Microorganism compositions for enhancing soil quality and plant growth

Abstract

The present disclosure relates to methods of improving soil nutrient composition and enhancing plant growth and plant productivity by contacting the soil and / or plants with a composition comprising one or more isolated Bacillus strains.

Description

MICROORGANISM COMPOSITIONS FOR ENHANCING SOIL QUALITY AND PLANT GROWTHCROSS REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to U.S. Provisional Patent Application No. 63 / 621,387 filed on January 16, 2024, the disclosure of which is expressly incorporated herein.BACKGROUND

[0002] A rising global population, climate change, and demand for sustainable farming practices are putting pressure on producers of commodity crops, fruits and vegetables, and other specialty crops to increase yields and reduce the use of chemical fertilizers. Drought, soil salinity, and poor nutrient availability are common stressors on plants resulting from a warming climate and overuse of land. Bacillus-bas&d biofertilizers are one potential solution to help crops overcome these challenges as well as help to reduce the use of chemical fertilizers.

[0003] One mechanism for growth promotion by Bacillus is their ability to increase the availably of nutrients for uptake by plants. Phosphorus is commonly found in soil in forms that cannot be easily utilized by plants, like calcium phosphate and phytase. Chemical fertilizers contribute to this problem by adding additional inorganic phosphate. Phosphorus in inorganic forms can be solubilized by Bacillus to make the phosphate available for uptake by plants. Bacillus can also produce the enzyme phytase, which removes phosphate from phytate making it available for uptake by plants. By transforming forms of phosphorus that are common in soil into forms usable by plants, a Bacillus -based biofertilizer can enhance plant growth and reduce the amount of chemical fertilizer needed. Reducing use of inorganic phosphorus fertilizer can reduce costs for producers as well as reduce runoff and eutrophication.SUMMARY

[0004] The present disclosure relates generally to microbial inoculants, comprising one or more microbial species or strains as described herein, particularly for use as biofertilizers and soil conditioners. The disclosure also relates to methods of promoting plant growth, increasing availability of nutrients in the soil and remediating degraded soils and pastures using microbial inoculants and fertilizer compositions of the present disclosure.

[0005] In one embodiment microbial inoculants for use in increasing plant growth, plant productivity and / or soil quality (in reference to supporting plant growth), are provided. In one embodiment, the compositions comprise one or more isolated Bacillus strains selected from thegroup consisting of Bacillus licheniformis strain 1000 (NRRL No. B-67888), a strain having all of the identifying characteristics of Bacillus strain 1000 (NRRL No. B-67888), Bacillus licheniformis strain 1001 (NRRL No. B-67889), a strain having all of the identifying characteristics of Bacillus strain 1001 (NRRL No. B-67889), Bacillus subtilis strain 8001 (NRRL No. B-67890), a strain having all of the identifying characteristics of Bacillus strain 8001 (NRRL No. B-67890), Bacillus strain Bacillus pumilus strain 1047 (NRRL No. B-67891), a strain having all of the identifying characteristics of Bacillus strain 1047 (NRRL No. B- 67891), Bacillus amyloliquefaciens strain 1607 (NRRL No. B-67666), a strain having all of the identifying characteristics of Bacillus strain 1607 (NRRL No. B-67666), and any combination thereof. The isolated Bacillus compositions disclosed herein can be applied as seed coating or applied to soil as a biofertilizer or biostimulant to enhance seed emergence, plant biomass and crop yields.

[0006] Bacillus strains Bacillus licheniformis MDG-1000 (NRRL No. B-67888), Bacillus licheniformis MDG- 1001 (NRRL No. B-67889), Bacillus subtilis / amyloliquefaciens MDG-8001 (NRRL No. B-67890), and Bacillus pumilus MDG- 1047 (NRRL No. B-67891), were deposited on November 26, 2019 at the Agricultural Research Service Culture Collection (NRRL), National Center for Agricultural Utilization Research, Agricultural Research Service, USDA, 1815 North University Street, Peoria, Illinois 61604-3999, and were given accession numbers B-67888, B-67889, B-67890, and B-67891, respectively. Bacillus strain Bacillus amyloliquefaciens MDG 1607 (NRRL No. B-67666) was deposited on August 14, 2018 at the Agricultural Research Service Culture Collection (NRRL), National Center for Agricultural Utilization Research, Agricultural Research Service, USDA, 1815 North University Street, Peoria, Illinois 61604-3999, and given accession number B-67666. The deposits were made under the provisions of the Budapest Treaty on the International Recognition of the Deposit of Microorganisms for the Purposes of Patent Procedure. Any and all restrictions imposed by Applicant on the availability to the public of the deposited material will be irrevocably removed upon the granting of the patent, and the deposits will be replaced if viable samples cannot be dispensed by the depository.

[0007] In one embodiment the present disclosure relates to treatment of a plant, or the soil used for growing plants, with one or more microorganisms for enhancing plant growth of the treated plants, or improving the plant nutrient content of the treated soil. More particularly, the disclosure relates to isolated Bacillus strains, and strains having all of the identifying characteristics of these strains, and combinations thereof, for a use comprising the above- mentioned use, including supplementing the soil with isolated Bacillus strains in an amount thatenhances seed germination, emergence, plant growth or plant productivity relative to plants grown in soil not supplemented with the isolated Bacillus strains. More particularly, in one embodiment the present disclosure is directed to isolated Bacillus strains, and strains having all of the identifying characteristics of these strains, and combinations thereof, for a use comprising the above-mentioned uses, including increasing the concentration of orthophosphates, H2PO4" and HPO42’ in soil contacted with the isolated Bacillus strains disclosed herein, and thus enhancing the growth of plants growing in such treated soil. In one embodiment seeds are coated or otherwise directly contacted with the isolated Bacillus strains disclosed herein, to enhance seed germination, or emergence, plant growth and / or plant productivity of the resulting germinated seedlings and plants.

[0008] Applicants have developed Bacillus strains, and combinations thereof, that are useful for improving plant growth and plant production. These strains comprise Bacillus licheniformis strain 1000 (NRRL No. B-67888), a strain having all of the identifying characteristics of Bacillus strain 1000 (NRRL No. B-67888), Bacillus licheniformis strain 1001 (NRRL No. B-67889), a strain having all of the identifying characteristics of Bacillus strain 1001 (NRRL No. B-67889), Bacillus suhtilis strain 8001 (NRRL No. B-67890), a strain having all of the identifying characteristics of Bacillus strain 8001 (NRRL No. B-67890), Bacillus strain Bacillus pumilus strain 1047 (NRRL No. B-67891), a strain having all of the identifying characteristics of Bacillus strain 1047 (NRRL No. B-67891), Bacillus amyloliquefaciens strain 1607 (NRRL No. B-67666), a strain having all of the identifying characteristics of Bacillus strain 1607 (NRRL No. B-67666), and any combination thereof.

[0009] In one embodiment, a method of enhancing the phosphate (PO4) content of soils is provided wherein the method comprises contacting soil with a composition comprising an effective amount of an isolated Bacillus strain selected from Bacillus licheniformis strain 1000 (NRRL No. B-67888), a strain having all of the identifying characteristics of Bacillus strain 1000 (NRRL No. B-67888), Bacillus licheniformis strain 1001 (NRRL No. B-67889), a strain having all of the identifying characteristics of Bacillus strain 1001 (NRRL No. B-67889), Bacillus subtilis strain 8001 (NRRL No. B-67890), a strain having all of the identifying characteristics of Bacillus strain 8001 (NRRL No. B-67890), Bacillus strain Bacillus pumilus strain 1047 (NRRL No. B-67891), a strain having all of the identifying characteristics of Bacillus strain 1047 (NRRL No. B-67891), Bacillus amyloliquefaciens strain 1607 (NRRL No. B-67666), a strain having all of the identifying characteristics of Bacillus strain 1607 (NRRL No. B-67666), and any combination thereof, under conditions conducive to the growth and proliferation of the Bacillus strain.

[0010] In one embodiment a composition comprising an isolated Bacillus strain selected from the group consisting of Bacillus licheniformis strain 1000 (NRRL No. B-67888), a strain having all of the identifying characteristics of Bacillus strain 1000 (NRRL No. B- 67888), Bacillus licheniformis strain 1001 (NRRL No. B-67889), a strain having all of the identifying characteristics of Bacillus strain 1001 (NRRL No. B-67889), Bacillus subtilis strain 8001 (NRRL No. B -67890), a strain having all of the identifying characteristics of Bacillus strain 8001 (NRRL No. B-67890), Bacillus strain Bacillus pumilus strain 1047 (NRRL No. B- 67891), a strain having all of the identifying characteristics of Bacillus strain 1047 (NRRL No. B-67891), Bacillus amyloliquefaciens strain 1607 (NRRL No. B-67666), a strain having all of the identifying characteristics of Bacillus strain 1607 (NRRL No. B-67666), and any combination thereof is provided, optionally in combination with additional plant nutrients and / or trace elements. In one embodiment the additional nutrients combined with the Bacillus strains of the present disclosure are selected from nitrogen (N), phosphorus (P), and potassium (K), magnesium (Mg), calcium (Ca), sulfur (S), iron (Fe), boron (B), molybdenum (Mo), manganese (Mn), copper (Cu), and zinc (Zn).

[0011] In another embodiment, a commercial package is provided. The commercial package comprises an isolated Bacillus strain selected from Bacillus licheniformis strain 1000 (NRRL No. B-67888), a strain having all of the identifying characteristics of Bacillus strain 1000 (NRRL No. B-67888), Bacillus licheniformis strain 1001 (NRRL No. B-67889), a strain having all of the identifying characteristics of Bacillus strain 1001 (NRRL No. B-67889), Bacillus subtilis strain 8001 (NRRL No. B-67890), a strain having all of the identifying characteristics of Bacillus strain 8001 (NRRL No. B-67890), Bacillus strain Bacillus pumilus strain 1047 (NRRL No. B-67891), a strain having all of the identifying characteristics of Bacillus strain 1047 (NRRL No. B-67891), Bacillus amyloliquefaciens strain 1607 (NRRL No. B-67666), a strain having all of the identifying characteristics of Bacillus strain 1607 (NRRL No. B-67666), and any combination thereof.

[0012] In yet another embodiment, a composition is provided comprising an isolatedBacillus strain selected from the group consisting of Bacillus licheniformis strain 1000 (NRRL No. B-67888), a strain having all of the identifying characteristics of Bacillus strain 1000 (NRRL No. B-67888), Bacillus licheniformis strain 1001 (NRRL No. B-67889), a strain having all of the identifying characteristics of Bacillus strain 1001 (NRRL No. B-67889), Bacillus subtilis strain 8001 (NRRL No. B-67890), a strain having all of the identifying characteristics of Bacillus strain 8001 (NRRL No. B-67890), Bacillus strain Bacillus pumilus strain 1047 (NRRL No. B-67891), a strain having all of the identifying characteristics of Bacillus strain1047 (NRRL No. B-67891), Bacillus amyloliquefaciens strain 1607 (NRRL No. B-67666), a strain having all of the identifying characteristics of Bacillus strain 1607 (NRRL No. B-67666), and any combination thereof, optionally in combination with additional plant nutrients and / or trace elements. In one embodiment the additional nutrients combined with the Bacillus strains of the present disclosure are selected from nitrogen (N), phosphorus (P), and potassium (K), magnesium (Mg), calcium (Ca), sulfur (S), iron (Fe), boron (B), molybdenum (Mo), manganese (Mn), copper (Cu), and zinc (Zn).BRIEF DESCRIPTION OF THE DRAWINGS

[0013] Fig. 1 provides data relating to the dry weight of 17-day old corn plants with and without treatment with the Bacillus blend comprising Bacillus licheniformis strain 1000, Bacillus licheniformis strain 1001, Bacillus subtilis strain 8001, Bacillus pumilus strain 1047 and Bacillus amyloliquefaciens strain 1607.

[0014] Fig. 2 shows resulting leaf greenness as measured by SPAD meter in 17-day old com plants with and without treatment with the Bacillus blend comprising Bacillus licheniformis strain 1000, Bacillus licheniformis strain 1001 , Bacillus subtilis strain 8001 , Bacillus pumilus strain 1047 and Bacillus amyloliquefaciens strain 1607.

[0015] Fig. 3 provides the results of treating com plants with the Bacillus compositions disclosed herein (the blend of 5 Bacillus strains). Com yield in bushels per acre when seeds were treated with the Bacillus blend is presented.

[0016] Fig. 4 is a graph presenting the concentration of free phosphate solubilized byBacillus strains from tricalcium phosphate in Pikovskaya broth over 48 hours.

[0017] Fig. 5 shows a photograph of a gel displaying RAPD-PCR profiles for Bacillus strain 1000. The leftmost and rightmost lanes, for each group of eight lanes, include markers.

[0018] Fig. 6 shows a photograph of a gel displaying RAPD-PCR profiles for Bacillus strain 1001. The leftmost and rightmost lanes, for each group of eight lanes, include markers.

[0019] Fig. 7 shows a photograph of a gel displaying RAPD-PCR profiles for Bacillus strain 8001. The leftmost and rightmost lanes, for each group of eight lanes, include markers.

[0020] Fig. 8 shows a photograph of a gel displaying RAPD-PCR profiles for Bacillus strain 1047. The leftmost and rightmost lanes, for each group of eight lanes, include markers.

[0021] Fig. 9 shows a photograph of a gel displaying RAPD-PCR profiles for Bacillus strain 1607. The leftmost and rightmost lanes include markers.DETAILED DESCRIPTIONDefinitions

[0022] The term "about" as used herein means greater or lesser than the value or range of values stated by 10 percent, but is not intended to designate any value or range of values to only this broader definition. Each value or range of values preceded by the term "about" is also intended to encompass the embodiment of the stated absolute value or range of values.

[0023] The term “isolated” as used herein means having been removed from its natural environment.

[0024] The term “purified,” as used herein relates to the isolation of a molecule or compound in a form that is substantially free of contaminants normally associated with the molecule or compound in a native or natural environment and means having been increased in purity as a result of being separated from other components of the original composition.

[0025] The term "plant productivity" as used herein refers to any aspect of growth or development of a plant, that is a reason for which the plant is grown. Thus, for purposes of the present disclosure, improved or increased plant productivity refers broadly to improvements in biomass or yield of leaves, stems, grain, fruit, vegetables, flowers, or other plant parts harvested or used for various purposes, and improvements in growth of plant parts, including stems, leaves and roots.

[0026] 'Plant growth" refers to the growth of any plant part, including stems, leaves and roots. Growth may refer to the rate of growth of any one of these plant parts.

[0027] The term "yield" refers to the amount of produced biological material. For crop plants, "yield" may also mean the amount of harvested material per unit of production or per area (e.g. hectare). Yield may be defined in terms of quantity or quality. The harvested material may vary from crop to crop, for example, it may be seeds, above-ground biomass, belowground biomass (e.g. potatoes), roots, fruits, or any other part of the plant which is of economic value.

[0028] The term "improving soil quality" defines a change in soil properties including an increase in the amount and / or availability of nutrients required by, or beneficial for seed germination, seedling emergence and plant growth. By way of example only, such nutrients include nitrogen, phosphorous, potassium, copper, zinc, boron and molybdenum.

[0029] As used herein, the term "effective amount" refers to an amount of microbial inoculant or fertilizer composition applied to a given area of soil or vegetation that is sufficient to effect one or more beneficial or desired outcomes, for example, in terms of plant growth rates, crop yields, or nutrient bio-availability in the soil. An "effective amount” can be provided in one or more administrations. The exact amount required will vary depending on factors suchas the identity and number of individual strains employed, the plant species being treated, the nature and condition of the soil to be treated, the exact nature of the microbial inoculant or fertilizer composition to be applied, the form in which the inoculant or fertilizer is applied and the means by which it is applied, and the stage of the plant growing season during which application takes place. Thus, it is not possible to specify an exact "effective amount". However, for any given case, an appropriate "effective amount” may be determined by one of ordinary skill in the art using only routine experimentation.

[0030] The term "crop" as used herein refers to any plant grown to be harvested or used for any economic purpose, including for example human foods, livestock fodder, fuel or pharmaceutical production.

[0031] Also contemplated by the present disclosure are variants of the microbial strains described herein. As used herein, the term "variant" refers to both naturally occurring and specifically developed variants or mutants of the microbial strains disclosed and exemplified herein that retain their ability to solubilize calcium phosphate and produce phytase.Embodiments

[0032] Applicants have developed Bacillus strains, and combinations thereof, that can be used to treat plants to increase plant growth and / or productivity. More particularly, the present disclosure relates to isolated Bacillus strains, and strains having all of the identifying characteristics of these strains, and combinations thereof, for a use comprising the above- mentioned use.

[0033] In accordance with one embodiment a microbial inoculant is provided for use in increasing plant growth, plant productivity and / or soil quality in reference to its ability to support plant growth. The inoculant can be directly applied to seeds or to plant parts, or the inoculant can be applied, mixed or injected into soil to improve the nutrient content of the soil. One or more of the strains in the inoculant may be encapsulated. Where multiple strains are encapsulated, the strains may be individually encapsulated or combined in a single encapsulation.

[0034] In one embodiment a method for increasing plant growth and / or productivity is provided wherein the method comprises contacting the plants, the seeds of said plant, or the soil in which said plants are grown, with an effective amount of a composition comprising an isolated Bacillus strain selected from the group consisting of Bacillus licheniformis strain 1000 (NRRL No. B-67888), a strain having all of the identifying characteristics of Bacillus strain 1000 (NRRL No. B-67888), Bacillus licheniformis strain 1001 (NRRL No. B-67889), a strainhaving all of the identifying characteristics of Bacillus strain 1001 (NRRL No. B-67889), Bacillus subtilis strain 8001 (NRRL No. B-67890), a strain having all of the identifying characteristics of Bacillus strain 8001 (NRRL No. B-67890), Bacillus strain Bacillus pumilus strain 1047 (NRRL No. B-67891), a strain having all of the identifying characteristics of Bacillus strain 1047 (NRRL No. B-67891), Bacillus amyloliquefaciens strain 1607 (NRRL No. B-67666), a strain having all of the identifying characteristics of Bacillus strain 1607 (NRRL No. B-67666), and any combination thereof.

[0035] In one embodiment the plants, the seeds of said plant, or the soil in which said plants are grown are contacted with an effective amount of a composition comprising two or more isolated Bacillus strains selected from

[0036] i) Bacillus licheniformis strain 1000 (NRRL No. B-67888) and Bacillus strain 1001 (NRRL No. B-67889); or

[0037] ii) Bacillus licheniformis strain 1000 (NRRL No. B-67888) and Bacillus subtilis strain 8001 (NRRL No. B-67890); or

[0038] iii) Bacillus licheniformis strain 1000 (NRRL No. B-67888) and Bacillus strain 1607 (NRRL No. B-67666); or

[0039] iv) Bacillus licheniformis strain 1000 (NRRL No. B-67888) and Bacillus strain 1047 (NRRL No. B-67891); or

[0040] v) Bacillus strain 1001 (NRRL No. B-67889) and Bacillus strain 8001 (NRRL No. B-67890); or

[0041] vi) Bacillus strain 1001 (NRRL No. B-67889) and Bacillus strain 1607 (NRRL No. B-67666); or

[0042] vii) Bacillus strain 1001 (NRRL No. B-67889) and Bacillus strain 1047 (NRRL No. B-67891); or

[0043] viii) Bacillus strain 8001 (NRRL No. B-67890); and Bacillus strain 1607 (NRRL No. B-67666); or

[0044] ix) Bacillus strain 8001 (NRRL No. B-67890); and Bacillus strain 1047 (NRRL No. B-67891); or

[0045] x) Bacillus strain 1607 (NRRL No. B-67666) and Bacillus strain 1047 (NRRL No. B-67891); or

[0046] xi) Bacillus licheniformis strain 1000 (NRRL No. B-67888), Bacillus strain 1001 (NRRL No. B-67889) and Bacillus strain 8001 (NRRL No. B-67890); or

[0047] xii) Bacillus licheniformis strain 1000 (NRRL No. B-67888) Bacillus strain 1001 (NRRL No. B-67889) and Bacillus strain 1047 (NRRL No. B-67891); or

[0048] xiii) Bacillus licheniformis strain 1000 (NRRL No. B-67888) Bacillus strain 1001 (NRRL No. B-67889) and Bacillus strain 1607 (NRRL No. B-67666); or

[0049] xiv) Bacillus licheniformis strain 1000 (NRRL No. B-67888), Bacillus strain 1607 (NRRL No. B-67666); and Bacillus strain 8001 (NRRL No. B-67890); or

[0050] xv) Bacillus licheniformis strain 1000 (NRRL No. B-67888), Bacillus strain 1607 (NRRL No. B-67666); and Bacillus strain 1047 (NRRL No. B-67891); or

[0051] xvi) Bacillus strain 1001 (NRRL No. B-67889), Bacillus strain 8001 (NRRL No. B-67890) and Bacillus strain 1607 (NRRL No. B-67666); or

[0052] xvii) Bacillus strain 1001 (NRRL No. B-67889), Bacillus strain 8001 (NRRL No. B-67890) and Bacillus strain 1047 (NRRL No. B-67891); or

[0053] xviii) Bacillus strain 8001 (NRRL No. B-67890), Bacillus strain 1047 (NRRL No. B-67891) and Bacillus strain 1607 (NRRL No. B-67666); or

[0054] xix) Bacillus licheniformis strain 1000 (NRRL No. B-67888) Bacillus strain 1001 (NRRL No. B-67889), Bacillus strain 1047 (NRRL No. B-67891) and Bacillus strain 1607 (NRRL No. B-67666); or

[0055] xx) Bacillus strain 1001 (NRRL No. B-67889), Bacillus strain 1047 (NRRL No. B-67891) Bacillus strain 8001 (NRRL No. B-67890), and Bacillus strain 1607 (NRRL No. B- 67666); or

[0056] xxi) Bacillus licheniformis strain 1000 (NRRL No. B-67888), Bacillus strain 1047 (NRRL No. B-67891), Bacillus strain 8001 (NRRL No. B-67890) and Bacillus strain 1607 (NRRL No. B-67666); or

[0057] xxii) Bacillus licheniformis strain 1000 (NRRL No. B-67888), Bacillus strain 1001 (NRRL No. B-67889), Bacillus strain 8001 (NRRL No. B-67890), and Bacillus strain 1607 (NRRL No. B-67666).

[0058] In accordance with one embodiment a fertilizer composition is provided comprising a microbial inoculant comprising a phytase expressing Bacillus strain of the present disclosure. The fertilizer composition may optionally comprise one or more additional components such as organic material, humic substances, penetrants, macronutrients, micronutrients and other soil and / or plant additives.

[0059] In one embodiment a composition is provided comprising an isolated Bacillus strain selected from the group consisting of Bacillus licheniformis strain 1000 (NRRL No. B- 67888), a strain having all of the identifying characteristics of Bacillus strain 1000 (NRRL No. B-67888), Bacillus licheniformis strain 1001 (NRRL No. B-67889), a strain having all of the identifying characteristics of Bacillus strain 1001 (NRRL No. B-67889), Bacillus subtilis strain8001 (NRRL No. B -67890), a strain having all of the identifying characteristics of Bacillus strain 8001 (NRRL No. B-67890), Bacillus strain Bacillus pumilus strain 1047 (NRRL No. B- 67891), a strain having all of the identifying characteristics di Bacillus strain 1047 (NRRL No. B-67891), Bacillus amyloliquefaciens strain 1607 (NRRL No. B-67666), a strain having all of the identifying characteristics of Bacillus strain 1607 (NRRL No. B-67666), and any combination thereof. In one embodiment the composition comprises an isolated Bacillus strain selected from the group consisting of Bacillus licheniformis strain 1000 (NRRL No. B-67888), Bacillus licheniformis strain 1001 (NRRL No. B-67889), Bacillus subtilis strain 8001 (NRRL No. B-67890), Bacillus strain Bacillus pumilus strain 1047 (NRRL No. B-67891), Bacillus amyloliquefaciens strain 1607 (NRRL No. B-67666), and any combination thereof. The composition may include additional bacterial or microbial strains that are known to be beneficial to the growth of plants. In addition the compositions may include addition nutrients beneficial to plant growth, including nitrate and potassium chloride.

[0060] The compositions of the present disclosure can be formulated as a liquid or dried, optionally freeze-dried, in the form of a powder, or in the form of a pellet.

[0061] Microbial inoculants and fertilizer compositions of the present disclosure may optionally further comprise one or more additional microbial organisms, for example, additional agronomically beneficial microorganisms. Such agronomically beneficial microorganisms may act in synergy or concert with, or otherwise cooperate with the organisms of the present disclosure in the inoculant or fertilizer. Examples of agronomically beneficial microorganisms include Pseudomonas sp., Rhizobium sp., Azospirillum sp., Azotobacter sp., phototrophic and cellulose degrading bacteria, Clostridium sp., Trichoderma sp. and the like. Those skilled in the art will appreciate that this list is merely exemplary only, and is not limited by reference to the specific examples here provided.

[0062] In the soil environment, inoculated bacteria can find survival difficult among naturally occurring competitor and predator organisms. To aid in survival of microorganisms, present in microbial inoculants and fertilizer compositions of the present disclosure, upon application in the environment, one or more of the strains may be encapsulated in, for example, a suitable polymeric matrix. In one example, encapsulation may comprise alginate beads such as has been described by Young et al, 2006, “Encapsulation Of Plant Growth-Promoting Bacteria In Alginate Beads Enriched With Humic Acid”, Biotechnology and Bioengineering 95:76-83, the disclosure of which is incorporated herein by reference in its entirety. Those skilled in the art will appreciate that any suitable encapsulation material or matrix may be used. Encapsulation may be achieved using methods and techniques known to those skilled in the art.Encapsulated microorganisms can include nutrients or other components of the inoculant or fertilizer composition in addition to the microorganisms.

[0063] Microbial inoculants and fertilizer compositions of the present disclosure may be applied directly to plants, plant parts (such as foliage) or seeds, or alternatively may be applied to soil in which the plants are growing or to be grown or in which seeds have been or are to be sown. Application may be by any suitable means and may be on any suitable scale. For example, application may comprise pouring, spreading or spraying, including broad scale or bulk spreading or spraying, soaking of seeds before planting, and / or drenching of seeds after planting or seedlings. Those skilled in the art will appreciate that multiple means of application may be used in combination (for example soaking of seeds prior to planting followed by drenching of planted seeds and / or application to seedlings or mature plants). Seeds, seedlings or mature plants may be treated as many times as appropriate. The number of applications required can readily be determined by those skilled in the art depending on, for example, the plant in question, the stage of development of the plant at which treatment is initiated, the state of health of the plant, the growth, environmental and / or climatic conditions in which the plant is grown and the purpose for which the plant is grown. For example, in the case of flowering crops such as tomatoes, it may be desirable to apply the microbial inoculant or fertilizer composition once or more than once during the flowering period.

[0064] Thus, in accordance with the present disclosure, microbial inoculants and fertilizer products as disclosed herein may be prepared in any suitable form depending on the means by which the inoculant or fertilizer composition is to be applied to the soil or to plant seeds or vegetation. Suitable forms can include, for example, slurries, liquids, and solid forms. Solid forms include powders, granules, larger particulate forms and pellets. Solid form fertilizer particles can be encapsulated in water soluble coatings (for example dyed or undyed gelatin spheres or capsules), extended release coatings, or by micro-encapsulation to a free flowing powder using one or more of, for example, gelatin, polyvinyl alcohol, ethylcellulose, cellulose acetate phthalate, or styrene maleic anhydride. Liquids may include aqueous solutions and aqueous suspensions, and emulsifiable concentrates.

[0065] In order to achieve effective dispersion, adhesion and / or conservation or stability within the environment of inoculants and fertilizer compositions disclosed herein, it may be advantageous to formulate the inoculants and compositions with suitable carrier components that aid dispersion, adhesion and conservation / stability. Suitable carriers will be known to those skilled in the art and include, for example, chitosan, vermiculite, compost, talc, milk powder, gels and the like.

[0066] Additional components may be incorporated into inoculants and fertilizer compositions of the present disclosure, such as humic substances, trace elements, organic material, penetrants, macronutrients, micronutrients and other soil and / or plant additives.

[0067] Humus or humic substances that may be incorporated may include, but are not limited to, humic acid derived from, for example oxidized lignite or leonardite, fulvic acid and humates such as potassium humate. Organic material added may include, but is not limited to, biosolids, animal manure, compost or composted organic byproducts, activated sludge or processed animal or vegetable byproducts (including blood meal, feather meal, cottonseed meal, ocean kelp meal, seaweed extract, fish emulsions and fish meal).

[0068] In one embodiment a method for improving soil quality or remediating degraded soil is provided, wherein the treated soil has an increased concentration of plant accessible nutrients. In one embodiment a soil conditioner composition is provided that coverts phytic acid and / or calcium phosphate to orthophosphates, H2PO4' and HPO42’. Accordingly, in one embodiment the soil is contacted with an effective amount of a composition comprising an isolated Bacillus strain selected from the group consisting of Bacillus licheniformis strain 1000 (NRRL No. B-67888), a strain having all of the identifying characteristics of Bacillus strain 1000 (NRRL No. B-67888), Bacillus licheniformis strain 1001 (NRRL No. B-67889), a strain having all of the identifying characteristics of Bacillus strain 1001 (NRRL No. B-67889), Bacillus subtilis strain 8001 (NRRL No. B-67890), a strain having all of the identifying characteristics of Bacillus strain 8001 (NRRL No. B-67890), Bacillus strain Bacillus pumilus strain 1047 (NRRL No. B-67891), a strain having all of the identifying characteristics of Bacillus strain 1047 (NRRL No. B-67891), Bacillus amyloliquefaciens strain 1607 (NRRL No. B-67666), a strain having all of the identifying characteristics of Bacillus strain 1607 (NRRL No. B-67666), and any combination thereof.

[0069] In one embodiment a method for increasing plant growth and / or productivity is provided, wherein the method comprises contacting the plant, plant seeds or to the soil in which the plant or plant seeds are grown, with an effective amount of any of the microbial compositions disclosed herein.

[0070] In accordance with the present disclosure the plant to be treated with the microbial compositions disclosed herein may be, for example, a pasture plant, crop plant (including fruit and vegetable plants) or ornamental plant. The crop may be, for example, any human or animal food crop or crop for use as fuel or for pharmaceutical production, including for example corn or soybean.Exemplary Embodiments

[0071] The following embodiments, provide various additional illustrative aspects of the invention described herein.

[0072] In embodiment 1 a method for increasing plant growth and / or productivity is provided wherein the method comprises contacting the plants, the seeds of said plant, or the soil in which said plants are grown, with an effective amount of a composition comprising an isolated Bacillus strain selected from the group consisting of Bacillus licheniformis strain 1000 (NRRL No. B-67888), a strain having all of the identifying characteristics of Bacillus strain 1000 (NRRL No. B-67888), Bacillus licheniformis strain 1001 (NRRL No. B-67889), a strain having all of the identifying characteristics of Bacillus strain 1001 (NRRL No. B-67889), Bacillus subtilis strain 8001 (NRRL No. B-67890), a strain having all of the identifying characteristics of Bacillus strain 8001 (NRRL No. B-67890), Bacillus strain Bacillus pumilus strain 1047 (NRRL No. B-67891), a strain having all of the identifying characteristics of Bacillus strain 1047 (NRRL No. B-67891), Bacillus amyloliquefaciens strain 1607 (NRRL No. B-67666), a strain having all of the identifying characteristics of Bacillus strain 1607 (NRRL No. B-67666), and any combination thereof.

[0073] In accordance with embodiment 2 the method of embodiment 1 is provided wherein the plants, plant seeds, or the soil in which said plants are grown, are contacted with an effective amount of a composition comprising two or more isolated Bacillus strain selected from the group consisting of Bacillus licheniformis strain 1000 (NRRL No. B-67888), a strain having all of the identifying characteristics of Bacillus strain 1000 (NRRL No. B-67888), Bacillus licheniformis strain 1001 (NRRL No. B-67889), a strain having all of the identifying characteristics of Bacillus strain 1001 (NRRL No. B-67889), Bacillus subtilis strain 8001 (NRRL No. B-67890), a strain having all of the identifying characteristics of Bacillus strain 8001 (NRRL No. B-67890), Bacillus strain Bacillus pumilus strain 1047 (NRRL No. B-67891), a strain having all of the identifying characteristics of Bacillus strain 1047 (NRRL No. B- 67891), Bacillus amyloliquefaciens strain 1607 (NRRL No. B-67666), a strain having all of the identifying characteristics of Bacillus strain 1607 (NRRL No. B-67666).

[0074] In accordance with embodiment 3 the method of embodiment 1 or 2 is provided wherein the plants, plant seeds or the soil in which said plants are grown, are contacted with an effective amount of a composition comprising three or more isolated Bacillus strain selected from the group consisting of Bacillus licheniformis strain 1000 (NRRL No. B-67888), a strain having all of the identifying characteristics of Bacillus strain 1000 (NRRL No. B-67888), Bacillus licheniformis strain 1001 (NRRL No. B-67889), a strain having all of the identifyingcharacteristics of Bacillus strain 1001 (NRRL No. B-67889), Bacillus subtilis strain 8001 (NRRL No. B-67890), a strain having all of the identifying characteristics of Bacillus strain 8001 (NRRL No. B-67890), Bacillus strain Bacillus pumilus strain 1047 (NRRL No. B-67891 ), a strain having all of the identifying characteristics of Bacillus strain f047 (NRRL No. B- 67891), Bacillus amyloliquefaciens strain 1607 (NRRL No. B-67666), a strain having all of the identifying characteristics of Bacillus strain 1607 (NRRL No. B-67666).

[0075] In accordance with embodiment 4 the method of any one of embodiments 1-3 is provided wherein the composition comprises isolated Bacillus licheniformis strain 1000 (NRRL No. B-67888).

[0076] In accordance with embodiment 5 the method of any one of embodiments 1-3 is provided wherein the composition comprises isolated Bacillus licheniformis strain 1001 (NRRL No. B-67889).

[0077] In accordance with embodiment 6 the method of any one of embodiments 3 is provided wherein the composition comprises isolated Bacillus subtilis strain 8001 (NRRL No. B-67890).

[0078] In accordance with embodiment 7 the method of any one of embodiments 1 -3 is provided wherein the composition comprises isolated Bacillus pumilus strain 1047 (NRRL No. B-67891).

[0079] In accordance with embodiment 8 the method of any one of embodiments 1-3 is provided wherein the composition comprises isolated Bacillus amyloliquefaciens strain 1607 (NRRL No. B-67666).

[0080] In accordance with embodiment 9 the method of any one of embodiments 1 -8 is provided wherein the plants, plant seeds or to the soil in which the plants are grown is contacted with an effective amount of a composition comprising

[0081] i) an isolated Bacillus licheniformis strain 1000 (NRRL No. B-67888), or a strain having all of the identifying characteristics of Bacillus strain W00 (NRRL No. B-67888);

[0082] ii) an isolated Bacillus licheniformis strain 1001 (NRRL No. B-67889), or a strain having all of the identifying characteristics of Bacillus strain 1001 (NRRL No. B-67889);

[0083] iii) an isolated Bacillus subtilis strain 8001 (NRRL No. B-67890), or a strain having all of the identifying characteristics of Bacillus strain 8001 (NRRL No. B-67890);

[0084] iv) an isolated Bacillus pumilus strain 1047 (NRRL No. B-67891), or a strain having all of the identifying characteristics of Bacillus strain 1047 (NRRL No. B-67891); and

[0085] v) an isolated Bacillus amyloliquefaciens strain 1607 (NRRL No. B -67666), or a strain having all of the identifying characteristics of Bacillus strain 1607 (NRRL No. B- 67666).

[0086] In accordance with embodiment 10 the method of any one of embodiments 1-9 is provided wherein said soil is contacted with said composition prior to planting said plant seeds.

[0087] In accordance with embodiment 11 the method of any one of embodiments 1-10 is provided wherein the seeds are contacted with said composition prior to planting the seeds.

[0088] In accordance with embodiment 12 the method of any one of embodiments 1-11 is provided wherein the soil in which the plants are grown is contacted with said composition prior to planting the seeds.

[0089] In accordance with embodiment 13 the method of any one of embodiments 1-12 is provided wherein the plants are pasture plants, crop plants or ornamental plants.

[0090] In accordance with embodiment 14 the method of any one of embodiments 1-13 is provided wherein the plants are crop plants, optionally wherein the crop plant is com or soybean.

[0091] In accordance with embodiment 15 a method for improving soil quality or remediating degraded soil in an agricultural field is provided, wherein the method comprises contacting the soil with an effective amount of a composition comprising an isolated Bacillus strain selected from the group consisting of Bacillus licheniformis strain 1000 (NRRL No. B- 67888), a strain having all of the identifying characteristics of Bacillus strain 1000 (NRRL No. B-67888), Bacillus licheniformis strain 1001 (NRRL No. B-67889), a strain having all of the identifying characteristics of Bacillus strain 1001 (NRRL No. B-67889), Bacillus subtilis strain 8001 (NRRL No. B-67890), a strain having all of the identifying characteristics of Bacillus strain 8001 (NRRL No. B-67890), Bacillus strain Bacillus pumilus strain 1047 (NRRL No. B- 67891), a strain having all of the identifying characteristics of Bacillus strain 1047 (NRRL No. B-67891), Bacillus amyloliquefaciens strain 1607 (NRRL No. B-67666), a strain having all of the identifying characteristics of Bacillus strain 1607 (NRRL No. B-67666), and any combination thereof.

[0092] In accordance with embodiment 16 the method of embodiments 15 is provided wherein said soil quality is improved or remediated based on an increased level of orthophosphates, H2PO4 and HPO42in said contacted soil.

[0093] In accordance with embodiment 17 the method of any one of embodiments 15- 16 is provided wherein the soil is contacted with an effective amount of a compositioncomprising two or more isolated Bacillus strains selected from the group consisting of Bacillus licheniformis strain 1000 (NRRL No. B-67888), a strain having all of the identifying characteristics of Bacillus strain 1000 (NRRL No. B-67888), Bacillus licheniformis strain 1001 (NRRL No. B-67889), a strain having all of the identifying characteristics of Bacillus strain 1001 (NRRL No. B-67889), Bacillus subtilis strain 8001 (NRRL No. B-67890), a strain having all of the identifying characteristics of Bacillus strain 8001 (NRRL No. B-67890), Bacillus strain Bacillus pumilus strain 1047 (NRRL No. B-67891), a strain having all of the identifying characteristics of Bacillus strain 1047 (NRRL No. B-67891), Bacillus amyloliquefaciens strain 1607 (NRRL No. B-67666), a strain having all of the identifying characteristics of Bacillus strain 1607 (NRRL No. B-67666).

[0094] In accordance with embodiment 18 the method of any one of embodiments 15-17 is provided wherein the soil in which the plant or plant seeds are grown is contacted with an effective amount of a composition comprising three or more isolated Bacillus strain selected from the group consisting of Bacillus licheniformis strain 1000 (NRRL No. B-67888), a strain having all of the identifying characteristics of Bacillus strain 1000 (NRRL No. B-67888), Bacillus licheniformis strain 1001 (NRRL No. B-67889), a strain having all of the identifying characteristics of Bacillus strain 1001 (NRRL No. B-67889), Bacillus subtilis strain 8001 (NRRL No. B-67890), a strain having all of the identifying characteristics of Bacillus strain 8001 (NRRL No. B-67890), Bacillus strain Bacillus pumilus strain 1047 (NRRL No. B-67891), a strain having all of the identifying characteristics of Bacillus strain 1047 (NRRL No. B- 67891), Bacillus amyloliquefaciens strain 1607 (NRRL No. B-67666), a strain having all of the identifying characteristics of Bacillus strain 1607 (NRRL No. B-67666).

[0095] In accordance with embodiment 19 the method of any one of embodiments 15-18 is provided wherein the composition comprises isolated Bacillus licheniformis strain 1000 (NRRL No. B-67888).

[0096] In accordance with embodiment 20 the method of any one of embodiments 15- 18 is provided wherein the composition comprises isolated Bacillus licheniformis strain 1001 (NRRL No. B-67889).

[0097] In accordance with embodiment 21 the method of any one of embodiments 15- 18 is provided wherein the composition comprises isolated Bacillus subtilis strain 8001 (NRRL No. B-67890).

[0098] In accordance with embodiment 22 the method of any one of embodiments 15- 18 is provided wherein the composition comprises isolated Bacillus pumilus strain 1047 (NRRL No. B-67891).

[0099] In accordance with embodiment 23 the method of any one of embodiments 15-18 is provided wherein the composition comprises isolated Bacillus amyloliquefaciens strain 1607 (NRRL No. B-67666).

[0100] In accordance with embodiment 24 the method of any one of embodiments 15-23 is provided wherein the soil is contacted with an effective amount of a composition comprising

[0101] i) an isolated Bacillus licheniformis strain 1000 (NRRL No. B-67888), or a strain having all of the identifying characteristics of Bacillus strain 1000 (NRRL No. B-67888);

[0102] ii) an isolated Bacillus licheniformis strain 1001 (NRRL No. B-67889), or a strain having all of the identifying characteristics of Bacillus strain 1001 (NRRL No. B-67889);

[0103] iii) an isolated Bacillus subtilis strain 8001 (NRRL No. B-67890), or a strain having all of the identifying characteristics of Bacillus strain 8001 (NRRL No. B-67890);

[0104] iv) an isolated Bacillus pumilus strain 1047 (NRRL No. B-67891), or a strain having all of the identifying characteristics of Bacillus strain 1047 (NRRL No. B-67891); and

[0105] v) an isolated Bacillus amyloliquefaciens strain 1607 (NRRL No. B-67666), a strain having all of the identifying characteristics of Bacillus strain 1607 (NRRL No. B-67666).

[0106] In accordance with embodiment 25 the method of any one of embodiments 15-24 is provided wherein the soil is present in a pasture.

[0107] In accordance with embodiment 26 the method of any one of embodiments 15- 24 is provided wherein the soil is present in a cultivated field.

[0108] In accordance with embodiment 27 the method of any one of embodiments 15- 26 is provided wherein the soil is contacted with said composition at least 1, 2, 3, 4, 5, 6, 7 or 14 days prior to the planting seeds in said contacted soil.

[0109] In accordance with embodiment 28 the method of any one of embodiments 1-27 is provided wherein said isolated Bacillus strains are placed in contact with plants or seeds in an amount that enhances seed germination, emergence, plant growth, and / or plant productivity relative to plants and seeds not contacted with the isolated Bacillus strains.

[0110] In accordance with embodiment 29 a composition or commercial package is provided comprising an isolated Bacillus strain selected from the group consisting of Bacillus licheniformis strain 1000 (NRRL No. B-67888), a strain having all of the identifying characteristics of Bacillus strain 1000 (NRRL No. B-67888), Bacillus licheniformis strain 1001 (NRRL No. B-67889), a strain having all of the identifying characteristics of Bacillus strain 1001 (NRRL No. B-67889), Bacillus subtilis strain 8001 (NRRL No. B-67890), a strain having all of the identifying characteristics of Bacillus strain 8001 (NRRL No. B-67890), Bacillusstrain Bacillus pumilus strain 1047 (NRRL No. B-67891), a strain having all of the identifying characteristics of Bacillus strain 1047 (NRRL No. B-67891), Bacillus amyloliquefaciens strain 1607 (NRRL No. B-67666), a strain having all of the identifying characteristics of Bacillus strain 1607 (NRRL No. B-67666), and any combination thereof.

[0111] In accordance with embodiment 30 the composition or commercial package of embodiment 29 is provided further comprising nitrate and potassium chloride.

[0112] In accordance with embodiment 31 the composition or commercial package of any one of embodiments 29-30 is provided wherein the composition is a liquid.

[0113] In accordance with embodiment 32 the composition or commercial package of any one of embodiments 29-30 is provided wherein the composition is dry.

[0114] In accordance with embodiment 33 the composition or commercial package of embodiment 32 is provided wherein the composition is freeze-dried, in the form of a powder, or in the form of a pellet.

[0115] In accordance with embodiment 34 the composition or commercial package of any one of embodiments 29-33 is provided wherein the Bacillus strain is in the form of a concentrate.

[0116] In accordance with embodiment 35 the composition or commercial package of any one of embodiments 29-34 is provided wherein the Bacillus strain is in the form of a superconcentrate.

[0117] In accordance with embodiment 36 the composition or commercial package of any one of embodiments 29-35 is provided further comprising a carrier for the Bacillus strain.

[0118] In accordance with embodiment 37 the composition or commercial package of embodiment 36 is provided wherein the carrier is selected from the group consisting of salt, a dextrin, and combinations thereof.

[0119] In accordance with embodiment 38 the composition or commercial package of any one of embodiments 29-37 is provided in a bag, optionally a paper or plastic bag.

[0120] In accordance with embodiment 39 the composition or commercial package of any one of embodiments 29-38 is provided further comprising instructions for use of one or more of the Bacillus strains.

[0121] In accordance with embodiment 40 the composition or commercial package of any one of embodiments 29-39 is provided in a 20-pound bag.

[0122] In accordance with embodiment 41 the composition or commercial package of any one of embodiments 29-39 is provided in a 50-pound bag.

[0123] In accordance with embodiment 42 the composition or commercial package of any one of embodiments 29-41 is provided further comprising a binder.

[0124] In accordance with embodiment 43 the composition or commercial package of embodiment 42 is provided wherein the binder is selected from the group consisting of clay, yeast cell wall components, aluminum silicate, and glucan, or combinations thereof.

[0125] In accordance with embodiment 44 the composition or commercial package of any one of embodiments 29-43 is provided wherein the Bacillus strains are encapsulated or embedded within a biodegradable matrix, optionally wherein the biodegradable matrix is a collagen based matrix, optionally wherein the biodegradable matrix is alginate.

[0126] In accordance with embodiment 45 the composition or commercial package of any one of embodiments 29-44 is provided wherein the composition or commercial package further comprises additional plant nutrients and / or trace elements, including for example, nutrients and trace elements selected from nitrogen (N), phosphorus (P), and potassium (K), magnesium (Mg), calcium (Ca), sulfur (S), iron (Fe), boron (B), molybdenum (Mo), manganese (Mn), copper (Cu), and zinc (Zn).

[0127] In accordance with embodiment 46 the composition or commercial package of any one of embodiments 29-45 is provided wherein the composition or commercial package further comprises seeds.

[0128] In accordance with embodiment 47 a composition is provided comprising seeds and one or more Bacillus strains selected from the group consisting of Bacillus licheniformis strain 1000 (NRRL No. B-67888), a strain having all of the identifying characteristics of Bacillus strain 1000 (NRRL No. B-67888), Bacillus licheniformis strain 1001 (NRRL No. B- 67889), a strain having all of the identifying characteristics of Bacillus strain 1001 (NRRL No. B-67889), Bacillus subtilis strain 8001 (NRRL No. B-67890), a strain having all of the identifying characteristics of Bacillus strain 8001 (NRRL No. B-67890), Bacillus strain Bacillus pumilus strain 1047 (NRRL No. B-67891), a strain having all of the identifying characteristics of Bacillus strain 1047 (NRRL No. B-67891), Bacillus amyloliquefaciens strain 1607 (NRRL No. B-67666), a strain having all of the identifying characteristics of Bacillus strain 1607 (NRRL No. B-67666), and any combination thereof.

[0129] In accordance with embodiment 48 the composition of claim 47 is provided wherein said seeds are coated or otherwise directly contacted with the isolated Bacillus strains disclosed herein, to enhance seed germination, or emergence, plant growth and / or plant productivity of the resulting germinated seedlings and plants.

[0130] In accordance with embodiment 49 the composition of claim 47 or 48 is provided wherein said isolated Bacillus strains are embedded in a biodegradable matrix and the matrix is contacted or coated onto said seeds.

[0131] In accordance with embodiment 50 the composition or commercial package of any one of embodiments 29-49 is provided for use in enhancing plant growth or plant productivity or enhancing soil quality. In various embodiments, the Bacillus strain for use in accordance with the methods, commercial packages, and compositions described herein can be selected from the group consisting of an isolated Bacillus strain selected from the group consisting of Bacillus licheniformis strain 1000 (NRRL No. B-67888), a strain having all of the identifying characteristics of Bacillus strain 1000 (NRRL No. B-67888), Bacillus licheniformis strain 1001 (NRRL No. B-67889), a strain having all of the identifying characteristics of Bacillus strain 1001 (NRRL No. B-67889), Bacillus subtilis strain 8001 (NRRL No. B-67890), a strain having all of the identifying characteristics of Bacillus strain 8001 (NRRL No. B- 67890), Bacillus strain Bacillus pumilus strain 1047 (NRRL No. B-67891), a strain having all of the identifying characteristics of Bacillus strain 1047 (NRRL No. B-67891), Bacillus amyloliquefaciens strain 1607 (NRRL No. B-67666), a strain having all of the identifying characteristics of Bacillus strain 1607 (NRRL No. B-67666), and any combination thereof.

[0132] In accordance with embodiment 51 the Bacillus strains of any one of embodiments 1-50 can optionally be encapsulated in a biocompatible polymer, optionally wherein the strains are encapsulated in alginate, optionally with humic acid. Where multiple strains are encapsulated, the strains may be individually encapsulated or combined in a single encapsulation.

[0133] In one illustrative aspect, any of these strains can be used alone or in combination in the form of a liquid or dry spray to be applied to the plant or to the soil from which the plant obtains nutrients, or in the form of a liquid or dry composition for manual application to the plant or to the soil from which the plant obtains nutrients. In one embodiment, multiple strains are used to treat the plants in combination in a single composition. In another embodiment, multiple strains are used to treat the plant in combination in separate compositions.

[0134] As used herein “a strain having all of the identifying characteristics of’ Bacillus strain an isolated Bacillus strain selected from the group consisting of Bacillus licheniformis strain 1000 (NRRL No. B-67888), a strain having all of the identifying characteristics of Bacillus strain 1000, 1001, 8001, 1047 or 1607 can be a mutant strain having all of the identifying characteristics of Bacillus strain 1000, 1001, 8001, 1047 or 1607 (e.g., a DNAfingerprint based on DNA analysis that corresponds to the DNA fingerprint of Bacillus strain1000, 1001, 8001, 1047 or 1607, enzyme activities that correspond to Bacillus strain 1000,1001 , 8001 , 1047 or 1607 (i.e., phytase activity), antimicrobial activity that corresponds to Bacillus strain 1000, 1001, 8001, 1047 or 1607, antibiotic sensitivity and tolerance profiles that correspond to Bacillus strain 1000, 1001, 8001, 1047 or 1607, or combinations thereof). In alternate embodiments, the mutation can be a natural mutation, or a genetically engineered mutation. In another embodiment, “a strain having all of the identifying characteristics of’ Bacillus strain 1000, 1001, 8001, 1047 or 1607 can be a strain, for example, produced by isolating one or more plasmids from Bacillus strain 1000, 1001, 8001, 1047 or 1607 and introducing the one or more plasmids into another bacterium, such as another Bacillus strain, as long as the one or more plasmids contain DNA that provides the identifying characteristics of Bacillus strain 1000, 1001, 8001, 1047 or 1607 (e.g., a DNA fingerprint based on DNA analysis that corresponds to the DNA fingerprint of Bacillus strain 1000, 1001, 8001, 1047 or 1607).

[0135] In another embodiment, one or more of the Bacillus strains described in the preceding paragraphs (e.g., Bacillus strain 1000, 1001 , 8001 , 1047 or 1607) can be used to treat plants along with another bacterial strain selected from the group consisting of a different Bacillus strain, a lactic acid bacterial strain, and combinations thereof. In still another embodiment, the additional Bacillus strain can be selected from the group consisting of Bacillus amyloliquefaciens, Bacillus licheniformis, Bacillus pumilus, other Bacillus strains, and combinations thereof. In yet another embodiment, one or more of the Bacillus strains described in the preceding paragraphs (e.g., Bacillus strain 1000, 1001, 8001, 1047 or 1607) can be used to treat plants along with any other bacterial strain effective to treat plants to inhibit a fungal or bacterial infection or a fungal or bacterial disease.

[0136] The compositions described herein can be used to treat plants for any period of time that is effective to induce the beneficial growth effects or soil remediation and includes multiple rounds of application and potential scheduled application to supplement the activity of the previously administered compositions. For example, in one embodiment treatment of the plants or application of the compositions to soils can occur daily, bi-weekly, three times a week, four times a week, five times a week, once weekly, monthly, or for any suitable time or using any suitable protocol. The time periods for treatment of the plants are non- limiting and it should be appreciated that any time period or treatment schedule determined to be effective to plant growth and / or plant productivity and / or soil remediation or improved soil nutrient quality may be used.

[0137] In various illustrative embodiments, the Bacillus strain (e.g., Bacillus strain 1000, 1001, 8001, 1047 or 1607), or any other bacterial strains added in addition to Bacillus strain 1000, 1001 , 8001 , 1047 or 1607, can be used to treat plants or improve soil quality by administrating at about 1.0 x 102CFU, about 1.0 x 102CFU to about 1.0 x 103CFU, about 1.0 x102CFU to about 1.0 x 104CFU, about 1.0 x 102CFU to about 1.0 x 105CFU, about 1.0 x 102CFU to about 1.0 x 106CFU, about 1.0 x 102CFU to about 1.0 x 107CFU. about 1.0 x 102CFU to about 1.0 x 108CFU, about 1.0 x 102CFU to about 1.0 x 109CFU, about 1.0 x 102CFU to about 1.0 x 1010CFU, about 1.0 x 103CFU to about 5.0 x 1012CFU, about 1.0 x 106CFU to about 5.0 x 1012CFU or at about 1.0 x 103CFU to about 1.0 x 1010CFU. In other embodiments, the Bacillus strain (e.g., Bacillus strain 1000, 1001, 8001, 1047 or 1607) can be used to treat the plants or soil at an amount greater than or about 1.0 x 102CFU, at greater than or about 1.0 x 103CFU, at greater than or about 1.1 x 103CFU, at greater than or about 1.25 x103CFU, at greater than or about 1.5 x 103CFU, at greater than or about 1.75 x 103CFU, at greater than or about 1.0 x 104CFU, at greater than or about 2.0 x 104CFU, at greater than or about 3.0 x 104CFU, at greater than or about 4.0 x 104CFU, at greater than or about 5.0 x 104CFU, at greater than or about 6.0 x 104CFU, at greater than or about 7.0 x 104CFU, at greater than or about 8.0 x 104CFU, at greater than or about 1.0 x 105CFU, at greater than or about 1.0 x 106CFU, at greater than or about 1.0 x 107CFU, at greater than or about 1.0 x 108CFU, at greater than or about 1.0 x 109CFU, at greater than or about 1.0 x 1010CFU, at greater than or about 1.0 x 1011CFU, or at greater than or about 1.0 x 1012CFU. In all of the embodiments using “CFU”, the CFU can be CFU / gram or CFU / ml of the liquid or powdered formulation applied to the plants.

[0138] In one embodiment the compositions comprising the Bacillus strains disclosed herein can be in the form of, for example, a liquid or a dry composition, such as a suspension or a lyophilized composition or a powder, a freeze-dried composition, a gel, or a pellet, for example. In one illustrative embodiment, the compositions comprising the Bacillus strain can be in the form of a powder or a liquid, and can be formulated to be sprayed on or manually applied to the plant or to the soil from which the plant obtains nutrients. In other embodiments, the Bacillus strain can be mixed with fertilizer, mixed with soil, or used as a seed coating for contacting the plant or a plant part. Any suitable method known in the art for contacting the plant or the soil with the Bacillus strain can be used to achieve any of the effective amounts of Bacillus strain 1000, 1001, 8001, 1047 or 1607, or a strain having all of the identifying characteristics of any of these Bacillus strains, or combinations thereof, described herein. In one aspect, the Bacillus strain can comprise live Bacillus cells or spores. In another illustrativeembodiment, the plant can be contacted with the Bacillus strain by contacting the Bacillus strain with a part of the plant selected from the group consisting of a leaf, a seed, a root, a flower, a shoot, a bud, and combinations thereof.

[0139] In illustrative aspects, Bacillus strain 1000, 1001, 8001, 1047 or 1607, or a strain having all of the identifying characteristics of any of these Bacillus strains, and combinations thereof, can be in the form of a commercial package, or any other suitable composition. In another illustrative embodiment, the Bacillus strain(s) in the commercial package, or other composition can be in the form of a concentrate (e.g., about 1 x 108to about 5 x 109CFU / g) or a superconcentrate (e.g., about 1 x 1010to about 5 x 1012CFU / g). In another embodiment, the Bacillus strain(s) in the commercial package or other composition can be in a dry form (e.g., a powder), a liquid form, a freeze-dried form, or in the form of a gel, or any other suitable form for application to plants or the soil from which the plant obtains nutrients.

[0140] In another illustrative embodiment, the Bacillus strain(s) in the commercial package or composition can further comprise a carrier for the Bacillus strain(s). In various embodiments, the carrier can be selected from the group consisting of bran, rice hulls, a salt, mineral oil, a dextrin (e.g., maltodextrin), whey, sugar, limestone, dried starch, sodium silico aluminate, vegetable oil, and combinations thereof. In another embodiment, the carrier can be any suitable carrier known in the art for a composition for application to plants to treat plants having a fungal or bacterial disease or infection. In another embodiment, the Bacillus strain(s) in the commercial package or composition can further comprise a binder such as clay, yeast cell wall components, aluminum silicate, glucan, or other known binders, and / or micronutrients, including but not limited to, nitrogen and phosphorus. Any of these components can be exogenously added (i.e., not naturally present in combination with the Bacillus strains) and any of the compositions described herein can be present in nutrient compositions for plants.

[0141] In another embodiment, the commercial package or composition can be in a container for commercial use. In various embodiments the container can be, for example, a bag (e.g., a 20-pound bag, a 50-pound bag, a 2-ounce bag, a 1-pound bag, or a 1-kilogram bag), a pouch, a drum, a bottle, or a box. In illustrative aspects, the container comprising the Bacillus strain(s) can comprise plastic, metal, foil, paper, fiber, or cardboard (e.g., a plastic pail, a paper bag, a foil bag, a fiber drum, etc.). The commercial package can further comprise instructions for use of one or more of the Bacillus strains.

[0142] The following examples are for illustrative purposes only. The examples are non-limiting, and are not intended to limit the invention in any way.EXAMPLE 1Rapid-PCR DNA Profiles

[0143] The Randomly Amplified Polymorphic DNA PCR method (herein referred to as RAPD-PCR) was used to identify genetic variability of each strain. Preparation of the DNA to be used in the RAPD-PCR reaction was done by using the QIAGEN® Tissue and Blood single column kit (QIAGEN®, Venlo, The Netherlands). Figs. 5 to 9 illustrate RAPD-PCR results for the strains described in this application. The results show that the strains are unique from each other.EXAMPLE 2Phosphorus availability

[0144] Each of the five strains Bacillus strain 1000 (NRRL No. B-67888), Bacillus licheniformis strain 1001 (NRRL No. B-67889), Bacillus subtilis strain 8001 (NRRL No. B- 67890), Bacillus strain Bacillus pumilus strain 1047 (NRRL No. B-67891), Bacillus amyloliquefaciens strain 1607 (NRRL No. B-67666)) was tested for the ability to solubilize phosphate and produce the enzyme phytase. In soil, solubilization of inorganic phosphate and digestion of phytate can convert phosphate into a form that is easier for plants to uptake. Each of the Bacillus strains were grown in tryptic soy broth for 24h. Bacillus (2ul) was spotted on Pikovskaya’s agar (PKA) and LB agar with 1% phytic acid and grown for 24h at 37°C. The PKA plates were examined for zones of clearing. The phytate agar was stained with a 2% cobalt chloride solution for five minutes, and destained with a 6.25% ammonium molybdate and 0.42% ammonium metavanadate solution for five minutes before being examined for zones of clearing. All five Bacillus strains showed zones of clearing on both types of agars indicating all five strains tested positive for their ability to solubilize calcium phosphate and produce phytase.

[0145] Table 1. Presence of phosphate solubilization and phytase activity in Bacillus strains.EXAMPLE 3:Growth promotion in corn plants

[0146] Com seeds were soaked in water with or without a suspension of Bacillus cells at 2 x 105CFU / ml for one hour. The suspension comprising a blend of B-67888, B-67889, B- 67890, B-67891 and B-67666. Treated and untreated seeds were planted in soil in pots (n=8) and placed in a growth chamber set to a daily cycle of 29°C for 16 hours, followed by 25 °C for 8 hours for 17 days. The pots were watered daily, and no chemical fertilizer was added. At 17 days leaf greenness was measured with a SPAD meter and the dry root weight, shoot weight, and overall plant weight were measured. There was a numerical increase in SPAD values and fresh weight in treated plants. Results are provided in Figs. 1 and 2.EXAMPLE 4:Improved yield in corn

[0147] A Bacillus blend, comprising B-67888, B-67889, B-67890, B-67891 and B- 67666, was applied in a seed coating to com seeds at 1.8 x 1012CFU per 100 lb of seed. Treated and untreated seeds were grown in four replicate 10 by 40 ft plots at 2 lbs of seed per plot. The com was grown using standard management practices and harvested at full maturity. The average yield increased from 165 Bu / acre to 176 Bu / acre when treated with the Bacillus blend. See the results presented in Fig. 3.EXAMPLE 5:Phosphate solubilization

[0148] Each strain was tested for the ability to solubilize phosphate using a quantitative method. Each strain was grown in TSB overnight, washed in sterile peptone, and adjusted to an optical density of 0.5 at 600nm. Pikovskaya broth containing 5g / L of insoluble tricalcium phosphate was inoculated with 200p I of culture. The broth was incubated at 32°C for 48 hours. The cultures were centrifuged for 10 minutes at 4000rpm. The supernatant was combined with 500pl of a 2.5% ammonium molybdate solution and 200pl of a 0.25% a-amino-naphthol solution in a total volume of 5ml. The solutions were incubated in the dark for 30 minutes, then absorbance was read at 660nm. The absorbance values were compared to a phosphate standard curve to quantify the amount of phosphate solubilized by each Bacillus strain. Each of the five strains was able to solubilize a portion of the tricalcium phosphate. Results shown in Fig. 4.

Claims

WHAT IS CLAIMED IS:1 . A method for increasing plant growth and / or productivity, the method comprising contacting the plants, the seeds of said plant, or the soil in which said plants are grown, with an effective amount of a composition comprising an isolated Bacillus strain selected from the group consisting of Bacillus licheniformis strain 1000 (NRRL No. B-67888), a strain having all of the identifying characteristics of Bacillus strain 1000 (NRRL No. B- 67888), Bacillus licheniformis strain 1001 (NRRL No. B-67889), a strain having all of the identifying characteristics of Bacillus strain 1001 (NRRL No. B-67889), Bacillus subtilis strain 8001 (NRRL No. B -67890), a strain having all of the identifying characteristics of Bacillus strain 8001 (NRRL No. B-67890), Bacillus strain Bacillus pumilus strain 1047 (NRRL No. B- 67891), a strain having all of the identifying characteristics of Bacillus strain 1047 (NRRL No. B-67891), Bacillus amyloliquefaciens strain 1607 (NRRL No. B-67666), a strain having all of the identifying characteristics of Bacillus strain 1607 (NRRL No. B-67666), and any combination thereof.

2. The method of claim 1 wherein the plants, plant seeds, or the soil in which said plants are grown, is contacted with an effective amount of a composition comprising two or more isolated Bacillus strains selected from the group consisting of Bacillus licheniformis strain 1000 (NRRL No. B-67888), a strain having all of the identifying characteristics of Bacillus strain 1000 (NRRL No. B-67888), Bacillus licheniformis strain 1001 (NRRL No. B-67889), a strain having all of the identifying characteristics of Bacillus strain 1001 (NRRL No. B-67889), Bacillus subtilis strain 8001 (NRRL No. B-67890), a strain having all of the identifying characteristics of Bacillus strain 8001 (NRRL No. B-67890), Bacillus strain Bacillus pumilus strain 1047 (NRRL No. B-67891), a strain having all of the identifying characteristics of Bacillus strain 1047 (NRRL No. B-67891), Bacillus amyloliquefaciens strain 1607 (NRRL No. B-67666), a strain having all of the identifying characteristics of Bacillus strain 1607 (NRRL No. B-67666).

3. The method of claim 1 wherein the plants, plant seeds or the soil in which said plants are grown, is contacted with an effective amount of a composition comprising three or more isolated Bacillus strains selected from the group consisting of Bacillus licheniformis strain 1000 (NRRL No. B-67888), a strain having all of the identifying characteristics of Bacillus strain 1000 (NRRL No. B-67888), Bacillus licheniformis strain 1001 (NRRL No. B-67889), a strain having all of the identifying characteristics of Bacillus strain 1001 (NRRL No. B-67889), Bacillus subtilis strain 8001 (NRRL No. B-67890), a strain having all of the identifyingcharacteristics of Bacillus strain 8001 (NRRL No. B-67890), Bacillus strain Bacillus pumilus strain 1047 (NRRL No. B-67891), a strain having all of the identifying characteristics of Bacillus strain 1047 (NRRL No. B-67891 ), Bacillus amyloliquefaciens strain 1607 (NRRL No. B-67666), a strain having all of the identifying characteristics of Bacillus strain 1607 (NRRL No. B-67666).

4. The method of any one of claims 1-3 wherein the composition comprises isolated Bacillus licheniformis strain 1000 (NRRL No. B-67888).

5. The method of any one of claims 1-3 wherein the composition comprises isolated Bacillus licheniformis strain 1001 (NRRL No. B-67889).

6. The method of any one of claims 1-3 wherein the composition comprises isolated Bacillus subtilis strain 8001 (NRRL No. B-67890).

7. The method of any one of claims 1-3 wherein the composition comprises isolated Bacillus pumilus strain 1047 (NRRL No. B-67891).

8. The method of any one of claims 1-3 wherein the composition comprises isolated Bacillus amyloliquefaciens strain 1607 (NRRL No. B-67666).

9. The method of claim 1 wherein the plants, plant seeds or to the soil in which the plants are grown is contacted with an effective amount of a composition comprising i) an isolated Bacillus licheniformis strain 1000 (NRRL No. B-67888), or a strain having all of the identifying characteristics of Bacillus strain 1000 (NRRL No. B-67888); ii) an isolated Bacillus licheniformis strain 1001 (NRRL No. B-67889), or a strain having all of the identifying characteristics of Bacillus strain 1001 (NRRL No. B-67889); iii) an isolated Bacillus subtilis strain 8001 (NRRL No. B-67890), or a strain having all of the identifying characteristics of Bacillus strain 8001 (NRRL No. B-67890); iv) an isolated Bacillus pumilus strain 1047 (NRRL No. B-67891), or a strain having all of the identifying characteristics of Bacillus strain 1047 (NRRL No. B-67891); and v) an isolated Bacillus amyloliquefaciens strain 1607 (NRRL No. B-67666), or a strain having all of the identifying characteristics of Bacillus strain 1607 (NRRL No. B-67666).

10. The method of any one of claims 1-9 wherein said soil is contacted with said composition prior to planting said plant seeds.

11. The method of claim 1 or 10 wherein the seeds are contacted with said composition prior to planting the seeds.

12. The method of any one of claims 1-11 wherein the soil in which the plants are grown is contacted with said composition prior to planting the seeds.

13. The method of any one of claims 1-12 wherein the plants are pasture plants, crop plants or ornamental plants.

14. The method of claim 13 wherein the plants are crop plants.

15. A method for increasing the level of orthophosphates, H2PO4" or HPO42" in an agricultural field, the method comprising contacting the agricultural field with an effective amount of a composition comprising an isolated Bacillus strain selected from the group consisting of Bacillus licheniformis strain 1000 (NRRL No. B-67888), a strain having all of the identifying characteristics of Bacillus strain 1000 (NRRL No. B-67888), Bacillus licheniformis strain 1001 (NRRL No. B-67889), a strain having all of the identifying characteristics of Bacillus strain 1001 (NRRL No. B-67889), Bacillus subtilis strain 8001 (NRRL No. B-67890), a strain having all of the identifying characteristics of Bacillus strain 8001 (NRRL No. B- 67890), Bacillus strain Bacillus pumilus strain 1047 (NRRL No. B-67891), a strain having all of the identifying characteristics of Bacillus strain 1047 (NRRL No. B-67891), Bacillus amyloliquefaciens strain 1607 (NRRL No. B -67666), a strain having all of the identifying characteristics of Bacillus strain 1607 (NRRL No. B-67666), and any combination thereof.

16. The method of claim 15 wherein said Bacillus strain is embedded within a biodegradable matrix.

17. The method of claim 15 or 16 wherein the soil is contacted with an effective amount of a composition comprising two or more isolated Bacillus strains selected from the group consisting of Bacillus licheniformis strain 1000 (NRRL No. B-67888), a strain having all of the identifying characteristics of Bacillus strain 1000 (NRRL No. B-67888), Bacillus licheniformis strain 1001 (NRRL No. B-67889), a strain having all of the identifying characteristics of Bacillus strain 1001 (NRRL No. B-67889), Bacillus subtilis strain 8001 (NRRL No. B-67890), a strain having all of the identifying characteristics of Bacillus strain 8001 (NRRL No. B-67890), Bacillus strain Bacillus pumilus strain 1047 (NRRL No. B-67891), a strain having all of the identifying characteristics of Bacillus strain 1047 (NRRL No. B- 67891), Bacillus amyloliquefaciens strain 1607 (NRRL No. B-67666), a strain having all of the identifying characteristics of Bacillus strain 1607 (NRRL No. B-67666).

18. The method of claim 15 wherein the soil in which the plant or plant seeds are grown is contacted with an effective amount of a composition comprising three or more isolated Bacillus strain selected from the group consisting of Bacillus licheniformis strain 1000 (NRRL No. B-67888), a strain having all of the identifying characteristics of Bacillus strain 1000 (NRRL No. B-67888), Bacillus licheniformis strain 1001 (NRRL No. B-67889), a strain having all of the identifying characteristics of Bacillus strain 1001 (NRRL No. B-67889), Bacillussubtilis strain 8001 (NRRL No. B-67890), a strain having all of the identifying characteristics of Bacillus strain 8001 (NRRL No. B-67890), Bacillus strain Bacillus pumilus strain 1047 (NRRL No. B-67891), a strain having all of the identifying characteristics of Bacillus strain 1047 (NRRL No. B-67891), Bacillus amyloliquefaciens strain 1607 (NRRL No. B-67666), a strain having all of the identifying characteristics of Bacillus strain f607 (NRRL No. B-67666).

19. The method of any one of claims 15-18 wherein the composition comprises isolated Bacillus licheniformis strain 1000 (NRRL No. B-67888).

20. The method of any one of claims 15-18 wherein the composition comprises isolated Bacillus licheniformis strain 1001 (NRRL No. B-67889).

21. The method of any one of claims 15-18 wherein the composition comprises isolated Bacillus subtilis strain 8001 (NRRL No. B-67890).

22. The method of any one of claims 15-18 wherein the composition comprises isolated Bacillus pumilus strain 1047 (NRRL No. B-67891).

23. The method of any one of claims 15-18 wherein the composition comprises isolated Bacillus amyloliquefaciens strain 1607 (NRRL No. B-67666).

24. The method of claim 15 or 16 wherein the soil is contacted with an effective amount of a composition comprising i) an isolated Bacillus licheniformis strain 1000 (NRRL No. B-67888), or a strain having all of the identifying characteristics of Bacillus strain 1000 (NRRL No. B-67888); ii) an isolated Bacillus licheniformis strain 1001 (NRRL No. B-67889), or a strain having all of the identifying characteristics of Bacillus strain 1001 (NRRL No. B-67889); iii) an isolated Bacillus subtilis strain 8001 (NRRL No. B-67890), or a strain having all of the identifying characteristics of Bacillus strain 8001 (NRRL No. B-67890); iv) an isolated Bacillus pumilus strain 1047 (NRRL No. B-67891), or a strain having all of the identifying characteristics of Bacillus strain 1047 (NRRL No. B-67891); and v) an isolated Bacillus amyloliquefaciens strain 1607 (NRRL No. B-67666), a strain having all of the identifying characteristics of Bacillus strain 1607 (NRRL No. B-67666).

25. The method of claim 15 wherein the agricultural field is a pasture.

26. The method of claim 15 wherein the agricultural field is a cultivated field.

27. The method of claim 26 wherein the agricultural field is contacted with said composition at least 7 days prior to planting seeds in said agricultural field.

28. A commercial package or a composition comprising an isolated Bacillus strain selected from the group consisting of Bacillus licheniformis strain 1000 (NRRL No. B-67888), a strain having all of the identifying characteristics of Bacillus strain 1000 (NRRL No. B-67888), Bacillus licheniformis strain 1001 (NRRL No. B-67889), a strain having all of the identifying characteristics of Bacillus strain 1001 (NRRL No. B-67889), Bacillus subtilis strain 8001 (NRRL No. B-67890), a strain having all of the identifying characteristics of Bacillus strain 8001 (NRRL No. B-67890), Bacillus strain Bacillus pumilus strain 1047 (NRRL No. B- 67891), a strain having all of the identifying characteristics of Bacillus strain 1047 (NRRL No. B-67891), Bacillus amyloliquefaciens strain 1607 (NRRL No. B-67666), a strain having all of the identifying characteristics of Bacillus strain 1607 (NRRL No. B-67666), and any combination thereof; and an exogenous source of purified nitrate and / or potassium chloride.

29. The composition of claim 28 wherein the composition is a liquid.

30. The composition of claim 28 wherein the composition is dry.

31. The composition of claim 30 wherein the composition is freeze-dried, in the form of a powder, or in the form of a pellet.

32. The composition of claim 28 further comprising seeds wherein the said Bacillus strain is coated onto said seeds.

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