Consortium of the bacillus megaterium кuк2а and paenibacillus sp. 02189 strains and use thereof

Abstract

The invention relates to agricultural biotechnology and microbiology, and more particularly to the biomodification of mineral fertilizers and soil improvers by the use of a bacterial consortium of Bacillus megaterium КУК2А and Paenibacillus sp. 02189, as well as to the production of biomodifiers and biomodified mineral fertilizers and soil improvers for use in crop cultivation for the purpose of stimulating plant growth, improving plant nutrition and increasing crop yield.

Description

[0001] CONSORTIUM OF BACILLUS MEGATERIUM KUK2A AND PAENIBACILLUS SP. 02189 STRAINS AND ITS APPLICATION

[0002] Field of technology

[0003] The invention relates to agricultural biotechnology and microbiology, in particular to the biomodification of mineral fertilizers and ameliorants and to the production of biomodifiers and biomodified mineral fertilizers and ameliorants for use in plant growing for the purpose of stimulating plant growth, improving nutrition and increasing the yield of agricultural crops.

[0004] State of the art

[0005] Bacillus megaterium bacteria live in the soil, decomposing organic matter and releasing the potassium and phosphorus they contain, converting them into soluble salts. The resulting compounds become available for absorption by plants. It has been established that the use of bacterial preparations is particularly effective in fertile soils. Since such soils are rare on our planet, it is especially important to identify microbial strains that are potentially active in mobilizing nutrients not only from organic but also from inorganic soil components, such as secondary minerals.

[0006] The biological product Phosphorobacterin is known for improving soil fertility, along with a method for its production (SU97943). In this method, bacteria are grown on a potato medium, dried, and mixed with powdered kaolin, which serves as a substrate for preserving the dried bacterial spores. Using Phosphorobacterin mineralizes complex organophosphorus compounds, leading to increased soil fertility.

[0007] It is known to use bacteria of the Bacillus megaterium species as a means of stimulating the growth of plants and / or their roots (for example, document US20130014434 is known, in which a nutritional composition for the roots of a transplanted plant contains, among other microorganisms, Bacillus megaterium bacteria) or as a means of combating plant diseases.

[0008] A strain of Bacillus megaterium bacteria is also known to mobilize phosphorus and silicon from lithospheric objects and is resistant to polyhexamethylene guanidine (RU2327737). The strain Bacillus megaterium var. phosphaticum BKM B-2357D is capable of leaching phosphorus and silicon from lithospheric objects and is resistant to polyhexamethylene guanidine. This strain can be used to produce a bacterial preparation that increases the survival rate in soil and the yield of agricultural crops, for the reclamation of man-made contaminated lands, and in biotechnological work in the field of geochemistry. The strain of Bacillus megaterium V3, deposited at the All-Russian Research Institute of Agricultural Microbiology under the number RCAM 04324, is known as a means of accelerating the growth and increasing the productivity of grain, vegetable, and tree crops (RU2649359).The Bacillus megaterium V3 strain has a phytostimulating effect on various agricultural crops, in particular oats, watercress, radishes, tomatoes, grapes, as well as woody crops, in particular spruce and pine.

[0009] The Bacillus megaterium 2-06-TS1 microbial strain, deposited in the All-Russian Collection of Industrial Microorganisms of the Federal State Unitary Enterprise "GosNIIGenetika" under the number VKPM B-12402, is known as an activator of photosynthesis and plant germination energy (RU2627608). Document RU2627608 describes the use of the Bacillus megaterium 2-06-TS1 strain for the development of biopreparations to enhance plant productivity (increasing seed germination energy, seed vigor, and stimulation of root system development and photosynthetic apparatus function).

[0010] A multifunctional plant growth product based on the bacterial strain Bacillus megaterium 501 GR (RU2558291) is available. This spore-forming bacterial strain, Bacillus megaterium 501 GR, has a growth-promoting effect, improves plant phosphorus nutrition, and promotes bioremediation of pesticide-contaminated soils. Using Bacillus megaterium 501 GR increases plant productivity and protects them from phytopathogenic fungi and microorganisms that cause root diseases.

[0011] The bacterium Paenibacillus mucilaginosus is a typical silicate bacterium and has proven itself as an effective biofertilizer in agriculture. Paenibacillus mucilaginosus is capable of decomposing insoluble soil minerals with the formation of potassium ions and water-soluble phosphorus necessary for plants [4-6]. Strains of P. mucilaginosus also contribute to an increase in plant yields due to the ability to synthesize auxins. A review retrospective analysis of the use of bacteria of the genus Paenibacillus in industrial biotechnology as a producer of biopreparations for agricultural purposes is given in the article by the authors Ha T.Z., Kanarsky A.V., Kanarskaya Z.A., Shcherbakov A.V., Shcherbakova E.N. "Prospects for the use of bacteria of the genus Paenibacillus in industrial biotechnology for the production of biopreparations for agricultural purposes" and Bulletin of the Volga State Technological University. Series: Forest. Ecology. Nature Management. 2020. No. 3 (47). P. 74-84.

[0012] A fertilizer based on the bacterial strain Bacillus mucilaginosus - Bac 1208, capable of producing an exopolysaccharide that creates beneficial microflora in the rhizosphere (RU Patent 2081867), and a fertilizer based on the bacterial strain Bacillus mucilaginosus, capable of transforming poorly soluble phosphorus and potassium compounds into a form accessible to plants, possessing phytopathogenic activity (RU Patent No. 2408722) are known.

[0013] Also known is the Paenibacillus mucilaginosus B-12259 strain, which stimulates plant growth and has an increased capacity to synthesize bactericins with a broad spectrum of suppression of phytopathogenic microflora. It is deposited in the All-Russian Collection of Industrial Microorganisms under the registration number VKPM B-12259. Paenibacillus mucilaginosus B-12259 is used as a fertilizer for agricultural crops and for the prevention and treatment of fungal plant diseases.

[0014] Patent RU2626543, dated July 28, 2017, "Paenibacillus mucilaginosus Strain Used as a Fertilizer and for Stimulating Plant Growth and Protecting Plants from Fungal Diseases," describes the Paenibacillus mucilaginosus Pm 2906 bacterial strain, deposited with the All-Russian Collection of Agricultural Products under No. B-12259. The strain was isolated from cultivated soils using a multistage selection method, and a fertilizer based on it. The fertilizer for growing agricultural crops is obtained by stepwise fermentation of the strain on Ashby's nutrient medium. Using this fertilizer increases crop yields and reduces the application rates of nitrogen, phosphorus, and potassium mineral fertilizers during cultivation. Paenibacillus mucilaginosus Pm 2906 can also be used as an antiphytopathogenic agent in the prevention and / or treatment of plant diseases.

[0015] The problem of increasing agricultural crop yields remains acute and relevant, so it is important to develop and find new approaches and means to solve this issue.

[0016] Despite many years of research on various plant growth-promoting rhizobacteria, including various strains of Bacillus megaterium and Paenibacillus sp., their combined action, which enhances and expands the potential for the beneficial use of these bacteria, has not been investigated, which allows us to address the current issues of increasing crop yields and their resistance to various diseases.

[0017] Disclosure of invention

[0018] The objective of the present invention is to achieve a highly effective, enhanced efficiency of mineral fertilizers and ameliorants, aimed at increasing their digestibility, enhancing plant growth and productivity, and increasing their resistance to diseases. This is achieved by applying a bacterial culture to the surface of mineral fertilizers and ameliorants. This culture consists of a consortium of bacterial strains that mutually enhance each other's beneficial effects on plants. Another objective of the present invention is to develop and create biomodified mineral (biomineral) fertilizers and ameliorants.

[0019] The task is being solved by developing a consortium of bacteria strains Bacillus megaterium KUK2A and Paenibacillus sp. 02189, intended for the biomodification of mineral fertilizers and ameliorants.

[0020] The stated task is also solved by using a consortium of Bacillus megaterium KUK2A and Paenibacillus sp. 02189 strains for the biomodification of mineral fertilizers and ameliorants.

[0021] To achieve this, the consortium of strains is applied to ameliorants or mineral fertilizers, used in both solid and liquid form. In some embodiments, the solid mineral fertilizer is granulated, prilled, microgranulated, powdered, or water-soluble.

[0022] In various embodiments of the invention, bacteria are applied directly to a mineral fertilizer or ameliorant, or using a carrier. In some embodiments, the carrier is a mineral or organic substance, natural or synthetic, suitable for agricultural use. In some particular embodiments, the carrier is a natural or non-natural substance based on siliceous rocks.

[0023] In some embodiments of the invention, an agent that enhances (increases) the adhesion of bacteria to a mineral fertilizer and / or carrier is used to immobilize bacteria. In some particular embodiments of the invention, the agent that enhances bacterial adhesion includes cationic starch, collagen glue, bone gelatin glue, albumin glue, fish glue, cherry glue, dextrin, molasses, gum arabic, or liquid glass.

[0024] In some embodiments of the invention, the mineral fertilizer is a nitrogen, phosphorus, potassium, boron, molybdenum, complex, complex-mixed, water-soluble, or micronutrient fertilizer, or a mixture of one of the fertilizers with micronutrients. In some particular embodiments of the invention, the mineral fertilizer is urea, ammonium nitrate, ammonium sulfate, calcium nitrate, ammophos, sodium nitrate, diammonium phosphate, nitroammophoska, phosphate rock, potassium sulfate, potassium salt, sulfoammophos, diammophos, azophoska, nitrophos, nitrophoska, potassium nitrate, potassium chloride, or potassium magnesium sulfate.

[0025] In some embodiments of the invention, the ameliorant is an ameliorant-sorbent, an ameliorant-structure-forming agent, a ameliorant fertilizer, including, for example, phosphogypsum, phosphate rock.

[0026] In various embodiments of the invention, bacteria are used in spore or vegetative form. In more preferred embodiments, the bacteria are used in their dormant (spore) form, as this ensures longer storage and survival of the bacteria.

[0027] The stated task is also solved by developing a biomodifier of mineral fertilizers and ameliorants (a bacterial preparation for modifying mineral fertilizers and ameliorants), including bacterial strains Bacillus megaterium KUK2A, Paenibacillus sp. 02189 and, optionally, at least one auxiliary substance.

[0028] In some particular embodiments of the invention, the excipient is a carrier and / or an agent that increases the adhesion of bacteria to a mineral fertilizer and / or ameliorant and / or carrier. In particular embodiments of the invention, the concentration of each bacterial strain on the carrier is at least 10 4 CFU / g carrier.

[0029] In particular embodiments of the invention, the concentration of each of the bacterial strains in the liquid biomodifier is at least 10 5 CFU / ml of bacterial suspension.

[0030] The stated task is also solved by developing a biomodified mineral (biomineral) fertilizer and ameliorant for improved nutrition and stimulation of plant growth, including mineral fertilizer and a consortium of bacteria strains Bacillus megaterium KUK2 and Paenibacillus sp. 02189.

[0031] In various embodiments of the invention, the mineral fertilizer is used in solid or liquid form. In some embodiments, the solid mineral fertilizer is in granular, prilled, microgranulated, or powder form.

[0032] In specific embodiments, the bacterial consortium is applied to the mineral fertilizer or ameliorant directly or using a carrier. In some embodiments of the invention, the concentration of each strain of the bacterial consortium in the biomodified mineral (biomineral) fertilizer (or, respectively, ameliorant) is at least 10 3 CFU per 1 g of solid mineral fertilizer. In special cases - at least 10 4 CFU per 1g of solid mineral fertilizer (ameliorant).

[0033] In some embodiments of the invention, the bacterial consortium is mixed with a liquid mineral fertilizer. In particular embodiments of the invention, the concentration of each strain of the bacterial consortium is at least 10 3 CFU per 1 ml of liquid mineral fertilizer.

[0034] The present invention also includes the use of a consortium of strains Bacillus megaterium KUK2A and Paenibacillus sp. 02189 for producing a biomodifier of mineral fertilizers and ameliorants.

[0035] The present invention also includes the use of a biomodifier based on a consortium of strains Bacillus megaterium KUK2A and Paenibacillus sp. 02189 for the production of a biomodified mineral (biomineral) fertilizer or ameliorant for improving nutrition and stimulating plant growth.

[0036] The problem is also solved by developing a method for biomodifying mineral fertilizers and ameliorants, comprising mixing bacteria of the Bacillus megaterium KUK2 and Paenibacillus sp. 02189 strains (a biomodifier according to the invention) with the mineral fertilizer or ameliorant, respectively. In some embodiments of the invention, said method comprises immobilizing bacteria of the Bacillus megaterium KUK2 and Paenibacillus sp. 02189 strains directly on the mineral fertilizer or ameliorant, as well as using a carrier.

[0037] In some particular embodiments of the invention, a bacterial suspension (or a bacterial suspension to which auxiliary substances have been pre-added) is mixed with a liquid mineral fertilizer or directly applied to the surface of a solid mineral fertilizer or ameliorant.

[0038] In other particular embodiments of the invention, a bacterial suspension (a consortium of Bacillus megaterium KUK2 and Paenibacillus sp. 02189 strains) is first applied to a carrier, after which the resulting biomodifier is applied to a mineral fertilizer (or ameliorant) by mixing. Since the CFU content of microorganisms in the bacterial suspension affects the number of CFU of microorganisms ultimately deposited on the carriers and the mineral fertilizer and ameliorant, their concentration is selected based on the amount that is the target content in the biomodified mineral (biomineral) fertilizer / ameliorant. The more CFU in the bacterial suspension, the greater the number of agronomically beneficial microorganisms deposited on the carrier and the mineral fertilizer / ameliorant. In particular embodiments of the invention, the bacterial consortium is applied to the carrier until the concentration of each of the strains on the carrier is at least 10 4CFU per 1 g of carrier. In specific embodiments of the invention, the resulting biomodifier is applied to mineral fertilizer / ameliorant at a ratio of at least 1 kg per ton of mineral fertilizer / ameliorant.

[0039] As a result of implementing the invention, the following technical results are achieved:

[0040] - a new effective method for biomodification of mineral fertilizers and ameliorants has been developed using a consortium of bacterial strains Bacillus megaterium KUK2 and Paenibacillus sp. 02189;

[0041] - a consortium of bacterial strains Bacillus megaterium KUK2 and Paenibacillus sp. 02189 is capable of increasing the assimilation of poorly soluble phosphorus and potassium compounds by directly influencing the solubilization (dissolution) of mineral fertilizers and increasing their assimilation by plants due to the production of various organic and inorganic acids by bacteria, and therefore the use of a consortium of Bacillus megaterium KUK2 and Paenibacillus sp. 02189 strains in biomineral fertilizers and biomodifiers ensures an increase in the assimilation of mineral fertilizers by plants; at the same time, due to the fact that the mechanism of solubilization of potassium and phosphorus minerals in the strains included in the consortium differs, their joint use ensures high activity in the mobilization of phosphorus and potassium compounds, as well as a prolonged effect;

[0042] - the ability of the consortium of Bacillus megaterium KUK2 and Paenibacillus sp. 02189 strains to stimulate plant growth ensures an increase in productivity, yield and quality of agricultural products when using the consortium as part of biomineral fertilizers and biomodifiers;

[0043] - the use of a consortium of bacteria strains Bacillus megaterium KUK2 and Paenibacillus sp. 02189 in the composition of biomineral fertilizers and biomodifiers mitigates the negative impact of contact with mineral fertilizers on plants by acidifying the rhizosphere, inducing plant resistance to stress, which in general also provides an additional positive effect on plants;

[0044] - biomodified mineral (biomineral) fertilizers and ameliorants according to the invention are effective for use in growing various plants - vegetable, grain and industrial crops in agriculture, plants used in floriculture, forestry, and others, without limitations;

[0045] - a biomodifier of mineral fertilizers based on a consortium of strains Bacillus megaterium KUK2 and Paenibacillus sp. 02189 allows to reduce the loss of mineral fertilizers by leaching and binding with complexing elements;

[0046] - the biomodified mineral (biomineral) fertilizer / ameliorant according to the invention is characterized by the long-term viability of bacteria in its composition, for at least 3 months (and in most cases much longer), in concentrations that ensure the effectiveness of biomineral fertilizers / ameliorants;

[0047] - The proposed new biomineral fertilizers / ameliorants and biomodifiers based on a consortium of Bacillus megaterium KUK2A and Paenibacillus sp. 02189 strains expand the range of products for increasing yields and improving the growth of agricultural crops and other plants and their resistance to various diseases.

[0048] Terms and definitions

[0049] Unless otherwise defined, all technical and scientific terms used in this application have the same meaning as understood by those skilled in the art. References to techniques used in the description of this invention relate to well-known methods, including variations of these methods and their replacement with equivalent methods known to those skilled in the art. The term "consortium" implies the use of two or more strains of microorganisms in a single composition. Various forms of relationships exist between the microorganisms in the consortium, which make it possible to maintain stable symbiosis and equilibrium within this consortium for a certain period of time. The consortium according to the invention includes the bacterial strains Bacillus megaterium KUK2A and Paenibacillus sp. 02189. Moreover, the ratio of strains in the consortium according to the invention during the biomodification of mineral fertilizers and / or ameliorants can be any (i.e.The amount of one of the strains can vary from 1 to 99%, more preferably from 10 to 90%, 20 to 80%, 30 to 70%, or 40 to 60%) and is selected based on plant needs, soil conditions, and other factors. In some non-limiting embodiments, strains within a consortium may be present in equal amounts. This ratio allows for the full utilization of the beneficial properties of each strain. Each strain has its own unique mechanisms for providing a positive effect on plants, so using these strains in a 1:1 ratio creates equal conditions for both strains. A shift in the ratio in one direction or the other will result in a shift in the predominance of one strain, although this does not necessarily mean a decrease in effectiveness.Moreover, in some cases, the predominance of one strain in a biomodified mineral (biomineral) fertilizer / ameliorant ensures maximum digestibility of the mineral fertilizer / ameliorant, and therefore, the maximum effectiveness of its positive impact on plants. The choice of ratio is determined during biomodification (or biomodifier preparation).

[0050] In the documents of this invention, the terms "includes," "including," and the like, as well as "contains," "comprising," and the like, are interpreted to mean "includes, among other things" (or "contains, among other things"). These terms are not intended to be construed as meaning "consists solely of."

[0051] The term "and / or" means one, more than one, or all of the listed elements.

[0052] Also here, listing numeric ranges by endpoints includes all numbers within that range.

[0053] The term "optional" or "optional" or "optionally" as used herein means that the subsequently described event or circumstance may, but does not necessarily, occur, and that the description includes instances in which the event or circumstance occurs and instances in which it does not occur.

[0054] According to the invention, a "carrier" is any substance permitted for use in agriculture. This may include, but is not limited to, a mineral or organic substance, both natural and synthetic. The primary requirements for the carriers according to the invention are a large specific surface area suitable for the application of the bacterial strain according to the invention, and safety for use in plant growing, particularly in agriculture. Above all, such carriers must not contain heavy metal cations or radioactive substances. In some non-limiting embodiments, the carrier is a natural or non-natural substance based on siliceous rocks. Non-limiting examples of carriers may include diatomite (kieselguhr), zeolite, opoka, tripoli, kaolin, glauconite, silica, perlite, metakaolin, wollastonite, bentonite, inorganic microsilica, belite, white carbon black, vermiculite (expanded), talc, and diabazite.

[0055] According to the invention, a "bacterial suspension" is understood to be a complex mixture obtained by in vitro cultivation of agronomically beneficial microorganisms and containing the cultured microorganisms, residual nutrients, and metabolic products of these microorganisms. The bacterial suspension is obtained as a result of fermentation—a series of sequential operations, from the introduction of a bacterial strain of agronomically beneficial microorganism into a pre-prepared nutrient medium heated to the required temperature to the completion of the cell growth process. The composition of the nutrient medium for cultivation (the content of substances necessary to meet nutritional and energy needs in an easily digestible form), its temperature, cultivation time, and other necessary conditions (including, for example, pH, the presence of growth factors, vitamins, buffering capacity, etc.)) are selected as optimal depending on the specific strain of cultivated agronomically beneficial microorganisms.

[0056] "Culture fluid" is a term used to describe the nutrient medium in which microorganisms were grown, which contains their metabolites and other target substances produced by microorganisms, as well as residual nutrients, as opposed to "bacterial suspension", which, in addition to the culture fluid, contains cells of agronomically beneficial microorganisms.

[0057] In the present invention, a "biomodifier" ("bacterial preparation for modifying mineral fertilizers") is understood to mean a composition comprising a consortium of bacteria of the strains Bacillus megaterium KUK2 and Paenibacillus sp. 02189 and, optionally, an auxiliary substance. In particular embodiments of the invention, the biomodifier according to the invention is a composition comprising a complex of a carrier with agronomically useful microorganisms of the strains Bacillus megaterium KUK2 and Paenibacillus sp. 02189 immobilized thereon. The said biomodifier is intended for modifying mineral fertilizers and ameliorants, in particular by mixing with a mineral fertilizer (liquid or solid) and an ameliorant. In particular embodiments of the invention, the biomodifier includes at least one auxiliary substance.

[0058] The term "adjuvant," as used herein, refers to substances that do not interfere with the biological activity and properties of bacteria and are acceptable for use in agriculture (safe for use in plant growing, particularly agriculture). Specific embodiments of adjuvants include dry carriers or liquid diluents, such as water. In some specific embodiments of the invention, the adjuvant is an agent that enhances the adhesion of bacteria to a mineral fertilizer and / or ameliorant and / or carrier.

[0059] According to the invention, "mineral fertilizer" refers to inorganic compounds that contain essential plant nutrients, i.e., a fertilizer of industrial or fossil origin containing nutrients in mineral form. The invention allows for the use of, without limitation, any mineral fertilizers in solid (dry) form (e.g., granules, powder, crystals in tablet form, etc.), including water-soluble fertilizers (solid fertilizers that are completely dissolved in water before use; they are primarily intended for use through plant irrigation systems) and liquid fertilizers.Among the fertilizers that can be used are simple mineral fertilizers—those guaranteed to contain only one essential nutrient (e.g., nitrogen, phosphorus, potassium, magnesium, boron, or molybdenum); complex (including complex-mixed) mineral fertilizers—those containing at least two essential nutrients (e.g., nitrogen-potassium, nitrogen-phosphorus-potassium (NPK), or nitrogen-phosphorus-potassium fertilizers with added sulfur (NPKS)); and micronutrient fertilizers—those in which the nutrients are trace elements in a form accessible to plants. Micronutrients can also be added to any mineral fertilizer.In some particular embodiments, for obtaining biomineral fertilizers according to the invention, the following may be used, for example, urea, ammonium nitrate (ammonium nitrate), ammonium sulfate, calcium nitrate, ammophos, sodium nitrate, diammonium phosphate, nitroammophoska, phosphate rock (phosphate rock), potassium sulfate, potassium salt, sulfoammophos, diammophos, azophoska, nitrophos, nitrophoska, potassium nitrate, potassium chloride, potassium magnesium sulfate, etc.

[0060] According to the invention, a "soil ameliorant" is a substance of industrial or fossil origin designed to improve the physicochemical properties and increase the fertility of acidic, alkaline, and other soils. According to the invention, any soil ameliorant can be biomodified, including, for example, sorbent soil ameliorants, structure-forming soil ameliorants, and fertilizer-ameliorants (see, for example, Petrova T.A. et al., Types of Soil Ameliorants for the Reclamation of Technogenically Disturbed Mining Territories, and Mining Information and Analysis Bulletin, 2021, No. 4, pp. 100-112). In some cases, the ameliorant according to the invention may be, for example, phosphogypsum, phosphate rock, etc.

[0061] The term "nutrient" ("active ingredient") refers to a chemical element in a fertilizer essential for plant growth and development. Nutrients are divided into three groups: major (essential) nutrients (N, P, K); macronutrients (N, P, K, Ca, Mg, S); and micronutrients (B, Mn, Cu, Zn, Co, Mo, Fe, etc.).

[0062] The term "biomodified mineral fertilizer" ("biomineral fertilizer") refers to a mineral fertilizer whose granules are coated with bacteria that improve their properties. This term also includes liquid mineral fertilizers (such as, but not limited to, urea-ammonia mixture (UAM)) in which bacteria are added to improve their properties.

[0063] The term "biomodified ameliorant" refers to an ameliorant whose granules or particles are coated with bacteria that improve its properties. This term also includes liquid ameliorants that are infused with bacteria to improve their properties.

[0064] The concentrations of bacteria in the biomineral fertilizer, in the biomodified ameliorant, and also in the biomodifier refer to those at the beginning of storage (i.e. immediately after application or no more than one month after application), unless otherwise specified, or not less than those specified for a certain period.

[0065] Unless otherwise defined, technical and scientific terms in this application have the standard meanings generally accepted in the scientific and technical literature.

[0066] Detailed disclosure of the invention

[0067] The Bacillus megaterium strain KUK2A (may also be identified as Bacillus pumilus or Bacillus safensis) (ЕВС / 22 AR5) was deposited on 20.10.2022 in the Network Bioresource Collection in the Field of Genetic Technologies for Agriculture (RCAM) of the All-Russian Research Institute of Agricultural Microbiology (VNIISKhM) under the number RCAM 05941, and also deposited on 15.10.2024 in the Collection of Eubiotic and Epiphytic Microorganisms (CEEM) under the number CEEM B-269 as part of the international patent deposit (the document on the international deposit is an appendix to this application).

[0068] Origin of the strain

[0069] The strain was isolated from soil samples in the Samara region.

[0070] Morphological, cultural and biochemical characteristics

[0071] Colonies are light beige, smooth, and shiny. Rod-shaped, they form spores. Cells are 3-4 µm long and 1-1.5 µm in diameter. Cells often occur in chains. Capable of dissolving potassium silicates. Method, conditions, and media composition for long-term storage of the strain.

[0072] For routine laboratory work, the culture is stored in a refrigerated MPA at +4°C, with subculture three times per year. Long-term storage is cryopreserved at -80°C.

[0073] Strain characteristics

[0074] Bacillus megaterium strain KUK2A is an active producer of organic acids and a mobilizer of poorly soluble potassium and phosphorus compounds. It can be recommended for the development of a bacterial preparation for use in plant growing.

[0075] The Bacillus megaterium strain KUK2A is non-toxic, non-toxigenic, does not disseminate in human internal organs, and is not pathogenic.

[0076] Bacillus megaterium strain KUK2A belongs to the PGPR (Plant Growth Promoting Rhizobacteria) group—rhizobacteria that promote plant growth. They have a broad host range, are capable of forming endospores, and dissolving calcium phosphate and calcium phytate, as well as the mineral phlogopite.

[0077] The key features of the Bacillus megaterium KUK2A strain, which ensure its high effectiveness in modifying mineral fertilizers, include its positive effects on plants: Bacillus megaterium KUK2A directly solubilizes mineral fertilizers and increases their availability to plants through the production of various organic and inorganic acids by bacteria, and it exhibits phosphate-mobilizing activity. Furthermore, the ability of Bacillus megaterium KUK2A to mobilize poorly soluble potassium compounds increases plant productivity and yield, as well as reduces potassium loss through leaching and binding.

[0078] The strain Paenibacillus sp. (may be identified as Paenibacillus mucilaginosus or Paenibacillus barengoltzii) 02189 was deposited on 06.06.2022 in the Network Bioresource Collection in the Field of Genetic Technologies for Agriculture (RCAM) of the All-Russian Research Institute of Agricultural Microbiology (VNIISKhM) under the number RCAM02189, and also deposited on 15.10.2024 in the Collection of Eubiotic and Epiphytic Microorganisms (CEEM) under the number CEEM B-270 as part of the international patent deposit (the document on the international deposit is an appendix to this application).

[0079] Origin of the strain

[0080] Isolated from sod-podzolic soil in the Leningrad region. Federal State Budgetary Scientific Institution All-Russian Research Institute of Agricultural Microbiology (VNIISKhM).

[0081] Morphological, cultural, and biochemical characteristics: Large, motile, single and double cells. On MPA medium, colonies are colorless, slimy, smooth, and spreading. Grows well on synthetic media with mineral nitrogen. Does not liquefy gelatin, does not decompose starch, and ferments sugars. Aerobic. Optimal culture temperature is +25°C.

[0082] Method, conditions and composition of media for long-term storage of the strain

[0083] For routine laboratory work, the culture is stored in MPA or Ashby medium in a refrigerator at +4°C, with subculture three times per year. Long-term storage is cryopreserved at -80°C.

[0084] Strain characteristics

[0085] Paenibacillus sp. 02189 is an active polysaccharide producer and mobilizer of poorly soluble potassium and phosphorus compounds. It can be recommended for the development of a bacterial preparation for use in plant growing.

[0086] The strain Paenibacillus sp. 02189 is non-toxic, non-toxigenic, does not disseminate in human internal organs, and is not pathogenic.

[0087] Paenibacillus sp. 02189 bacteria belong to the PGPR (Plant Growth Promoting Rhizobacteria) group—rhizobacteria that promote plant growth. They have a broad host range, are capable of forming endospores, and dissolving calcium phosphate and calcium phytate, as well as the mineral phlogopite.

[0088] The key features of the Paenibacillus sp. 02189 strain, which ensure its high efficiency in modifying mineral fertilizers, are the mechanisms by which it positively influences plants: the Paenibacillus sp. 02189 strain directly solubilizes (dissolves) mineral fertilizers and increases their digestibility by plants through the production of various organic and inorganic acids by bacteria. Furthermore, the ability of the Paenibacillus sp. 02189 bacterial strain to mobilize poorly soluble potassium compounds increases plant productivity and yield, as well as reduces potassium loss through leaching and binding.

[0089] The aforementioned properties of Bacillus megaterium KUK2A and Paenibacillus sp. 02189 bacteria to directly enhance mineral absorption by plants distinguish their use from existing biological fertilizer modification technology based on the Bacillus Subtilis 4-13 strain. While an effective growth-promoting agent, the latter only indirectly influences the absorption of nutrients from fertilizer granules by stimulating root growth, including increasing the number of root hairs. Using a consortium of strains according to the invention increases the effectiveness of fertilizers through cross-action: if external conditions adversely affect the development of one strain, the second strain provides a beneficial effect. Moreover, the strains in the consortium have different mechanisms for their beneficial effects on plants.Thus, the Bacillus megaterium KUK2A strain is responsible for the prolonged action of the consortium: when entering the soil, the bacteria in spore form are able to maintain their viability for a long time, until they reach the root zone, and the plant's root exudates activate these bacteria ("exit from the spores"). This property allows for an extended period of beneficial effects on the plant. In addition, the Bacillus megaterium KUK2A strain predominantly solubilizes potassium and phosphorus minerals by acidifying the environment, thereby ensuring the availability of these elements to the plant. In turn, the Paenibacillus sp. 02189 strain is characterized by the active production of polysaccharide shells, which ensure the safety of the bacteria. It has been established that polysaccharides play an important role in phosphorus mobilization, they allow the retention of free phosphorus anions (PO4. 3 ), which results in the release of phosphorus (HPC 2-, H2PO4) from insoluble phosphate. Therefore, the polysaccharides of the strain Paenibacillus sp. 02189 serve as an additional factor in the dissolution of insoluble mineral phosphates by bacteria, in addition to organic acids and H ions. + The Paenibacillus sp. 02189 strain provides an immediate positive effect on plants: unlike the Bacillus megaterium KUK2a strain, Paenibacillus sp. 02189 does not require "awakening"; its polysaccharide shell contains the necessary nutrients. Due to these properties, the use of these strains in a consortium ensures high activity in mobilizing phosphorus and potassium compounds, expanding their effectiveness, and also provides a prolonged effect on plants.

[0090] The possibility of objectively demonstrating the technical result when implementing the invention is confirmed by reliable data given below in the examples, containing experimental information obtained in the process of conducting research using methods accepted in this field.

[0091] It should be understood that the examples given in the application materials are not limiting and are given only to illustrate the present invention.

[0092] Example 1. Determination of the ability of Bacillus megaterium KUK2A and Paenibacillus sp. 02189 strains to dissolve poorly soluble inorganic potassium compounds

[0093] The experiment determined the potash utilization of soil minerals. The ability of the studied microorganisms to dissolve poorly soluble inorganic potassium compounds was determined by growth activity on potassium silicate substrates: phlogopite, bentonite, vermiculite, and zeolite. Alexandrov's medium with the following composition (g / L) was used as the base medium:

[0094] Glucose - 5.0 MgSC - 0.5

[0095] CaCO3 - 0.1

[0096] FeCI3- 0.006

[0097] Ca3(PO4)2 — 2.0

[0098] Potassium mineral - 3.0

[0099] Agar - 20.0

[0100] In the experiment, these minerals served as the sole source of potassium, allowing us to assess the microorganisms' ability to utilize potassium compounds from the studied sources. The bacterial strains were inoculated onto the surface of solid Alexandrov medium and incubated for 5-7 days at 28°C. The ability of the bacteria to mobilize poorly soluble potassium compounds was assessed based on their growth activity and the diameter of the mineral clearing zones.

[0101] To quantitatively determine the ability of strains to release potassium compounds from inorganic minerals, a study was conducted on the content of chemical elements in the culture fluid of the studied samples with the addition of phlogopite, determined by the method of atomic emission spectroscopy on an ISP iCAP 6300 DUO AES.

[0102] The experiment revealed that Bacillus megaterium strain KUK2A is capable of utilizing silicate substrates as the sole source of potassium. Growth rate on vermiculite was rated at 5 out of 5. Paenibacillus sp. 02189 also exhibits the ability to utilize all four silicate substrates as the sole source of potassium. Growth rate on these substrates was rated at 4 out of 5.

[0103] The appearance of sesquioxides and silicon in solution is a criterion for the profound transformation of silicate minerals. The results of many studies confirm the ability of microorganisms to utilize potassium and other elements from aluminosilicates. Therefore, the second part of the study on the ability of microorganisms to mobilize potassium involved analyzing the chemical element content in the culture fluid of the test samples using atomic emission spectroscopy on an ICP iCAP 6300 DUO NPP.

[0104] As a result of the experiment, it was established that the content of potassium elements in the culture liquid of Bacillus megaterium KUK2A is 1.24 mg / l, while the value of the control variant (nutrient medium without colonization with bacteria) is 0.63 mg / l. The amount of other elements in the culture medium also increased: Ca - by 17.4% more than the control variant, Na - by 17% more than the control variant. Determination of Mg compounds showed an increase of 16.5%, in comparison with the control variant. The content of potassium elements in the culture liquid of Paenibacillus sp. 02189 is 1.06 mg / l. An increase in Ca content was also found - by 14.7% more than the control variant. A similar trend was observed in the detection of Na compounds: an increase of 14.4% more than the control variant. Determination of Mg compounds showed an increase of 17.6% more than in the control variant. This indicates the release of these elements by the strain into the nutrient solution.

[0105] Thus, as the experiment showed, the studied bacteria are capable of mobilizing potassium and other chemical elements from poorly soluble inorganic compounds in the soil.

[0106] Example 2. Application of bacterial cultures to mineral fertilizer granules using a carrier and an agent that increases the adhesion of bacteria to the mineral fertilizer

[0107] Preparation of components:

[0108] - bacterial cultures of the strains Bacillus megaterium KUK2A and Paenibacillus sp. 02189 are grown separately in a liquid nutrient medium at 34°C for 2-5 days on a shaker at 180 rpm until a bacterial titer of 10 is obtained 7 -10 8 CFU / ml suspension;

[0109] - the developed bacterial cultures are mixed in equal proportions (1:1) and used for application;

[0110] - a substance that increases bacterial adhesion (corn dextrin) is mixed with a saline solution in a ratio of 1:5. The mixture is gelatinized in a water bath for 10 minutes and then sterilized at 120°C for 60 minutes;

[0111] - the carrier (zeolite) is sterilized in a drying oven at 200°C for 8 hours;

[0112] - granules of nitroammophoska mineral fertilizer are sterilized in a drying oven at 100°C for 10 hours.

[0113] Preparation of a mixture for biomodification (biomodifier):

[0114] - corn dextrin is added to 2 ml of bacterial suspension (a 1:1 mixture of strains) in a ratio of 1:3 (1 part dextrin to 3 parts suspension). The mixture is thoroughly mixed on a vortex for 10 seconds;

[0115] Place 5 g of zeolite in a separate sterile 50 ml flask. Add 5 ml of the resulting mixture. Mix the resulting mixture thoroughly with a sterile glass rod.

[0116] Application of bacteria and titer determination:

[0117] - Add 15 g of mineral fertilizer to sterile 50 ml falcons. Add 0.06 g of the resulting biomodifier to the fertilizer. Mix thoroughly with a vortex mixer for 30 seconds.

[0118] - The bacterial titer on mineral fertilizer granules is determined 24 hours after application by serial dilutions with seeding on solid nutrient media and cultivation at 28°C for 72-120 hours, after which the grown colonies are counted. The established titer on mineral fertilizer granules was 1.86*10 4 CFU / g of fertilizers of the Bacillus megaterium strain KUK2A, and 1.82*10 4 CFU / g of strain Paenibacillus sp. 02189. Example 3. Application of bacterial cultures to mineral fertilizer granules and determination of numbers

[0119] Since the Bacillus megaterium KUK2A and Paenibacillus sp. 02189 microorganism strains are intended to improve the efficiency of mineral fertilizer application, the ability of bacterial cells to maintain their viability on the surface of nitroammophoska granules and influence plant growth and productivity was determined.

[0120] To treat granules of mineral fertilizers with nitroammophoska, a bacterial mixture of strains Bacillus megaterium KUK2A and Paenibacillus sp. 02189 was used, containing bacteria in a titer of at least 2.5*10 8CFU / ml of each strain. Mineral fertilizer granules were treated with a bacterial suspension by applying it to the granule surface at a rate of 1 liter per ton (or 1 ml per kg). After 24 hours, the bacterial count on the granules was determined by serial dilution with inoculation on solid nutrient media and cultivation at 28°C for 72-120 hours, after which the grown colonies were counted. Repeated determination of the colony-forming units (CFU) of bacteria on the surface of the mineral fertilizers was performed 3 and 12 months after application.

[0121] Determining the titer of Bacillus megaterium KUK2A bacteria on the surface of mineral fertilizer granules allowed us to establish the ability of the bacteria to maintain their viability after their application to mineral fertilizers. The titer of the Bacillus megaterium KUK2A strain on the granule surface, with direct application, at the beginning of storage (24 hours after application) was 7.5 x 10 6CFU / g, after 12 months of storage the bacterial titer was 1.2*10 5 CFU / g.

[0122] The established titer of the strain Paenibacillus sp. 02189 on the surface of the granules, with direct application, at the beginning of storage (24 hours after application) was 2.7*10 6 CFU / g, after 3 months of storage the bacterial titer was 2.2*10 4 CFU / g. Thus, it was demonstrated that the Paenibacillus sp. 02189 strain can remain viable on the surface of mineral fertilizer granules at concentrations that ensure the effectiveness of biomineral fertilizers for over 3 months. In turn, the Bacillus megaterium KUK2A strain can remain viable on the surface of mineral fertilizer granules at concentrations that ensure the effectiveness of biomineral fertilizers for over 12 months.

[0123] As the experiment demonstrated, when co-cultivated, the strains do not exhibit antagonism toward each other and are able to maintain equilibrium within the consortium. Moreover, due to differences in the life cycle and mechanism of solubilization of potassium and phosphorus minerals in the strains within the consortium, their combined use can ensure high activity in mobilizing phosphorus and potassium compounds, as well as prolonged action.

[0124] Example 4. Determination of the phytostimulating activity of Bacillus megaterium KUK2A and Paenibacillus sp. 02189 strains when applied with mineral fertilizer granules

[0125] It was experimentally established that Bacillus megaterium strains KUK2A and Paenibacillus sp. 02189 have a phytostimulating effect on various agricultural crops. A pot experiment was conducted to determine this.

[0126] Laboratory studies were conducted to determine the number of microorganisms on nitroammophoska mineral fertilizer granules. Quantitative determination of microorganisms on the granules was performed using serial dilutions followed by inoculation on solid nutrient media. The study utilized biomodified mineral fertilizers with an optimal bacterial cell count on the surface of 10 4 CFU / g of fertilizer.

[0127] The following plant seeds were used in the work:

[0128] 1. Corn, hybrid P8521

[0129] 2. Sunflower, hybrid EC AGORA

[0130] 3. Potatoes, variety "Hussar".

[0131] The 3000 ml containers were filled with soil to 1 / 3 of their volume and then weight-balanced. The application rate of mineral fertilizers in all treatments, except the control without fertilizer, was 30 g / m3. 2(0.72 g per 3-liter vessel). The seeding rate was adjusted according to the crop type and generally accepted seeding rate recommendations. The experiment was repeated three times. Plants were grown in a polycarbonate greenhouse with automatic ventilation for 5 months. The following biometric parameters were recorded: number of plants and stems, plant height, and root parameters.

[0132] Analysis of the data from the study of corn plants showed that the Bacillus megaterium strain KUK2A increased plant weight by 14.1%, plant height by 18.6%, and leaf area by 9.1% compared to the control variant (nitroammophoska).

[0133] Analysis of sunflower plants also showed the ability of the Bacillus megaterium KUK2A strain to stimulate plant growth, as the plant weight increased by 8.7% and the leaf area by 6.2% compared to the control variant.

[0134] In a study of the effect of the biomineral fertilizer (BMF) nitroammophoska with the Bacillus megaterium strain KUK2A on potatoes, a reliable effect on the increase in the number of tubers +66.4% compared to the control variant, as well as on the weight of tubers +258.2% compared to the control variant was established. The strain Paenibacillus sp. 02189 increased plant weight by 22%, the number of leaves - by 11.3%, leaf area - by 15.1% compared to the control variant (nitroammophoska).

[0135] Analysis of sunflower plants also demonstrated the ability of Paenibacillus sp. 02189 to stimulate plant growth: plant weight increased by 12.6%, leaf count by 5.9%, and leaf area by 20.1% compared to the control. In a study examining the effect of nitroammophoska with Paenibacillus sp. 02189 on potatoes, a significant increase in tuber count was observed: +18.9% compared to the control.

[0136] The data indicate that Bacillus megaterium KUK2A and Paenibacillus sp. 02189 microbial strains can be successfully used for the biomodification of mineral fertilizers to accelerate plant growth and maturation, increase yields, and improve the absorption efficiency of mineral fertilizers.

[0137] Example 5. Study of the effectiveness of a consortium of Bacillus megaterium KUK2A and Paenibacillus sp. 02189 strains when applied together for the biomodification of mineral fertilizers in field experiments

[0138] To study the effect of a consortium of Bacillus megaterium KUK2A and Paenibacillus sp. 02189 bacterial strains on crop yields, field trials of biomodified fertilizers based on this consortium were conducted. The testing geography included the Leningrad and Volgograd regions. Acron PJSC mineral fertilizers NPK 16:16:16 (total nitrogen (N)* 16%, phosphorus (P2O5) 16%, potassium (K2O) 16%) were used for the tests. The studies were conducted on spring wheat, potato, pepper, and tomato crops.

[0139] A study of the effect of a biofertilizer based on a consortium of Bacillus megaterium KUK2A and Paenibacillus sp. 02189 on spring wheat was conducted at the Belogorka Agricultural Research Institute, Leningrad Region. Fertilizers were applied during primary cultivation at a rate of 100 kg / ha. The study found that the use of a biofertilizer based on a consortium of Bacillus megaterium KUK2A and Paenibacillus sp. 02189 resulted in a 35% higher spring wheat yield than the control variant, which used the same mineral fertilizers at the same rate without modification. Thus, the yield in the variant using the consortium was 2.93 t / ha, while in the control it was 2.17 t / ha.

[0140] A study of the effect of a biofertilizer based on the bacterial consortium Bacillus megaterium KUK2A and Paenibacillus sp. 02189 on potatoes was also conducted at the Belogorka Agricultural Research Institute in the Leningrad Region. Fertilizers were applied during primary cultivation at a rate of 300 kg / ha. Results showed that the use of a biofertilizer based on the bacterial consortium Bacillus megaterium KUK2A and Paenibacillus sp. 02189 resulted in a 13.5% higher potato yield than the control, which used the same mineral fertilizers at the same rate without modification. Thus, the yield in the variant using the consortium was 41.6 t / ha (marketable yield (yield of marketable potatoes) - 37.94 t / ha), while in the control it was 36.64 t / ha (marketable yield - 33.48 t / ha). A 17% increase in ascorbic acid was also noted.

[0141] A study of the effect of biomodification fertilizers based on the bacterial consortium Bacillus megaterium KUK2A and Paenibacillus sp. 02189 on pepper was conducted at the Bykovskaya melon breeding experimental station, Volgograd region. Fertilizers were applied during primary cultivation at a rate of 300 kg / ha. The study found that the use of biomodification fertilizers based on the bacterial consortium Bacillus megaterium KUK2A and Paenibacillus sp. 02189 contributes to pepper yields 18.8% higher than in the control variant, which used the same mineral fertilizers at the same rate without biomodification. Thus, the yield in the variant using the consortium was 10.13 kg / m3. 2 , while in the control it was 4.63 kg / m3 2 The marketability of pepper also increases - 91%, while in the control - 72%.

[0142] A study of the effect of a biofertilizer based on a consortium of bacteria Bacillus megaterium KUK2A and Paenibacillus sp. 02189 on tomato was also conducted at the Bykovskaya melon breeding experimental station in the Volgograd region. Fertilizers were applied during primary cultivation at a rate of 300 kg / ha. The study found that the use of a biofertilizer based on a consortium of bacteria Bacillus megaterium KUK2A and Paenibacillus sp. 02189 contributes to a 12% higher tomato yield than in the control variant, which used the same mineral fertilizers at the same rate without modification. Thus, the yield in the variant using the consortium was 8.2 kg / m3. 2 , while in the control it was 7.3 kg / m3 2 .

[0143] Analyzing the conducted studies, it was established that the use of Bacillus megaterium KUK2A and Paenibacillus sp. 02189 strains in a consortium allows for the maximum potential of both strains to be realized.

[0144] Although the invention has been described with reference to the disclosed embodiments, it will be apparent to those skilled in the art that the specific, detailed examples are provided merely for the purpose of illustrating the present invention and should not be construed as limiting the scope of the invention in any way. It should be understood that various modifications are possible without departing from the spirit of the present invention.

Claims

CLAUSES OF THE INVENTION 1. A consortium of bacteria including the bacterial strains Bacillus megaterium KUK2A (ROAM 05941) and Paenibacillus sp. 02189 (RCAM02189), intended for the biomodification of mineral fertilizers and / or ameliorants.

2. Use of a consortium of bacterial strains Bacillus megaterium KUK2A and Paenibacillus sp. 02189 for biomodification of mineral fertilizers and / or ameliorants.

3. The use according to paragraph 2, in which the biomodification provides improved nutrition and / or stimulation of plant growth, increased resistance to diseases, and increased efficiency of digestibility of, respectively, mineral fertilizers and / or ameliorants.

4. The use according to any of paragraphs 2 or 3, in which the mineral fertilizer or ameliorant is used in solid or liquid form.

5. The use according to claim 4, wherein the mineral fertilizer in solid form is a fertilizer in granulated, prilled, microgranulated, powdered form or in the form of a water-soluble mineral fertilizer.

6. The use according to any of paragraphs 2-5, in which the bacterial consortium is applied directly to the mineral fertilizer or ameliorant by direct application of the bacterial suspension or using a carrier and / or an agent that increases the adhesion of the bacteria to the mineral fertilizer and / or ameliorant and / or carrier.

7. The use according to claim 6, wherein the carrier is a mineral or organic natural or synthetic substance acceptable in agriculture.

8. The use according to claim 7, wherein the carrier is diatomite, zeolite, opoka, tripoli, kaolin, glauconite, silica, perlite, metakaolin, wollastonite, bentonite, inorganic microsilica, belite, white carbon black, vermiculite (expanded), talc or diabasite.

9. The use according to paragraph 6, in which cationic starch, dextrin, collagen glue, bone gelatin glue, albumin glue, fish glue, cherry glue, molasses, gum arabic or liquid glass are used as the agent that increases bacterial adhesion.

10. Use according to any of paragraphs 2-9, wherein the mineral fertilizer is a nitrogen, phosphorus, potassium, boron, molybdenum, complex, complex-mixed or micronutrient fertilizer, or a mixture of one of the fertilizers with micronutrients.

11. The use according to claim 10, wherein the mineral fertilizer is urea, ammonium nitrate, ammonium sulfate, calcium nitrate, ammophos, sodium nitrate, diammonium phosphate, nitroammophoska, phosphate rock, sulfate potassium, potassium salt, sulfoammophos, diammonium phosphate, azophoska, nitrophos, nitrophoska, potassium nitrate, potassium chloride or potassium magnesium sulfate.

12. Use according to any of paragraphs 2-4, 6-9, in which the ameliorant is an ameliorant-sorbent, an ameliorant-structure-forming agent, an ameliorant fertilizer, including phosphogypsum or phosphate rock.

13. Use according to any of paragraphs 2-12, in which the bacteria are used in spore and / or vegetative form.

14. A biomodifier of mineral fertilizers or ameliorants, comprising a consortium of bacteria Bacillus megaterium KUK2A and Paenibacillus sp. 02189 and, optionally, at least one auxiliary substance.

15. A biomodifier according to claim 14, wherein the auxiliary substance is a carrier and / or an agent that increases the adhesion of bacteria to a mineral fertilizer and / or an ameliorant and / or a carrier.

16. A biomodifier of mineral fertilizers according to claim 15, wherein the carrier is diatomite, zeolite, opoka, tripoli, kaolin, glauconite, silica, perlite, metakaolin, wollastonite, bentonite, inorganic microsilica, belite, white carbon black, vermiculite (expanded), talc or diabazite.

17. A biomodified mineral fertilizer or a biomodified ameliorant, comprising, respectively, a mineral fertilizer or ameliorant, and a consortium of bacteria Bacillus megaterium KUK2A and Paenibacillus sp. 02189.

18. A biomodified mineral fertilizer or a biomodified ameliorant according to paragraph 17, obtained by biomodifying, respectively, a mineral fertilizer or ameliorant using a biomodifier according to any of paragraphs 14-16.

19. A biomodified mineral fertilizer or a biomodified ameliorant according to any of paragraphs 17-18, in which, respectively, the mineral fertilizer or ameliorant is used in solid or liquid form.

20. A biomodified mineral fertilizer or a biomodified ameliorant according to claim 19, wherein the mineral fertilizer in solid form is a fertilizer in granulated, prilled, microgranulated, powdered form, or a water-soluble mineral fertilizer.

21. A biomodified mineral fertilizer or a biomodified ameliorant according to any of paragraphs 17-20, in which the mineral fertilizer is a nitrogen, phosphorus, potassium, boron, molybdenum, complex, complex-mixed or micro fertilizer or a mixture of one of the fertilizers with microelements, and the ameliorant is an ameliorant-sorbent, an ameliorant-structure-forming, or a fertilizer-ameliorant.