Compositions and methods for treating a plant or seed thereof with bacterial strains for nitrogen acquistion in crop plants

Abstract

Disclosed herein are compositions for application to plants, plant seeds, or a plant growth medium including an agriculturally acceptable carrier and an effective amount of a biologically pure bacterial culture, wherein the bacteria in the bacterial culture includes Bacillus subtilis MH1 (ATCC Accession No. PTA- 127648), Bacillus pseudomycoides MH2 (ATCC Accession No. PTA-127649) or a combination thereof. Methods for treating plants with Bacillus subtilis MH1, Bacillus pseudomycoides MH2 or combinations thereof are also disclosed.

Description

[0001] COMPOSITIONS AND METHODS FOR TREATING A PLANT OR SEED THEREOF WITH BACTERIAL STRAINS FOR NITROGEN ACQUISTION IN CROP PLANTS

[0002] CROSS-REFERENCE TO RELATED APPLICATION

[0003] This application claims priority to U.S. Provisional Patent Application No. 63 / 613,348 filed on December 21 , 2023, the contents of which are incorporated herein by reference in their entirety for all purposes.

[0004] FIELD

[0005] The present disclosure relates to compositions for application to plants, plant seeds, or plant growth medium that include an effective amount of a biologically pure bacterial culture containing Bacillus subtilis MH1 (ATCC Accession No. PTA- 127648, deposited October 13, 2023), Bacillus pseudomycoides MH2 (ATCC Accession No. PTA-127649, deposited October 13, 2023) or a combination thereof. BACKGROUND

[0006] The medical practice of fecal microbiota transplantation has gained tremendous traction in recent years, particularly for treating conditions such as metabolic syndrome by transferring gut microbiota from healthy individuals to affected recipients. This simple transplant in the gut microbiome can profoundly improve an individual's overall health and metabolic function. While fecal microbiome transplant (FMT) is a well-established procedure in human medicine, similar applications in agriculture have been limited, largely due to the lack of analogous ‘healthy’ soils defined by a rich diversity of microbial communities. The concept of swapping or rewilding microbiomes in plants is a new frontier and few studies have shown that swapping of topsoil microbiomes can alter the aboveground vegetation.

[0007] Plant derived compounds, mainly root secretions, play a critical role in recruiting a microbiome. Plant secretions influence the selection of bacterial symbionts and commensals that thrive in association with them. Anthropogenic practices, such as fertilizer application and nutrient management, also affect soil microbial composition significantly. Over time, global practices like monoculture and excessive chemical use have led to a decline in microbial diversity in arable soils, negatively impacting soil health and plant growth. For example, the excessive use of synthetic nitrogen (N) fertilizers in agriculture has been shown to be unhealthy, unsustainable, and costly. The synthetic production of N consumes fossil fuels and N fertilizer contributes to greenhouse gas emissions and water pollution. One percent of global fossil energy is required to perform the Haber-Bosch reaction in producing N fertilizer, and the use efficiency of N-fertilizer from production to incorporation into plants is extremely poor. Of the N created, only 2-10% is present in plants and crops that reach the market. Beyond the issues with energy, N fertilizer is also extremely labile, and its over-application to fields is frequently the cause of ‘dead zones’ in lakes and bays from run-off. In addition, global nitrogen fertilizer consumption is estimated at 108 million Metric Tons for 2024 and is expected to rise through 2030. One of the biggest issues with synthetic N fixation relates to its detrimental effect on soil health. Synthetic N applications in soil can add to more plant-microbe competitive pressures and lead to growth promotion of N microbial competitors. These N competitors evolved to take up N faster than plants, thus leading to N deficiency in soil. In addition to the N microbial competitors, synthetic N application leads to eutrophication issues.

[0008] Leguminous plants, such as soy and alfalfa are known to mitigate these N fertilizer liabilities through their ability to form symbiosis with N-fixing Rhizobia bacteria in their roots. That is, leguminous plants are able to form a mutualistic plant-microbe partnership with symbiotic nitrogen-fixing bacteria in the family Rhizobia. In a process referred to as ‘biological nitrogen fixation’ (BNF), Rhizobia can fix atmospheric N for the plant in exchange for carbon-rich photosynthates within specialized structures formed on the plant root called nodules. N-fixation by crop and forage legumes globally through this symbiosis is roughly estimated to be 50 Tg per year. Older estimates of non-agricultural BNF may be up to 100 Tg N fixed per year. Through their ability to form symbiosis with N-fixing Rhizobia bacteria in their roots, legumes can reduce the requirement for N application, decrease greenhouse gas emissions and reduce the need for fossil energy inputs. Commercial formulations of Rhizobia have already been in use in agricultural settings to boost N-fixation for valuable legume crops as well as enriching the soil N in crop rotational systems. Unfortunately, legumes represent only 2-2.5% of the current staple crops market. The majority of the current staple crops market are non-legumes, such as cereals. Cereal consumption, including that of rice and wheat, accounts for over -1000 million metric tons globally. These non-legume crops require synthetic fertilizer to grow, which poses a huge challenge for sustainable agricultural practices. Presently little is known about microbes or consortia of microbes that can help non-leguminous plants to grow under low fertilizer treatments.

[0009] While human activities and modern farming practices have substantially altered topsoil environments and have depleted microbial diversity, there are soils that still retain microbiomes from a pre-agricultural era. For example, along midAtlantic creeks and streams in the US, dark, organic, and hydric soil horizons are often visible at the base of the streambanks. These hydric soils date back to a precolonial era where the landscape was very different from present-day agricultural environments. These buried, organic horizons are remnants or “living fossils” of surficial marshes, bogs, or wetlands, that were once widespread in valley-bottoms of the eastern US.

[0010] The art’s understanding of these buried historic soils (BHS) (> 300 years old) and pre-colonial organic soils (POS), and their associated microbiomes for plant growth and nutrient availability, is limited and non-existent. Current understanding of these soils suggests that they were likely highly diverse and resilient. BHS and POS also predate modern, harmful, anthropogenic practices (e.g., intensive agriculture with pesticides and fertilizer) and could contain distinct microbiota that have unique plant associations missing from modern soils. Almost all BHS and POS environments represent a period before intense agriculture or synthetic N amendments. Indeed, preliminary work has showed that these soils supported plant growth and could harbor distinct microbes, thus suggesting that the nutrient availability and mobilization in these soils were different compared to the agricultural modern soils.

[0011] Harnessing BHS and POS to isolate novel microbes that could enhance plant growth and nutrient fortification in present-day agricultural soil systems has been a challenging endeavor given the limited understanding of their microbiome. The rewilding of these BHS / POS microbiomes, via their successful incorporation into current agriculture environments and soils, could provide a novel approach for improving soil health, plant growth and microbiome diversity in agricultural.

[0012] Thus, to address the foregoing issues and provide a novel approach to improving plant growth and sustainable agricultural practices, we disclose the use of POS-derived and BHS-derived bacterial strains in agricultural compositions and methods for improving plant growth.

[0013] SUMMARY

[0014] An aspect of the present disclosure is a composition for application to plants, plant seeds, or a plant growth medium that includes ( / .e., comprises) one or more of an agriculturally acceptable carrier and / or an effective amount of a biologically pure bacterial culture, wherein the bacteria in the bacterial culture includes Bacillus subtilis MH1 (ATCC Accession No. PTA-127648, deposited October 13, 2023), Bacillus pseudomycoides MH2 (ATCC Accession No. PTA-127649, deposited October 13, 2023) or a combination thereof.

[0015] Another aspect of the present disclosure is a method for treating a plant or a seed thereof including one or more of transplanting the plant or the seed thereof into a soil composition; administering to the plant or the seed thereof an effective amount of an isolated bacterial strain selected from Bacillus subtilis MH1 (ATCC Accession No. PTA-127648, deposited October 13, 2023), Bacillus pseudomycoides MH2 (ATCC Accession No. PTA-127649, deposited October 13, 2023) or a combination thereof; and / or cultivating the plant or seed thereof in an environment after the administering of the effective amount of the isolated bacterial strain.

[0016] Another aspect of the present disclosure is a plant seed coated with any one of the compositions disclosed herein.

[0017] Another aspect of the present disclosure is a kit for treating a plant or a seed thereof that includes any one of the compositions disclosed herein.

[0018] BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Other features and advantages of the compositions, devices and methods disclosed herein will be apparent to those skilled in the art reading the following detailed description in conjugation with the exemplary embodiments illustrated in the drawings, wherein:

[0020] FIG. 1 depicts a representative illustration of the isolation of POS, legacy sediments (LS) and modern top-soil (TS) (e.g., soils less than 100 years old).

[0021] FIG. 2 depicts a representative illustration of the effects that POS, sterilized POS and agricultural soil have on the growth of a tomato plant.

[0022] FIG. 3 depicts a representative illustration of the effects that POS, sterilized POS and agricultural soil have on the root length (RL), shoot length (SL), fresh root weight (FRW), fresh shoot weight (FSW), dry root weight (DRW), and dry shoot weight (DSW) of tomato plants. The notations A, AB, a, b, B and c are used to represent whether there is a significant difference between the RL, SL, FRW, FSW, DRW and DSW of the various treatment groups. If two groups have the same letter (e.g., A and A), then no significant difference was detected. If two groups have different letters (e.g., A and B), then a significant difference was detected. Significance was determined via a two-way analysis of variance (ANOVA).

[0023] FIG. 4 depicts a representative illustration of a serial dilution method used to dilute samples of modern topsoil, middle legacy sediments, POS and sterilized POS.

[0024] FIG. 5 depicts the growth of microbial communities present in serial diluted modern topsoil, middle legacy sediments, POS and sterilized POS solutions.

[0025] FIG. 6 depicts a representative illustration of the Shannon Index and Chaol values of historic soils (i.e., POS soils), wetland soils and modern agriculture soils. The notations a and b are used to represent whether there is a significant difference between the Shannon index and Chaol values of the various soils. If two groups have the same letter (e.g., a and a), then no significant difference was detected. If two groups have different letters (e.g., a and b), then a significant difference was detected. Significance was determined via a two-way ANOVA.

[0026] FIG. 7 depicts a representative illustration of the nifH gene copy numbers found in the microbial communities of agricultural soils without crop growth, agricultural soils with crop growth, historic (i.e., POS) soils without crop growth, and historic (i.e., POS) soils with crop growth. FIG. 8 depicts NMD plots for the community structures of bacteria / archaea in POS soils (represented by HIS soil), sterile POS soils (represented by Sterile soil) and modern topsoil (represented by Modern soil). The NMD plot on the left was prepared with rarefied data and the NMD plot on the right was prepared with nonrarefied data.

[0027] FIG. 9 depicts NMD plots for the community structures of fungi in POS soils (represented by HIS soil), sterile POS soils (represented by Sterile soil) and modern topsoil (represented by Modern soil). The NMD plot on the left was prepared with rarefied data and the NMD plot on the right was prepared with nonrarefied data.

[0028] FIG. 10 depicts a representative illustration of the anti-fungal properties of Bacillus subtilis MH1 , Bacillus pseudomycoides MH2 and B. subtilis LID1022 have against Clarireedia jacksonii, Sclerotinia sclerotiorum, Diaporthae ueckerae, Rhizoctonia solani and Bipolaris zeicola.

[0029] FIG. 11 depicts a representative illustration of the mycelial inhibition percent that Bacillus subtilis MH1 , Bacillus pseudomycoides MH2 and B. subtilis UD1022 have on Clarireedia jacksonii, Sclerotinia sclerotiorum, Diaporthae ueckerae, Rhizoctonia solani and Bipolaris zeicola. The effect that Bacillus subtilis MH1 , Bacillus pseudomycoides MH2 and B. subtilis LID1022 have on mycelail growth of Clarireedia jacksonii, Sclerotinia sclerotiorum, Diaporthae ueckerae, Rhizoctonia solani and Bipolaris zeicola is also depicted. The notations A, B, BC, and C are used to represent whether there is a significant difference between the mycelail growth of the various plant pathogenic fungi for the different treatment groups. If two groups have the same letter (e.g., A and A), then no significant difference was detected. If two groups have different letters (e.g., A and C), then a significant difference was detected. Significance was determined via a two-way ANOVA.

[0030] FIG. 12 depicts a representative illustration of the anti-fungal properties of heat-killed Bacillus subtilis MH1 cultures, heat-killed Bacillus pseudomycoides MH2 cultures and heat-killed B. subtilis UD1022 cultures have against Clarireedia jacksonii, Sclerotinia sclerotiorum, Diaporthae ueckerae, Rhizoctonia solani and Bipolaris zeicola. FIG. 13 depicts a representative illustration of the effect that various concentrations of Bacillus subtilis MH1 has on tomato plant growth.

[0031] FIG. 14 depicts a representative illustration of the effect that various concentrations of Bacillus subtilis MH1 has on the root length (RL), shoot length (SL), fresh root weight (FRW), fresh shoot weight (FSW), dry root weight (DRW), and dry shoot weight (DSW) of tomato plants. The notations A, b, a, ab, CD, be, D, c, and B are used to represent whether there is a significant difference between the RL, SL, FRW, FSW, DRW, and DSW of the various treatment groups. If two groups have the same letter (e.g., A and A), then no significant difference was detected. If two groups have different letters (e.g., A and B), then a significant difference was detected. Significance was determined via a two-way ANOVA.

[0032] FIG. 15 depicts a representative illustration of the effect that various concentrations of Bacillus pseudomycoides MH2 has on tomato plant growth.

[0033] FIG. 16 depicts a representative illustration of the effect that various concentrations of Bacillus pseudomycoides MH2 has on the root length (RL), shoot length (SL), fresh root weight (FRW), fresh shoot weight (FSW), dry root weight (DRW), and dry shoot weight (DSW) of tomato plants. The notations BC, be, B, ab, A, a, C, c, CD, D, b and AB are used to represent whether there is a significant difference between the RL, SL, FRW, FSW, DRW, and DSW of the various treatment groups. If two groups have the same letter (e.g., A and A), then no significant difference was detected. If two groups have different letters (e.g., A and B), then a significant difference was detected. Significance was determined via a two-way ANOVA.

[0034] FIG. 17 depicts a representative illustration of tomato plant root colonization of Bacillus subtilis MH1 , Bacillus pseudomycoides MH2 and B. subtilis UD1022.

[0035] FIG. 18 depicts a representative illustration of the effects that Bacillus subtilis MH1 has on tomato plant fresh total biomass (FTB), fresh weight of roots (FRW) and fresh weight of stem (FSW) under various nitrogen poor conditions. The notations A, B, C, ab, a, abc, be, c, cd and d are used to represent whether there is a significant difference between the FTB, FRW and FSW of the various treatment groups. If two groups have the same letter (e.g., A and A), then no significant difference was detected. If two groups have different letters (e.g., A and B), then a significant difference was detected. Significance was determined via a two-way ANOVA.

[0036] FIG. 19 depicts a representative illustration of the effects that Bacillus subtilis MH1 has on tomato plant dry total biomass (DTB), dry weight of roots (DRW) and dry weight of stem (DSW) under various nitrogen poor conditions. The notations a, be, ab, c, a, d, b and abc are used to represent whether there is a significant difference between the DTB, DRW and DSW of the various treatment groups. If two groups have the same letter (e.g., a and a), then no significant difference was detected. If two groups have different letters (e.g., a and c), then a significant difference was detected. Significance was determined via a two-way ANOVA.

[0037] FIG. 20 depicts a representative illustration of the effects that Bacillus subtilis MH1 has on tomato plant fruit yield and yield per hectare under various nitrogen poor conditions. The notations a, ab, be, and c are used to represent whether there is a significant difference between the fruit yield and yield per hectare of the various treatment groups. If two groups have the same letter (e.g., a and a), then no significant difference was detected. If two groups have different letters (e.g., a and be), then a significant difference was detected. Significance was determined via a two-way ANOVA.

[0038] FIG. 21 depicts the overall growth effects that Bacillus subtilis MH1 has on a group of tomato plants under various nitrogen poor conditions.

[0039] FIG. 22 depicts the effects that Bacillus subtilis MH1 has on plant growth for individual tomato plants under various nitrogen poor conditions.

[0040] FIG. 23 depicts the effects that Bacillus subtilis MH1 has on the fruit yield for tomato plants grown under various nitrogen poor conditions.

[0041] FIG. 24 depicts a representative illustration of the nitrogen content percentage in tomato plants grown under various nitrogen poor conditions with Bacillus subtilis MH1 . The notations b, be, a and c are used to represent whether there is a significant difference between the nitrogen content percentage of the various treatment groups. If two groups have the same letter (e.g., b and b), then no significant difference was detected. If two groups have different letters (e.g., b and c), then a significant difference was detected. Significance was determined via a two-way ANOVA.

[0042] FIG. 25 depicts a representative illustration of the effects that Bacillus pseudomycoides MH2 has on tomato plant fresh total biomass (FTB), fresh weight of roots (FRW) and fresh weight of stem (FSW) under various nitrogen poor conditions. The notations b, c, a, d, de, e, and be are used to represent whether there is a significant difference between the FTB, FRW and FSW of the various treatment groups. If two groups have the same letter (e.g., c and c), then no significant difference was detected. If two groups have different letters (e.g., c and e), then a significant difference was detected. Significance was determined via a two-way ANOVA.

[0043] FIG. 26 depicts a representative illustration of the effects that Bacillus pseudomycoides MH2 has on tomato plant dry total biomass (DTB), dry weight of roots (DRW) and dry weight of stem (DSW) under various nitrogen poor conditions. The notations b, c, a, d, ab, and be are used to represent whether there is a significant difference between the DTB and DRW of the various treatment groups. If two groups have the same letter (e.g., b and b), then no significant difference was detected. If two groups have different letters (e.g., b and be), then a significant difference was detected. Significance was determined via a two-way ANOVA.

[0044] FIG. 27 depicts a representative illustration of the effects that Bacillus pseudomycoides MH2 has on tomato plant fruit yield and yield per hectare under various nitrogen poor conditions. The notations ab, b, a, and c are used to represent whether there is a significant difference between the yield of each plant and the yield per hectare of the various treatment groups. If two groups have the same letter (e.g., b and b), then no significant difference was detected. If two groups have different letters (e.g., b and c), then a significant difference was detected. Significance was determined via a two-way ANOVA.

[0045] FIG. 28 depicts the overall growth effects that Bacillus pseudomycoides MH2 has on a group of tomato plants under various nitrogen poor conditions.

[0046] FIG. 29 depicts the effects that Bacillus pseudomycoides MH2 has on the fruit yield for tomato plants grown under various nitrogen poor conditions. FIG. 30 depicts the effects that Bacillus subtilis MH1 and Bacillus pseudomycoides MH2 have on the root depth or root length of a group of tomato plants grown under various nitrogen poor conditions.

[0047] FIG. 31 depicts a representative illustration of the effects that Bacillus subtilis MH1 has on onion plant fresh total biomass (FTB), fresh weight of roots (FRW) and fresh weight of stem (FSW) under various nitrogen poor conditions. The notations cd, be, abc, a, ab, d, and b are used to represent whether there is a significant difference between the FTB, FRW and FSW of the various treatment groups. If two groups have the same letter (e.g., ab and ab), then no significant difference was detected. If two groups have different letters (e.g., d and a), then a significant difference was detected. Significance was determined via a two-way ANOVA.

[0048] FIG. 32 depicts a representative illustration of the effects that Bacillus subtilis MH1 has on onion plant dry total biomass (DTB), dry weight of roots (DRW) and dry weight of stem (DSW) under various nitrogen poor conditions. The notations ab, be, a, abc, and c are used to represent whether there is a significant difference between the DTB and DSW of the various treatment groups. If two groups have the same letter (e.g., be and be), then no significant difference was detected. If two groups have different letters (e.g., a and c), then a significant difference was detected. Significance was determined via a two-way ANOVA.

[0049] FIG. 33 depicts a representative illustration of the effects that Bacillus pseudomycoides MH2 has on onion plant fresh total biomass (FTB), fresh weight of roots (FRW) and fresh weight of stem (FSW) under various nitrogen poor conditions. The notations a, abc, ab, be, and c are used to represent whether there is a significant difference between the FTB and FRW of the various treatment groups. If two groups have the same letter (e.g., a and a), then no significant difference was detected. If two groups have different letters (e.g., a and c), then a significant difference was detected. Significance was determined via a two-way ANOVA.

[0050] FIG. 34 depicts a representative illustration of the effects that Bacillus pseudomycoides MH2 has on onion plant dry total biomass (DTB), dry weight of roots (DRW) and dry weight of stem (DSW) under various nitrogen poor conditions. The notations be, abc, a, c, b, and ab are used to represent whether there is a significant difference between the DTB, DRW and DSW of the various treatment groups. If two groups have the same letter (e.g., a and a), then no significant difference was detected. If two groups have different letters (e.g., a and c), then a significant difference was detected. Significance was determined via a two-way ANOVA.

[0051] FIG. 35 depicts the effects that Bacillus subtilis MH1 , Bacillus pseudomycoides MH2 or a combination thereof has on corn plant growth under various nitrogen poor conditions.

[0052] FIG. 36 depicts the overall effects that Bacillus subtilis MH1 , Bacillus pseudomycoides MH2 or a combination thereof has on corn plant growth under various nitrogen poor conditions.

[0053] FIG. 37 depicts the nitrogen percentage in leaves of corn plants grown in contact with Bacillus subtilis MH1 , Bacillus pseudomycoides MH2 or a combination thereof. The notations A and B are used to represent whether there is a significant difference between the nitrogen percentage in the leaves of the various treatment groups. If two groups have the same letter (e.g., A and A), then no significant difference was detected. If two groups have different letters (e.g., A and B), then a significant difference was detected. Significance was determined via a two-way ANOVA.

[0054] DETAILED DESCRIPTION

[0055] All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In case of conflict, the present specification, including definitions, will control.

[0056] The present disclosure relates to bacterial strains isolated from a -1000- year-old (950±30 BP, Beta Analytic sample# Beta - 510411) POS and the uses thereof, for example, in compositions for plant application (e.g., root application) to promote both plant growth and disease protection. One aspect of the present disclosure is a composition for application to plants, plant seeds, or a plant growth medium that includes (i.e., comprises) one or more of an agriculturally acceptable carrier and / or an effective amount of a biologically pure bacterial culture, wherein the bacteria in the bacterial culture includes Bacillus subtilis MH1 (ATCC Accession No. PTA-127648, deposited October 13, 2023), Bacillus pseudomycoides MH2 (ATCC Accession No. PTA- 127649, deposited October 13, 2023) or a combination thereof.

[0057] In exemplary embodiments, the effective amount of the biologically pure bacterial culture is at most 2 x 108CFU / mL. In other embodiments, the effective amount of the biologically pure bacterial culture is any amount or range of amounts falling within the range of 1 x 104CFU / mL to 2 x 108CFU / mL.

[0058] In exemplary embodiments, wherein the composition has a nitrogen content ranging from 0% to 75% of a recommend nitrogen level for a growth stage of the plant.

[0059] As used herein, “a recommend nitrogen level for a growth stage of the plant” means an amount of nitrogen the plant is recommended or required to receive during a growth stage to ensure the plant reaches the harvesting stage of its growth cycle. A growth stage of the plant can include any stage falling within the sowing of the seed of the plant to the harvesting of the plant. Possible plant growth stages include, but are not limited to, sowing (at day 0), early growth (e.g., 25 to 30 days after sowing), vegetative (e.g., 20-25 days after early growth), flowering (e.g., 20-30 days after vegetative), fruit formation (e.g., 20-30 days after flowering), mature fruiting (e.g., 15-20 after fruit formation) and harvesting. Those of ordinary skill in the art are readily capable of determining the recommend nitrogen amounts for a variety of plants during their different growth stages. For example, the University of Delaware’s Agriculture and Natural Resources Department’s Soil Testing Program discloses that nutrient recommendations can be determined with soil test calibration studies that relate the probability of a profitable plant response to nutrient addition to the plant nutrient concentration in the soil as measured by a soil test. The University of Delaware’s Agriculture and Natural Resources Department provides various soil test methods that those of ordinary skill in the art can readily perform to determine the recommended level of nitrogen to provide a plant. Those of ordinary skill in the art will appreciate that nitrogen level recommendations for various plants and crops will vary with soil productivity and are readily capable of determining recommended nitrogen levels in view of soil productivity.

[0060] In exemplary embodiments, the composition includes an effective amount of an auxiliary plant growth promoting rhizobacteria (PGPR). Possible auxiliary PGPR that can be include in the composition are, but not limited to, Bacillus subtilis SJ- 101 , Pseudomonas species, Bradyrhizobium species, Sinorhizobium species, Brevibacillus species, Bacillus species, Rhizobacteria species, Rhizobium species, Rhizobacterium species, and any bacterial species known to possess one or more of the following characteristics: live freely and colonize plant roots, metabolize phosphates, produce hormones, fix nitrogen, promote plant water and nutrient uptake, enhance root development, increase plant enzymatic activity, synthesize phytohormones and their analogs, produce siderophores, produce antioxidant enzymes, or produce and release cytokinins.

[0061] In exemplary embodiments, the composition includes a phosphorus content ranging from 90% to 100% of a recommend phosphorus level for the growth stage of the plant.

[0062] As used herein, “a recommend phosphorus level for a growth stage of the plant” means an amount of phosphorus the plant is recommended or required to receive during a growth stage to ensure the plant reaches the harvesting stage of its growth cycle. A growth stage of the plant can include any stage falling within the sowing of the seed of the plant to the harvesting of the plant. Possible plant growth stages include, but are not limited to, sowing (at day 0), early growth (e.g., 25 to 30 days after sowing), vegetative (e.g., 20-25 days after early growth), flowering (e.g., 20-30 days after vegetative), fruit formation (e.g., 20-30 days after flowering), mature fruiting (e.g., 15-20 after fruit formation) and harvesting. Those of ordinary skill in the art are readily capable of determining the recommend phosphorus amounts for a variety of plants during their different growth stages and will appreciate that phosphorus level recommendations for various plants and crops will vary with soil productivity. Accordingly, those of ordinary skill in the art are readily capable of determining recommended phosphorus levels in view of soil productivity. In exemplary embodiments, the composition includes a potassium content ranging from 90% to 100% of a recommend potassium level for the growth stage of the plant.

[0063] As used herein, “a recommend potassium level for a growth stage of the plant” means an amount of potassium the plant is recommended or required to receive during a growth stage to ensure the plant reaches the harvesting stage of its growth cycle. A growth stage of the plant can include any stage falling within the sowing of the seed of the plant to the harvesting of the plant. Possible plant growth stages include, but are not limited to, sowing (at day 0), early growth (e.g., 25 to 30 days after sowing), vegetative (e.g., 20-25 days after early growth), flowering (e.g., 20-30 days after vegetative), fruit formation (e.g., 20-30 days after flowering), mature fruiting (e.g., 15-20 after fruit formation) and harvesting. Those of ordinary skill in the art are readily capable of determining the recommend potassium amounts for a variety of plants during their different growth stages and will appreciate that potassium level recommendations for various plants and crops will vary with soil productivity. Accordingly, those of ordinary skill in the art are readily capable of determining recommended potassium levels in view of soil productivity.

[0064] In exemplary embodiments, the agriculturally acceptable carrier includes one or more of a dispersant, a surfactant, an additive, water, a thickener, an anti-caking agent, residue breakdown, a composting formulation, a granular application, diatomaceous earth, an oil, a coloring agent, a stabilizer, a preservative, a polymer, or a coating.

[0065] Possible additives include, but are not limited to, an oil, a gum, a resin, a clay, a polyoxyethylene glycol, a terpene, a viscid organic, a fatty acid ester, a sulfated alcohol, an alkyl sulfonate, a petroleum sulfonate, an alcohol sulfate, a sodium alkyl butane diamate, a polyester of sodium thiobutant dioate, a proteinaceous material, or a combination thereof.

[0066] Possible thickeners include, but are not limited to, a long chain alkylsulfonate of polyethylene glycol, polyoxyethylene oleate, or a combination thereof.

[0067] Possible surfactants include, but are not limited to, a heavy petroleum oil, a heavy petroleum distillate, a polyol fatty acid ester, a polyethoxylated fatty acid ester, an aryl alkyl polyoxyethylene glycol, an alkyl amine acetate, an alkyl aryl sulfonate, a polyhydric alcohol, an alkyl phosphate, or a combination thereof.

[0068] Possible anti-caking agents include, but are not limited to, a sodium salt, a calcium carbonate, diatomaceous earth, or a combination thereof.

[0069] In exemplary embodiments, the agriculturally acceptable carrier includes vermiculite, charcoal, sugar factory carbonation press mud, rice husk, carboxymethyl cellulose, peat, perlite, fine sand, calcium carbonate, flour, alum, a starch, talc, polyvinyl pyrrolidone, or a combination thereof.

[0070] In exemplary embodiments, the composition is formulated as a seed coating formulation, a liquid formulation for application to plants or to a plant growth medium, or a solid formulation for application to plants or to a plant growth medium.

[0071] In exemplary embodiments, the composition is formulated as a seed coating formulation, the seed coating formulation being either an aqueous or oil-based solution for application to seeds, or a powder or granular formulation for application to seeds.

[0072] In exemplary embodiments, the composition is formulated as a concentrated or a ready-to-use liquid formulation.

[0073] In exemplary embodiments, the composition is formulated as a granular or a powder agent solid formulation.

[0074] In exemplary embodiments, the composition includes a fertilizer, a micronutrient fertilizer material, an insecticide, a herbicide, a plant growth amendment, a molluscicide, an algicide, a bacterial inoculant, a fungal inoculant, or a combination thereof.

[0075] Possible fertilizers include, but are not limited to, ammonium sulfate, ammonium nitrate, ammonium sulfate nitrate, ammonium chloride, ammonium bisulfate, ammonium polysulfide, ammonium thiosulfate, aqueous ammonia, anhydrous ammonia, ammonium polyphosphate, aluminum sulfate, calcium nitrate, calcium ammonium nitrate, calcium sulfate, calcined magnesite, calcitic limestone, calcium oxide, calcium nitrate, dolomitic limestone, hydrated lime, calcium carbonate, diammonium phosphate, monoammonium phosphate, magnesium nitrate, magnesium sulfate, potassium nitrate, potassium chloride, potassium magnesium sulfate, potassium sulfate, sodium nitrates, magnesian limestone, magnesia, urea, urea-formaldehydes, urea ammonium nitrate, sulfur-coated urea, polymer-coated urea, isobutylidene diurea, K2SO4-2MgSC>4, kainite, sylvinite, kieserite, Epsom salts, elemental sulfur, marl, ground oyster shells, fish meal, oil cakes, fish manure, blood meal, rock phosphate, super phosphates, slag, bone meal, wood ash, manure, bat guano, peat moss, compost, green sand, cottonseed meal, feather meal, crab meal, fish emulsion, or a combination thereof.

[0076] Possible micronutrient fertilizer materials include, but are not limited to, boric acid, a borate, a boron frit, copper sulfate, a copper frit, a copper chelate, a sodium tetraborate decahydrate, an iron sulfate, an iron oxide, iron ammonium sulfate, an iron frit, an iron chelate, a manganese sulfate, a manganese oxide, a manganese chelate, a manganese chloride, a manganese frit, a sodium molybdate, molybdic acid, a zinc sulfate, a zinc oxide, a zinc carbonate, a zinc frit, zinc phosphate, a zinc chelate, or a combination thereof.

[0077] Possible insecticides include, but are not limited to, an organophosphate, a carbamate, a pyrethroid, an acaricide, an alkyl phthalate, boric acid, a borate, a fluoride, sulfur, a haloaromatic substituted urea, a hydrocarbon ester, a biologically- based insecticide, or a combination thereof.

[0078] Possible herbicides include, but are not limited to, a chlorophenoxy compound, a nitrophenolic compound, a nitrocresolic compound, a dipyridyl compound, an acetamide, an aliphatic acid, an anilide, a benzamide, a benzoic acid, anisic acid, a benzonitrile, benzothiadiazinone dioxide, a thiocarbamate, a carbamate, a carbanilate, chloropyridinyl, a fluorodinitrotoluidine compound, isoxazolidinone, nicotinic acid, isopropylamine, oxadiazolinone, a phosphate, a phthalate, a picolinic acid compound, a triazine, a triazole, a uracil, endothall, sodium chlorate, or a combination thereof.

[0079] Possible fungal inoculants include, but are not limited to, a fungal inoculant of the family Glomeraceae, a fungal inoculant of the family Claroidoglomeraceae, a fungal inoculant of the family Gigasporaceae, a fungal inoculant of the family Acaulosporaceae, a fungal inoculant of the family Sacculosporaceae, a fungal inoculant of the family Entrophosporaceae, a fungal inoculant of the family Pacidsporaceae, a fungal inoculant of the family Diversisporaceae, a fungal inoculant of the family Paraglomeraceae, a fungal inoculant of the family Archaeosporaceae, a fungal inoculant of the family Geosiphonaceae, a fungal inoculant of the family Ambisporaceae, a fungal inoculant of the family Scutellosporaceae, a fungal inoculant of the family Dentiscultataceae, a fungal inoculant of the family Racocetraceae, a fungal inoculant of the phylum Basidiomycota, a fungal inoculant of the phylum Ascomycota, a fungal inoculant of the phylum Zygomycota, or a combination thereof.

[0080] Possible bacterial inoculants include, but are not limited to, a bacterial inoculant of the genus Rhizobium, a bacterial inoculant of the genus Bradyrhizobium, a bacterial inoculant of the genus Mesorhizobium, a bacterial inoculant of the genus Azorhizobium, a bacterial inoculant of the genus Allorhizobium, a bacterial inoculant of the genus Sinorhizobium, a bacterial inoculant of the genus Kluyvera, a bacterial inoculant of the genus Azotobacter, a bacterial inoculant of the genus Pseudomonas, a bacterial inoculant of the genus Azospirillium, a bacterial inoculant of the genus Bacillus, a bacterial inoculant of the genus Streptomyces, a bacterial inoculant of the genus Paenibacillus, a bacterial inoculant of the genus Paracoccus, a bacterial inoculant of the genus Enterobacter, a bacterial inoculant of the genus Alcaligenes, a bacterial inoculant of the genus Mycobacterium, a bacterial inoculant of the genus Trichoderma, a bacterial inoculant of the genus Gliocladium, a bacterial inoculant of the genus Glomus, a bacterial inoculant of the genus Klebsiella, or a combination thereof.

[0081] Another aspect of the present disclosure is a method for treating a plant or a seed thereof including one or more of transplanting the plant or the seed thereof into a soil composition; administering to the plant or the seed thereof an effective amount of an isolated bacterial strain selected from Bacillus subtilis MH1 , Bacillus pseudomycoides MH2 or a combination thereof; and / or cultivating the plant or seed thereof in an environment after the administering of the effective amount of the isolated bacterial strain.

[0082] In exemplary embodiments, the effective amount of the isolated bacterial strain is at most 2 x 108CFU / mL. In other embodiments, the effective amount of the isolated bacterial strain is any amount or range of amounts falling within the range of 1 x 104CFU / mL to 2 x 108CFU / mL. In exemplary embodiments, the isolated bacterial strain is administered under one or more of the following conditions: a nitrogen content ranging from 0% to 100%, 0% to 80%, 0% to 75%, 0% to 60%, 0% to 50%, 0% to 40%, 0% to 30%, 0% to 20%, 0% to 10% or any percentage or range of percentages falling within 0% to 100% of a recommend nitrogen level for a growth stage of the plant or seed thereof; a phosphorus content ranging from 80% to 100%, 85% to 100%, 90% to 100%, 95% to 100% or any percentage or range of percentages falling within 80% to 100% of a recommend phosphorus level for the growth stage of the plant or seed thereof; and / or a potassium content ranging from 80% to 100%, 85% to 100%, 90% to 100%, 95% to 100% or any percentage or range of percentages falling within 80% to 100% of a recommend potassium level for the growth stage of the plant or seed thereof.

[0083] In exemplary embodiments, the cultivating of the plant or seed thereof occurs in environments having from 0 to 120 lbs of nitrogen per acre, 0 to 110 lbs of nitrogen per acre, 0 to 100 lb of nitrogen per acre, 0 to 90 lbs of nitrogen per acre, 0 to 80 lbs of nitrogen per acre, 0-70 lbs of nitrogen per acre, 0 to 60 lbs of nitrogen per acre, 0 to 50 lbs of nitrogen per acre, 0 to 40 lbs of nitrogen per acre, 0 to 30 lbs of nitrogen per acre, 0 to 20 lbs of nitrogen per acre, 0 to 10 lbs of nitrogen per acre, or an amount or range of amounts for nitrogen per acre falling within the range of 0 to 120 lbs of nitrogen per acre.

[0084] In exemplary embodiments, the cultivating of the plant or seed thereof occurs in environments having 44 to 55 lbs of phosphorus per acre, 46 to 55 lbs of phosphorus per acre, 48 to 55 lbs of phosphorus per acre, 50 to 55 lbs of phosphorus per acre, 52 to 55 lbs of phosphorus per acre, or any amount of range of amounts falling within the range of 44 to 55 lbs of phosphorus per acre.

[0085] In exemplary embodiments, the cultivating of the plant or seed thereof occurs in environments having 26 to 33 lbs of potassium per acre, 28 to 33 lbs of potassium per acre, 30 to 33 lbs of potassium per acre, or any amount or range of amounts falling within the range of 26 to 33 lbs of potassium per acre.

[0086] In exemplary embodiments, the cultivating of the plant or seed thereof occurs in environments having 26 to 33 lbs of zinc per acre, 28 to 33 lbs of zinc per acre, 30 to 33 lbs of zinc per acre, or any amount or range of amounts falling within the range of 26 to 33 lbs of zinc per acre.

[0087] In exemplary embodiments, the isolated bacterial strain is administered to an aerial part of the plant.

[0088] In exemplary embodiments, the isolated bacterial strain is administered to at least a portion of a root of the plant.

[0089] In exemplary embodiments, the method includes providing foliar resistance to a pathogen to the plant after the administering of the isolated bacterial strain. Possible pathogens include, but are not limited to, Clarireedia jacksonii, Sclerotinia sclerotiorum, Diaporthae ueckerae, Rhizoctonia solani, and Bipolaris zeicola.

[0090] In exemplary embodiments, the effective amount of the isolated bacterial strain selected from Bacillus subtilis MH1 , Bacillus pseudomycoides MH2 or a combination thereof inhibits the mycelail growth of Clarireedia jacksonii by at least 25%, by at least 20%, by at least 15%, by at least 10%, by at least 5%, by 5 to 25% or any percentage or range of percentages falling within the range of 5 to 25% when compared to the mycelail growth of Clarireedia jacksonii that are not exposed or subjected to the effective amount of Bacillus subtilis MH1 , Bacillus pseudomycoides MH2 or a combination thereof.

[0091] In exemplary embodiments, the effective amount of the isolated bacterial strain selected from Bacillus subtilis MH1 , Bacillus pseudomycoides MH2 or a combination thereof inhibits the mycelail growth of Sclerotinia sclerotiorum by at least 30%, at least 25%, by at least 20%, by at least 15%, by at least 10%, by at least 5%, by 5 to 30% or any percentage or range of percentages falling within the range of 5 to 30% when compared to the mycelail growth of Sclerotinia sclerotiorum that are not exposed or subjected to the effective amount of Bacillus subtilis MH1 , Bacillus pseudomycoides MH2 or a combination thereof.

[0092] In exemplary embodiments, the effective amount of the isolated bacterial strain selected from Bacillus subtilis MH1 , Bacillus pseudomycoides MH2 or a combination thereof inhibits the mycelail growth of Diaporthae ueckerae by at least 40%, by at least 35%, by at least 30%, at least 25%, by at least 20%, by at least 15%, by at least 10%, by at least 5%, by 5 to 40% or any percentage or range of percentages falling within the range of 5 to 40% when compared to the mycelail growth of Diaporthae ueckerae that are not exposed or subjected to the effective amount of Bacillus subtilis MH1 , Bacillus pseudomycoides MH2 or a combination thereof.

[0093] In exemplary embodiments, the effective amount of the isolated bacterial strain selected from Bacillus subtilis MH1 , Bacillus pseudomycoides MH2 or a combination thereof inhibits the mycelail growth of Rhizoctonia solani by at least 40%, by at least 35%, by at least 30%, at least 25%, by at least 20%, by at least 15%, by at least 10%, by at least 5%, by 5 to 40% or any percentage or range of percentages falling within the range of 5 to 40% when compared to the mycelail growth of Rhizoctonia solani that are not exposed or subjected to the effective amount of Bacillus subtilis MH1 , Bacillus pseudomycoides MH2 or a combination thereof.

[0094] In exemplary embodiments, the effective amount of the isolated bacterial strain selected from Bacillus subtilis MH1 , Bacillus pseudomycoides MH2 or a combination thereof inhibits the mycelail growth of Bipolaris zeicola by at least 35%, by at least 30%, at least 25%, by at least 20%, by at least 15%, by at least 10%, by at least 5%, by 5 to 35% or any percentage or range of percentages falling within the range of 5 to 35% when compared to the mycelail growth of Bipolaris zeicola that are not exposed or subjected to the effective amount of Bacillus subtilis MH1 , Bacillus pseudomycoides MH2 or a combination thereof.

[0095] In exemplary embodiments, the plant or the seed thereof is a monocotyledon crop plant selected from, but not limited to, rice, wheat, corn, barley, rye, pineapples, dates, bananas, sugarcane, onions, garlic, palms, bamboo, asparagus, ginger, turmeric, cardamom or any plant having one or more of the following characteristics: a seed with a single cotyledon, or seed leaf, in their embryo; leaves having parallel veins that run the length of the leaf; leaves that are long and slender with smooth margins; an adventitious root system (meaning their roots can grow from almost any part of the plant that touches the soil); roots that are fibrous and web out in many directions, occupying the upper level of the soil; flowers with parts in multiples of three, such as three or six petals; stems with scattered vascular bundles; an underground storage organ (e.g., a bulb, corm, or rhizome); and / or pollen grains with a single aperture, or furrow. In exemplary embodiments, the plant or seed thereof is a dicotyledon crop plant selected from, but not limited to, soybean, cotton, potato, beans, sugar beet, melons, squash, pumpkins, flax, peppers, amaranth, lentils, peas, peanuts, roses, geraniums, carrots, tomato, lettuce, spinach, mint or any plant having one or more of the following characteristics: two seed leaves, or cotyledons, within the seed; a tap root system (which means they have one main root with smaller roots branching off); leaves with net venation (meaning the veins branch out in a network pattern); leaves with a ground tissue made up of two types of mesophyll (palisade and spongy); stems with vascular bundles arranged in a ring; stems having a layer of actively growing cells called the cambium, which allows the stem to grow laterally and increase in diameter; flowers with whorls made up of four or five parts; and / or pollen with three pores or furrows.

[0096] In exemplary embodiments, the growing occurs under anaerobic conditions and the isolated bacterial strain is Bacillus subtilis MH1 , Bacillus pseudomycoides MH2 or a combination of both.

[0097] In exemplary embodiments, the method includes one or more of: increasing root colonization in the plant or the seed thereof after the administering of the isolated bacterial strain; increasing resistance to a pathogen in an aerial part of the plant after the administering of the isolated bacterial strain; increasing growth of the plant or the seed thereof after the administering of the isolated bacterial strain; and / or increasing nitrogen uptake in the plant or the seed thereof after the administering of the isolated bacterial strain when compared to the root colonization, aerial pathogen resistance, plant growth, and / or nitrogen uptake of a plant that was not administered the isolated bacterial strain.

[0098] In exemplary embodiments, the cultivating occurs from 1 to 160 days, 1 day to 130 days, 1 day to 110 days, 1 day to 100 days, 1 day to 80 days, 1 day to 70 days, 1 day to 60 days, 1 day to 30 days, or any amount of time or range of times falling with 1 day to 160 days. In exemplary embodiments, the cultivating occurs until the plant reaches the harvesting stage of its growth cycle. Those of ordinary skill in the art readily capable of determining when a plant has reached its harvesting stage by, for example, the number of mature fruits present on the plant. Other characteristics, such as plant height or stem thickness, can also be used to determine whether a plant has reached its harvesting stage.

[0099] In exemplary embodiments, the method includes coating the seed, before transplanting, with an effective amount of a biologically pure bacterial culture, wherein the bacterium in the bacterial culture includes Bacillus subtilis MH1 , Bacillus pseudomycoides MH2 or a combination thereof. The effective amount can be at most 2 x 108CFU / mL, or any amount or range of amounts falling within the range of 1 x 104CFU / mL to 2 x 108CFU / mL.

[0100] In exemplary embodiments, the effective amount of the isolated bacterial strain recruits plant growth promoting rhizobacteria (PGPR) from the environment to the plant or the seed thereof.

[0101] In exemplary embodiments, the biologically pure bacterial culture includes an effective amount of Bacillus subtilis MH1 and the effective amount of Bacillus subtilis MH1 improves the root length of the plant by at least 45%, by at least 40%, by at least 35%, by at least 30%, by at least 20%, by at least 15%, by at least 10%, by at least 5%, by 5 to 45%, or by any percentage or range of percentages falling within the range of 5 to 45% when compared to the root length of the plant not exposed or subjected to the effective amount of Bacillus subtilis MH1.

[0102] In exemplary embodiments, the biologically pure bacterial culture includes an effective amount of Bacillus subtilis MH1 and the effective amount of Bacillus subtilis MH1 improves the shoot length of the plant by at least 55%, by at least 50%, by at least 45%, by at least 40%, by at least 35%, by at least 30%, by at least 20%, by at least 15%, by at least 10%, by at least 5%, by 5 to 55%, or by any percentage or range of percentages falling within the range of 5 to 55% when compared to the shoot length of the plant not exposed or subjected to the effective amount of Bacillus subtilis MH1 .

[0103] In exemplary embodiments, the biologically pure bacterial culture includes an effective amount of Bacillus subtilis MH1 and the effective amount of Bacillus subtilis MH1 improves the fresh shoot weight of the plant by at least 170%, by at least 150%, by at least 130%, by at least 110%, by at least 90%, by at least 70%, by at least 50%, by at least 40%, by at least 35%, by at least 30%, by at least 20%, by at least 15%, by at least 10%, by at least 5%, by 5 to 170%, or by any percentage or range of percentages falling within the range of 5 to 170% when compared to the fresh shoot weight of the plant not exposed or subjected to the effective amount of Bacillus subtilis MH1 .

[0104] In exemplary embodiments, the biologically pure bacterial culture includes an effective amount of Bacillus subtilis MH1 and the effective amount of Bacillus subtilis MH1 improves the fresh root weight of the plant by at least 140%, by at least 130%, by at least 1 10%, by at least 90%, by at least 70%, by at least 50%, by at least 40%, by at least 35%, by at least 30%, by at least 20%, by at least 15%, by at least 10%, by at least 5%, by 5 to 140%, or by any percentage or range of percentages falling within the range of 5 to 140% when compared to the fresh root weight of the plant not exposed or subjected to the effective amount of Bacillus subtilis MH1 .

[0105] In exemplary embodiments, the biologically pure bacterial culture includes an effective amount of Bacillus subtilis MH1 and the effective amount of Bacillus subtilis MH1 improves the dry root weight of the plant by at least 320%, by at least 300%, by at least 250%, by at least 200%, by at least 150%, by at least 100%, by at least 80%, by at least 40%, by at least 30%, by at least 20%, by at least 15%, by at least 10%, by at least 5%, by 5 to 320%, or by any percentage or range of percentages falling within the range of 5 to 320% when compared to the dry root weight of the plant not exposed or subjected to the effective amount of Bacillus subtilis MH1 .

[0106] In exemplary embodiments, the biologically pure bacterial culture includes an effective amount of Bacillus subtilis MH1 and the effective amount of Bacillus subtilis MH1 improves the dry shoot weight of the plant by at least 420%, by at least 400%, by at least 350%, by at least 300%, by at least 250%, by at least 200%, by at least 150%, by at least 100%, by at least 80%, by at least 40%, by at least 30%, by at least 20%, by at least 15%, by at least 10%, by at least 5%, by 5 to 420%, or by any percentage or range of percentages falling within the range of 5 to 420% when compared to the dry shoot weight of the plant not exposed or subjected to the effective amount of Bacillus subtilis MH1 .

[0107] In exemplary embodiments, the biologically pure bacterial culture includes an effective amount of Bacillus pseudomycoides MH2 and the effective amount of Bacillus pseudomycoides MH2 improves the root length of the plant by at least 120%, by at least 100%, by at least 80%, by at least 60%, by at least 50%, by at least 45%, by at least 40%, by at least 35%, by at least 30%, by at least 20%, by at least 15%, by at least 10%, by at least 5%, by 5 to 120%, or by any percentage or range of percentages falling within the range of 5 to 120% when compared to the root length of the plant not exposed or subjected to the effective amount of Bacillus pseudomycoides MH2.

[0108] In exemplary embodiments, the biologically pure bacterial culture includes an effective amount of Bacillus pseudomycoides MH2 and the effective amount of Bacillus pseudomycoides MH2 improves the shoot length of the plant by at least 45%, by at least 40%, by at least 35%, by at least 30%, by at least 20%, by at least 15%, by at least 10%, by at least 5%, by 5 to 45%, or by any percentage or range of percentages falling within the range of 5 to 45% when compared to the shoot length of the plant not exposed or subjected to the effective amount of Bacillus pseudomycoides MH2.

[0109] In exemplary embodiments, the biologically pure bacterial culture includes an effective amount of Bacillus pseudomycoides MH2 and the effective amount of Bacillus pseudomycoides MH2 improves the fresh shoot weight of the plant by at least 750%, by at least 700%, by at least 650%, by at least 600%, by at least 550%, by at least 500%, by at least 450%, by at least 400%, by 5 to 750%, or by any percentage or range of percentages falling within the range of 5 to 750% when compared to the fresh shoot weight of the plant not exposed or subjected to the effective amount of Bacillus pseudomycoides MH2.

[0110] In exemplary embodiments, the biologically pure bacterial culture includes an effective amount of Bacillus pseudomycoides MH2 and the effective amount of Bacillus pseudomycoides MH2 improves the dry shoot weight of the plant by at least 3000%, by at least 2000%, by at least 1000%, by at least 750%, by at least 700%, by at least 650%, by at least 600%, by at least 550%, by at least 500%, by at least 450%, by at least 400%, by 5 to 3000%, or by any percentage or range of percentages falling within the range of 5 to 3000% when compared to the dry shoot weight of the plant not exposed or subjected to the effective amount of Bacillus pseudomycoides MH2. In exemplary embodiments, the biologically pure bacterial culture includes an effective amount of Bacillus pseudomycoides MH2 and the effective amount of Bacillus pseudomycoides MH2 improves the fresh root weight of the plant by at least 750%, by at least 700%, by at least 650%, by at least 600%, by at least 550%, by at least 500%, by at least 450%, by at least 400%, by 5 to 750%, or by any percentage or range of percentages falling within the range of 5 to 750% when compared to the fresh root weight of the plant not exposed or subjected to the effective amount of Bacillus pseudomycoides MH2.

[0111] In exemplary embodiments, the biologically pure bacterial culture includes an effective amount of Bacillus pseudomycoides MH2 and the effective amount of Bacillus pseudomycoides MH2 improves the dry root weight of the plant by at least 2500%, by at least 2000%, by at least 1500%, by at least 1000%, by at least 750%, by at least 700%, by at least 650%, by at least 600%, by at least 550%, by at least 500%, by at least 450%, by at least 400%, by 5 to 2500%, or by any percentage or range of percentages falling within the range of 5 to 2500% when compared to the dry root weight of the plant not exposed or subjected to the effective amount of Bacillus pseudomycoides MH2.

[0112] In exemplary embodiments, the biologically pure bacterial culture includes an effective amount of Bacillus subtilis MH1 and the effective amount of Bacillus subtilis MH1 improves the fresh total biomass of the plant, when cultivated with the plant at 0 to 75% of the plant’s recommend nitrogen level, by at least 400%, by at least 350%, by at least 300%, by at least 200%, by at least 100%, by at least 50%, by at least 45%, by at least 40%, by at least 35%, by at least 30%, by at least 20%, by at least 15%, by at least 10%, by at least 5%, by 5 to 400%, or by any percentage or range of percentages falling within the range of 5 to 400% when compared to the fresh total biomass of the plant when the plant is cultivated at 100% of the recommended nitrogen level and without exposure to the effective amount of Bacillus subtilis MH1 .

[0113] In exemplary embodiments, the biologically pure bacterial culture includes an effective amount of Bacillus subtilis MH1 and the effective amount of Bacillus subtilis MH1 improves the fresh total biomass of the plant, when cultivated with the plant at 0 to 75% of the plant’s recommend nitrogen level, by at least 220%, by at least 200%, by at least 100%, by at least 50%, by at least 45%, by at least 40%, by at least 35%, by at least 30%, by at least 20%, by at least 15%, by at least 10%, by at least 5%, by 5 to 220%, or by any percentage or range of percentages falling within the range of 5 to 220% when compared to the fresh total biomass of the plant when the plant is cultivated at 100% of the recommended nitrogen level and with the effective amount of Bacillus subtilis MH1 .

[0114] In exemplary embodiments, the biologically pure bacterial culture includes an effective amount of Bacillus subtilis MH1 and the effective amount of Bacillus subtilis MH1 improves the fresh root weight of the plant, when cultivated with the plant at 0 to 75% of the plant’s recommend nitrogen level, by at least 240%, by at least 200%, by at least 100%, by at least 50%, by at least 45%, by at least 40%, by at least 35%, by at least 30%, by at least 20%, by at least 15%, by at least 10%, by at least 5%, by 5 to 240%, or by any percentage or range of percentages falling within the range of 5 to 240% when compared to the fresh root weight of the plant when the plant is cultivated at 100% of the recommended nitrogen level and without exposure to the effective amount of Bacillus subtilis MH1 .

[0115] In exemplary embodiments, the biologically pure bacterial culture includes an effective amount of Bacillus subtilis MH1 and the effective amount of Bacillus subtilis MH1 improves the fresh root weight of the plant, when cultivated with the plant at 0 to 75% of the plant’s recommend nitrogen level, by at least 130%, by at least 110%, by at least 100%, by at least 50%, by at least 45%, by at least 40%, by at least 35%, by at least 30%, by at least 20%, by at least 15%, by at least 10%, by at least 5%, by 5 to 130%, or by any percentage or range of percentages falling within the range of 5 to 130% when compared to the fresh root weight of the plant when the plant is cultivated at 100% of the recommended nitrogen level and with the effective amount of Bacillus subtilis MH1.

[0116] In exemplary embodiments, the biologically pure bacterial culture includes an effective amount of Bacillus subtilis MH1 and the effective amount of Bacillus subtilis MH1 improves the fresh shoot weight of the plant, when cultivated with the plant at 0 to 75% of the plant’s recommend nitrogen level, by at least 440%, by at least 400%, by at least 350%, by at least 300%, by at least 250%, by at least 200%, by at least 100%, by at least 50%, by at least 45%, by at least 40%, by at least 35%, by at least 30%, by at least 20%, by at least 15%, by at least 10%, by at least 5%, by 5 to 440%, or by any percentage or range of percentages falling within the range of 5 to 440% when compared to the fresh shoot weight of the plant when the plant is cultivated at 100% of the recommended nitrogen level and without exposure to the effective amount of Bacillus subtilis MH1.

[0117] In exemplary embodiments, the biologically pure bacterial culture includes an effective amount of Bacillus subtilis MH1 and the effective amount of Bacillus subtilis MH1 improves the fresh shoot weight of the plant, when cultivated with the plant at 0 to 75% of the plant’s recommend nitrogen level, by at least 245%, by at least 220%, by at least 200%, by at least 150%, by at least 130%, by at least 110%, by at least 100%, by at least 50%, by at least 45%, by at least 40%, by at least 35%, by at least 30%, by at least 20%, by at least 15%, by at least 10%, by at least 5%, by 5 to 245%, or by any percentage or range of percentages falling within the range of 5 to 245% when compared to the fresh shoot weight of the plant when the plant is cultivated at 100% of the recommended nitrogen level and with the effective amount of Bacillus subtilis MH1 .

[0118] In exemplary embodiments, the biologically pure bacterial culture includes an effective amount of Bacillus subtilis MH1 and the effective amount of Bacillus subtilis MH1 improves the dry total biomass of the plant, when cultivated with the plant at 0 to 75% of the plant’s recommend nitrogen level, by at least 350%, by at least 300%, by at least 250%, by at least 200%, by at least 100%, by at least 50%, by at least 45%, by at least 40%, by at least 35%, by at least 30%, by at least 20%, by at least 15%, by at least 10%, by at least 5%, by 5 to 350%, or by any percentage or range of percentages falling within the range of 5 to 350% when compared to the dry total biomass of the plant when the plant is cultivated at 100% of the recommended nitrogen level and without exposure to the effective amount of Bacillus subtilis MH1 .

[0119] In exemplary embodiments, the biologically pure bacterial culture includes an effective amount of Bacillus subtilis MH1 and the effective amount of Bacillus subtilis MH1 improves the dry total biomass of the plant, when cultivated with the plant at 0 to 75% of the plant’s recommend nitrogen level, by at least 100%, by at least 95%, by at least 50%, by at least 45%, by at least 40%, by at least 35%, by at least 30%, by at least 20%, by at least 15%, by at least 10%, by at least 5%, by 5 to 100%, or by any percentage or range of percentages falling within the range of 5 to 100% when compared to the dry total biomass of the plant when the plant is cultivated at 100% of the recommended nitrogen level and with the effective amount of Bacillus subtilis MH1 .

[0120] In exemplary embodiments, the biologically pure bacterial culture includes an effective amount of Bacillus subtilis MH1 and the effective amount of Bacillus subtilis MH1 improves the dry root weight of the plant, when cultivated with the plant at 0 to 75% of the plant’s recommend nitrogen level, by at least 320%, by at least 300%, by at least 250%, by at least 200%, by at least 100%, by at least 50%, by at least 45%, by at least 40%, by at least 35%, by at least 30%, by at least 20%, by at least 15%, by at least 10%, by at least 5%, by 5 to 320%, or by any percentage or range of percentages falling within the range of 5 to 320% when compared to the dry root weight of the plant when the plant is cultivated at 100% of the recommended nitrogen level and without exposure to the effective amount of Bacillus subtilis MH1 .

[0121] In exemplary embodiments, the biologically pure bacterial culture includes an effective amount of Bacillus subtilis MH1 and the effective amount of Bacillus subtilis MH1 improves the dry root weight of the plant, when cultivated with the plant at 0 to 75% of the plant’s recommend nitrogen level, by at least 90%, by at least 85%, by at least 50%, by at least 45%, by at least 40%, by at least 35%, by at least 30%, by at least 20%, by at least 15%, by at least 10%, by at least 5%, by 5 to 90%, or by any percentage or range of percentages falling within the range of 5 to 90% when compared to the dry root weight of the plant when the plant is cultivated at 100% of the recommended nitrogen level and with the effective amount of Bacillus subtilis MH1 .

[0122] In exemplary embodiments, the biologically pure bacterial culture includes an effective amount of Bacillus subtilis MH1 and the effective amount of Bacillus subtilis MH1 improves the dry shoot weight of the plant, when cultivated with the plant at 0 to 75% of the plant’s recommend nitrogen level, by at least 360%, by at least 320%, by at least 300%, by at least 250%, by at least 200%, by at least 100%, by at least 50%, by at least 45%, by at least 40%, by at least 35%, by at least 30%, by at least 20%, by at least 15%, by at least 10%, by at least 5%, by 5 to 360%, or by any percentage or range of percentages falling within the range of 5 to 360% when compared to the dry shoot weight of the plant when the plant is cultivated at 100% of the recommended nitrogen level and without exposure to the effective amount of Bacillus subtilis MH1 .

[0123] In exemplary embodiments, the biologically pure bacterial culture includes an effective amount of Bacillus subtilis MH1 and the effective amount of Bacillus subtilis MH1 improves the dry shoot weight of the plant, when cultivated with the plant at 0 to 75% of the plant’s recommend nitrogen level, by at least 100%, by at least 90%, by at least 85%, by at least 50%, by at least 45%, by at least 40%, by at least 35%, by at least 30%, by at least 20%, by at least 15%, by at least 10%, by at least 5%, by 5 to 100%, or by any percentage or range of percentages falling within the range of 5 to 100% when compared to the dry shoot weight of the plant when the plant is cultivated at 100% of the recommended nitrogen level and with the effective amount of Bacillus subtilis MH1 .

[0124] In exemplary embodiments, the biologically pure bacterial culture includes an effective amount of Bacillus subtilis MH1 and the effective amount of Bacillus subtilis MH1 improves the fruit yield (grams of fruit / plant) of the plant, when cultivated with the plant at 0 to 75% of the plant’s recommend nitrogen level, by at least 160%, by at least 120%, by at least 100%, by at least 50%, by at least 45%, by at least 40%, by at least 35%, by at least 30%, by at least 20%, by at least 15%, by at least 10%, by at least 5%, by 5 to 160%, or by any percentage or range of percentages falling within the range of 5 to 160% when compared to the fruit yield (grams of fruit / plant) of the plant when the plant is cultivated at 100% of the recommended nitrogen level and without exposure to the effective amount of Bacillus subtilis MH1 .

[0125] In exemplary embodiments, the biologically pure bacterial culture includes an effective amount of Bacillus subtilis MH1 and the effective amount of Bacillus subtilis MH1 improves the fruit yield (grams of fruit / plant) of the plant, when cultivated with the plant at 0 to 75% of the plant’s recommend nitrogen level, by at least 80%, by at least 75%, by at least 70%, by at least 50%, by at least 45%, by at least 40%, by at least 35%, by at least 30%, by at least 20%, by at least 15%, by at least 10%, by at least 5%, by 5 to 80%, or by any percentage or range of percentages falling within the range of 5 to 80% when compared to the fruit yield (grams of fruit / pla nt) of the plant when the plant is cultivated at 100% of the recommended nitrogen level and with the effective amount of Bacillus subtilis MH1 .

[0126] In exemplary embodiments, the biologically pure bacterial culture includes an effective amount of Bacillus subtilis MH1 and the effective amount of Bacillus subtilis MH1 improves the yield (tons / hectare) of the plant, when cultivated with the plant at 0 to 75% of the plant’s recommend nitrogen level, by at least 160%, by at least 120%, by at least 100%, by at least 50%, by at least 45%, by at least 40%, by at least 35%, by at least 30%, by at least 20%, by at least 15%, by at least 10%, by at least 5%, by 5 to 160%, or by any percentage or range of percentages falling within the range of 5 to 160% when compared to the yield (tons / hectare) of the plant when the plant is cultivated at 100% of the recommended nitrogen level and without exposure to the effective amount of Bacillus subtilis MH1 .

[0127] In exemplary embodiments, the biologically pure bacterial culture includes an effective amount of Bacillus subtilis MH1 and the effective amount of Bacillus subtilis MH1 improves the yield (tons / hectare) of the plant, when cultivated with the plant at 0 to 75% of the plant’s recommend nitrogen level, by at least 80%, by at least 75%, by at least 70%, by at least 50%, by at least 45%, by at least 40%, by at least 35%, by at least 30%, by at least 20%, by at least 15%, by at least 10%, by at least 5%, by 5 to 80%, or by any percentage or range of percentages falling within the range of 5 to 80% when compared to the yield (tons / hectare) of the plant when the plant is cultivated at 100% of the recommended nitrogen level and with the effective amount of Bacillus subtilis MH1 .

[0128] In exemplary embodiments, the biologically pure bacterial culture includes an effective amount of Bacillus subtilis MH1 and the effective amount of Bacillus subtilis MH1 improves the nitrogen content percentage of the plant, when cultivated with the plant at 0 to 75% of the plant’s recommend nitrogen level, by at least 55%, by at least 50%, by at least 45%, by at least 40%, by at least 35%, by at least 30%, by at least 20%, by at least 15%, by at least 10%, by at least 5%, by 5 to 55%, or by any percentage or range of percentages falling within the range of 5 to 55% when compared to the nitrogen content percentage of the plant when the plant is cultivated at 100% of the recommended nitrogen level and without exposure to the effective amount of Bacillus subtilis MH1 .

[0129] In exemplary embodiments, the biologically pure bacterial culture includes an effective amount of Bacillus subtilis MH1 and the effective amount of Bacillus subtilis MH1 improves the nitrogen content percentage of the plant, when cultivated with the plant at 0 to 75% of the plant’s recommend nitrogen level, by at least 55%, by at least 50%, by at least 45%, by at least 40%, by at least 35%, by at least 30%, by at least 20%, by at least 15%, by at least 10%, by at least 5%, by 5 to 55%, or by any percentage or range of percentages falling within the range of 5 to 55% when compared to the nitrogen content percentage of the plant when the plant is cultivated at 100% of the recommended nitrogen level and with the effective amount of Bacillus subtilis MH1 .

[0130] In exemplary embodiments, the biologically pure bacterial culture includes an effective amount of Bacillus pseudomycoides MH2 and the effective amount of Bacillus pseudomycoides MH2 improves the fresh total biomass of the plant, when cultivated with the plant at 0 to 75% of the plant’s recommend nitrogen level, by at least 320%, by at least 300%, by at least 250%, by at least 200%, by at least 150%, by at least 100%, by at least 55%, by at least 50%, by at least 45%, by at least 40%, by at least 35%, by at least 30%, by at least 20%, by at least 15%, by at least 10%, by at least 5%, by 5 to 320%, or by any percentage or range of percentages falling within the range of 5 to 320% when compared to the fresh total biomass of the plant when the plant is cultivated at 100% of the recommended nitrogen level and without exposure to the effective amount of Bacillus pseudomycoides MH2.

[0131] In exemplary embodiments, the biologically pure bacterial culture includes an effective amount of Bacillus pseudomycoides MH2 and the effective amount of Bacillus pseudomycoides MH2 improves the fresh total biomass of the plant, when cultivated with the plant at 0 to 75% of the plant’s recommend nitrogen level, by at least 200%, by at least 150%, by at least 100%, by at least 55%, by at least 50%, by at least 45%, by at least 40%, by at least 35%, by at least 30%, by at least 20%, by at least 15%, by at least 10%, by at least 5%, by 5 to 200%, or by any percentage or range of percentages falling within the range of 5 to 200% when compared to the fresh total biomass of the plant when the plant is cultivated at 100% of the recommended nitrogen level and with the effective amount of Bacillus pseudomycoides MH2.

[0132] In exemplary embodiments, the biologically pure bacterial culture includes an effective amount of Bacillus pseudomycoides MH2 and the effective amount of Bacillus pseudomycoides MH2 improves the fresh root weight of the plant, when cultivated with the plant at 0 to 75% of the plant’s recommend nitrogen level, by at least 240%, by at least 230%, by at least 200%, by at least 150%, by at least 100%, by at least 55%, by at least 50%, by at least 45%, by at least 40%, by at least 35%, by at least 30%, by at least 20%, by at least 15%, by at least 10%, by at least 5%, by 5 to 240%, or by any percentage or range of percentages falling within the range of 5 to 240% when compared to the fresh root weight of the plant when the plant is cultivated at 100% of the recommended nitrogen level and without exposure to the effective amount of Bacillus pseudomycoides MH2.

[0133] In exemplary embodiments, the biologically pure bacterial culture includes an effective amount of Bacillus pseudomycoides MH2 and the effective amount of Bacillus pseudomycoides MH2 improves the fresh root weight of the plant, when cultivated with the plant at 0 to 75% of the plant’s recommend nitrogen level, by at least 160%, by at least 130%, by at least 100%, by at least 55%, by at least 50%, by at least 45%, by at least 40%, by at least 35%, by at least 30%, by at least 20%, by at least 15%, by at least 10%, by at least 5%, by 5 to 160%, or by any percentage or range of percentages falling within the range of 5 to 160% when compared to the fresh root weight of the plant when the plant is cultivated at 100% of the recommended nitrogen level and with the effective amount of Bacillus pseudomycoides MH2.

[0134] In exemplary embodiments, the biologically pure bacterial culture includes an effective amount of Bacillus pseudomycoides MH2 and the effective amount of Bacillus pseudomycoides MH2 improves the fresh shoot weight of the plant, when cultivated with the plant at 0 to 75% of the plant’s recommend nitrogen level, by at least 330%, by at least 300%, by at least 250%, by at least 230%, by at least 200%, by at least 150%, by at least 100%, by at least 55%, by at least 50%, by at least 45%, by at least 40%, by at least 35%, by at least 30%, by at least 20%, by at least 15%, by at least 10%, by at least 5%, by 5 to 330%, or by any percentage or range of percentages falling within the range of 5 to 330% when compared to the fresh shoot weight of the plant when the plant is cultivated at 100% of the recommended nitrogen level and without exposure to the effective amount of Bacillus pseudomycoides MH2.

[0135] In exemplary embodiments, the biologically pure bacterial culture includes an effective amount of Bacillus pseudomycoides MH2 and the effective amount of Bacillus pseudomycoides MH2 improves the fresh shoot weight of the plant, when cultivated with the plant at 0 to 75% of the plant’s recommend nitrogen level, by at least 200%, by at least 190%, by at least 160%, by at least 130%, by at least 100%, by at least 55%, by at least 50%, by at least 45%, by at least 40%, by at least 35%, by at least 30%, by at least 20%, by at least 15%, by at least 10%, by at least 5%, by 5 to 200%, or by any percentage or range of percentages falling within the range of 5 to 200% when compared to the fresh shoot weight of the plant when the plant is cultivated at 100% of the recommended nitrogen level and with the effective amount of Bacillus pseudomycoides MH2.

[0136] In exemplary embodiments, the biologically pure bacterial culture includes an effective amount of Bacillus pseudomycoides MH2 and the effective amount of Bacillus pseudomycoides MH2 improves the dry total biomass of the plant, when cultivated with the plant at 0 to 75% of the plant’s recommend nitrogen level, by at least 340%, by at least 300%, by at least 250%, by at least 230%, by at least 200%, by at least 150%, by at least 100%, by at least 55%, by at least 50%, by at least 45%, by at least 40%, by at least 35%, by at least 30%, by at least 20%, by at least 15%, by at least 10%, by at least 5%, by 5 to 340%, or by any percentage or range of percentages falling within the range of 5 to 340% when compared to the dry total biomass of the plant when the plant is cultivated at 100% of the recommended nitrogen level and without exposure to the effective amount of Bacillus pseudomycoides MH2.

[0137] In exemplary embodiments, the biologically pure bacterial culture includes an effective amount of Bacillus pseudomycoides MH2 and the effective amount of Bacillus pseudomycoides MH2 improves the dry total biomass of the plant, when cultivated with the plant at 0 to 75% of the plant’s recommend nitrogen level, by at least 210%, by at least 190%, by at least 160%, by at least 130%, by at least 100%, by at least 55%, by at least 50%, by at least 45%, by at least 40%, by at least 35%, by at least 30%, by at least 20%, by at least 15%, by at least 10%, by at least 5%, by 5 to 210%, or by any percentage or range of percentages falling within the range of 5 to 210% when compared to the dry total biomass of the plant when the plant is cultivated at 100% of the recommended nitrogen level and with the effective amount of Bacillus pseudomycoides MH2.

[0138] In exemplary embodiments, the biologically pure bacterial culture includes an effective amount of Bacillus pseudomycoides MH2 and the effective amount of Bacillus pseudomycoides MH2 improves the dry root weight of the plant, when cultivated with the plant at 0 to 75% of the plant’s recommend nitrogen level, by at least 120%, by at least 100%, by at least 55%, by at least 50%, by at least 45%, by at least 40%, by at least 35%, by at least 30%, by at least 20%, by at least 15%, by at least 10%, by at least 5%, by 5 to 120%, or by any percentage or range of percentages falling within the range of 5 to 120% when compared to the dry root weight of the plant when the plant is cultivated at 100% of the recommended nitrogen level and without exposure to the effective amount of Bacillus pseudomycoides MH2.

[0139] In exemplary embodiments, the biologically pure bacterial culture includes an effective amount of Bacillus pseudomycoides MH2 and the effective amount of Bacillus pseudomycoides MH2 improves the dry root weight of the plant, when cultivated with the plant at 0 to 75% of the plant’s recommend nitrogen level, by at least 65%, by at least 55%, by at least 50%, by at least 45%, by at least 40%, by at least 35%, by at least 30%, by at least 20%, by at least 15%, by at least 10%, by at least 5%, by 5 to 65%, or by any percentage or range of percentages falling within the range of 5 to 65% when compared to the dry root weight of the plant when the plant is cultivated at 100% of the recommended nitrogen level and with the effective amount of Bacillus pseudomycoides MH2.

[0140] In exemplary embodiments, the biologically pure bacterial culture includes an effective amount of Bacillus pseudomycoides MH2 and the effective amount of Bacillus pseudomycoides MH2 improves the dry shoot weight of the plant, when cultivated with the plant at 0 to 75% of the plant’s recommend nitrogen level, by at least 360%, by at least 300%, by at least 250%, by at least 200%, by at least 150%, by at least 120%, by at least 100%, by at least 55%, by at least 50%, by at least 45%, by at least 40%, by at least 35%, by at least 30%, by at least 20%, by at least 15%, by at least 10%, by at least 5%, by 5 to 360%, or by any percentage or range of percentages falling within the range of 5 to 360% when compared to the dry shoot weight of the plant when the plant is cultivated at 100% of the recommended nitrogen level and without exposure to the effective amount of Bacillus pseudomycoides MH2.

[0141] In exemplary embodiments, the biologically pure bacterial culture includes an effective amount of Bacillus pseudomycoides MH2 and the effective amount of Bacillus pseudomycoides MH2 improves the dry shoot weight of the plant, when cultivated with the plant at 0 to 75% of the plant’s recommend nitrogen level, by at least 220%, by at least 200%, by at least 150%, by at least 100%, by at least 65%, by at least 55%, by at least 50%, by at least 45%, by at least 40%, by at least 35%, by at least 30%, by at least 20%, by at least 15%, by at least 10%, by at least 5%, by 5 to 220%, or by any percentage or range of percentages falling within the range of 5 to 220% when compared to the dry shoot weight of the plant when the plant is cultivated at 100% of the recommended nitrogen level and with the effective amount of Bacillus pseudomycoides MH2.

[0142] In exemplary embodiments, the biologically pure bacterial culture includes an effective amount of Bacillus pseudomycoides MH2 and the effective amount of Bacillus pseudomycoides MH2 improves the fruit yield (grams of fruit / plant) of the plant, when cultivated with the plant at 0 to 75% of the plant’s recommend nitrogen level, by at least 410%, by at least 360%, by at least 300%, by at least 250%, by at least 200%, by at least 150%, by at least 120%, by at least 100%, by at least 55%, by at least 50%, by at least 45%, by at least 40%, by at least 35%, by at least 30%, by at least 20%, by at least 15%, by at least 10%, by at least 5%, by 5 to 410%, or by any percentage or range of percentages falling within the range of 5 to 410% when compared to the fruit yield (grams of fruit / plant) of the plant when the plant is cultivated at 100% of the recommended nitrogen level and without exposure to the effective amount of Bacillus pseudomycoides MH2. In exemplary embodiments, the biologically pure bacterial culture includes an effective amount of Bacillus pseudomycoides MH2 and the effective amount of Bacillus pseudomycoides MH2 improves the fruit yield (grams of fruit / plant) of the plant, when cultivated with the plant at 0 to 75% of the plant’s recommend nitrogen level, by at least 170%, by at least 150%, by at least 100%, by at least 65%, by at least 55%, by at least 50%, by at least 45%, by at least 40%, by at least 35%, by at least 30%, by at least 20%, by at least 15%, by at least 10%, by at least 5%, by 5 to 170%, or by any percentage or range of percentages falling within the range of 5 to 170% when compared to the fruit yield (grams of fruit / plant) of the plant when the plant is cultivated at 100% of the recommended nitrogen level and with the effective amount of Bacillus pseudomycoides MH2.

[0143] In exemplary embodiments, the biologically pure bacterial culture includes an effective amount of Bacillus pseudomycoides MH2 and the effective amount of Bacillus pseudomycoides MH2 improves the yield (tons / hectare) of the plant, when cultivated with the plant at 0 to 75% of the plant’s recommend nitrogen level, by at least 410%, by at least 360%, by at least 300%, by at least 250%, by at least 200%, by at least 150%, by at least 120%, by at least 100%, by at least 55%, by at least 50%, by at least 45%, by at least 40%, by at least 35%, by at least 30%, by at least 20%, by at least 15%, by at least 10%, by at least 5%, by 5 to 410%, or by any percentage or range of percentages falling within the range of 5 to 410% when compared to the yield (tons / hectare) of the plant when the plant is cultivated at 100% of the recommended nitrogen level and without exposure to the effective amount of Bacillus pseudomycoides MH2.

[0144] In exemplary embodiments, the biologically pure bacterial culture includes an effective amount of Bacillus pseudomycoides MH2 and the effective amount of Bacillus pseudomycoides MH2 improves the yield (tons / hectare) of the plant, when cultivated with the plant at 0 to 75% of the plant’s recommend nitrogen level, by at least 175%, by at least 150%, by at least 100%, by at least 65%, by at least 55%, by at least 50%, by at least 45%, by at least 40%, by at least 35%, by at least 30%, by at least 20%, by at least 15%, by at least 10%, by at least 5%, by 5 to 175%, or by any percentage or range of percentages falling within the range of 5 to 175% when compared to the yield (tons / hectare) of the plant when the plant is cultivated at 100% of the recommended nitrogen level and with the effective amount of Bacillus pseudomycoides MH2.

[0145] In exemplary embodiments, the biologically pure bacterial culture includes an effective amount of Bacillus pseudomycoides MH2 and the effective amount of Bacillus pseudomycoides MH2 improves the nitrogen content percentage of the plant, when cultivated with the plant at 0 to 75% of the plant’s recommend nitrogen level, by at least 55%, by at least 50%, by at least 45%, by at least 40%, by at least 35%, by at least 30%, by at least 20%, by at least 15%, by at least 10%, by at least 5%, by 5 to 55%, or by any percentage or range of percentages falling within the range of 5 to 55% when compared to the nitrogen content percentage of the plant when the plant is cultivated at 100% of the recommended nitrogen level and without exposure to the effective amount of Bacillus pseudomycoides MH2.

[0146] In exemplary embodiments, the biologically pure bacterial culture includes an effective amount of Bacillus pseudomycoides MH2 and the effective amount of Bacillus pseudomycoides MH2 improves the nitrogen content percentage of the plant, when cultivated with the plant at 0 to 75% of the plant’s recommend nitrogen level, by at least 55%, by at least 50%, by at least 45%, by at least 40%, by at least 35%, by at least 30%, by at least 20%, by at least 15%, by at least 10%, by at least 5%, by 5 to 55%, or by any percentage or range of percentages falling within the range of 5 to 55% when compared to the nitrogen content percentage of the plant when the plant is cultivated at 100% of the recommended nitrogen level and with the effective amount of Bacillus pseudomycoides MH2.

[0147] Another aspect of the present disclosure is a method for treating a plant or a seed thereof. The method can include growing the plant or the seed thereof in a soil composition. The soil composition can be or include an organic layer of a precolonial soil. The method can include administering to the plant or the seed thereof one or more bacterial strains isolated from the POS. The one or more isolated bacterial strains can be selected from the group of Bacillus subtilis MH1 (ATCC Accession No. PTA-127648), Bacillus pseudomycoides MH2 (ATCC Accession No. PTA-127649), or a combination thereof. The one or more isolated bacterial strains can be administered to a root interface or an aerial part of the plant. The method can include administering one or more isolated bacterial strains in an amount effective for recruiting plant growth promoting rhizobacteria (PGPR) to the plant or the seed thereof.

[0148] Beneficial soil bacteria confer immunity against a wide range of foliar diseases by activating plant defenses, thereby reducing a plant's susceptibility to pathogen attack. Several biocontrol bacteria, also known as plant growth promoting rhizobacteria (PGPR), protect plants from soil-borne pathogens. Such bacteria colonizing on the roots can also induce systemic pathogen resistance in aerial plant parts, such as leaves, which are spatially separated from the inducing PGPR. This mechanism of induction of systemic resistance by root colonizing rhizobacteria in aerial plant parts is referred to as induced systemic resistance (ISR). Various PGPR strains are known to induce ISR in plants against various soil and air borne bacterial and fungal pathogens.

[0149] In exemplary embodiments, the method includes administering the one or more isolated bacterial strains in an amount effective for promoting growth of the plant or the seed thereof. The method can include growing the plant or the seed thereof under limited nitrogen, phosphorous and / or potassium (NPK) conditions. Limited NPK conditions can include reduced concentrations of inorganic NPK when compared to the NPK conditions of an agricultural soil and / or the recommend NPK levels a plant requires at various growth stages.

[0150] The method can also include administering the one or more isolated bacterial strains in an amount effective for promoting root colonization in the plant or the seed thereof. In exemplary embodiments, the effective amount is at most 2 x 108CFU / mL. In other embodiments, the effective amount of the isolated bacterial strain is any amount or range of amounts falling within the range of 1 x 104CFU / mL to 2 x 108CFU / mL.

[0151] The terms "inoculating with", "administering to" and "applying to" are used herein interchangeably.

[0152] The method can include administering the one or more isolated bacterial strains in an amount effective for increasing resistance to a pathogen in an aerial part of the plant. The resistance in the aerial part of the plant can include foliar resistance to the pathogen. The pathogen can be, but is not limited to, a fungal pathogen selected from the group consisting of Clarireedia jacksonii, Sclerotinia sclerotiorum, Diaporthae ueckerae, Rhizoctonia solani, and Bipolaris zeicola.

[0153] In exemplary embodiments, the plant is a crop plant. The crop plant can be selected from, but is not limited to, tomato, onion, corn, any dicotyledon or any monocotyledon.

[0154] Another aspect of the present disclosure is a method for promoting the health of a plant grown in a soil composition, the method including administering an isolated PGPR from a POS. The PGPR can be selected from, but are not limited to, Bacillus subtilis MH1 , Bacillus pseudomycoides MH2 or a combination thereof.

[0155] Another aspect of the present disclosure is a method for promoting the health of a plant grown in a soil composition, the method including administering Bacillus subtilis MH1 and / or Bacillus pseudomycoides MH2 to the soil composition in an amount effective to promote growth and restrict pathogens. In exemplary embodiments, the plant is root inoculated or seed inoculated with Bacillus subtilis MH1 and / or Bacillus pseudomycoides MH2. Root or seed inoculation can provide advantages over alternative methods of applying bacterial strains directly to aerial parts of the plants (e.g., plant leaves). Applying bioinoculums directly to the aerial parts of the plant, such as by aerially spraying bioinoculums onto the plant, can have harmful or toxic effects on the surrounding environment. Thus, root or seed inoculation provides a safer method for administering the beneficial microbe to a plant, as the surrounding air and foliage are less likely to meet the bioinoculums.

[0156] In exemplary embodiments, the methods disclosed herein include administering Bacillus subtilis MH1 and / or Bacillus pseudomycoides MH2 to a plant under nitrogen poor and / or limited fertilizer conditions.

[0157] As used herein, “nitrogen poor” and / or “limited fertilizer conditions” mean conditions that contain amounts of nitrogen, phosphorus, and / or potassium that are below a recommended level for the plant. Nitrogen poor and limited fertilizer conditions can be present throughout the entire growth cycle of the plant or can be present at one or more growth stages of the plant.

[0158] The inventors have surprisingly discovered that isolated strains of Bacillus subtilis MH1 and Bacillus pseudomycoides MH2 promote plant growth under limited fertilizer application, indicating that these strains can support uptake of nutrients from soil under NPK limited conditions. Another aspect of the present disclosure, therefore, is a method for promoting the health of a plant grown in a soil composition, the method including administering Bacillus subtilis MH1 and / or Bacillus pseudomycoides MH2 to the plant or the soil composition of the plant in an amount effective to increase pathogen resistance and nutrient-limiting conditions.

[0159] Bacillus subtilis MH1 and Bacillus pseudomycoides MH2 have been discovered to effectively colonize on the roots of plants. This trait of effective root colonization is prudent for a bioinoculum’s ability to persist in soil and be able to promote growth and protect plants against any biotic stressors.

[0160] Another aspect of the present disclosure is a plant seed coated with a composition including: an effective amount of a biologically pure bacterial culture, wherein the bacteria in the bacterial culture includes Bacillus subtilis MH1 (ATCC Accession No. PTA-127648), Bacillus pseudomycoides MH2 (ATCC Accession No. PTA-127649) or a combination thereof, and wherein the composition has a nitrogen content ranging from 0% to 75% of a recommend nitrogen level for growth of the seed.

[0161] In exemplary embodiments, the composition coated on the seed comprises an effective amount of an auxiliary plant growth promoting rhizobacteria (PGPR). Possible auxiliary PGPR that can be include in the composition are, but not limited to, Bacillus subtilis SJ-101 , Pseudomonas species, Bradyrizobium species, Sinorhizobium species, Brevibacillus species, Bacillus species, Rhizobacteria species, Rhizobium species, Rhizobacterium species, and any bacterial species known to possess one or more of the following characteristics: live freely and colonize plant roots, metabolize phosphates, produce hormones, fix nitrogen, promote plant water and nutrient uptake, enhance root development, increase plant enzymatic activity, synthesize phytohormones and their analogs, produce siderophores, produce antioxidant enzymes, or produce and release cytokinins.

[0162] In exemplary embodiments, composition coated on the seed includes one or more of: a phosphorus content ranging from 90% to 100% of a recommend phosphorus level for growth of the seed, a potassium content ranging from 90% to 100% of a recommend potassium level for growth stage of the seed, or any combination thereof. In exemplary embodiments, the composition coated on the seed comprises an agriculturally acceptable carrier selected from one or more of a dispersant, a surfactant, an additive, water, a thickener, an anti-caking agent, residue breakdown, a composting formulation, a granular application, diatomaceous earth, an oil, a coloring agent, a stabilizer, a preservative, a polymer, or an additional coating.

[0163] In exemplary embodiments, the composition coated on the seed is a liquid formulation or a solid formulation, wherein the liquid formulation is an aqueous or oil-based solution, and wherein the solid formulation is a powder agent or granular formulation.

[0164] Another aspect of the present disclosure is a kit for treating a plant or a seed thereof including any one of the compositions disclosed herein.

[0165] In exemplary embodiments, the kit contains at least one container storing any one of the compositions disclosed herein.

[0166] In exemplary embodiments, the container is configured to allow application of any composition disclosed herein onto an aerial part or at least a portion of a root of the plant.

[0167] Examples

[0168] The present disclosure will be described in more detail with reference to the following Examples, which shows exemplary embodiments in accordance with the present disclosure. The present disclosure is not limited to these exemplary embodiments.

[0169] The following examples examine the effects that exemplary compositions containing Bacillus subtilis MH1 and Bacillus pseudomycoides MH2 have on plant growth promotion, antifungal resistance, nitrogen fixation and root colonization.

[0170] Example 1 : Evaluation of POS, sterilized POS and TS on Tomato Plant Growth

[0171] Soil was collected from pre-colonial organic soils and sediments from Gramies Run, Cecil County, Maryland (Coordinates: 39.689153, -75.852737). The collected soils and sediments are referred to as pre-colonial organic soil (POS), legacy sediments (LS) and modern topsoil (TS). These soils were used to cultivate Bacillus subtilis MH1 and Bacillus pseudomycoides MH2 serial dilutions. FIG. 1 illustrates the collection of these soils and their respective locations.

[0172] The collected POS was homogenized, and a separate subset was sampled and autoclaved. Accordingly, the three soils used in this example were namely POS, autoclaved POS ( / .e., sterilized POS) and agriculture soil ( / .e., TS). Tomato seeds were sown into pots containing these three different soils. The seedlings were grown under controlled conditions with regular irrigation and nutrient supply through a Hoagland solution. The Hoagland solution contains the 115.03 mg / L of ammonium phosphate monobasic, 2.86 mg / L of boric acid, 656.4 mg / L of calcium nitrate, 0.08 mg / L of cupric sulfate 5H2O, 5.32 mg / L of ferric tartrate 2 H2O, 240.76 mg / L of magnesium sulfate, 1 .81 mg / L of manganese chloride-4H2O, 0.016 mg / L of molybdenum trioxide, 606.6 mg / L potassium nitrate, and 0.22 mg / L zinc sulfate-7H2O.

[0173] One month old tomato seedlings were used for recording biomass parameters. The tomato seedlings were shown to exhibit improved plant growth when grown in POS when compared to POS sterilized and agriculture soil (see FIG. 2). Indeed, the tomato seed grown in POS exhibited a higher shoot length, root length, fresh and dry shoot weight, and fresh and dry root weight when compared to the tomato seeds grown in sterilized POS and agriculture soil (see FIG. 3).

[0174] The results of this example indicated that the collected POS could contain a microbial community capable of supporting and improving plant growth in current agricultural soils.

[0175] Example 2: Detection of microbial communities in POS, LS, sterilized POS and TS

[0176] The presence of a microbial community in each of the soil samples (i.e., in the POS, LS, TS and sterilized POS) was examined via serially diluting each sample and plating each serial dilution. The serial dilution method used in this example is illustrated in FIG. 4. Each serial dilution was cultured for 3-4 days under 30°C in incubator.

[0177] The results of the serial dilutions are depicted in FIG. 5. These results indicated that POS, LS and TS all contained microbial communities. Little-to-no microbial growth was seen with the serial dilutions of the sterilized POS, thereby demonstrating the effectiveness of the autoclave sterilization process.

[0178] Example 3: Investigation of microbial communities in POS

[0179] In view of the improved plant growth observed in tomato plants grown in POS and the detection of microbial growth in serial dilution samples of the collected POS, the microbial biome of the POS was next investigated.

[0180] Compared to concurrent wetland soil and modern agricultural soils, the POS have lower microbial diversity. Both Shannon Index and Chaol showed the alpha diversity is significantly lower than the other two types of soils (see FIG. 6). Analysis of the microbial diversity also showed that POS soil contained distinct microbiomes than other types of soils, with higher abundance of Chloroflexi, Crenarchaeota, Nitrospirota and Sva0485, but lower abundance of Proteobacteria, Verrucomicrobiota, Planctomycetota, Actinobacteriota, Bacteroidota, and Myxococcota. Although low or limiting N concentrations were observed in POS, significant groups of N transforming microbes were found. Specifically, both non- symbiotic and symbiotic nitrogen fixing bacteria were observed in POS. Non- symbiotic bacteria include Beijerinckiaceae and Clostridium, while symbiotic counterparts included a variety of group of microbes, such as Allorhizobium, Neorhizobium, Pararhizobium, Rhizobium, Bradyrhizobium, and Frankiaceae. Corresponding qPCR quantification of nitrogen fixing genes (n / fH) confirmed that POS did contain a higher abundance of nifH genes than other soils (see FIG. 7). The POS soil, both sterilized and unsterilized, was also shown to contain bacterial and fungal community structures vastly different from modern soil community structures (see NMD plots of FIG. 8 and FIG. 9). Overall, compared to modern agricultural soil, both POS and sterilized POS supported plant growth and enriched nitrogen fixing genes in bulk soil and rhizosphere soils, although unsterile POS had the maximum impact on plant growth.

[0181] To further investigate the microbial communities, present in POS, POS soils were homogenized separately under aseptic conditions and were serially diluted using sterile water. The serial dilutions were plated in Luria Bertani media and grown under at 30°C until single bacterial colonies were formed. The single colonies were picked and later purified. DNA from the purified colonies were sequenced through Illumina MiSeq platform at the Delaware Biotechnology Institute in Newark, Delaware, USA and then blasted on NCBI nucleotide blast server to determine their homology via the 16S ribosomal RNA. One bacterial culture had 100% homology with Bacillus subtilis, and thereafter was classified as Bacillus subtilis MH1 . Another bacterial culture had 99% homology with Bacillus pseudomycoides, and thereafter was classified as Bacillus pseudomycoides MH2.

[0182] Example 4: Investigation of Bacillus pseudomycoides MH2 Anaerobic Growth Capability

[0183] The identification of Bacillus pseudomycoides MH2 in the microbial communities of the POS suggested that this bacterial strain could act as a facultative anaerobe given the low oxygen conditions of the POS. To investigate whether Bacillus pseudomycoides MH2 could function as a facultative anaerobe, Bacillus pseudomycoides MH2 was cultured under both aerobic and anerobic conditions (see Behbehani et al, “Simple and convenient method for culturing anaerobic bacteria”, Appl. Environ. Microbiol. 1982 Jan; 43(1):255-256). Specifically, samples containing Bacillus pseudomycoides MH2 were cultured in the presence of oxygen and in the absence of oxygen. The Bacillus pseudomycoides MH2 strains were cultured in duplicates (i.e., two cultures for each condition) and the optical densities (OD) of the bacterial cultures were measured at an absorbance of 600nm. The resulting OD values are provided below in Table 1.

[0184] Table 1 : Anaerobic and Aerobic Growth of Bacillus pseudomycoides MH2

[0185] In view of the similar OD values obtained under both anaerobic and aerobic conditions, there is evidence that Bacillus pseudomycoides MH2 does function as a facultative anaerobe.

[0186] Example 5: Anti-fungal Action of Bacillus subtilis MH1 and Bacillus pseudomycoides MH 2 The anti-fungal properties of both Bacillus subtilis MH1 and Bacillus pseudomycoides MH2 were next investigated. The antifungal properties of Bacillus subtilis MH1 and Bacillus pseudomycoides MH2 were tested against five fungal different plant fungal pathogens (Clarireedia jacksonii, Sclerotinia sclerotiorum, Diaporthae ueckerae, Rhizoctonia solani and Bipolaris zeicola) through paired test using PDA medium. The bacterial isolate B. subtilis UD1022 was used as positive control, as it has demonstrated broad antifungal activity against various plant pathogens (see, e.g., S. Markland, et al. Foodborne Pathog Dis 2015 Oct;12(10):828-35). A 5 mm mycelial disc of fungal pathogen was sub-cultured on a PDA medium in a Petri dish (90 mm) and each bacterial culture was streaked on the same plate. The control plate was inoculated with only fungal pathogen. The results of the co-culturing of the fungal pathogens with the above bacterial strains are depicted in FIG. 10.

[0187] The mycelial growth in control and bacteria inoculated Petri dishes were measured and percent inhibition in mycelial growth was calculated. Five replications were maintained in each case and the entire experiment was repeated twice. The mycelial inhibition percentage of the bacterial strains involved in this experiment and the mycelial growth of the fungal pathogens in the presence of these bacterial strains are depicted in FIG. 11.

[0188] The results from this example overall demonstrate that there was strong antifungal action of both Bacillus subtilis MH1 and Bacillus pseudomycoides MH2 against these fungal pathogens.

[0189] Heat killed cultures of Bacillus subtilis MH1 and Bacillus pseudomycoides MH2 were also evaluated for anti-fungal properties against Clarireedia jacksonii, Sclerotinia sclerotiorum, Diaporthae ueckerae, Rhizoctonia solani and Bipolaris zeicola. The effectiveness of the heat killed cultures is depicted in FIG. 12. Contrary to live bacterial cells, the heat killed bacterial cells displayed reduced anti-fungal activity.

[0190] Example 6: The Effect of Various Bacillus subtilis MH1 Concentrations on Tomato Plant Growth

[0191] In this example, various concentrations of Bacillus subtilis MH1 and their effect on tomato plant growth were investigated. Tomato plants were root-treated with different concentrations of Bacillus subtilis MH1 . The tomato seeds were surface sterilized and then sown in pots using Promix under greenhouse conditions. The pots were subsequently inoculated with bacterial suspensions of respective concentrations three times at weekly interval. The bacterial suspensions contained different concentrations of Bacillus subtilis MH1 isolate multiplied in LB medium. The seedlings were harvested at 35 days after sowing and the root length, shoot length, root weight, and shoot weight (fresh and dry) was documented in each replication. The concentrations that were investigated are as follows: T1 = 1 - 2x104CFU / mL; T2 = 1-2x105CFU / mL; T3 = 1-2x106CFU / mL; T4 = 1-2x107CFU / mL; T5 = 1-2x108CFU / mL; T6 = Control.

[0192] The visual effects that each concentration of Bacillus subtilis MH1 had on tomato plant growth are depicted in FIG. 13. The effects that each concentration of Bacillus subtilis MH1 had on root length, shoot length, fresh root weight, fresh shoot weight, dry root weight, and dry shoot weight can be seen in FIG. 14.

[0193] Example 7: The Effect of Various Bacillus pseudomycoides MH2 Concentrations on Tomato Plant Growth

[0194] In this example, various concentrations of Bacillus pseudomycoides MH2 and their effect on tomato plant growth were investigated. Tomato plants were root- treated with different concentrations of Bacillus pseudomycoides MH2. The tomato seeds were surface sterilized and then sown in pots using Promix under greenhouse conditions. The pots were subsequently inoculated with bacterial suspensions of respective concentrations three times at weekly interval. The bacterial suspensions contained different concentrations of Bacillus pseudomycoides MH2 isolate multiplied in LB medium. The seedlings were harvested at 35 days after sowing and the root length, shoot length, root weight, and shoot weight (fresh and dry) was documented in each replication. The concentrations that were investigated are as follows: T1 = 1 -2x104CFU / mL; T2 = 1- 2x105CFU / mL; T3 = 1-2x106CFU / mL; T4 = 1-2x107CFU / mL; T5 = 1-2x108CFU / mL; T6 = Control.

[0195] The visual effects that each concentration of Bacillus subtilis MH1 had on tomato plant growth are depicted in FIG. 15. The effects that each concentration of Bacillus subtilis MH1 had on root length, shoot length, fresh root weight, fresh shoot weight, dry root weight, and dry shoot weight can be seen in FIG. 16.

[0196] Example 8: Root Colonization of Bacillus subtilis MH1 and Bacillus pseudomycoides MH 2

[0197] The ability of Bacillus subtilis MH1 and Bacillus pseudomycoides MH2 to colonize around the roots of tomato plants was next investigated. Tomato roots were treated with 1x106cells / mL of Bacillus subtilis MH1 , Bacillus pseudomycoides MH2 and B. subtilis UD1022. A negative control wherein the tomato roots were not treated with a bacterial strain was also included. The roots were stained with SYTO13 and analyzed using confocal microscopy for bacterial colonization. The colonization of the above bacterial strains is depicted in FIG. 17. The arrows indicate clusters of bacteria on tomato roots.

[0198] Example 9: Effect of Bacillus subtilis MH1 and Bacillus pseudomycoides MH2 Treatments on Tomato Plants in Various Low Nitrogen Conditions.

[0199] The N requirement for tomato plants during fermentation is 200:250:250 kg of nitrogen:phosphorus:potassium (i.e., NPK) per hectare. 75% of the required P (i.e., 187.5 kg, which comes to 1172 kg of superphosphate) is applied as basal. The remaining quantity of 200:62.5:250 kg of NPK per hectare is applied through fertigation. Every day irrigation should be given for one hour. Along with this, water soluble fertilizers have to be given. The dose is split and given once in 3 days for the entire crop period through fertigation. The typical dosage of NPK fertilizers at the various crop stages of tomato plants can be found in Table 2.

[0200] Table 2: NPK Requirements for Tomato Plants at Various Growth Stages

[0201] F = Fertilizer and D = Duration in days

[0202] The total requirement for a 19:19:19 fertilizer during the crop stages of a tomato plant is 132 kg / ha, for a 12:61 :0 fertilizer the total is 62 kg / ha, for a 13:0:45 fertilizer the total is 500 kg / ha, and for urea the total is 223 kg / ha.

[0203] In view of the total nitrogen requirement for tomato plants during their crop stages, this example investigated the effect that various nitrogen poor conditions have on tomato plant growth when Bacillus subtilis MH1 and Bacillus pseudomycoides MH2 is present in the soil. For this example, the following treatment groups were examined: T1 = 0% of recommended N, T2 = 10% of recommended N, T3 = 25% of recommended N, T4 = 50% of recommended N, T5 = 75% of recommended N, T6 = 100% of recommended N, and T7 = Control.

[0204] Tomato seeds were first surface sterilized and then imbibed with known concentrations of Bacillus subtilis MH1 and Bacillus pseudomycoides MH2 (OD Value = 2). The seeds were then sowed in 1 kg of promix or soil. There were 5 seedlings per pot of soil and 2 pot for each treatment group. Two weeks after sowing, the soil was drenched with a suspension of either Bacillus subtilis MH1 or Bacillus pseudomycoides MH2 (OD Value - 2). 30 days after sowing (sowing stage 1 ) a first dosage of fertilizer was applied to each treatment group. The recommend amount of N (i.e, 100% N) that tomato plants require at this sowing stage, in 1 kg of soil, is 4.91 mg.

[0205] The fertilizer used in this example was a 1 .63 g / L Hoagland stock solution. 1 L of the Hoagland stock solution provides 115.03 mg of N. The achieve the target recommended N percentages of each treatment group at this sowing stage, the Hoagland stock solution was diluted down. The dilutions formed for each treatment group, and the amounts of those dilutions administered to each treatment group at this sowing stage, are shown in Table 3.

[0206] Table 3: Amount of Fertilizer Applied Per Treatment Group at Sowing Stage 1

[0207] Four weeks after the first dosage of fertilizer, the soil of each treatment group was drenched with solutions of Bacillus subtilis MH1 and Bacillus pseudomycoides MH2 (OD Value = 2). 60 days after sowing (sowing stage 2), a second dosage of fertilizer was administered to each treatment group. The recommend amount of N (i.e, 100% N) that tomato plants require at this sowing stage, in 1 kg of soil, is 1.932 mg. The achieve the target recommended N percentages of each treatment group at this sowing stage, the Hoagland stock solution was again diluted down. The dilutions formed for each treatment group, and the amounts of those dilutions administered to each treatment group at this sowing stage, are shown in Table 4.

[0208] Table 4: Amount of Fertilizer Applied Per Treatment Group at Sowing Stage 2 Six weeks after the second dosage of fertilizer, the soil of each treatment group was drenched with solutions of Bacillus subtilis MH1 and Bacillus pseudomycoides MH2 (OD Value = 2). 90 days after sowing (sowing stage 3), a third dosage of fertilizer was administered to each treatment group. The recommend amount of N (i.e, 100% N) that tomato plants require at this sowing stage, in 1 kg of soil, is 4.94 mg. The achieve the target recommended N percentages of each treatment group at this sowing stage, the Hoagland stock solution was again diluted down. The dilutions formed for each treatment group, and the amounts of those dilutions administered to each treatment group at this sowing stage, are shown in Table 5. Table 5: Amount of Fertilizer Applied Per Treatment Group at Sowing Stage 3

[0209] Eight weeks after the third dosage of fertilizer, the soil of each treatment group was drenched with solutions of Bacillus subtilis MH1 and Bacillus pseudomycoides MH2 (OD Value = 2). 110 days after sowing (sowing stage 4), a fourth dosage of fertilizer was administered to each treatment group. The recommend amount of N ( / .e, 100% N) that tomato plants require at this sowing stage, in 1 kg of soil, is 1.531 mg. The achieve the target recommended N percentages of each treatment group at this sowing stage, the Hoagland stock solution was again diluted down. The dilutions formed for each treatment group, and the amounts of those dilutions administered to each treatment group at this sowing stage, are shown in Table 6.

[0210] Table 6: Amount of Fertilizer Applied Per Treatment Group at Sowing Stage 4

[0211] After the fourth dosage of fertilizer, the tomato plants were harvested.

[0212] The fresh total biomass, fresh weight of roots, and fresh weight of stem of the tomato plants treated with Bacillus subtilis MH1 , for each treatment group, are depicted in FIG. 18. The dry total biomass, the dry weight, and the dry weight on stem of the tomato plants treated with Bacillus subtilis MH1 , for each treatment group, are depicted in FIG. 19. The fruit yield of each plant and yield per hectare for the tomato plants treated with Bacillus subtilis MH1 , for each treatment group, are depicted in FIG. 20. The visual effects that Bacillus subtilis MH1 had on the tomato plants, when subjected to the treatment conditions of this example, are depicted in FIGs. 21-23. The nitrogen content of each tomato plant treated with Bacillus subtilis MH1 , for each treatment group, is shown in FIG. 24.

[0213] The fresh total biomass, fresh weight of roots, and fresh weight of stem of the tomato plants treated with Bacillus pseudomycoides MH2, for each treatment group, are depicted in FIG. 25. The dry total biomass, the dry weight, and the dry weight on stem of the tomato plants treated with Bacillus pseudomycoides MH2, for each treatment group, are depicted in FIG. 26. The fruit yield of each plant and yield per hectare for the tomato plants treated with Bacillus pseudomycoides MH2, for each treatment group, are depicted in FIG. 27. The visual effects that Bacillus pseudomycoides MH2 had on the tomato plants, when subjected to the treatment conditions of this example, are depicted in FIGs. 28 and 29.

[0214] The effect that Bacillus subtilis MH1 and Bacillus pseudomycoides MH2 had on the root length or depth of tomato plants grown under various nitrogen poor conditions is depicted in FIG. 30. From this data, the inventors of the present disclosure discovered that Bacillus subtilis MH1 and Bacillus pseudomycoides MH2 can promote increased root length or depth when subjected to nitrogen poor soil conditions.

[0215] Overall, the results from this example demonstrate that both Bacillus subtilis MH1 and Bacillus pseudomycoides MH2 can promote and improve tomato plant growth under nitrogen poor conditions.

[0216] Example 10: Effect of Bacillus subtilis MH1 and Bacillus pseudomycoides MH2 Treatments on Onion Plants in Various Low Nitrogen Conditions

[0217] The nitrogen conditions used to evaluate the effects that Bacillus subtilis MH1 and Bacillus pseudomycoides MH2 treatments have on onion plant growth are identical to those of Example 9. The same nitrogen treatment groups of Example 9 were used in this Example and the amount of nitrogen each treatment group received for each sowing phase was identical to those administered in Example 9. The process used in Example 9 to grow the tomato plants (i.e., the process involving the repeated dosing of fertilizer and Bacillus subtilis MH1 / Bacillus pseudomycoides MH2 solutions) was also used to grow the onion plants in this Example.

[0218] The fresh total biomass, fresh weight of roots, and fresh weight of stem of the onion plants treated with Bacillus subtilis MH1 , for each treatment group, are depicted in FIG. 31 . The dry total biomass, the dry weight, and the dry weight on stem of the onion plants treated with Bacillus subtilis MH1 , for each treatment group, are depicted in FIG. 32.

[0219] The fresh total biomass, fresh weight of roots, and fresh weight of stem of the onion plants treated with Bacillus pseudomycoides MH2, for each treatment group, are depicted in FIG. 33. The dry total biomass, the dry weight, and the dry weight on stem of the onion plants treated with Bacillus pseudomycoides MH2, for each treatment group, are depicted in FIG. 34.

[0220] Overall, the results from this example demonstrate that both Bacillus subtilis MH1 and Bacillus pseudomycoides MH2 can promote and improve onion plant growth under nitrogen poor conditions.

[0221] Example 11 : Effect of Bacillus subtilis MH1 and Bacillus pseudomycoides MH2 Treatments on Corn Plants in Various Low Nitrogen Conditions The nitrogen conditions used to evaluate the effects that Bacillus subtilis MH1 and Bacillus pseudomycoides MH2 treatments have on corn plant growth are outlined in Table 7.

[0222] Table 7: Treatment Groups of Example 11

[0223] Treatment groups T1 to T12 had bacterial strains administered at the time of sowing (i. e. , seed treatment with the bacterial strains), at the 25thday of sowing (bacterial strains were administered via through the soil application and the foliar of the com plants), and at the 45thday of sowing (bacterial strains were administered via through the soil application and the foliar of the corn plants). T reatment at the time of sowing involved imbibing the corn plant seeds with a solution of Bacillus subtilis MH1 and / or Bacillus pseudomycoides MH2 (OD Value = 2) for 10 minutes before sowing. Treatment at the 25thday and 45thday after sowing involved administering a solution of Bacillus subtilis MH1 and / or Bacillus pseudomycoides MH2 (OD Value = 2) at 10 mL of solution / L of water in the soil and on the foliar of the corn plants.

[0224] The recommend dosages of nitrogen, phosphorus and potassium for corn plants are 135 kg / ha (about 120 Ib / acre), 62.5 kg / ha (about 55 Ib / acre), and 37.50 kg / ha (about 33 Ib / acre), respectively. Accordingly, treatment groups grown with 10% of the recommended nitrogen level were cultivated in plots having about 12 Ib / acre or about 13 kg / ha of nitrogen. Treatment groups grown with 50% of the recommended nitrogen level were cultivated in plots having about 60 Ib / acre or about 67 kg / ha of nitrogen. Each treatment group was grown under 55 Ib / acre or about 61 kg / ha of phosphorus, 33 Ib / acre or about 37 kg / ha of potassium, and 33 Ib / acre or about 37 kg / ha of zinc.

[0225] Each treatment group was cultivated on a 3x4 m2plot with 0.75 m x 0.25 m spacing. The total number of corn plants in each plot was 64. Accordingly, one replication had 64 plants. The total replications performed in this example were 3.

[0226] The visual effects that each treatment group had on corn plant growth can be seen in FIG. 35 and FIG. 36. The amount of nitrogen in the corn plant for each treatment group is shown in FIG. 37.

[0227] Overall, the results from this example demonstrate that both Bacillus subtilis MH1 and Bacillus pseudomycoides MH2 can promote and improve corn plant growth under nitrogen poor conditions.

[0228] It will be appreciated by those skilled in the art that the present disclosure can be embodied in other specific forms without departing from the spirit or essential characteristics thereof. The presently disclosed embodiments are therefore considered in all respects to be illustrative and not restricted. The scope of the disclosure is indicated by the appended claims rather than the foregoing description and all changes that come within the meaning and range and equivalence thereof are intended to be embraced therein.

Claims

CLAIMS:1 . A composition for application to plants, plant seeds, or a plant growth medium, wherein the composition comprises: an agriculturally acceptable carrier and an effective amount of a biologically pure bacterial culture, wherein the bacteria in the bacterial culture includes Bacillus subtilis MH1 (ATCC Accession No. PTA-127648), Bacillus pseudomycoides MH2 (ATCC Accession No. PTA-127649) or a combination thereof, and wherein the composition has a nitrogen content ranging from 0% to 75% of a recommend nitrogen level for a growth stage of the plant.

2. The composition of claim 1 , wherein the composition comprises an effective amount of an auxiliary plant growth promoting rhizobacteria (PGPR).

3. The composition of claim 1 , wherein the composition comprises: a phosphorus content ranging from 90% to 100% of a recommend phosphorus level for the growth stage of the plant, a potassium content ranging from 90% to 100% of a recommend potassium level for the growth stage of the plant, or any combination thereof.

4. The composition of claim 1 , wherein the agriculturally acceptable carrier comprises one or more of a dispersant, a surfactant, an additive, water, a thickener, an anti-caking agent, residue breakdown, a composting formulation, a granular application, diatomaceous earth, an oil, a coloring agent, a stabilizer, a preservative, a polymer, or a coating.

5. The composition of claim 4, wherein: the additive comprises an oil, a gum, a resin, a clay, a polyoxyethylene glycol, a terpene, a viscid organic, a fatty acid ester, a sulfated alcohol, an alkyl sulfonate, a petroleum sulfonate, an alcohol sulfate, a sodium alkyl butane diamate, a polyester of sodium thiobutant dioate, a proteinaceous material, or a combination thereof;the thickener comprises a long chain alkylsulfonate of polyethylene glycol, polyoxyethylene oleate, or a combination thereof; the surfactant comprises a heavy petroleum oil, a heavy petroleum distillate, a polyol fatty acid ester, a polyethoxylated fatty acid ester, an aryl alkyl polyoxyethylene glycol, an alkyl amine acetate, an alkyl aryl sulfonate, a polyhydric alcohol, an alkyl phosphate, or a combination thereof; or the anti-caking agent comprises a sodium salt, a calcium carbonate, diatomaceous earth, or a combination thereof.

6. The composition of claim 1 , wherein the agriculturally acceptable carrier comprises vermiculite, charcoal, sugar factory carbonation press mud, rice husk, carboxymethyl cellulose, peat, perlite, fine sand, calcium carbonate, flour, alum, a starch, talc, polyvinyl pyrrolidone, or a combination thereof.

7. The composition of claim 1 , wherein the composition is formulated as a seed coating formulation, a liquid formulation for application to plants or to a plant growth medium, or a solid formulation for application to plants or to a plant growth medium.

8. The composition of claim 7, wherein: the seed coating formulation is an aqueous or oil-based solution for application to seeds or a powder or granular formulation for application to seeds; the liquid formulation for application to plants or to a plant growth medium is in a concentrated formulation or a ready-to-use formulation; or the solid formulation for application to plants or to a plant growth medium is a granular formulation or a powder agent.

9. The composition of claim 1 , wherein the composition comprises: a fertilizer, a micronutrient fertilizer material, an insecticide, a herbicide, a plant growth amendment, a molluscicide, an algicide, a bacterial inoculant, a fungal inoculant, or a combination thereof.

10. The composition of claim 9, wherein:the fertilizer comprises ammonium sulfate, ammonium nitrate, ammonium sulfate nitrate, ammonium chloride, ammonium bisulfate, ammonium polysulfide, ammonium thiosulfate, aqueous ammonia, anhydrous ammonia, ammonium polyphosphate, aluminum sulfate, calcium nitrate, calcium ammonium nitrate, calcium sulfate, calcined magnesite, calcitic limestone, calcium oxide, calcium nitrate, dolomitic limestone, hydrated lime, calcium carbonate, diammonium phosphate, monoammonium phosphate, magnesium nitrate, magnesium sulfate, potassium nitrate, potassium chloride, potassium magnesium sulfate, potassium sulfate, sodium nitrates, magnesian limestone, magnesia, urea, ureaformaldehydes, urea ammonium nitrate, sulfur-coated urea, polymer-coated urea, isobutylidene diurea, K2SO4-2MgSC>4, kainite, sylvinite, kieserite, Epsom salts, elemental sulfur, marl, ground oyster shells, fish meal, oil cakes, fish manure, blood meal, rock phosphate, super phosphates, slag, bone meal, wood ash, manure, bat guano, peat moss, compost, green sand, cottonseed meal, feather meal, crab meal, fish emulsion, or a combination thereof; the micronutrient fertilizer material comprises boric acid, a borate, a boron frit, copper sulfate, a copper frit, a copper chelate, a sodium tetraborate decahydrate, an iron sulfate, an iron oxide, iron ammonium sulfate, an iron frit, an iron chelate, a manganese sulfate, a manganese oxide, a manganese chelate, a manganese chloride, a manganese frit, a sodium molybdate, molybdic acid, a zinc sulfate, a zinc oxide, a zinc carbonate, a zinc frit, zinc phosphate, a zinc chelate, or a combination thereof; the insecticide comprises an organophosphate, a carbamate, a pyrethroid, an acaricide, an alkyl phthalate, boric acid, a borate, a fluoride, sulfur, a haloaromatic substituted urea, a hydrocarbon ester, a biologically-based insecticide, or a combination thereof; the herbicide comprises a chlorophenoxy compound, a nitrophenolic compound, a nitrocresolic compound, a dipyridyl compound, an acetamide, an aliphatic acid, an anilide, a benzamide, a benzoic acid, anisic acid, a benzonitrile, benzothiadiazinone dioxide, a thiocarbamate, a carbamate, a carbanilate, chloropyridinyl, a fluorodinitrotoluidine compound, isoxazolidinone, nicotinic acid, isopropylamine, oxadiazolinone, a phosphate, a phthalate, a picolinic acidcompound, a triazine, a triazole, a uracil, endothall, sodium chlorate, or a combination thereof; the fungal inoculant comprises a fungal inoculant of the family Glomeraceae, a fungal inoculant of the family Claroidoglomeraceae, a fungal inoculant of the family Gigasporaceae, a fungal inoculant of the family Acaulosporaceae, a fungal inoculant of the family Sacculosporaceae, a fungal inoculant of the family Entrophosporaceae, a fungal inoculant of the family Pacidsporaceae, a fungal inoculant of the family Diversisporaceae, a fungal inoculant of the family Paraglomeraceae, a fungal inoculant of the family Archaeosporaceae, a fungal inoculant of the family Geosiphonaceae, a fungal inoculant of the family Ambisporaceae, a fungal inoculant of the family Scutellosporaceae, a fungal inoculant of the family Dentiscultataceae, a fungal inoculant of the family Racocetraceae, a fungal inoculant of the phylum Basidiomycota, a fungal inoculant of the phylum Ascomycota, a fungal inoculant of the phylum Zygomycota, or a combination thereof; or the bacterial inoculant comprises a bacterial inoculant of the genus Rhizobium, a bacterial inoculant of the genus Bradyrhizobium, a bacterial inoculant of the genus Mesorhizobium, a bacterial inoculant of the genus Azorhizobium, a bacterial inoculant of the genus Allorhizobium, a bacterial inoculant of the genus Sinorhizobium, a bacterial inoculant of the genus Kluyvera, a bacterial inoculant of the genus Azotobacter, a bacterial inoculant of the genus Pseudomonas, a bacterial inoculant of the genus Azospirillium, a bacterial inoculant of the genus Bacillus, a bacterial inoculant of the genus Streptomyces, a bacterial inoculant of the genus Paenibacillus, a bacterial inoculant of the genus Paracoccus, a bacterial inoculant of the genus Enterobacter, a bacterial inoculant of the genus Alcaligenes, a bacterial inoculant of the genus Mycobacterium, a bacterial inoculant of the genus Trichoderma, a bacterial inoculant of the genus Gliocladium, a bacterial inoculant of the genus Glomus, a bacterial inoculant of the genus Klebsiella, or a combination thereof.

11. A method for treating a plant or a seed thereof, comprising: transplanting the plant or the seed thereof into a soil composition;administering to the plant or the seed thereof an effective amount of an isolated bacterial strain selected from the group consisting of Bacillus subtilis MH1 (ATCC Accession No. PTA-127648), Bacillus pseudomycoides MH2 (ATCC Accession No. PTA-127649) or a combination thereof; and cultivating the plant or seed thereof in an environment after the administering of the effective amount of the isolated bacterial strain.

12. The method of claim 11 , wherein the isolated bacterial strain is administered under one or more of the following conditions: a nitrogen content ranging from 0% to 75% of a recommend nitrogen level for a growth stage of the plant or seed thereof; a phosphorus content ranging from 90% to 100% of a recommend phosphorus level for the growth stage of the plant or seed thereof; or a potassium content ranging from 90% to 100% of a recommend potassium level for the growth stage of the plant or seed thereof.

13. The method of claim 11 , wherein the isolated bacterial strain is administered to an aerial part of the plant.

14. The method of claim 11 , wherein the isolated bacterial strain is administered to at least a portion of a root of the plant.

15. The method of claim 11 , wherein the administering of the effective amount of the isolated bacterial strain comprises: providing foliar resistance to a fungal pathogen to the plant or seed thereof.

16. The method of claim 15, wherein the fungal pathogen is selected from the group consisting of Clarireedia jacksonii, Sclerotinia sclerotiorum, Diaporthae ueckerae, Rhizoctonia solani, and Bipolaris zeicola.

17. The method of claim 11 , wherein the plant or the seed thereof is a monocotyledon crop plant selected from the group consisting of rice, wheat, corn,barley, rye, pineapples, dates, bananas, sugarcane, onions, garlic, palms, bamboo, asparagus, ginger, turmeric and cardamom.

18. The method of claim 11 , wherein the plant or seed thereof is a dicotyledon crop plant selected from the group consisting of soybean, cotton, potato, beans, sugar beet, melons, squash, pumpkins, flax, peppers, amaranth, lentils, peas, peanuts, roses, geraniums, carrots, tomato, lettuce, spinach, and mint.

19. The method of claim 11 , wherein the growing occurs under anaerobic conditions.

20. The method of claim 11 , wherein the method comprises one or more of: increasing root colonization in the plant or the seed thereof after the administering of the isolated bacterial strain; increasing resistance to a pathogen in an aerial part of the plant after the administering of the isolated bacterial strain; increasing growth of the plant or the seed thereof after the administering of the isolated bacterial strain; or increasing nitrogen uptake in the plant or the seed thereof after the administering of the isolated bacterial strain, when compared to a root colonization, an aerial pathogen resistance, a plant growth, and / or a nitrogen uptake of the plant when the isolated bacterial strain is not administered to the plant.

21. The method of claim 11 , wherein the cultivating occurs from 1 to 110 days.

22. The method of claim 11 , wherein the effective amount of the isolated bacterial strain recruits plant growth promoting rhizobacteria (PGPR) from the environment to the plant or the seed thereof.

23. The method of claim 11 , comprising:coating the seed, before transplanting, with an effective amount of a biologically pure bacterial culture, wherein the bacteria in the bacterial culture includes Bacillus subtilis MH1 , Bacillus pseudomycoides MH2 or a combination thereof.

24. The method of claim 11 , wherein the effective amount of the isolated bacterial strain improves a root depth or root length of the plant during the cultivating when the environment has a nitrogen content ranging from 0% to 75% of a recommend nitrogen level for a growth stage of the plant, when compared to a root depth or root length of the plant when:- cultivated in an environment having a nitrogen content ranging from 0% to 75% of a recommend nitrogen level for a growth stage of the plant, and- when cultivated without any administration of the effective amount of the isolated bacterial strain.

25. A plant seed coated with a composition comprising: an effective amount of a biologically pure bacterial culture, wherein the bacteria in the bacterial culture includes Bacillus subtilis MH1 (ATCC Accession No. PT A-127648), Bacillus pseudomycoides MH2 (ATCC Accession No. PTA- 127649) or a combination thereof, and wherein the composition has a nitrogen content ranging from 0% to 75% of a recommend nitrogen level for growth of the seed.

26. The plant seed of claim 25, wherein the composition coated on the seed comprises an effective amount of an auxiliary plant growth promoting rhizobacteria (PGPR).

27. The plant seed of claim 25, wherein the composition coated on the seed comprises: a phosphorus content ranging from 90% to 100% of a recommend phosphorus level for growth of the seed, a potassium content ranging from 90% to100% of a recommend potassium level for growth stage of the seed, or any combination thereof.

28. The plant seed of claim 25, wherein the composition coated on the seed comprises an agriculturally acceptable carrier selected from one or more of a dispersant, a surfactant, an additive, water, a thickener, an anti-caking agent, residue breakdown, a composting formulation, a granular application, diatomaceous earth, an oil, a coloring agent, a stabilizer, a preservative, a polymer, or an additional coating.

29. The plant seed of claim 25, wherein the composition coated on the seed is a liquid formulation or a solid formulation, wherein the liquid formulation is an aqueous or oil-based solution, and wherein the solid formulation is a powder agent or granular formulation.

30. A kit for treating a plant or a seed thereof, comprising the composition of claim 1.

31. The kit of claim 30, wherein the kit contains at least one container storing the composition of claim 1 .

32. The kit of claim 31 , wherein the container is configured to allow application of the composition of claim 1 onto an aerial part or at least a portion of a root of the plant.

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