybean-based protein polymers and copolymers, lignosulfonates, acrylic copolymers, starches, polyvinylacrylates, zeins, gelatin, carboxymethylcellulose, chitosan, polyethylene oxide, acrylamide polymers and copolymers, polyhydroxyethyl acrylate, methylacrylamide monomers, alginate, ethylcellulose, polychloroprene and syrups or mixtures thereof. Other useful agriculturally acceptable adjuvants that can promote coating include, but are not limited to, polymers and copolymers of vinyl acetate, polyvinylpyrrolidone-vinyl acetate copolymer and water-soluble waxes. Various surfactants,dispersants, anticaking-agents, foam-control agents, and dyes disclosed herein and in US Patent No. 8,181,388 can be adapted for use with compositions comprising a suitable Methylobacterium strain. In certain embodiments, the seed and / or seedling is exposed to the composition by providing the Methylobacterium strain in soil in which the plant or a plant arising from the seed are grown, or other plant growth media in which the plant or a plant arising from the seed are grown. Examples of methods where the Methylobacterium strain is provided in the soil include in furrow applications, soil drenches, and the like. Thus, adjuvants or excipients that act as lubricants or flowing agents to increase flowability at planting, including dry powders, such as talc and graphite, also find use in the compositions and methods described herein.

[0057] In certain embodiments of any of the aforementioned compositions, uses thereof, or methods related thereto, the Methylobacterium compositions may further comprise one or more fertilizers. In certain embodiments, the fertilizer can be a single nutrient nitrogen fertilizer, such as urea, ammonia or ammonia solutions (including ammonium nitrate, ammonium sulfate, calcium ammonium nitrate, and urea ammonium nitrate). In certain embodiments, the fertilizer can be a single nutrient phosphate fertilizer, such as a superphosphate or triple superphosphate or mixtures thereof, including double superphosphate. In certain embodiments, the fertilizer can be a single nutrient potassium-based fertilizer, such as muriate of potash. In certain embodiments, the compositions comprise multinutrient fertilizers including binary fertilizers (NP, NK, PK), including, for example monoammonium phosphate, diammonium phosphate, potassium nitrate and potassium chloride. In further embodiments, three- component fertilizers (NPK) providing nitrogen, phosphorus, and potassium are present in the aqueous compositions. In still further embodiments, the fertilizer comprises micronutrients, which may be chelated or non-chelated. In some embodiments, combinations of various fertilizers can be present in the compositions, including combinations of nitrogen, phosphorus and / or micronutrient fertilizers.

[0058] In certain embodiments of any of the aforementioned compositions, uses thereof, or methods related thereto, the Methylobacterium is adhered to a solid substance. In certain embodiments of the methods, t e Methylobacterium that is adhered to the solid substance is combined with a liquid to form a composition that is a colloid. In certain embodiments of the methods, the colloid is a gel. In certain embodiments of the methods, the Methylobacterium adhered to the solid substance is provided by culturing the Methylobacterium in the presence of the solid substance. In certain embodiments of the methods, the composition comprises an emulsion. In certain embodiments of the methods, the Methylobacterium is provided by culturing the Methylobacterium in an emulsion.

[0059] Also provided are methods for inhibiting a plant pathogenic fungus or oomycete that comprise applying any of the aforementioned compositions as a first composition in a treatment to a plant or a plant part, to soil where a plant is grown, to a nutrient solution in which a plant is grown, to soil where a plant part such as a seed is deposited, in aeroponic applications, such as in a root mist, or anycombination thereof, in an amount that inhibits or reduces infection by a plant pathogenic Fusarium or Pythium species and / or reduces severity of a disease caused by a Fusarium, Pythium or other plant pathogenic fungal or oomycete species. Treatment of plants can be by application to the plant, seed or other plant part in a dry powder form, application as a liquid spray, in furrow application to seeds, dipping of plants or plant parts into a liquid solution comprising the Methylobacterium, and the like. Treatments or applications can include, but are not limited to, spraying, coating, partially coating, immersing, and / or imbibing the plant or plant parts with the compositions provided herein. In certain embodiments, a seed, a leaf, a vegetative cutting, a fruit, a stem, a root, a tuber, or a coleoptile can be immersed, dipped in, and / or imbibed with a liquid, semi-liquid, emulsion, or slurry of a composition provided herein. In this manner, inhibition of infection or reduction of disease severity by the plant pathogenic fungus or oomycete in the plant, plant part, or a plant obtained therefrom relative to infection of a control plant, plant part, or plant obtained therefrom that had not received an application of the composition or been grown in soil or nutrient solution that was not treated with the composition, is achieved.

[0060] In certain embodiments of any of the aforementioned compositions or methods for inhibiting a plant pathogen, the Methylobacterium is selected from the group consisting of NLS0059, NLS0165, NLS0214, NLS0419, NLS0433, NLS0498, NLS0532, NLS0632, NLS0952, NLS0729, NLS1310, NLS0502, NLS0575, NLS0704, NLS0707, NLS0934 and variants thereof. In certain embodiments, the pathogen is a Fusarium species and the Methylobacterium is selected from the group consisting of NLS0419, NLS0433, NLS0498, NLS0952, NLS0729, NLS1310, NLS0502, NLS0575, NLS0704, NLS0707, NLS0934 and variants thereof. In certain embodiments, the pathogen is a Pythium species and the Methylobacterium is selected from the group consisting of NLS0059, NLS0165, NLS0214, NLS0532, NLS0632, NLS0704, and variants thereof.

[0061] In certain embodiments of methods disclosed herein, the fungal pathogen is a Fusarium species and the Methylobacterium is NLS0017 or a variant thereof having in its genome one or more polynucleotide marker fragments of at least 50, 60, 100, 120, 180, 200, 240, or 300 nucleotides of SEQ ID NOS: 1-3. In certain embodiments of such methods, the fungal pathogen is a. Pythium species and the Methylobacterium is NLS0064 or a variant thereof having in its genome one or more polynucleotide marker fragments of at least 50, 60, 100, 120, 180, 200, 240, or 300 nucleotides of SEQ ID NOS: 13-15. In certain embodiments, the Methylobacterium has in its genome one or more marker fragments comprising a sequence having at least 98%, 99%, or 99.5% sequence identity across the entire length of SEQ ID NOS: 1-3 or 13-15. In certain embodiments of any of the aforementioned compositions, the Methylobacterium is NLS0017 or NLS0064. In certain embodiments, variants of a Methylobacterium strain are identified by the presence of specific marker fragments identified as described herein. NLS0017 or a Methylobacterium strain related thereto can be identified in by the presence of one ormore marker fragments selected from the group consisting of SEQ ID NO: 1-3. NLS0064 or a Methylobacterium strain related thereto can be identified in by the presence of one or more marker fragments selected from the group consisting of SEQ ID NO: 13-15. The marker fragments can be identified by PCR analysis using specific DNA detection assays that include, but are not limited to, DNA primer and probe combinations provided herein.

[0062] Also provided herein are methods of identifying and / or isolating Methylobacterium strains that can inhibit disease caused by Fusarium or Pythium by assaying for the presence of SEQ ID NO: 1-3 or SEQ ID NO: 13-15 in the Methylobacterium. In certain embodiments, such methods can comprise identifying a candidate Methylobacterium, performing a nucleic acid analysis technique on a genetic sample from said candidate, and determining that the sample contains nucleic acids containing a sequence of at least about 50, 100, 200, or 300 nucleotides that is identical to one or more or SEQ ID NO: 1-3 or SEQ ID NO: 13-15. Such nucleic acid analyses include, but are not limited to, techniques based on nucleic acid hybridization, polymerase chain reactions, mass spectroscopy, nanopore based detection, branched DNA analyses, combinations thereof, and the like. In certain embodiments of any of the aforementioned methods or uses, the application of the composition provides for at least 10%, 20%, 30%, 40%, 50%, 75%, at least 85%, or at least 95% inhibition or reduction in severity of symptoms of a disease caused by Fusarium or Pythium in the plant, plant part, or a plant derived therefrom relative to infection or reduction in severity of symptoms in the control plant, plant part, or plant obtained therefrom. The ability of a Methylobacterium strain to prevent infection of plants by a plant pathogenic Fusarium or Pythium species can be measured by various means including by measuring or visually assessing seedling emergence rate, stand count, early plant vigor, visible lesions, the presence of hyphae on seedlings, root rot incidence and severity, root biomass or length, and percent dead seed. Any measurable decrease in disease severity may also be measured including by assessing plant tissue damage or necrosis, plant yield reduction, reduction in the value of the crop plant product or failure of transplanted seedlings to thrive in field soil.

[0063] In certain embodiments of the methods, the plant part is selected from the group consisting of a leaf, a stem, a fruit, a vegetative cutting, a flower, a root, a seedling, a tuber, or a seed. In certain embodiments of the methods, the method further comprises the step of harvesting at least one plant part selected from the treated plant or plant grown from the treated plant part. In certain embodiments of the aforementioned methods, the method further comprises obtaining a processed food or feed composition from the plant or plant part. In certain embodiments, an amount of the Methylobacterium sufficient to prevent or reduce infection by a Fusarium or Pythium species is applied to the plant or plant part. In certain embodiments, the amount of t e Methylobacterium applied to a plant or plant part (e.g., a seed) is about l.OxlO2, l.OxlO3, l.OxlO4, or l.OxlO5to about l.OxlO7, l.OxlO8, l.OxlO9, or 1.0xl010CFUs of Methylobacterium per plant or plant part (e.g., a seed). In certain embodiments, the Methylobacteriumis heterologous to the plant or plant part. In certain embodiments of any of the aforementioned methods, the plant part is a leaf, a stem, a fruit, a vegetative cutting, a flower, a root, a seedling, a tuber, or a seed. In certain embodiments of the methods, the Methyl obacterium is NLS0059, NLS0165, NLS0214, N NLS0419, NLS0433, NLS0498, LS0532, NLS0632, NLS0952, NLS0729, NLS1310, NLS0502, NLS0575, NLS0704, NLS0707 or NLS0934. In certain embodiments of any of the aforementioned methods, the composition further comprises Methylobacterium strain NLS0020, NLS0021, NLS0037, NLS0042, NLS0066, NLS0089, or NLS0109, NLS0934 or a variant of NLS0020, NLS0021, NLS0037, NLS0042, NLS0066, NLS0089, NLS0109, or NLS0934. In certain embodiments, the plant or plant part is a soybean, wheat, com, hops, cannabis, grape, leafy green, lettuce, tomato, pepper, vegetable, or floriculture / ornamental plant or plant part.

[0064] In some embodiments, the compositions or methods disclosed herein may comprise one or more additional components. In some embodiments a second component can be an additional active ingredient, for example, a pesticide or a second biological. The pesticide may be, for example, an insecticide, a fungicide, an herbicide, or a nematicide. The second biological can be a biocontrol agent. The second biological can also comprise a plant-beneficial microorganism. Plant beneficial microorganisms include Rhizobium sp., Bradyrhizobium sp., or other nitrogen-fixing microorganisms.

[0065] Non-limiting examples of insecticides and nematicides include carbamates, diamides, macrocyclic lactones, neonicotinoids, organophosphates, phenylpyrazoles, pyrethrins, spinosyns, synthetic pyrethroids, tetronic and tetramic acids. In particular embodiments insecticides and nematicides include abamectin, aldicarb, aldoxycarb, bifenthrin, carbofuran, chlorantraniliporle, chlothianidin, cyfluthrin, cyhalothrin, cypermethrin, deltamethrin, dinotefuran, emamectin, ethiprole, fenamiphos, fipronil, flubendiamide, fosthiazate, imidacloprid, ivermectin, lambda-cyhalothrin, milbemectin, nitenpyram, oxamyl, permethrin, tioxazafen, spinetoram, spinosad, spirodichlofen, spirotetramat, tefluthrin, thiacl oprid, thiamethoxam, and thiodicarb.

[0066] Non-limiting examples of useful fungicides include aromatic hydrocarbons, azoxystrobin, benzimidazoles, benzthiadi azole, carboxamides, carboxylic acid amides, morpholines, phenylamides, phosphonates, quinone outside inhibitors (e.g. strobilurins), thiazolidines, thiophanates, thiophene carboxamides, and triazoles. Particular examples of fungicides include acibenzolar-S-methyl, azoxystrobin, benalaxyl, bixafen, boscalid, carbendazim, cy proconazole, dimethomorph, epoxiconazole, fluopyram, fluoxastrobin, flutianil, flutolanil, fluxapyroxad, fosetyl-Al, ipconazole, isopyrazam, kresoxim-methyl, mefenoxam, metalaxyl, metconazole, myclobutanil, orysastrobin, penflufen, penthiopyrad, picoxystrobin, propi conazole, prothioconazole, pyraclostrobin, sedaxane, silthiofam, tebuconazole, thifluzamide, thiophanate, tolclofos-methyl, trifloxystrobin, and tri ti conazole.

[0067] Non-limiting examples of herbicides include ACCase inhibitors, acetanilides, AHAS inhibitors, carotenoid biosynthesis inhibitors, EPSPS inhibitors, glutamine synthetase inhibitors, PPO inhibitors,PS II inhibitors, and synthetic auxins, Particular examples of herbicides include acetochlor, clethodim, dicamba, flumioxazin, fomesafen, glyphosate, glufosinate, mesotrione, quizalofop, saflufenacil, sulcotrione, and 2,4-D. In some embodiments, the compositions or methods disclosed herein may include a fungicide selected from the group consisting of a strobilurin, azoxystrobin, Trichoderma, mefenoxam, and metalaxyl. In some embodiments, the compositions or methods disclosed herein may include a synergistic effect between a methylobaceteria and a fungicide selected from the group consisting of a strobilurin, azoxystrobin, Trichoderma, mefenoxam, and metalaxyl. In some embodiments, the compositions or methods disclosed herein may include a synergistic effect between NLS0089 and a fungicide selected front the group consisting of a strobilurin, azoxystrobin, Trichoderma, mefenoxam, and metalaxyl.

[0068] In some embodiments, compositions or methods disclosed herein may comprise an additional active ingredient which may be a second biological. The second biological could be a biocontrol agent, other beneficial microorganisms, microbial extracts, natural products, plant growth activators or a plant defense agent. Non-limiting examples of biocontrol agents include bacteria, fungi, beneficial nematodes, and viruses.

[0069] In certain embodiments, the second biological can be Methylobacterium. In certain embodiments, the second biological is a Methylobacterium listed in Table 1 or a variant thereof. In certain embodiments, the second biological can be a bacterium of the genus Actinomycetes, Agrobacterium, Arthrobacter, Alcaligenes, Aureobacterium, Azobacter, Beijerinckia, Brevibacillus, Burkholderia, Chromobacterium, Clostridium, Clavibacter, Comomonas, Corynebacterium, Curtobacterium, Enterobacter, Flavobacterium, Gluconobacter, Hydrogenophage, Klebsiella, Methylobacterium, Paenibacillus, Pasteuria, Phingobacterium, Photorhabdus, Phyllobacterium, Pseudomonas, Rhizobium, Bradyrhizobium, Serratia, Stenotrophomonas, Trichoderma, Variovorax, and Xenorhadbus. In particular embodiments the bacteria is selected from the group consisting of Bacillus amyloliquefaciens, Bacillus cereus, Bacillus firmus, Bacillus, lichenformis, Bacillus pumilus, Bacillus sphaericus, Bacillus subtilis, Bacillus thuringiensis, Chromobacterium suttsuga, Pasteuria penetrans, Pasteuria usage, and Pseudomona jluorescens.

[0070] In certain embodiments the second biological can be a fungus of the genus Alternaria, Ampelomyces, Aspergillus, Aureobasidium, Beauveria, Colletotrichum, Coniothyrium, Gliocladium, Metarhisium, Muscodor, Paecilonyces, Trichoderma, Typhula, Ulocladium, and Verticilium. In particular embodiments the fungus is Beauveria bassiana, Coniothyrium minitans, Gliocladium vixens, Muscodor albus, Paecilomyces lilacinus, or Trichoderma polysporum.

[0071] In further embodiments the second biological can be a plant growth activator or plant defense agent including, but not limited to harpin, Reynoutria sachalinensis, j asm onate, lipochitooligosaccharides, and isoflavones.

[0072] In further embodiments, the second biological can include, but is not limited to, various Bacillus sp., Pseudomonas sp., Coniothyrium sp., Pantoea sp., Streptomyces sp., and Trichoderma sp. Microbial biopesticides can be a bacterium, fungus, virus, or protozoan. Particularly useful biopesticidal microorganisms include various Bacillus subtilis, Bacillus thuringiensis, Bacillus pumilis, Pseudomonas syringae, Trichoderma harzianum, Trichoderma virens. and Streptomyces lydicus strains. Other microorganisms that are added can be genetically engineered or wild-type isolates that are available as pure cultures. In certain embodiments, it is anticipated that the biological or biocontrol agent can be provided in the fermentation broth, fermentation broth product, or composition in the form of a spore. In certain embodiments, the methods can further comprise applying a fungicide to a plant, plant part or soil. In such embodiments, the fungicide can be applied simultaneously with one or more of the Methylobacterium sp. or may be applied as a separate composition to a plant or plant part. In certain embodiments, the fungicide is metalaxyl, mefenoxam, ethaboxam or oxathiapiprolin. In certain embodiments, the fungicide is a strobilurin, a phenylamide, a thiazole-carboxamide, or a piperidinyl thiazole isoxazoline fungicide. In certain embodiments, the compositions provided herein can further comprise one or more of the aforementioned fungicides.

[0073] The compositions and methods disclosed herein find use in agricultural row crops, as well as in various other crops, including for example specialty crops. In certain embodiments of any of the foregoing compositions and methods, the plant is corn, soybean, Brassica sp. (e.g., B. napus, B. rapa, B. juncea . alfalfa, rice, rye, wheat, barley, oats, sorghum, millet (e.g., pearl millet (Pennisetum glaucum) proso millet (Panicum miliaceum . foxtail millet (Setaria ilalica). finger millet Eleusine coracana), sunflower, safflower, tobacco, potato, peanuts, cotton, species in the genus Cannabis (including, but not limited to, Cannabis sativa and industrial hemp varieties), sweet potato (Ipomoea batatus), cassava, coffee, coconut, pineapple, citrus trees, cocoa, tea, date palm, banana, apple, pear, grape, berry plants (including, but not limited to blackberry, raspberry, strawberry or blueberry plants), avocado, fig, guava, kiwi, mango, olive, papaya, tree nuts, (including cashew, macadamia, almond, pistachios, hazelnut, Brazil nut, walnut and pecan), sugar beets, sugarcane, tomatoes, peppers, lettuce, leafy greens (including, but not limited to, spinach, kale, microgreens, collard greens, cabbage, beet greens, watercress, romaine lettuce, swiss chard, arugula, endive, bok choy and turnip greens, cruciferous vegetables ), green beans, lima beans, peas, lentils, cucurbits (including, but not limited to cucumber, cantaloupe, melons, squash, pumpkin, and zucchini), and herbs (including, but not limited to basil, oregano, chives, parsley, cilantro, catnip, chamomile, lavender and St. John’s wort). In other embodiments, treated plants include ornamentals (including, but not limited to, azalea, hydrangea, hibiscus, roses, tulips, daffodils, petunias, carnation, poinsettia, and chrysanthemum), conifers (including, but not limited to pines such as loblolly pine, slash pine, ponderosa pine, lodge pole pine, and Monterey pine; Douglas-fir; Western hemlock; Sitka spruce; redwood; true firs such as silver firand balsam fir; and cedars such as Western red cedar and Alaska yellow-cedar) and turfgrass (including, but are not limited to, annual bluegrass, annual ryegrass, Canada bluegrass, fescue, bentgrass, wheatgrass, Kentucky bluegrass, orchard grass, ryegrass, redtop, Bermuda grass, St. Augustine grass, and zoysia grass).

[0074] In certain embodiments of any of the foregoing compositions and methods, fermentation products and compositions with plant pathogenic fungus or oomycete-inhibiting Methylobacterium at a titer of greater than about 5 x 107colony-forming units per milliliter, at a titer of greater than about 1 x 108colony -forming units per milliliter, at a titer of greater than about 5 x 108colony-forming units per milliliter, at a titer of greater than about 1 x 109colony-forming units per milliliter, at a titer of greater than about 1 x IO10colony-forming units per milliliter, at a titer of at least about 3 x IO10colony-forming units per milliliter are provided herein. In certain embodiments, fermentation products and compositions provided herein can comprise Methylobacterium useful for inhibition of plant pathogenic fungi and / or oomycetes at a titer of at least about 5 x 107, 1 x 108, or 5 x 108colony-forming units per milliliter to at least about 4 x IO10colony -forming units per milliliter, at least about 5 x 108colony-forming units per milliliter to at least about 4 x IO10colony -forming units per milliliter, or at least about 5 x 108colonyforming units per milliliter to at least about 6 x IO10colony -forming units per milliliter. In certain embodiments, fermentation products and compositions provided herein can comprise Methylobacterium at a titer of at least about 1 x 109colony -forming units per milliliter to at least about 3 x IO10colonyforming units per milliliter, at least about 1 x 109colony-forming units per milliliter to at least about 4 x IO10colony-forming units per milliliter, or at least about 1 x 109colony -forming units per milliliter to at least about 6 x IO10colony-forming units per milliliter. In certain embodiments, fermentation products and compositions provided herein will comprise Methylobacterium at a titer of at least about 1 x IO10colony-forming units per milliliter to at least about 3 x IO10colony-forming units per milliliter, at least about 1 x IO10colony-forming units per milliliter to at least about 4 x IO10colony-forming units per milliliter, or at least about 1 x IO10colony -forming units per milliliter to at least about 6 x IO10colonyforming units per milliliter. In certain embodiments, fermentation products and compositions provided herein will comprise Methylobacterium at a titer of, at least about 3 x IO10colony-forming units per milliliter to at least about 4 x IO10colony-forming units per milliliter, or at least about 3 x IO10colonyforming units per milliliter to at least about 6 x IO10colony -forming units per milliliter. In any of the aforementioned fermentation products or compositions, the indicated concentrations can be at concentrations sufficient to inhibit a fungal and / or oomycete-induced plant disease. In any of the aforementioned fermentation products or compositions, the fermentation products or compositions can be essentially free of contaminating microorganisms, can comprise Methylobacterium that are adhered to and / or associated with materials that the Methylobacterium are not are adhered to and / or associated with in nature, or any combination thereof.

[0075] In certain embodiments of any of the foregoing compositions and methods, fermentation products and compositions with Methylobacterium that are useful for inhibition of plant pathogenic fungi and / or oomycetes at a titer of greater than about 5 x 107, 1 x 108, or 5 x 108colony-forming units per gram, at a titer of greater than about 1 x 109colony -forming units per gram, at a titer of greater than about 1 x IO10colony-forming units per gram, at a titer of at least about 3 x IO10colony -forming units per gram are provided herein. In certain embodiments, fermentation products and compositions provided herein can comprise Methylobacterium at a titer of at least about 5 x 107, 1 x 108, or 5 x 108colony-forming units per gram to at least about 3 x IO10colony -forming units per gram, at least about 5 x 107, 1 x 108, or 5 x 108colony-forming units per gram to at least about 4 x IO10colony-forming units per gram, or at least about 5 x 107, 1 x 108, or 5 x 108colony -forming units per gram to at least about 6 x IO10colony-forming units per gram. In certain embodiments, fermentation products and compositions provided herein can comprise Methylobacterium at a titer of at least about 1 x 109colonyforming units per gram to at least about 3 x IO10colony-forming units per gram, at least about 1 x 109colony-forming units per gram to at least about 4 x IO10colony-forming units per gram, or at least about 1 x 109colony-forming units per gram to at least about 6 x IO10colony-forming units per gram. In certain embodiments, fermentation products and compositions provided herein will comprise Methylobacterium at a titer of at least about 1 x IO10colony-forming units per gram to at least about 3 x IO10colony-forming units per gram, at least about 1 x IO10colony-forming units per gram to at least about 4 x IO10colony -forming units per gram, or at least about 1 x IO10colony-forming units per gram to at least about 6 x IO10colony -forming units per gram. In certain embodiments, fermentation products and compositions provided herein will comprise Methylobacterium at a titer of at least about 3 x IO10colony-forming units per gram to at least about 4 x IO10colony-forming units per gram, or at least about 3 x IO10colony-forming units per gram to at least about 6 x IO10, 1 x 1011, 1 x 1012, IxlO13, or 5xl013colony-forming units per gram. In any of the aforementioned fermentation products or compositions, the fermentation or composition can comprise a mono- or co-culture of Methylobacterium that is adhered to a solid substance. In any of the aforementioned fermentation products or compositions, the indicated concentrations can be concentrations sufficient to inhibit a fungal or oomycete-induced plant disease. In any of the aforementioned fermentation products or compositions, the fermentation products or compositions can be essentially free of contaminating microorganisms, can comprise Methylobacterium that are adhered to and / or associated with materials that the Methylobacterium are not are adhered to and / or associated with in nature, or any combination thereof.

[0076] V arious Methylobacterium sp. isolates provided herein are disclosed in Table 1 and Table 1A.Table Y Methylobacterium sp. isolates1Deposit number for strain deposited with the AGRICULTURAL RESEARCH SERVICE CULTURE COLLECTION (NRRL) of the National Center for Agricultural Utilization Research, Agricultural Research Service, U.S. Department of Agriculture, 1815 North University Street, Peoria, Illinois 61604 U.S.A, under the terms of the Budapest Treaty on the International Recognition of the Deposit of Microorganisms for the Purposes of Patent Procedure. Subject to 37 CFR §1.808(b), all restrictions imposed by the depositor on the availability to the public of the deposited material will be irrevocably removed upon the granting of any patent from this patent application.Deposit Information

[0077] Samples of the following Methylobacterium sp. strains have been deposited with the AGRICULTURAL RESEARCH SERVICE CULTURE COLLECTION (NRRL) of the National Center for Agricultural Utilization Research, Agricultural Research Service, U.S. Department of Agriculture, 1815 North University Street, Peoria, Illinois 61604 U.S.A, under the terms of the Budapest Treaty on the International Recognition of the Deposit of Microorganisms for the Purposes of Patent Procedure. Methylobacterium sp. NRRL B-50930, NRRL B-50931, NRRL B-50932, NRRL B-50933, NRRL B-50938, NRRL B-50939, NRRL B-50940, and NRRL B-50941 were deposited with NRRL on March 12, 2014. Methylobacterium sp. NRRL B-67340 and NRRL B-67341 were deposited with NRRL on November 18, 2016. Methylobacterium sp. Methylobacterium sp. NRRL B- 68032 was deposited with NRRL on May 20, 2021. NRRL-B-68195 was deposited with NRRL on August 30, 2022. NRRL B-68217 was deposited with NRRL on November 2, 2022.T able 1A. Methylotrophs Methylobacterium sp. isolates

[0078] Various Methanotroph isolates provided herein are disclosed in Table IB.T able IB. MethanotrophsDeposit Statement.

[0079] The NRRL numbers referenced herein are the deposit identification numbers for strains deposited with the AGRICULTURAL RESEARCH SERVICE CULTURE COLLECTION (NRRL) of the National Center for Agricultural Utilization Research, Agricultural Research Service, U.S. Department of Agriculture, 1815 North University Street, Peoria, Illinois 61604 U.S.A, under the terms of the Budapest Treaty on the International Recognition of the Deposit of Microorganisms for the Purposes of Patent Procedure. Subject to 37 CFR §1.808(b), all restrictions imposed by the depositor on the availability to the public of the deposited material will be irrevocably removed upon the granting of any patent from this patent application.

[0080] Various Methanotroph consortia strains are provided herein are disclosed in Table 1C.Table 1C. Example Consortium StrainsDefinitions

[0081] The term "and / or" where used herein is to be taken as specific disclosure of each of the two specified features or components with or without the other. Thus, the term “and / or" as used in a phrase such as "A and / or B" herein is intended to include "A and B," "A or B," "A" (alone), and "B" (alone). Likewise, the term "and / or" as used in a phrase such as "A, B, and / or C" is intended to encompass each of the following embodiments: A, B, and C; A, B, or C; A or C; A or B; B or C; A and C; A and B; B and C; A (alone); B (alone); and C (alone).

[0082] Where a term is provided in the singular, embodiments comprising the plural of that term are also provided.

[0083] As used herein, the terms “include,” “includes,” and “including” are to be construed as at least having the features or encompassing the items to which they refer while not excluding any additional unspecified features or unspecified items.

[0084] As used herein, the phrases “adhered thereto” and “adherent” refer to Methylobacterium that are associated with a solid substance by growing, or having been grown, on a solid substance.

[0085] As used herein, the phrase “active ingredient” refers to a biological or pesticide in a composition for treatment of plants and / or plant parts.

[0086] As used herein, the term “biological” refers to a component of a composition for treatment of plants or plant parts comprised of or derived from a microorganism. Biologicals include biocontrol agents, other beneficial microorganisms, microbial extracts, natural products, plant growth activators or plant defense agents. Non-limiting examples of biocontrol agents include bacteria, fungi, beneficial nematodes, and viruses.

[0087] As used herein, the phrase “agriculturally acceptable adjuvant” refers to a substance that enhances the performance of a biological or pesticide in a composition for treatment of plants and / or plant parts. In certain compositions, a biological can comprise a mono-culture or co-culture of Methylobacterium.

[0088] As used herein, the phrase “agriculturally acceptable excipient” refers to an essentially inert substance that can be used as a diluent and / or carrier for a biological or pesticide in a composition for treatment of plants and / or plant parts. In certain compositions, a biological can comprise a mono-culture or co-culture of Methylobacterium.

[0089] As used herein, the phrase “Methylobacterium concentration sufficient to inhibit a fungal or oomycete-induced plant disease” is a concentration that provides for at least a 10%, 20%, 30%, 40%, 50%, 60%, 70%, at least 80%, or at least 90% inhibition of infection and / or reduction in severity of symptoms of a disease induced by Fusarium or Pythium in a plant, plant part, or a plant derived therefrom relative to infection of the control plant or plant part.

[0090] As used herein, the term “strain” shall include all isolates of such strain.

[0091] As used herein, the phrase “control plant” refers to a plant that did not receive treatment with a Methylobacterium effective for control of fungal or oomycete-induced plant disease or composition comprising the same at either the seed or any subsequent stage of the control plant’s development. In certain embodiments, a control plant can be a plant that was treated with an additional active ingredient or a. Methylobacterium that is not effective for control of the subject plant disease.

[0092] As used herein, the phrase “co-culture of Methylobacterium" refers to a Methylobacterium culture comprising at least two strains of Methylobacterium or at least two species of Methylobacterium.

[0093] As used herein, the phrase “contaminating microorganism” refers to microorganisms in a culture, fermentation broth, fermentation broth product, or composition that were not identified prior to introduction into the culture, fermentation broth, fermentation broth product, or composition.

[0094] As used herein, “variant” when used in the context of a Methylobacterium isolate, refers to any isolate that has chromosomal genomic DNA with at least 99%, 99.9, 99.8, 99.7, 99.6%, or 99.5% sequence identity to chromosomal genomic DNA of a deposited Methylobacterium isolate provided herein. A variant of an isolate can be obtained from various sources including soil, plants or plant material, and water, particularly water associated with plants and / or agriculture. Variants include derivatives obtained from deposited isolates. Methylobacterium isolates or strains can be sequenced (for example as taught by Sanger et al. (1977), Bentley et al. (2008) or Caporaso et al. (2012)) and genomescale comparison of the sequences conducted (Konstantinos etal. (2005)) using sequence analysis tools (for example, BLAST, as taught by Altschul et al. (1990)).

[0095] As used herein, “derivative” when used in the context of a Methylobacterium isolate, refers to any Methylobacterium that is obtained from a deposited Methylobacterium isolate provided herein. Derivatives of a Methylobacterium isolate include, but are not limited to, derivatives obtained by selection, derivatives selected by mutagenesis and selection, and genetically transformed Methylobacterium obtained from a Methylobacterium isolate. A “derivative” can be identified, for example based on genetic identity to the strain or isolate from which it was obtained and will generally exhibit chromosomal genomic DNA with at least 99%, 99.9, 99.8, 99.7, 99.6%, or 99.5% sequence identity to chromosomal genomic DNA of the strain or isolate from which it was derived.

[0096] As used herein, the term “emulsion” refers to a colloidal mixture of two immiscible liquids wherein one liquid is the continuous phase and the other liquid is the dispersed phase. In certain embodiments, the continuous phase is an aqueous liquid and the dispersed phase is liquid that is not miscible, or partially miscible, in the aqueous liquid.

[0097] As used herein, the phrase “essentially free of contaminating microorganisms” refers to a culture, fermentation broth, fermentation product, or composition where at least about 95% of the microorganisms present by amount or type in the culture, fermentation broth, fermentation product, or composition are the desired Methylobacterium or other desired microorganisms of pre-determined identity.

[0098] As used herein, the term “heterologous”, when used in the context of Methylobacterium that at least partially coats a plant or plant part, refers to a Methylobacterium that is not naturally associated with a plant or plant part of the same species as the plant or plant part that is at least partially coated with the Methylobacterium . In certain embodiments, the heterologous Methylobacterium that is used to at least partially coat a plant or plant part of a first plant species is a. Methylobacterium that was isolated, or can be isolated, from a second and distinct plant species.

[0099] As used herein, the phrase “inanimate solid substance” refers to a substance which is insoluble or partially soluble in water or aqueous solutions and which is either non-living or which is not a part of a still-living organism from which it was derived.

[0100] As used herein, the phrase “mono-culture of Melhylobaclerium" refers to a Methylobacterium culture consisting of a single strain of Methylobacterium.

[0101] As used herein, a “pesticide” refers to an insecticide, fungicide, nematocide, bacteriocide, or any combination thereof.

[0102] As used herein, the phrase “bacteriostatic agent” refers to agents that inhibit growth of bacteria but do not kill the bacteria.

[0103] As used herein, the phrase “pesticide does not substantially inhibit growth of said Methylobacterium ” refers to any pesticide that when provided in a composition comprising a fermentation product comprising a solid substance wherein a mono-culture or co-culture of Methylobacterium is adhered thereto, results in no more than a 50% inhibition of Methylobacterium growth when the composition is applied to a plant or plant part in comparison to a composition lacking the pesticide. In certain embodiments, the pesticide results in no more than a 40%, 30%, 20%, 10%, 5%, or 1% inhibition of Methylobacterium growth when the composition is applied to a plant or plant part in comparison to a composition lacking the pesticide.

[0104] As used herein, the term “Methylobacterium” refers to genera and species in the methylobacteriaceae family, including bacterial species in the Methylobacterium genus and proposed Methylorubrum genus (Green and Ardley (2018)). Methylobacterium includes pink-pigmented facultative methylotrophic bacteria (PPFM) and also encompasses the non-pink-pigmented Methylobacterium nodulans, as well as colorless mutants of Methylobacterium isolates. For example, and not by way of limitation, “Methylobacterium” refers to bacteria of the species listed below as well as any new Methylobacterium species that have not yet been reported or described that can be characterized as Methylobacterium or Methylorubrum based on phylogenetic analysis: Methylobacterium adhaesivum; Methylobacterium oryzae; Methylobacterium aerolatum; Methylobacterium oxalidis; Methylobacterium aquaticum; Methylobacterium persicinum; Methylobacterium brachiatum; Methylobacterium phyllosphaerae; Methylobacterium brachythecii; Methylobacterium phyllostachyos; Methylobacterium bullatum; Methylobacterium platani; Methylobacterium cerastii; Methylobacterium pseudosasicola; Methylobacterium currus; Methylobacterium radiotolerans; Methylobacterium dankookense; Methylobacterium soli; Methylobacterium frigidaeris; Methylobacterium specialis; Methylobacterium fujisawaense; Methylobacterium tardum; Methylobacterium gnaphalii; Methylobacterium tarhaniae; Methylobacterium goesingense; Methylobacterium thuringiense ; Methylobacterium gossipiicola; Methylobacterium trifolii; Methylobacterium gregans; Methylobacterium variabile; Methylobacterium haplocladii; Methylobacterium aminovorans (Methylorubrum aminovorans); Methylobacteriumhispanicum; Methylobacterium extorquens (Methylorubrum extorquens); Methylobacterium indicum; Methylobacterium podarium (Methylorubrum podarium); Methylobacterium iners; Methylobacterium populi (Methylorubrum populi); Methylobacterium isbiliense; Methylobacterium pseudosasae (Methylorubrum pseudosasae); Methylobacterium jeotgali; Methylobacterium rhodesianum (Methylorubrum rhodesianum); Methylobacterium komagatae; Methylobacterium rhodinum (Methylorubrum rhodinum); Methylobacterium longum; Methylobacterium salsuginis (Methylorubrum salsuginis); Methylobacterium marchantiae; Methylobacterium suomiense (Methylorubrum suomiense; Methylobacterium mesophilicum; Methylobacterium thiocyanatum (Methylorubrum thiocyanatum) ; Methylobacterium nodulans; Methylobacterium zatmanii (Methylorubrum zatmanii); Methylobacterium organophilum.

[0105] As used herein, the phrase “solid substance” refers to a substance which is insoluble or partially soluble in water or aqueous solutions.

[0106] As used herein, the phrase “solid phase that can be suspended therein” refers to a solid substance that can be distributed throughout a liquid by agitation.

[0107] As used herein, the term “non-regenerable” refers to either a plant part or processed plant product that cannot be regenerated into a whole plant.

[0108] As used herein, the phrase “substantially all of the solid phase is suspended in the liquid phase” refers to media wherein at least 95%, 98%, or 99% of solid substance(s) comprising the solid phase are distributed throughout the liquid by agitation.

[0109] As used herein, the phrase “substantially all of the solid phase is not suspended in the liquid phase” refers to media where less than 5%, 2%, or 1% of the solid is in a particulate form that is distributed throughout the media by agitation.Additional Components.

[0110] In certain embodiments of the methods provided herein, plants, plant seeds and / or plant parts are treated with both a strain and at least one additional component. In some embodiments an additional component can be an additional active ingredient, for example, a pesticide or a second biological. In certain embodiments, the pesticide can be an insecticide, a fungicide, an herbicide, a nematicide or other biocide. The second biological could be a strain that improves yield or controls an insect, pest, fungi, weed, or nematode. In some embodiments, a second biological is an additional methanotroph strain. In some embodiments, a second biological is a Methylobacterium strain. In some embodiments, an additional strain in the methods and compositions provided herein is selected from the Methylotrophs listed in Table 1 A and / or a Methanotroph in Table IB.

[0111] Insecticides and Nemacides. Non-limiting examples of insecticides and nematicides include Biological insecticides and nematicides (Bacillus thuringiensis, Beauveria bassiana. Metarhizium anisopliae. Paecilomyces lilacinus. Purpureocillium lilacinum. Steinernema carpocapsae. Heterorhabditis bacleriophora. azadirachtin, neem oil, pyrethrins), Carbamates (aldicarb, aldoxycarb, carbofuran, methomyl, oxamyl), Diamides (chlorantraniliprole, cyantraniliprole, flubendiamide), Formamidines (amitraz), Insect growth regulators (IGRs) (diflubenzuron, lufenuron, methoprene, pyriproxyfen, tebufenozide), Macrocyclic lactones (avermectins and milbemycins) (abamectin, emamectin, ivermectin, milbemectin), Metaflumizone (metaflumizone), Neonicotinoids (clothianidin, dinotefuran, imidacloprid, nitenpyram, thiacloprid, thiamethoxam), Organophosphates (chlorpyrifos, dichlorvos, fosthiazate, malathion, phorate, phosmet, terbufos), Oxadiazines (indoxacarb), Phenylpyrazoles (ethiprole, fipronil), Pyrethroids (synthetic) (bifenthrin, cyfluthrin, cyhalothrin, cypermethrin, deltamethrin, lambda-cyhalothrin, permethrin, tefluthrin), Spinosyns (spinetoram, spinosad), Sulfoximines (sulfoxaflor), Tetronic and tetramic acid derivatives (spirodi cl of en, spiromesifen, spirotetramat), Uncouplers of oxidative phosphorylation (chlorfenapyr), Other nematicides (fluensulfone, tioxazafen).

[0112] Fungicides. Non-limiting examples of useful fungicides include Anilinopyrimidines (pyrimethanil), Aromatic Hydrocarbons (chlorothalonil), Benzimidazoles (carbendazim, thiophanate- methyl), Biological Fungicides (bacillus subtilis, trichoderma), Carboxamides (benalaxyl, bixafen, boscalid, flutolanil, fluxapyroxad, isopyrazam, penthiopyrad, sedaxane), Dicarboximides (iprodione), Dithiocarbamates (mancozeb, thiram, ziram), Inorganics (copper hydroxide, copper oxychloride, sulfur), Morpholines (dimethomorph), Other or Miscellaneous Classes (acibenzolar-S-methyl, captan, cyazofamid, cymoxanil, fludioxonil, fluopyram, flutianil, propamocarb, silthiofam, thifluzamide, tolclofos-methyl), Phenylamides (mefenoxam, metalaxyl), Phosphonates (fosetyl-al), Strobilurins (azoxystrobin, fluoxastrobin, kresoxim-methyl, orysastrobin, picoxystrobin, pyraclostrobin,trifloxystrobin), Triazoles (cyproconazole, difenoconazole, epoxiconazole, ipconazole, metconazole, myclobutanil, propi conazole, prothioconazole, tebuconazole, triti conazole).

[0113] Herbicides. Non-limiting examples of herbicides include Amino acid synthesis inhibitors (glufosinate, glyphosate, imazapyr, imazethapyr, metsulfuron-methyl), Aryloxyphenoxypropionates (fluazifop-P-butyl, haloxyfop-R-methyl, quizalofop-P-ethyl), Benzonitriles (bromoxynil, ioxynil), Biological herbicides (acetic acid, caprylic acid, citric acid, clove oil, eugenol, Burkholderia rinojensis. Leptospermum scoparium oil, lemongrass oil, Phoma macrostoma, pelargonic acid, Sorghum bicolor extract, Streptomyces viridochromogenes), Bipyridyliums (diquat, paraquat), Carotenoid biosynthesis inhibitors (clomazone, norflurazon), Cellulose biosynthesis inhibitors (isoxaben), Chloroacetamides (acetochlor, alachlor, dimethenamid-P, metolachlor, pretilachlor), Growth regulators (chlormequat, mepiquat), HPPD inhibitors (isoxaflutole, mesotrione, tembotrione, topramezone), Photosystem I inhibitors (diquat, paraquat), Photosystem II inhibitors (atrazine, bentazon, diuron, linuron, metribuzin, propanil, simazine), PPO inhibitors (acifluorfen, fomesafen, lactofen, oxyfluorfen, sulfentrazone), Synthetic auxins (2,4-D, clopyralid, dicamba, MCPA, picloram, triclopyr), Triazines (atrazine, simazine, terbuthylazine), Ureas (diuron, fluometuron, linuron), VLCFA inhibitors (ethalfluralin, napropamide, pendimethalin, prodiamine, trifluralin).

[0114] Active Ingredient. In some embodiments, the composition or method disclosed herein may comprise a methanotroph strain and an additional active ingredient selected from the group consisting of clothianidin, ipconazole, imidacloprid, metalaxyl, mefenoxam, tioxazafen, azoxystrobin, thiomethoxam, fluopyram, prothioconazole, pyraclostrobin, and sedaxane.

[0115] Additional biological.

[0116] The second biological could be a biological control agent, other beneficial microorganisms, microbial extracts, plant extracts, yeast extracts, vegetal chitosan, natural products, plant growth activators or plant defense agent. Non-limiting examples of the second biological could include bacteria, fungi, beneficial nematodes, and viruses. In certain embodiments, the second biological can be a Methylotroph and / or a Methanotroph. In certain embodiments, the second biological is a strain listed in Table 1A and / or Table IB.

[0117] Methylotrophs. In certain embodiments, the second biological can be a Methylobacterium selected from M. gregans, M. radiotolerans, M. extorquens, M. populi, M. salsuginis, M. brachiatum, andM. komagatae.

[0118] Methanotrophs. In certain embodiments, the second biological can be a Methanotroph selected from Methyloacidimicrobium, Methyloacidiplilum, Methylobacter, Methylocaldum, Methylocapsa, Methylocella, Methylococcus, Methylocystis, Methyloferula, Methylogaea, Methyloglobus, Methylohalobius, Methylomagnum, Methylomarinum, Methylomicrobium, Methylomonas, Methyloparacoccus, Methyloperedens, Methyloprofundus, Methylosarcina, Methylosinus,Methylosoma, Methylosphaera, Melhylolhermus, and Methylovulum. In some embodiments, a methanotroph provided herein is a Methylocystis species selected from M. hirsuta, M. rosea and M. parvus. In some embodiments, a methanotroph provided herein is Methylosinus species selected from M. trichosporium and M. sporium. In some embodiments, a methanotroph provided herein is a Methylomicrobium lacus or Methylosarcina fibrate strain.

[0119] Bacterium. In certain embodiments, the second biological can be a bacterium of the genus Actinomycetes, Agrobacterium, Arthrobacter, Alcaligenes, Aureobacterium, Azobacter, Azorhizobium, Azospirillum, Azotobacter, Beijerinckia, Bacillus, Brevibacillus, Burkholderia, Chromobacterium, Clostridium, Clavibacter, Comomonas, Corynebacterium, Curtobacterium, Enterobacter, Flavobacterium, Gluconacetobacter, Gluconobacter, Herbaspirillum, Hydrogenophage, Klebsiella, Luteibacter, Lysinibacillus, Mesorhizobium, Methylobacterium, Microbacterium, Ochrobactrum, Paenibacillus, Pantoea, Pasteuria, Phingobacterium, Photorhabdus, Phyllobacterium, Pseudomonas, Rhizobium, Rhodococcus, Bradyrhizobium, Serratia, Sinorhizobium, Sphingomonas, Streptomyces, Stenotrophomonas, Variovorax, Xanthomonas and Xenorhadbus. In particular embodiments the bacteria is selected from the group consisting of Bacillus amyloliquefaciens, Bacillus cereus, Bacillus firmus, Bacillus, lichenformis, Bacillus pumilus, Bacillus sphaericus, Bacillus subtilis, Bacillus thuringiensis, Chromobacterium suttsuga, Pasteuria penetrans, Pasteuria usage, and Pseudomona fluorescens.

[0120] Fungus. In certain embodiments the second biological can be a fungus of the genus Acremonium, Alternaria, Ampelomyces, Aspergillus, Aureobasidium, Beauveria, Botryosphaeria, Cladosporium, Cochliobolus, Colletotrichum, Coniothyrium, Embellisia, Epicoccum, Fusarium, Gigaspora, Gliocladium, Glomus, Laccaria, Metarhisium, Muscodor, Nigrospora, Paecilonyces, Paraglomus, Penicillium, Phoma, Pisolithus, Podospora, Rhizopogon, Scleroderma, Trichoderma, Typhula, Ulocladium, and Verticilium. In particular embodiments, the fungus is Beauveria bassiana, Coniothyrium minitans, Gliocladium vixens, Muscodor albus, Paecilomyces lilacinus, or Trichoderma polysporum.

[0121] Biostimulant. In further embodiments the second biological can be a biostimulant, including but not limited to seaweed extract or hummates, plant growth activators or plant defense agents including, but not limited to harpin, Reynoutria sachalinensis, jasmonate, lipochitooligosaccharides, and isoflavones.

[0122] Biopesticides. In further embodiments, the second biological can include, but are not limited to, various Bacillus sp., Pseudomonas sp., Coniothyrium sp., Pantoea sp., Streptomyces sp., and Trichoderma sp. Microbial biopesticides can be a bacterium, fungus, virus, or protozoan. Particularly useful biopesticidal microorganisms include various Bacillus subtilis, Bacillus thuringiensis, Bacillus pumilis, Pseudomonas syringae, Trichoderma harzianum, Trichoderma virens, and Streptomyceslydicus strains. Other microorganisms that are added can be genetically engineered or wild-type isolates that are available as pure cultures. In certain embodiments, it is anticipated that the second biological can be provided in the composition in the form of a spore.

[0123] Lubricants. In certain embodiments where plant seeds are treated with methanotroph compositions provided herein, the compositions further comprise one or more lubricants to ensure smooth flow and separation (singulation) of seeds in the seeding mechanism, for example a planter box. Lubricants for use in such compositions include talc, graphite, polyethylene wax based powders (such as Fluency Agent), protein powders, for example soybean protein powders, or a combination of protein powders and a lipid, for example lecithin or a vegetable oil. Lubricants can be applied to seeds simultaneously with application of a methanotroph, or may be mixed with a methanotroph prior to application of the compositions to the seeds.

[0124] Excipients. Agriculturally acceptable excipients include, but are not limited to, woodflours, clays, activated carbon, diatomaceous earth, fine-grain inorganic solids, calcium carbonate and the like. Clays and inorganic solids that can be used with the include, but are not limited to, calcium bentonite, kaolin, china clay, talc, perlite, mica, vermiculite, silicas, quartz powder, montmorillonite and mixtures thereof. Agriculturally acceptable excipients also include various lubricants such as talc, graphite, polyethylene wax based powders (such as Fluency Agent), protein powders, for example soybean protein powders, or a combination of protein powders and a lipid, for example lecithin or a vegetable oil.

[0125] Adjuvants. Preferably, the agriculturally acceptable adjuvant comprises kaolin, talc, graphite, mica, vermiculite, soyobean protein powder, or a combination thereof. Agriculturally acceptable adjuvants that promote sticking to the seed that can be used include, but are not limited to, polyvinyl acetates, polyvinyl acetate copolymers, hydrolyzed polyvinyl acetates, polyvinylpyrrolidone-vinyl acetate copolymer, polyvinyl alcohols, polyvinyl alcohol copolymers, polyvinyl methyl ether, polyvinyl methyl ether-maleic anhydride copolymer, waxes, latex polymers, celluloses including ethylcelluloses and methylcelluloses, hydroxy methylcelluloses, hydroxypropylcellulose, hydroxymethylpropylcelluloses, polyvinyl pyrrolidones, alginates, dextrins, malto-dextrins, polysaccharides, fats, oils, proteins, karaya gum, jaguar gum, tragacanth gum, polysaccharide gums, mucilage, gum arabics, shellacs, vinylidene chloride polymers and copolymers, soybean-based protein polymers and copolymers, lignosulfonates, acrylic copolymers, starches, polyvinylacrylates, zeins, gelatin, carboxymethylcellulose, chitosan, polyethylene oxide, acrylamide polymers and copolymers, polyhydroxyethyl acrylate, methyl acrylamide monomers, alginate, ethylcellulose, polychloroprene and syrups or mixtures thereof. Other useful agriculturally acceptable adjuvants that can promote coating include, but are not limited to, polymers and copolymers of vinyl acetate, polyvinylpyrrolidone-vinyl acetate copolymer and water-soluble waxes. Further, agriculturally acceptable adjuvants also includevarious lubricants (which can provide for smooth flow and separation (singulation) of seeds) such as talc, graphite, polyethylene wax based powders (such as Fluency Agent), protein powders, for example soybean protein powders, or a combination of protein powders and a lipid, for example lecithin or a vegetable oil. Various surfactants, dispersants, anticaking-agents, foam-control agents, and dyes disclosed herein and in US Patent No. 8,181,388 can be adapted for use with compositions comprising a suitable methanotroph strain. In certain embodiments, the seed and / or seedling is exposed to the composition by providing the methanotroph strain in soil in which the plant or a plant arising from the seed are grown, or other plant growth media in which the plant or a plant arising from the seed are grown. Examples of methods where the methanotroph strain is provided in the field and soil include in furrow applications, soil drenches, and the like. Preferably, agriculturally acceptable adjuvants that promote sticking to the seed are celluloses dextrins, maltodextrins, polysaccharides, polysaccharide gums, or a combination thereof.

[0126] The agriculturally acceptable adjuvant, excipient, lubricant, and / or other ingredients, can be present in the composition at a concentration of from 0 wt.% to about 95 wt.%, from about 0.1 wt.% to about 95 wt.%, from about 0.5 wt.% to about 95 wt.%, from about 1 wt.% to about 95 wt.%, from about2 wt.% to about 95 wt.%, from about 3 wt.% to about 95 wt.%, from about 4 wt.% to about 95 wt.%, from about 5 wt.% to about 95 wt.%, from about 0.1 wt.% to about 90 wt.%, from about 0.5 wt.% to about 90 wt.%, from about 1 wt.% to about 90 wt.%, from about 2 wt.% to about 90 wt.%, from about3 wt.% to about 90 wt.%, from about 4 wt.% to about 90 wt.%, from about 5 wt.% to about 90 wt.%, from about 0.1 wt.% to about 85 wt.%, from about 0.5 wt.% to about 85 wt.%, from about 1 wt.% to about 85 wt.%, from about 2 wt.% to about 85 wt.%, from about 3 wt.% to about 85 wt.%, from about4 wt.% to about 85 wt.%, from about 5 wt.% to about 85 wt.%, from about 0.1 wt.% to about 80 wt.%, from about 0.5 wt.% to about 80 wt.%, from about 1 wt.% to about 80 wt.%, from about 2 wt.% to about 80 wt.%, from about 3 wt.% to about 80 wt.%, from about 4 wt.% to about 80 wt.%, or more preferably, from about 5 wt.% to about 80 wt.%.

[0127] Treatments.

[0128] The compositions and methods herein are useful for treatment of various environments, and can be applied in a variety of locations and timing intervals.

[0129] Environments. The invention is useful in various environments, for example: in wetlands, landfills, and agricultural applications, including plant production in flooded fields.

[0130] Application. Compositions comprising the strains herein and optionally one or more additional strain may be applied to soil or other growth medium where plants are grown. Soil treatments or applications can include, but are not limited to, fields (e.g. flooded or irrigated fields), in-furrow applications (e.g., before, during, and / or after seed deposition), soil drenches, distribution of granular or other dried formulations to the soil (e.g., before, during, and / or after seed deposition or plant growth).Treatments for plants grown in hydroponic systems can include seed treatments prior to germination, foliar applications to germinated plants or parts thereof, and applications in a liquid solution used in the hydroponic system. In certain embodiments, treatment of a plant can include application to the seed, plant, and / or a part of the plant and can thus comprise any treatment or application resulting in colonization of the plant by the strain. In some embodiments, application of one or more methanotrophs and optionally one or more methylotrophs to crops that are propagated by cutting can enhance growth and / or rooting of such plants. Field transplants of such treated and rooted cuttings may demonstrate decreased cycling time, and / or improved biomass and / or yield as a result of such treatments.

[0131] Treatments or applications to plants described herein can include, but are not limited to, spraying, coating, partially coating, immersing, drenching, and / or imbibing the field, seed, plant or plant parts with the methanotroph, and optionally one or more methylotroph, strains, or compositions comprising such strains. In certain embodiments, soil, a seed, a leaf, a stem, a root, a tuber, or a shoot can be sprayed, immersed drenched and / or imbibed with a liquid, semi-liquid, emulsion, or slurry of a composition provided herein. In some embodiments, one or more methanotroph strains may be applied together or separately with one or more methylotroph strains. In some embodiments, methanotroph, and optionally methylotroph strains, are applied to multiple plant parts and / or at multiple stages of plant growth. In certain embodiments, methane oxidizing methanotrophs described herein are applied as foliar sprays or seed treatments to row crops. In some embodiments, the crop is com and a methanotroph is applied as a seed treatment. In some embodiments, the corn crop is grown under nitrogen limited conditions and the ability of the applied methanotroph to enhance nutrient uptake efficiency (NUE) is observed.

[0132] Timing and Conditions. In some embodiments, com seeds are treated in a planter box application. In some embodiments, the crop is rice, and plants are treated with an initial foliar application at a flooded stage. In some embodiments, foliar applications are made when a rice paddy is at full flood stage.

[0133] Such treatments, applications, seed immersion, or imbibition can be sufficient to provide for, enhanced early growth and / or increased levels of one or more mineral nutrients and / or vitamins content in harvestable tissue from a treated plant or plant grown from a treated seed in comparison to an untreated plant or plant grown from an untreated seed. Enhanced early growth can lead to further improvements in plant production including an increase in biomass of treated plants, such as increased shoot, root, or whole seedling biomass. Enhanced early growth can result in various additional improvements in plant production, including for example increased yield of harvested plants or harvested plant parts, increased and / or more uniform fruit production, faster seed set, earlier maturation, increased rate of leaf growth, increased rate of root growth, increased seed yield, and decreased cycle time.

[0134] In certain embodiments, plant seeds or cuttings can be immersed and / or imbibed for at least 1, 2, 3, 4, 5, or 6 hours. Such immersion and / or imbibition can, in certain embodiments, be conducted attemperatures that are not deleterious to the plant seed or the methanotroph. In certain embodiments, the seeds can be treated at about 15 to about 30 degrees Centigrade or at about 20 to about 25 degrees Centigrade. In certain embodiments, seed imbibition and / or immersion can be performed with gentle agitation. Seed treatments can be effected with both continuous and / or batch seed treaters. In certain embodiments, the coated seeds can be prepared by slurrying seeds with a coating composition comprising a methanotroph strain that increases the levels of one or more mineral nutrients and / or vitamins and air-drying the resulting product. Air-drying can be accomplished at any temperature that is not deleterious to the seed or the methanotroph, but will typically not be greater than 30 degrees Centigrade. The proportion of coating that comprises the methanotroph strain includes, but is not limited to, a range of 0.1% to 25% by weight of the seed or other plant part, 0.5 to 5% by weight of the seed or other plant part, and 0.5 to 2.5% by weight of the seed or other plant part. In certain embodiments, a solid substance used in the seed coating or treatment will have a methanotroph strain that increases mineral nutrient and or vitamin content adhered to a solid substance as a result of being grown in biphasic media comprising the methanotroph strain, solid substance, and liquid media.

[0135] Hydroponics. In certain embodiments, treated plants are cultivated in a hydroponic system. In some embodiments, plant seeds are treated and plants are grown from the treated seeds continuously in the same cultivation system. In some embodiments, plant seeds are treated and cultivated in a hydroponic nursery to produce seedlings. The seedlings transferred to a different hydroponic system, for example for commercial production of leafy greens. In some embodiments, a methanotroph strain that enhances early growth or increases the levels of one or more mineral nutrients and / or vitamins persists in the seedlings transferred to a greenhouse production system and continues to provide advantages such as improved micronutrient and / or vitamin content and / or biomass production, through the further growth of the leafy green plant.

[0136] In some embodiments, plant seedlings transferred to a greenhouse production system may be further treated with NLS0064, NLS1026, NLS1084, NLS0433, NLS0940 or variants thereof, or with one or more other Methylobacterium strains that increase the levels of one or more mineral nutrients and / or vitamins prior to, during or after transfer to the production system.

[0137] Plants.

[0138] Plant compositions and methods to treat plants include, for example, alfalfa; barley; brassica sp.; cannabis sp.; carrot; cassava; coconut; coffee; conifers; com; cotton; cucurbits; cucumber; fruit plants (including fruit trees); green bean; herbs; leafy greens; lettuce; microgreens; millet; oat; onion; ornamental; pea; peanut; pepper; potato; rice; rye; safflower; sorghum; soybean; squash; sugar beet; sunflower; sweet potato; tobacco; tomato; turfgrass; and wheat.

[0139] For instance, the following plants are commercially improved by the present invention and included in the present compositions and methods:

[0140] Cereal grain crops: amaranth (pseudocereal), arborio rice, barley, basmati rice, black rice, brown rice, buckwheat (pseudocereal), bulgur, corn (maize), durum wheat, emmer, farro, fonio, glutinous rice, jasmine rice, millet, oats, pearl millet, quinoa (pseudocereal), red rice, rye, sorghum, spelt, teff, triticale, wheat, white rice, and wild rice.

[0141] Fruiting vegetables: acorn squash, avocado, bell pepper, bitter melon, calabash, cantaloupe, caper berries, chayote, cherry tomato, chili pepper, cucumber, eggplant, gourds, ground cherry, honeydew melon, jicama, kabocha, luffa, okra, pattypan squash, pepino melon, pumpkin, roselle (Hibiscus sabdariffa), snap peas, spaghetti squash, squash, string beans, taro fruit, tomatillo, tomato, watermelon, winter melon, and zucchini.

[0142] Legume vegetables: adzuki bean, black bean, black-eyed pea (cowpea), butter bean, cannellini bean, chickpea (garbanzo bean), cranberry bean, edamame (young soybean), fava bean (broad bean), green bean, hyacinth bean, kidney bean, lentil, lima bean, mung bean, navy bean, pigeon pea, pinto bean, runner bean, snap pea, snow pea, sugar snap pea, winged bean, and yardlong bean.

[0143] Pome fruit: apple, loquat, medlar, nashi pear (Asian pear), pear, and quince.

[0144] Seed crops: amaranth, basil, black cumin, buckwheat, canola, caraway, chia, coriander, cumin, flax, hemp, millet, mustard, nigella, poppy, pumpkin, quinoa, safflower, sesame, squash, sunflower, and watermelon.

[0145] Small fruit crops and berries: aronia (chokeberry), barberry, blackberry, blackcurrant, blueberry, boy senberry, buffaloberry, cloudberry, cranberry, currant, elderberry, feijoa (pineapple guava), fig, gooseberry, grape, huckleberry, jostaberry, juneberry (serviceberry), kiwi, lingonberry, loganberry, mulberry, olallieberry, pomegranate, raspberry, redcurrant, salal berry, saskatoon berry, sea buckthorn, serviceberry (Juneberry), strawberry, tayberry, and white currant.

[0146] Lastly, the following plants are also within the scope of the present invention: For instance, plants include: arugula; azalea; B. juncea; B. napus; B. rapa; basil; beans; beet greens; bentgrass; Bermuda grass; bok choy; broccoli; cabbage; Canada bluegrass; carnation; cauliflower; celery; chickpeas; chicory; clover; collard greens; cover-crops; daffodils; dill; Douglas-fir; endive; escarole; fennel; fescue; finger millet; foxtail millet; French tarragon; fruit; garlic; golf grass; hibiscus; hydrangea; iceberg lettuce; kale; Kentucky bluegrass; leek; loblolly pine; lodgepole pine; melon; Monterey pine; nuts; oats; orchard grass; ornamentals; peanuts; pearl millet; peas; Pennisetum; petunias; pines; poinsettia; pome fruit; ponderosa pine; proso millet; radicchio; radish; redtop; redwood; romaine lettuce; rosemary; roses; ryegrass; Sitka spruce; slash pine; spinach; St. Augustine grass; swiss chard; tea; trees; tropical fruit; turnip greens; tulips; watercress; watermelon; wheatgrass; zoysia grass.

[0147] To the extent to which any of the preceding definitions is inconsistent with definitions provided in any patent or non-patent reference incorporated herein by reference, any patent or non-patentreference cited herein, or in any patent or non-patent reference found elsewhere, it is understood that the preceding definition will be used herein.EXAMPLES

[0148] The following examples are included to demonstrate various embodiments. It will be appreciated by those of skill in the art that the techniques disclosed in the following examples represent techniques discovered by the Applicants to function well. However, those of skill in the art should, in light of the instant disclosure, appreciate that many changes can be made in the specific embodiments that are disclosed, while still obtaining like or similar results, without departing from the scope of the disclosure.Example 1 Pythium field 2019

[0149] Small plot soybean field trials with randomized complete block (RCB) design and six replications per location were conducted at six locations to assess the ability of Methyl obacterium spp. and Methylorubrum spp. isolates to suppress disease caused by Pythium spp. Four of the six locations had sufficient disease pressure to include in the analysis.

[0150] Soybean seed was treated with 27 individual Methylobacterium spp. isolates to achieve a target titer of 1 x 105to 1 x 106CFU per seed. The control treatment (UTC) received no Methylobacterium treatment and the commercial control treatment received Axyl Shield ST (active ingredient metalaxyl) at the high end of the label rate. Four row plots 10 ft wide by 10 ft long with row spacing of 30 in were seeded with a regionally adapted soybean variety at a planting density of 140,000 plants per acre. Sorghum grain spawn inoculum prepared separately with Pythium ultimum var. ullimum. Pythium irregulare and Pythium sylvaticum was combined at a 1 : 1 : 1 ratio and applied in furrow at planting at a rate of 1 g per row foot. Plots were irrigated for two days after planting and then for an additional two days one week after planting to encourage the establishment of viable pathogen inoculum in the soil. Stand counts were recorded at 21 days after emergence (DAE) to assess pre-emergence damping off and final yield (bushels per acre) was measured.

[0151] Table 2. Emergence (%) compared to UTC. Average UTC emergence was 53%.Table 3. Yield (bushels per acre) compared to UTCExample 2. Pythium full size plot field 2019

[0152] Small plot field trials with randomized complete block (RCB) design and six replications per location were conducted at six locations to assess the ability of Methylobacterium spp. and Methylorubrum spp. isolates to suppress disease caused by Pythium spp. Five of the six locations had sufficient disease pressure to include in the analysis.

[0153] Soybean seed was treated with Methylobacterium spp. isolates to achieve a target titer of 1 x 105to 1 x 106CFU per seed. The control treatment (UTC) received no Methylobacterium treatment and the commercial control treatments received Axyl Shield ST (active ingredient metalaxyl) or oxathiopiprolin at the high end of the label rate. Four row plots 10 ft wide by 40 ft long with row spacing of 30 in were seeded with a regionally adapted soybean variety at a planting density of 140,000 plants per acre. Sorghum grain spawn inoculum prepared separately with Pythium ultimum var. ullimum. Pythium irregulare and Pythium sylvaticum was combined at a 1 : 1 : 1 ratio and applied in furrow at planting at a rate of 1 g per row foot. Plots were irrigated for two days after planting and then for an additional two days one week after planting to encourage the establishment of viable pathogen inoculum in the soil. Stand counts were recorded at 21 days after emergence (DAE) to assess pre-emergence damping off and final yield (bushels per acre) was measured. Data were analyzed using a linear mixed effects model with the lme4 (Bates et al. (2015) Journal of Statistical Software; vol 67, issue 1) and ImerTest (Kuznetsova et al. (2017) Journal of Statistical Software; vol 82, issue 13) packages in R (R Core Team (2019), “R: A language and environment for statistical computing”, R Foundation for Statistical Computing, Vienna, Austria).Table 4. Yield (bushels per acre) for 5 locations inoculated with a Pythium mixture.significantly different than UTC treatment at p < 0.001.Example 3 Pythium aphanidermatum growth chamber assay on soy 2019

[0154] A growth chamber assay with randomized complete block (RCB) design and 11 replicates was conducted to evaluate the ability of Methylobacterium spp. to prevent pre-emergence damping off caused by the pathogen Pythium aphanidermatum. Soybean seed was treated with Methylobacterium spp. and Methylorubrum spp. isolates to achieve a target titer of 1 x 105to 1 x 106CFU per seed. The control treatment (UTC) received no Methylobacterium or Methylorubrum treatment, and the commercial biocontrol treatment received Integral® (active ingredient Bacillus subtilis strain MBI 600; BASF Ag Products, NC, USA). Seeds were sown in autoclaved sand in aluminum tins, 5 seeds per tin, 11 tins per treatment, and the growth medium was brought up to 30% volumetric moisture. Five mL P.aphanidermatum zoospore suspension (8 x 105zoospores per mL) was added to the center of each tin, a clear lid was placed over the tin and tins were placed into clear plastic boxes and incubated at 27°C. At 3 days after planting (DAP), clear lids were removed from each tin, a thin layer of water was added to the plastic box, box lids were replaced to create a humid chamber and incubated at 27°C. At 6 DAP seedlings were rated for emergence and visible lesions and / or hyphae present on the aboveground portions of the seedling.Table 5. Seedling disease incidence compared to the untreated control (n = 55 seeds per treatment)Example 4 Pythium aphanidermatum growth chamber assay on Cannabis

[0155] A growth chamber assay as described in Example 3 will be conducted to evaluate the ability of Methylobacterium spp., including NLS0017, NLS0064 and NLS0432, to prevent pre-emergence damping off caused by the pathogen Pythium aphanidermatum. Hemp (Cannabis sativa L.) seed will be treated with Methylobacterium spp. and Methylorubrum spp. isolates to achieve a titer of 1 x 105to 1 x 106CFU per seed. The control treatment (UTC) will receive no Methylobacterium o Methylorubrum treatment, and the commercial biocontrol treatment will be a commercial seed treatment. Seeds will be sown as described in Example 3, and 5 mL of a P. aphanidermatum zoospore suspension (8 x 105zoospores per mL) will be added to the center of each tin. A clear lid will be placed over the tin and tins will be placed into clear plastic boxes and incubated at 27°C. At 3 days after planting (DAP), the lids will be removed from each tin, a thin layer of water was added to the plastic box, and box lids will be replaced to create a humid chamber followed by incubation at 27°C. At 6 DAP seedlings will be rated for emergence and visible lesions and / or hyphae present on the aboveground portions of the seedling. Methylobacterium and Methylorubrum strains that result in reduced seedling disease indicence in the growth chamber assay will be advanced to field testing to further analyze effects of the strains on yt / zzwm-induced diseases on hemp, and to identify strains that result in improved yield in hemp plants grown under Pythium disease pressure.Example 5 Pythium Field 2020

[0156] Methylobacterium isolates that demonstrated activity against Pythium aphanidermatum in the growth chamber assay were analyzed in a small plot field trial essentially as describe in Example 2. Results are shown in Tables 6 and 7 below.

[0157] All isolates tested demonstrated improved yield versus the untreated control and activity comparable to or greater than that of metalaxyl.Table 6 Emergence at V3-V4 (Connecting letters report using Student’s t test)Table 7 Yield (Connecting letters report using Student’s t test)Example 6 Soybean Sudden Death Syndrome (SDS) Fusarium virguliforme greenhouse 2018 / 2019

[0158] A combination growth chamber and greenhouse assay with randomized complete block (RCB) design and 25 replicates was conducted to evaluate the ability of Methylobacterium isolates to prevent disease caused by Fusarium virguliforme . Soybean seed was treated with Methylobacterium to achieve a titer of 1 x 106CFU per seed. The control treatment (UTC) received no Methylobacterium and the commercial control treatment received ILEVO (2018 at the high end of the label rate, 2019 at the low end of the label rate). Soybean seeds were sown in a 1 : 1 field soil: sand growth medium (partially steam pasteurized by heating for 30 min at 85°C) in cone-tainers with a 4.0 cubic inch (66 mL) volume. Pathogen inoculated treatments received a band of 3 mL of red sorghum grain spawn inoculum with Fusarium virguliforme Mont-1 directly below the seed. Plants were grown in a growth chamber for 14 days at 20°C, then transferred to a greenhouse for an additional 14 days. Plants were then rated for foliar disease symptoms using a 0-9 rating system where 0 is no disease and 9 is a dead plant, harvested and separated into below and aboveground tissue. Aboveground tissue was oven dried and weighed. Belowground tissue was imaged using WinRHIZO software from Regent Instruments Inc. then oven dried and weighed. The first series of isolates was run through the experiment at 25 individual pots per run for two separate runs with a total n = 50 pots per isolate while the second series of isolates was run through the experiment at 25 individual pots in a single run with a total of n = 25 pots per isolate. Results of root analyses and visual disease rating are presented in the below tables and shown using connecting letters report format using Student’s t test.Table 82018 greenhouse screen Average root dry biomass (g) 28 DAPTable 9 2018 greenhouse screen Average total root length (cm) 28 DAPTable 10 2018 greenhouse screen Average length of roots (cm) in 0 - 0.5 mm diameter size class 28DAPTable 11 2018 greenhouse screen Average foliar disease visual rating score 28 DAPTable 122019 greenhouse screen Average root dry biomass (g) 28 DAPTable 13 2019 greenhouse screen Average total root length (cm) 28 DAPTable 14 2019 greenhouse screen Average length of roots (cm) in 0 - 0.5 mm diameter size class 28DAPTable 15 2019 greenhouse screen Average foliar disease visual rating score 28 DAP

[0159] Two of the isolates tested significantly reduced foliar disease by one level in the visual rating scale. NLS0952 reduced the average foliar disease rating to 2.6 from 3.5 in the control (Table 11), andNLS0017 reduced the average foliar disease rating to 3.2 from 4.5 in the control (Table 15). For reference, a visual rating of 3 is equivalent to 20 - 40% chlorosis and 10 - 20% necrosis and a visual rating of 4 is equivalent to 40 - 60% chlorosis and 20 - 40% necrosis (Hashmi et al. 2005). Five of the isolates tested showed significantly higher root dry biomass compared to the control under high disease pressure; NLS0502, NLS0704, NLS0729, NLS0952, and NLS1310 (Table 8). Seven isolates; NLS0575, NLS0704, NLS0707, NLS0729, NLS0952, Methylobacterium SFGC33, and NLS1310 showed significantly larger root systems compared to the control under high disease pressure, as measured by total aggregated root length per plant via WinRHIZO (Table 9). In addition, three of these isolates, NLS0704, NLS0729 and NLS0952 showed higher total aggregate length of fine roots within the 0 - 0.5 mm diameter size class as measured by WinRHIZO (Table 10).Example 7 Detection or Identification of Methylobacterium Strains, Variants and Derivatives

[0160] Assays are developed for detection or identification of specific Methylobacterium strains and closely related derivatives. Genomic DNA fragments unique to a. Methylobacterium strain are identified and qPCR Locked Nucleic Acid (LNA) based assays are developed.

[0161] Genomic DNA sequences of Methylobacterium strains are compared by BLAST analysis of approximately 300bp fragments using a sliding window of from 1-25 nucleotides to whole genome sequences of over 1000 public and proprietary Methylobacterium isolates. Genomic DNA fragments were identified that had weak BLAST alignments, indicative of approximately 60-95% identity over the entire fragment, to corresponding fragments from a target Methylobacterium isolate. Fragments from the target Methylobacterium isolate that corresponded to the identified weak alignment regions were selected for assay development. Methylobacterium NLS0017 specific fragments are provided as SEQ ID NOS: 1-3. Primer and probe sequences for specific detection of the NLS0017 fragments are provided as SEQ ID NOS: 4-12. Methylobacterium NLS0064 specific fragments are provided as SEQ ID NOS: 13-15. Primer and probe sequences for specific detection of the NLS0064 fragments are provided as SEQ ID NOS: 16-24. Each of the probes contains a 5' FAM reporter dye and a 3’ Iowa Black FQ quencher.

[0162] For detection of fragments specific to Methylobacterium NLS0017 or NLS0064 isolate in isolated DNA, a qPCR reaction is conducted in 20 ul and contains 10 ul of 2x KiCqStartTM Probe qPCR ReadyMixTM, Low ROXTM from Sigma (Cat# KCQS05-1250RXN), lul of 20x primer-probe mix (final concentration of primers is 0.5 uM each and final concentration of probe is 0.25 uM), and 9ul of DNA template / water. Approximately 30-40 ng of DNA template is used per reaction. The reaction is conducted in a Stratagene Mx3005P qPCR machine with the following program: 95°C for 3 min, then 40 cycles of 95°C for 15 sec and 60°C for 1 min. The MxPro software on the machine calculates athreshold and Ct value for each sample. Each sample was run in triplicate on the same qPCR plate. A positive result is indicated where the delta Ct between positive and negative controls is at least 5.

[0163] For detection of Methylobacterium NLS0017 or NLS0064 on treated above-ground plant material, the plant material is washed and the bacteria pelleted by centrifugation. DNA is extracted from the pelleted sample using a MOBio UltraClean Microbial DNA Extraction kit Cat#12224-250 and analyzed using specific primers and probes as described above. For detection of Methylobacterium NLS0017 or NLS0064 on roots from treated plants, roots are harvested and bulk soil not physically adhered to the root surface is shaken off. The root system is then vortexed in 15 ml of saline solution and placed in an ultrasonic bath for 10 minutes. The roots are then removed and the tube contents combined with the soil contents from the vortexing step. The soil pellet is allowed to dry completely, and a soil PCR kit is used to detect the presence of NLS0017 or NLS0064 using specific primers and probes.

[0164] Table 16 Target Fragment Sequences of NLS0017

[0165] Table 17 Primer and Probe Sequences for Specific Detection of NLS0017*Bold and underlined letters represent the position of an LNA base

[0166] Table 18 Target Fragment Sequences of NLS0064

[0167] Table 19 Primer and Probe Sequences for Specific Detection of NLS0064and underlined letters represent the position of an LNA baseExample 8 Protection of Lettuce Roots and Shoots from Fusarium oxysporum f. sp. lactucae

[0168] A growth chamber assay was conducted using a randomized complete block (RCB) design to evaluate the ability of Methylobacterium isolates to prevent disease caused by Fusarium oxysporum f. sp. lactucae on lettuce seedlings. Pathogen and Methylobacterium isolates were added at planting. Control treatments were included that had no pathogen o Methylobacterium isolates (UTC) and another control treatment included the pathogen but no Methylobacterium isolates (UTC-path).

[0169] Lettuce seeds were planted in containers containing 1 cup of vermiculite planting medium plus fertilizer. Pathogen and Methylobacterium isolate treatments (1 x 106CFU per seed) were added at planting. Plants were randomized and transferred to a growth chamber (16-hour day, 25C, 0% humidity). Green leaf tissue area was measured weekly using PlantCV software. Root characteristics weremeasured at harvest (3-5 weeks after planting) using WinRhizo software. Results are shown in Table 20 below.

[0170] Table 20

[0171] Six of the isolates tested showed increased leaf area compared to the UTC-path control, NLS0498, NLS0433, NLS0934, NLS0419, NLS0575 and NLS0502. NLS0498, NLS0433, and NLS0934 demonstrated significantly different increases in leaf area versus control. Increased leaf area is indicative of redueced foliar disease resulting in a reduction of chlorotic and necrotic areas of leaf tissue.

[0172] REFERENCES1) Altschul S.F., Gish W., Miller W., Myers E.W., Lipman D.J. (1990) Basic local alignment search tool. J. Mol. Biol. 215:403-410 Bentley D.R., Balasubramanian S., Swerdlow H.P., Smith G.P., Milton J., Brown D. G., Hall K.P., Evers D.J., Barnes C.L., Bignell H.R. et al. (2008) Accurate whole human genome sequencing using reversible terminator chemistry. Nature 456:53-59.2) Caporaso J.G., Lauber C.L., Walters W. A., Berg-Lyons D., Huntley J., Fierer N., Owens S.M., Betley J., Fraser L., Bauer M., Gormley N., Gilbert J. A., Smith G., Knight R. (2012) Ultra-high- throughput microbial community analysis on the Illumina HiSeq and MiSeq platforms. ISME J 6: 1621-1624.3) Green, P.N. 2005. Methylobacterium. In Brenner, D.J. , N.R. Krieg, and J.T. Staley (eds.). “Bergey’s Manual of Systematic Bacteriology. Volume two, The Proteobacteria. Part C, The alpha-, beta-, delta-, and epsilonproteobacteria.“ Second edition. Springer, New York. Pages 567-571.4) Green, P.N. 2006. Methylobacterium. In Dworkin, M., S. Falkow, E. Rosenberg, K.-H. Schleifer, and E. Stackebrandt (eds.). “The Prokaryotes. A Handbook on the Biology of Bacteria. Volume 5. Proteobacteria: Alpha and Beta Subclasses. “ Third edition. Springer, New York. Pages 257-265.5) Konstantinos T. K. and Tiedje J.M. (2005) Proc. Nat. Acad. Sci. USA 102:2567-2572.6) Lidstrom, M.E. 2006. Aerobic methylotrophic prokaryotes. In Dworkin, M., S. Falkow, E. Rosenberg, K.-H. Schleifer, and E. Stackebrandt (eds.). “The Prokaryotes. A Handbook on the Biology of Bacteria. Volume 2. Ecophysiology and biochemistry. “ Third edition. Springer, New York. Pages 618-634.7) Lidstrom, M.E.. 2006. Aerobic methylotrophic prokaryotes. In Dworkin, M., S. Falkow, E. Rosenberg, K.-H. Schleifer, and E. Stackebrandt (eds.). “The Prokaryotes. A Handbook on the Biology of Bacteria. Volume 2. Ecophysiology and biochemistry. “ Third edition. Springer, New York. Pages 618-634.8) Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 74(12):5463-5467.9) Sy, A., Giraud, E., Jourand, P., Garcia, N., Willems, A., De Lajudie,P., Prin, Y., Neyra, M., Gillis, M., Boivin-Masson,C., and Dreyfus, B. 2001. Methylotrophic Methylobacterium Bacteria Nodulate and Fix Nitrogen in Symbiosis with Legumes. Jour. Bacteriol. 183(l):214-220.

[0173] The inclusion of various references herein is not to be construed as any admission by the Applicant that the references constitute prior art. Applicants expressly reserve their right to challenge any allegations of unpatentability of inventions disclosed herein over the references included herein.

[0174] Having illustrated and described the principles of the present disclosure, it should be apparent to persons skilled in the art that the disclosure can be modified in arrangement and detail without departing from such principles.

[0175] Although the materials and methods of this disclosure have been described in terms of various embodiments and illustrative examples, it will be apparent to those of skill in the art that variations can be applied to the materials and methods described herein without departing from the concept, spirit and scope of the disclosure. All such similar substitutes and modifications apparent to those skilled in the art are deemed to be within the spirit, scope and concept of the disclosure as defined by the appended claims or otherwise disclosed herein.

Claims

CLAIMS1. A composition comprising a Methylobacterium that prevents infection of plants and / or reduces severity of disease associated with one or more oomycete or fungal plant pathogens, wherein the Methylobacterium is selected from the group consisting of: NLS0059, NLS0165, NLS0214, NLS0419, NLS0433, NLS0498, NLS0532, NLS0632, NLS0952, NLS0729, NLS1310, NLS0502, NLS0575, NLS0704, NLS0707, NLS0934, and variants thereof2 The composition of claim 1, wherein the plant pathogen is a Pythium species or a Fusarium species.3 The composition of claim 1, wherein said plant pathogen is a species of Pythium, Peronospora, Phytophthora, Plasmopara, Haliotidica, Saprolegnia, Albugo, Pustula, Wilsoniana, Hyaloperonospora, Pseudoper onospora, Globosporangium or Aphanomyces.4 The composition of claim 2, wherein said plant pathogen is a Pythium species and said Methylobacterium is selected from the group consisting of: NLS0059, NLS0165, NLS0214, NLS0532, NLS0632, NLS0704, and variants thereof.5 The composition of claim 2, wherein said plant pathogen is a Fusarium species and said Methylobacterium is selected from the group consisting of: NLS0419, NLS0433, NLS0498, NLS0952, NLS0729, NLS1310, NLS0502, NLS0575, NLS0704, NLS0707, NLS0934, and variants thereof.6 The composition of any one of claims 1-5, wherein the composition further comprises Methylobacterium strain selected from the group consisting of: one or more strain in Table 1, one or more strain in Table 1A, one or more strain in Table IB, NLS0017, NLS0064, NLS0020, NLS0021, NLS0042, NLS0066, NLS0089, NLS0109, and variants thereof.7 The composition of claim 5, wherein the composition further comprises a Methylobacterium strain that is a variant of NLS0017 or NLS0064 has a sequence of any one of SEQ ID NOS: 1-3 or 13- 158 The composition of any one of claims 1-7, wherein the fungal plant pathogen is a soilbome pathogen.9 The composition of any one of claims 1-8, wherein the composition further comprises a fungicide.10 Thea composition of claim 9, wherein the fungicide is strobilurin, azoxystrobin, Trichoderma, metalaxyl, mefenoxam, ethaboxam, or oxathiapiprolin.

11. A method for reducing the adverse effects on a plant caused by an oomycete or fungal plant pathogen, wherein said method comprises applying a composition comprising Methylobacterium selected from the group consisting of: NLS0017, NLS0064, NLS0059, NLS0165, NLS0214, NLS0419, NLS0433, NLS0498, NLS0532, NLS0632, NLS0952, NLS0729, NLS1310, NLS0502, NLS0575, NLS0704, NLS0707, NLS0934, and variants thereof to a plant, a plant part, to soil or to a nutrient solution where a plant is grown, or any combination thereof in an amount that provides for i) inhibition of infection by said plant pathogen in said plant, plant part, or a plant obtained therefrom, or ii) reduction in severity of symptoms of said plant pathogen; wherein said inhibition of infection and / or reduction of severity of symptoms are relative to infection of a control plant, plant part, or plant that had not received an application of said composition or been grown in soil or nutrient solution treated with the composition.

12. The method of claim 11, wherein application of said composition provides for at least about 10%, at least about 20%, at least about 25%, at least about 30%, 40%, 50%, 75%, at least 85%, or at least 95% inhibition of infection or reduction in severity of symptoms in said plant, plant part, or a plant derived therefrom relative to the control plant, plant part, or plant.

13. The method of claim 11 or 12, wherein said plant part is selected from the group consisting of: a leaf, a stem, a vegetative cutting, a fruit, a flower, a root, a tuber, and a seed.

14. The method of any one of claims 11-13, wherein the plant or plant is selected the group consisting of: berry, cannabis, corn, fl ori culture / ornamental, grape, herb, hops, leafy green, lettuce, microgreen, peanut, pepper, pome fruit, rice, soybean, tomato, tree nut, turf grass, vegetable, and wheat.

15. The method of any one of claims 11-14, wherein said plant pathogen is a Pythium species and said Methylobacterium is selected from the group consisting of: NLS0059, NLS0165, NLS0214, NLS0532, NLS0632, NLS0704, and variants thereof.

16. The method of any one of claims 11-15, wherein said plant pathogen is a Fusarium species and said Methylobacterium is selected from the group consisting of: NLS0419, NLS0433, NLS0498, NLS0952, NLS0729, NLS1310, NLS0502, NLS0575, NLS0704, NLS0707, NLS0934, and variants thereof.

17. The method of any one of claims 11-14, wherein the plant pathogen is a Fusarium species and the Methylobacterium is NLS0017 or a variant of NLS0017 comprising any one of SEQ ID NOS: 1-3, or the plant pathogen is a Pythium species and the Methylobacterium is NLS0064, or a variant of NLS0064 comprising any one of SEQ ID NOS: 13-15.

18. Thea method of any one of claims 15-17, further comprising applying a second Methylobacterium to a plant, a plant part, to soil where a plant is grown, or any combination thereof, wherein said second Methylobacterium is selected from the group consisting of: one or more strain in Table 1, one or more strain in Table 1 A, one or more strain in Table IB, NLS0017, NLS0064, NLS0020, NLS0021, NLS0042, NLS0066, NLS0089, NLS0109, and variants thereof19. The method of claim 18, wherein the second Methylobacterium is applied to the soil in furrow with the plant part, on a seed from which the plant is grown, to the plant foliage, as a liquid to a vegetative cutting, or in a nutrient solution in which the plant is grown.

20. The method of claim 18, wherein Methylobacterium and said second Methylobacterium are applied simultaneously.

21. The method of any one of claims 11-20, wherein said method further comprises applying a fungicide in a composition comprising said Methylobacterium or following application of said Methylobacterium .

22. The method of claim 21, wherein the fungicide is strobilurin, azoxystrobin, Trichoderma. metalaxyl, mefenoxam, ethaboxam, or oxathiapiprolin.

23. The method of claim 21, wherein the fungicide is active against Pythium species.

24. The method of claim 21 wherein the fungicide is active against Fusarium species.

25. A plant or plant part that is at least partially coated with the composition of any one of claims 1- 10.

26. The plant or plant part of claim 25, wherein the plant or plant part is selected from the group consisting of: berry, cannabis, corn, fl ori culture / ornamental grape, herb, hops, leafy green, lettuce, microgreen, peanut, pepper, pome fruit, rice, soybean, seed crop, tomato, tree nut, turf grass, vegetable, and wheat.

27. The plant or plant part of claims 25 or 26, wherein the plant or plant part is isolated.

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