Bacterial composition and inoculation method

Abstract

A bacterial composition for substrate inoculation to increase substrate fertility, comprising: a predetermined concentration of at least one strain of autotrophic bacteria wherein the autotrophic bacteria are characterized by having the ability to produce organic compounds in a predetermined substrate; and a predetermined concentration of at least one mineral nutrition-producing bacterial strain selected from the group consisting of nitrogen-fixing bacteria, potassium-solubilizing bacteria and phosphate-solubilizing bacteria wherein the bacteria are characterized by having the ability to produce mineral nutrition in such a predetermined substrate. Inoculating the autotrophic bacterial portion of the composition into the substrate increases the yield of the metabolizable form of organic compounds and inoculating the mineral nutrition-producing bacterial portion of the composition increases the metabolism of the organic compounds, thereby increasing the yield of the metabolizable form of mineral nutrition in the predetermined substrate.

Description

Technical Field

[0001] The present invention relates to a bacterial composition and inoculation method. More particularly, but not exclusively, the present invention relates to a bacterial composition and method for improving the fertility of a biota substrate, such as large-scale plant substrates (including agricultural soils and other plant substrates) and large-scale animal substrates (including large water bodies, such as marine and freshwater resources). Background Art

[0002] The growth and health of all plants in the plant matrix are affected by an adequate supply of water and mineral nutrients. These mineral nutrients can be divided into major or macronutrients such as nitrogen, potassium and phosphate, and minor or trace nutrients such as boron, zinc, iron, manganese, copper and chlorine. Plants have a large demand for mineral nutrients. Mineral nutrient deficiencies, especially macronutrient deficiencies, usually cause plants to lose green color, yellow stems and leaves, stunted development, and overall weakness; while an adequate supply of mineral nutrients helps plants grow healthily. Therefore, nitrogen, potassium and phosphate are the main nutrients required for plant growth and agricultural production.

[0003] The above macronutrients are present in the environment, however, a large part of them are in forms that are not easily absorbed and / or assimilated by plants and other organisms. For example, nitrogen is mainly present in the atmosphere as N 2 It exists in elemental form, in which two nitrogen atoms are tightly bound in a so-called triple bond, resulting in relatively low reactivity, making it relatively difficult for humans, plants and microorganisms to convert or absorb elemental nitrogen into useful compounds. In addition, potassium exists in soil in a variety of forms, including mineral potassium, non-exchangeable potassium, exchangeable potassium and solution potassium. Depending on the soil type, 90% to 98% of soil potassium exists in the form of mineral potassium, most of which cannot be absorbed / assimilated by plants. Similarly, although soil contains abundant phosphorus, both organic and inorganic, its availability is limited because it is mostly in insoluble form. The phosphorus content in soil is generally 0.05%, but because phosphorus is poorly soluble and fixed in the soil, the phosphorus content that plants can absorb and utilize is only 0.1% of the total phosphorus.

[0004] Therefore, some consider the inclusion and use of macronutrients in synthetic fertilizers to be a fundamental practice in modern agriculture and essential for increasing crop productivity. However, synthetic fertilization and watering such as irrigation may together lead to unwanted leaching and deterioration of soils and other plant substrates, resulting in salinization, salination and acidification of soils and other plant substrates. Intensive agricultural production and the use of synthetic fertilizers over the past decades have led to the degradation, impoverishment and / or salinization of a significant proportion of arable soils worldwide. The inevitable consequence of this degradation, impoverishment and salinization of soils (or other plant substrates) is a further negative impact on the microbial flora of the resulting soils, especially dry soils or soils with large pH variations, leading to further disintegration of the soil structure or so-called soil destruction, while hindering the humification of organic matter and the formation of soil organic matter. Therefore, the reduction in the supply of macronutrients over the past decades has had a major negative impact on the fertility of arable lands worldwide.

[0005] Furthermore, the reduction of microbial flora in alkaline saline, saline and / or acidic soils (or other substrates) further leads to an increase in the dominance of mold and other less desirable microbial flora. These increases further adversely affect the quality and quantity of the crop.

[0006] Another negative impact of using relatively inefficient synthetic fertilizers is that excess fertilizers can leach out of the soil, causing nitrate contamination of surface and groundwater.

[0007] Furthermore, the widespread use of synthetic fertilizers around the world has been shown to have indirect, significant, harmful effects on the environment. Fossil fuels are the basis for the production of synthetic fertilizers. For example, liquid ammonia can be extracted by burning natural gas. This powerful substance has a high nitrogen content and can be used as an effective fertilizer, however burning natural gas produces greenhouse gases, including methane and carbon dioxide as waste products, which contribute to climate change. When used in excess, nitrogen fertilizers are oxidized and lost to the air in the form of nitrous oxide, an even more powerful greenhouse gas that can have a negative impact on the environment. Finally, since agriculture requires large amounts of synthetic fertilizers, transporting them to farmland leads to further greenhouse gas emissions.

[0008] Some plants, such as legumes, require relatively little nitrogen due to the presence of nitrogen-fixing structures or so-called root nodules, in which nitrogen-fixing microorganisms (also called diazotrophs) form a mutually beneficial symbiosis with the plant by providing metabolizable nitrogen, which the plant can take up in the form of ammonium, nitrite and nitrate, while the microorganisms take up carbon and other input nutrients from the plant cells. The diazotrophic population provides the plant with a constant, balanced supply of nitrogen, thereby alleviating the need for synthetic fertilizers compared to synthetic fertilizers. The diazotrophic population can also play a growth-promoting role by producing plant hormone-like substances that are of particular value in agriculture. However, this symbiotic root nodule nitrogen fixation is almost exclusively found in legumes.

[0009] Diazotrophs, such as Azotobacter and Azospirrilum, are found among the free-living bacteria naturally present in agricultural soils. However, because these bacterial populations are free-living in the soil and do not form direct associations with plant roots, the nitrogen produced by these bacteria during nitrogen fixation and taken up by the plants is insufficient to meet the plants' overall nitrogen needs. Furthermore, conventional agricultural processes, such as the use of biocides, have been shown to kill free-living bacteria in the soil, including diazotrophs. Therefore, these bacteria alone are unable to provide sufficient nitrogen to the roots of economically valuable cereals such as wheat, corn, rice, barley, millet, and sorghum in conventional agricultural practices.

[0010] In addition, the nitrogen-fixing bacteria found in agricultural soils are often present in low concentrations among the free-living bacteria that supply enough nitrogen to the roots of surrounding plants. The carbon and other input nutrients provided by plant roots are also often insufficient for the nitrogen-fixing bacteria to produce enough nitrogen to supply the roots of surrounding plants.

[0011] In addition, K-solubilizing bacteria secrete organic acids and enzymes that act on insoluble potassium and convert it into a mineralized form, thus providing a source of potassium that can be absorbed / assimilated by plants. K-solubilizing bacteria also produce amino acids, vitamins, and growth-promoting substances that promote plant growth and help increase crop yields. In addition, K-solubilizing bacteria are able to mineralize rock potassium, such as mica, illite, and orthoclase, and can also bring potassium into solution by producing and secreting organic acids or chelating silicon ions.

[0012] Phosphate solubilizing bacteria can increase the bioavailability of phosphorus in the soil to plants. They solubilize insoluble inorganic (mineral) phosphorus and mineralize insoluble organic phosphates. Salt-tolerant or halophilic soil microorganisms can solubilize insoluble phosphates, thus promoting the development of agriculture in saline-alkali soils.

[0013] Likewise, the growth and health of all plants and animals in large-scale matrices such as large bodies of water are influenced by the availability of adequate oxygen and mineral nutrients. The availability of mineral nutrients, including nitrogen, phosphate, and potassium, in aquatic ecosystems is critical as they together form key elements for the maintenance and vitality of aquatic plants and animals. Nitrogen, in various forms, is a major nutrient required for the growth of aquatic plants, including phytoplankton and submerged plants, which form the basis of aquatic food webs. Phosphate is equally important as it promotes the growth of these primary producers and regulates various metabolic processes in aquatic organisms. In turn, although potassium is required in lesser quantities, it plays a vital role in maintaining osmotic balance and enzyme function in aquatic plants. Therefore, careful management and maintenance of these micronutrients in aquatic environments is essential to protect and enhance aquatic biodiversity, support sustainable fisheries, and ensure the overall ecological health of water bodies on which countless ecosystems and human life depend.

[0014] One of the greatest threats to the fertility of water bodies such as ocean water today is oil spills. Oil often causes oxygen depletion in seawater, destroys the insulating ability of fur-bearing mammals such as sea otters and the waterproofing of bird feathers, causing these creatures to become hypothermic, trapping small animals and other marine life, being mistaken for food, being inhaled by dolphins and whales, where it affects their lungs, immune function, and reproduction, and being ingested by fish. In turn, while not immediately harmed, shellfish and corals become contaminated when the oil mixes into the water column, and shellfish are exposed in intertidal zones. For example, adult fish can experience symptoms such as reduced growth, enlarged livers, changes in heart and respiratory rates, fin erosion, and reproductive disorders, while eggs and juveniles are often sensitive to lethal and sublethal effects. Ultimately, oil can render marine life such as fish and shellfish unfit for human consumption.

[0015] The potential for petroleum biodegradation or the ability of bacteria to degrade hydrocarbons is well known. However, a key limiting factor that has always hampered the efficiency of hydrocarbon-degrading organisms is the availability of nitrogen in metabolizable form in the surrounding environment. Nitrogen is therefore a decisive factor in the performance of hydrocarbon-degrading microorganisms. Traditional methods of supplementing synthetic nitrogen have proven to be not only costly but also harmful to the surrounding environment, causing pollution problems.

[0016] In this manual, the terms: i. "Substrate" shall include plant substrates and animal substrates, where plant substrates include solid-based root growth media (e.g. soil), liquid-based root growth media (e.g. media used in hydroponic systems), and air-based (i.e. gas) root growth media (e.g. media used in aeroponic systems); animal substrates include large bodies of water (e.g. ocean or sea water, natural and artificial freshwater lakes, rivers, dams, etc.); and ii. “Fertility” refers to the ability of a substrate to support and sustain the growth, reproduction and flourishing of organisms (including animals and plants). Purpose of the Invention

[0017] Therefore, the object of the present invention is to provide a novel and relatively inexpensive and effective bacterial composition for substrate inoculation, which can improve the fertility of the substrate. Summary of the invention

[0018] According to a first aspect of the present invention, there is provided a bacterial composition for substrate inoculation to improve substrate fertility, the composition comprising: at least one autotrophic bacterial strain at a predetermined concentration, wherein the autotrophic bacteria is characterized by the ability to produce an organic compound in a predetermined substrate; and a predetermined concentration of at least one mineral nutrient producing bacterial strain selected from the group consisting of nitrogen-fixing bacteria, potassium-solubilizing bacteria, and phosphate-solubilizing bacteria, wherein the bacteria is characterized by having the ability to produce mineral nutrients in such a predetermined substrate; The autotrophic bacteria part of the composition is inoculated into the substrate to increase the yield of metabolizable organic compounds, and the bacteria part of the composition that produces mineral nutrients is inoculated into the substrate to increase the metabolism of organic compounds, thereby increasing the yield of metabolizable mineral nutrients in the predetermined substrate.

[0019] The bacterial composition may comprise the mineral nutrient producing bacteria in a concentration of 30% to 70%, preferably 50%, based on the total weight of the composition.

[0020] According to a second aspect of the present invention, there is provided a bacterial composition for substrate inoculation to improve nitrogen fixation in the substrate, the composition comprising: at least one autotrophic bacterial strain at a predetermined concentration, wherein the autotrophic bacteria is characterized by the ability to produce an organic compound in a predetermined substrate; and a predetermined concentration of at least one strain of nitrogen-fixing bacteria, wherein the nitrogen-fixing bacteria is characterized by the ability to biologically fix atmospheric nitrogen in such predetermined substrate; The autotrophic bacteria part of the composition is inoculated into the substrate to increase the yield of metabolizable organic compounds, and the nitrogen-fixing bacteria part of the composition is inoculated into the substrate to increase the metabolism of organic compounds, thereby increasing the yield of metabolizable nitrogen in the predetermined substrate.

[0021] The bacterial composition may comprise nitrogen-fixing bacteria in a concentration of 30% to 70%, preferably 50%, based on the total weight of the composition.

[0022] The nitrogen-fixing bacteria may include a bacterial culture of one or more strains selected from the group consisting of Azobacter species, Azospirillum species, Bacillus Pumilus species, Paenibacillus Polymyxa species, and the like.

[0023] The bacterial composition may also have the following characteristics: after the substrate is inoculated, it can improve the aeration of the plant substrate, the plant substrate is selected from: solid-based root growth medium, such as soil, sand, gravel, volcanic ash and calcined clay; liquid-based root growth medium, such as medium for hydroponic system; and air-based (i.e. gas) root growth medium, such as medium for aeroponic system. More preferably, the bacterial composition may also have the characteristics of improving the aeration of solid-based root growth medium (such as anaerobic soil).

[0024] The bacterial composition may be further characterized in that it absorbs carbon dioxide from the atmosphere, thereby reducing carbon dioxide in the atmosphere.

[0025] According to a third aspect of the present invention, there is provided a bacterial composition for substrate inoculation to increase the content of metabolizable potassium in the substrate, the composition comprising: at least one autotrophic bacterial strain at a predetermined concentration, wherein the autotrophic bacteria is characterized by the ability to produce an organic compound in a predetermined substrate; and a predetermined concentration of at least one strain of potassium-solubilizing bacteria characterized by the ability to solubilize potassium-containing minerals, converting insoluble potassium into soluble potassium, wherein the bacteria is characterized by the ability to produce metabolizable potassium in such predetermined substrate; The autotrophic bacteria part of the composition is inoculated into the substrate to increase the yield of metabolizable organic compounds, and the potassium-solubilizing bacteria part of the composition is inoculated into the substrate to increase the metabolism of organic compounds, thereby increasing the yield of metabolizable potassium in the predetermined substrate.

[0026] The potassium-dissolving bacteria may include a bacterial culture of one or more strains selected from Pseudomonas species, Burkholderia species, Acidothiobacillus ferrooxidans species, Bacillus species, and Paenibacillus species.

[0027] According to a fourth aspect of the present invention, there is provided a bacterial composition for substrate inoculation to increase the content of metabolizable phosphate in the substrate, the composition comprising: at least one autotrophic bacterial strain at a predetermined concentration, wherein the autotrophic bacteria is characterized by the ability to produce an organic compound in a predetermined substrate; and a predetermined concentration of at least one strain of phosphate solubilizing bacteria characterized by the ability to solubilize phosphate-containing minerals, converting insoluble phosphates into soluble phosphates, wherein the bacteria is characterized by the ability to produce metabolizable phosphates in such predetermined substrate; The autotrophic bacteria part of the composition is inoculated into the substrate to increase the yield of metabolizable organic compounds, and the phosphate-solubilizing bacteria part of the composition is inoculated into the substrate to increase the metabolism of organic compounds, thereby increasing the yield of metabolizable phosphate in the predetermined substrate.

[0028] The phosphate-solubilizing bacteria may include a bacterial culture of one or more strains selected from Bacillus Coagulans species, Bacillus Megaterium species, Psuedomonas Flourocens species, Rhodococcus species, Arthrobacter species, Serratia species, Chryseobacterium species, Gordonia species, Phyllobacterium species, Delftia species, Azotobacter species, Xanthomonas species, Enterobacter species, Pantoea species, and Xanthobacteragilis species.

[0029] The substrate may comprise any substrate suitable for supporting the growth of the intended plant species, the substrate preferably being selected from the group comprising: solid-based root growth media (e.g., soil, sand, gravel, volcanic ash and calcined clay), liquid-based root growth media (e.g., media used in hydroponic systems) and air-based (i.e., gaseous) root growth media (e.g., media used in aeroponic systems).

[0030] The bacterial composition may comprise the autotrophic bacteria in a concentration of 30% to 70%, preferably 50%, based on the total weight of the composition.

[0031] The autotrophic bacteria may include photoautotrophic bacteria capable of producing organic compounds by photosynthesis. Preferably, the autotrophic bacteria include a bacterial culture of one or more strains selected from Rhodopsuedomonas species and Rhodospirrilum species.

[0032] The bacterial composition may be selected from the group consisting of a fermented liquid preparation (with or without an incubation period), a freeze-dried preparation (rehydrated before application to the soil, with or without a short incubation period), a spray-dried preparation (rehydrated before application to the soil, with or without a short incubation period), and a refractory dried preparation (rehydrated before application to the soil, with or without a short incubation period).

[0033] The bacterial composition may be applied to the soil at a concentration of 1 billion to 1 trillion colony forming units (CFU) per hectare of soil, depending on the type of soil being treated, preferably about 500 billion CFU per hectare of soil.

[0034] The predetermined solid-based root growth medium may be selected from sandy soil, clay soil, loam soil, low carbon soil, and high carbon soil.

[0035] According to a fifth aspect of the present invention, there is provided a method for inoculating a substrate by bacteria to improve fertility, the method comprising the following steps: inoculating at least one autotrophic bacterial strain at a predetermined concentration, wherein the autotrophic bacteria is characterized by the ability to produce an organic compound in a predetermined substrate; and inoculating a predetermined concentration of at least one mineral nutrient producing bacterium selected from the group consisting of nitrogen fixing bacteria, potassium solubilizing bacteria, and phosphate solubilizing bacteria, wherein the bacterium is characterized by having the ability to produce mineral nutrients in such a predetermined substrate; The production of metabolizable organic compounds is increased by inoculating the autotrophic bacteria portion of the composition, and the metabolism of organic compounds is increased by inoculating the mineral nutrient-producing bacteria portion of the composition, thereby increasing the production of metabolizable mineral nutrients in the predetermined substrate to a predetermined level.

[0036] The bacterial composition may comprise the mineral nutrient producing bacteria in a concentration of 30% to 70%, preferably 50%, based on the total weight of the composition.

[0037] According to a sixth aspect of the present invention, there is provided a method for inoculation of bacteria to improve nitrogen fixation in a substrate, the method comprising the following steps: inoculating at least one autotrophic bacterial strain at a predetermined concentration, wherein the autotrophic bacteria is characterized by the ability to produce an organic compound in a predetermined substrate; and inoculating with a predetermined concentration of at least one strain of nitrogen-fixing bacteria, wherein the nitrogen-fixing bacteria is characterized by having the ability to biologically fix atmospheric nitrogen in such predetermined substrate; The production of metabolizable organic compounds is increased by inoculating the autotrophic bacteria portion of the composition, and the metabolism of organic compounds is increased by inoculating the nitrogen-fixing bacteria portion of the composition, thereby increasing the production of metabolizable nitrogen in the predetermined substrate to a predetermined level.

[0038] The bacterial composition may comprise nitrogen-fixing bacteria in a concentration of 30% to 70%, preferably 50%, based on the total weight of the composition.

[0039] The nitrogen-fixing bacteria may include a bacterial culture of one or more strains selected from the group consisting of Azobacter species, Azospirillum species, Bacillus Pumilus species, and Paenibacillus Polymyxa species.

[0040] The bacterial composition may further be characterized in that it improves the aeration of the soil after soil inoculation. More preferably, the bacterial composition may also have the characteristic of improving the aeration of anaerobic soil.

[0041] The bacterial composition may also be characterized in that it absorbs carbon dioxide from the atmosphere, thereby reducing carbon dioxide in the atmosphere.

[0042] According to a seventh aspect of the present invention, there is provided a method for inoculating a substrate with bacteria to increase the content of metabolizable potassium, the method comprising the following steps: inoculating at least one autotrophic bacterial strain at a predetermined concentration, wherein the autotrophic bacteria is characterized by the ability to produce an organic compound in a predetermined substrate; and inoculating a predetermined concentration of at least one strain of potassium-solubilizing bacteria, the potassium-solubilizing bacteria being characterized by the ability to dissolve potassium-containing minerals, converting insoluble potassium into soluble potassium, wherein the bacteria is characterized by the ability to produce metabolizable potassium in such a predetermined substrate; The production of metabolizable organic compounds is increased by inoculating the autotrophic bacteria in the composition, and the metabolism of organic compounds is increased by inoculating the potassium-solubilizing bacteria in the composition, thereby increasing the production of metabolizable potassium in the predetermined substrate to a predetermined level.

[0043] The potassium-dissolving bacteria may include a bacterial culture of one or more strains selected from Pseudomonas species, Burkholderia species, Acidothiobacillus ferrooxidans species, Bacillus species, and Paenibacillus species.

[0044] According to an eighth aspect of the present invention, there is provided a method for inoculating a substrate with bacteria to increase the content of metabolizable phosphate, the method comprising the following steps: inoculating at least one autotrophic bacterial strain at a predetermined concentration, wherein the autotrophic bacteria is characterized by the ability to produce an organic compound in a predetermined substrate; and inoculating a predetermined concentration of at least one strain of phosphate-solubilizing bacteria, the phosphate-solubilizing bacteria being characterized by the ability to solubilize phosphate-containing minerals, converting insoluble phosphates into soluble phosphates, wherein the bacteria is characterized by the ability to produce metabolizable phosphates in such a predetermined substrate; The production of metabolizable organic compounds is increased by inoculating the autotrophic bacteria in the composition, and the metabolism of organic compounds is increased by inoculating the phosphate-solubilizing bacteria in the composition, thereby increasing the production of metabolizable phosphate in the predetermined substrate to a predetermined level.

[0045] The phosphate-solubilizing bacteria may include a bacterial culture of one or more strains selected from the group consisting of Rhodococcus species, Arthrobacter species, Serratia species, Chryseobacterium species, Gordonia species, Phyllobacterium species, Delftia species, Azotobacter species, Xanthomonas species, Enterobacter species, Pantoea species, and Xanthobacter agilis species.

[0046] The inoculation step may be selected from the group consisting of: applying the bacterial composition to the planting trench; applying the bacterial composition to the planting furrow and watering the furrow; and The existing vegetation or its surroundings are sprayed with a liquid solution containing the composition in a predetermined volume and at predetermined intervals.

[0047] The substrate may comprise any substrate suitable for supporting the growth of the intended plant species, the substrate preferably being selected from the group comprising: solid-based root growth media (e.g., soil, sand, gravel, volcanic ash and calcined clay), liquid-based root growth media (e.g., media used in hydroponic systems) and air-based (i.e., gaseous) root growth media (e.g., media used in aeroponic systems).

[0048] The bacterial composition may comprise the autotrophic bacteria in a concentration of 30% to 70%, preferably 50%, based on the total weight of the composition.

[0049] The autotrophic bacteria may include photoautotrophic bacteria capable of producing organic compounds by photosynthesis. Preferably, the autotrophic bacteria include a bacterial culture of one or more strains selected from Rhodopsuedomonas species and Rhodospirrilum species.

[0050] The bacterial composition may be selected from the group consisting of a fermented liquid preparation (with or without an incubation period), a freeze-dried preparation (rehydrated before application to the soil, with or without a short incubation period), a spray-dried preparation (rehydrated before application to the soil, with or without a short incubation period), and a refractory dried preparation (rehydrated before application to the soil, with or without a short incubation period).

[0051] The bacterial composition may be applied to the soil at a concentration of 1 billion to 1 trillion colony forming units (CFU) per hectare of soil, preferably about 500 billion CFU per hectare of soil.

[0052] The predetermined solid-based root growth medium may be selected from sandy soil, clay soil, loam soil, low carbon soil, and high carbon soil. DETAILED DESCRIPTION

[0053] Non-limiting embodiments of the composition according to the present invention include at least one autotrophic bacterial strain and at least one mineral nutrient producing bacterial strain.

[0054] The characteristic of autotrophic bacteria is that they can produce organic compounds such as carbohydrates, proteins, etc. More specifically, the autotrophic bacteria in the composition include photoautotrophic bacterial species that produce organic compounds by photosynthesis.

[0055] The mineral nutrient producing bacteria include at least one mineral nutrient producing bacterial strain selected from the group consisting of nitrogen-fixing bacteria, potassium-solubilizing bacteria and phosphate-solubilizing bacteria, wherein the bacteria are characterized in that they are capable of producing mineral nutrients in a predetermined substrate.

[0056] The characteristic of nitrogen-fixing bacteria is that they can fix atmospheric nitrogen into a form that is more easily metabolized and taken up by organisms such as plants. These metabolizable forms of atmospheric nitrogen include nitrate, nitrite and ammonia.

[0057] Potassium-solubilizing bacteria are characterized by their ability to dissolve potassium-containing minerals and convert insoluble potassium into soluble potassium, among which the bacteria are characterized by their ability to produce metabolizable potassium. Efficient potassium-solubilizing bacteria secrete organic acids and enzymes that act on insoluble potassium present in the substrate and convert it into a mineralized form, thereby providing available potassium to plants. Potassium-solubilizing bacteria also produce amino acids, vitamins, and growth-promoting substances that promote plant growth and help increase crop yields.

[0058] Phosphate solubilizing bacteria are characterized by their ability to solubilize phosphate-containing minerals, convert insoluble phosphates into soluble phosphates, and among these bacteria are characterized by their ability to produce metabolizable phosphates. Phosphate solubilizing bacteria secrete mineral-soluble compounds, such as organic acid anions, iron-binding compounds, protons and hydroxyl ions, which act on insoluble phosphates present in the substrate and convert them into mineralized forms, thus providing phosphate forms that are available to plants.

[0059] A non-limiting embodiment of the inoculation method according to the present invention comprises spraying selected existing plants and their surroundings in a selected soil type with a liquid solution containing the composition at a predetermined volume and at predetermined intervals.

[0060] When selected plants and their soil types are sprayed with the bacterial composition, a symbiotic relationship is formed between the autotrophic bacteria and the mineral nutrient producing bacteria, wherein organic compounds produced by the autotrophic bacteria, more specifically carbohydrates such as glucose, are metabolized by the mineral nutrient producing bacteria, thereby greatly enhancing the ability of the mineral nutrient producing bacteria to produce metabolizable mineral nutrients.

[0061] The symbiotic relationship between autotrophic bacteria and nitrogen-fixing bacteria results in a substantial increase in the free metabolizable forms of nitrogen (e.g., nitrates, nitrites, and ammonia) in the soil. The increased nitrogen content in the soil surrounding the plant roots is then metabolized by the plant roots, thereby increasing the productivity and growth of the plant. The nitrogen-fixing bacteria include cultures of autotrophic bacterial species, including one or more strains of the genus Azobacterspecies, Azospirillum species, Bacillus Pumilusspecies, and / or Paenibacillus Polymyxa species.

[0062] The symbiotic relationship between the autotrophic bacteria and potassium-solubilizing bacteria results in a substantial increase in the free metabolizable form of potassium in the soil. The increased potassium content in the soil surrounding the plant roots is then metabolized by the plant roots, thereby increasing plant productivity and growth. The potassium-solubilizing bacteria include cultures of autotrophic bacterial species, including one or more strains of Pseudomonas species, Burkholderia species, Acidothiobacillusferrooxidans species, Bacillus species, and Paenibacillus species.

[0063] The symbiotic relationship between autotrophic bacteria and phosphate solubilizing bacteria results in a substantial increase in the free metabolizable form of phosphate in the soil. The increased phosphate content in the soil surrounding the plant roots is then metabolized by the plant roots, thereby increasing the productivity and growth of the plant. The phosphate solubilizing bacteria include cultures of autotrophic bacterial species, including one or more strains of Rhodococcus species, Arthrobacter species, Serratia species, Chryseobacterium species, Gordonia species, Phyllobacterium species, Delftia species, Azotobacterspecies, Xanthomonas species, Enterobacter species, Pantoea species, and Xanthobacter agilis species.

[0064] Autotrophic bacteria include cultures of autotrophic bacterial species, including one or more strains of Rhodopsuedomonas species and Rhodospirrilum species.

[0065] It is contemplated that the bacterial composition comprises a freeze-dried formulation that is rehydrated prior to application to the soil, with or without a brief incubation period; or a spray-dried formulation that is rehydrated prior to application to the soil, with or without a brief incubation period; or a refractory dried formulation that is rehydrated prior to application to the soil, with or without a brief incubation period.

[0066] Cultures of each strain may be prepared separately and then combined to form a stock solution. It is further envisioned that such a stock solution will be appropriately diluted with water or a suitable medium and incubated as a batch culture until the microorganisms reach a predetermined growth stage. The bacterial composition may include mineral nutrient producing bacteria at a concentration of 30% to 70% of the total weight of the composition, with the remainder of the composition comprising autotrophic bacteria. Alternatively, it is envisioned that the bacterial composition may include autotrophic bacteria at a concentration of 30% to 70% of the total weight of the composition, with the remainder of the composition comprising autotrophic bacteria.

[0067] Depending on the plant species selected for the inoculated soil, the increased plant productivity includes faster plant growth, maintenance of higher chlorophyll content in leaves, increased fruit and / or seed number, increased fruit and / or seed unit weight, or any combination of these characteristics. In addition, the increased plant productivity will also reduce nitrogen dioxide emissions due to reduced nitrogen fertilizer use.

[0068] The bacterial composition can be applied in a variety of ways, depending on soil type, soil conditions, plant species and / or climatic conditions. For example, it is contemplated that while the preferred method may involve spraying a liquid mixture of the bacterial composition onto the exposed parts of existing plants and their surroundings at regular intervals, the bacterial composition may also be applied to the roots by inserting the bacterial composition into the furrow where the plant seeds are planted, watering the soil and / or dipping the roots into a suspension of the bacterial composition. Alternatively, the bacterial composition may be applied as a seed coating.

[0069] Generally, the density of the bacterial composition inoculated on the seed, in the trench or in the root wet should be sufficient to colonize the lower soil area adjacent to the plant roots with an active bacterial community. Similarly, the inoculation density of the bacterial composition sprayed on the plant should be sufficient to form an effective bacterial colonization on the leaves of the plant. However, it should be noted that an effective concentration or dosage of the bacterial inoculant should be used.

[0070] The effective concentration should be sufficient to establish sufficient bacterial growth to increase plant productivity to a preselected level. It is expected that the effective concentration of the bacterial composition can reach 1 billion to 1 trillion CFU per hectare of soil.

[0071] Although autotrophic bacteria primarily produce their own enzymes and biomolecules to meet their metabolic needs, they can indirectly influence enzyme production by surrounding bacteria (e.g., nitrogen-fixing, potassium-solubilizing, and phosphate-solubilizing bacteria) in a variety of ways. It has been shown that autotrophic bacteria can influence the availability of inorganic nutrients in their environment. For example, they can take up and metabolize inorganic carbon sources, which can alter the CO 2or the concentration of bicarbonate ions. Changes in the availability of these inorganic compounds can affect the metabolism and enzyme production of neighboring bacteria. In addition, the metabolic activities of autotrophic bacteria produce metabolic byproducts, such as organic acids or bases. These byproducts can change the pH and chemical composition of the surrounding environment. Changes in pH or the presence of specific compounds can affect the enzyme activity and gene expression of other bacteria in the same habitat, while reducing the need to increase soil pH through liming. In addition, autotrophic bacteria can produce secondary metabolites, such as antibiotics or signaling molecules, which can affect the growth and enzyme production of neighboring bacteria. These secondary metabolites can act as signaling molecules and affect gene expression of surrounding microorganisms. Finally, autotrophic bacteria can become part of complex microbial communities, such as biofilms. Within biofilms, different bacterial species interact and share resources. The metabolic activities of autotrophic bacteria in biofilms can create microenvironments with unique chemical and physical conditions, affecting enzyme production and gene expression of other community members.

[0072] It is further contemplated that the bacterial compositions described herein will have several significant advantages over synthetic fertilizers and / or growth hormones or similar agents commonly used in agriculture today. Due to the very nature of the bacterial compositions, once the soil is inoculated by any of the methods described herein, the bacterial species will be self-sustaining in a continuous manner. Thus, there is little or no need to re-treat the soil during the crop growing season. The bacteria grow with the plants during the cultivation process and continue to exert beneficial effects on the plants throughout the agricultural season. This is in stark contrast to chemical or synthetic growth agents or fungicides, which must be re-applied periodically to continue to suppress the relevant fungi or help improve the growth of the plant throughout its life cycle.

[0073] It is also envisioned that since the bacterial composition can be introduced into seeds using a dry or wet formulation, or inoculated into the soil prior to planting, or sprayed onto existing plants and their surroundings, the compositions and methods according to the present invention can be relatively simple and freely available for agricultural applications, thus having significant economic advantages.

[0074] It should be understood that many changes in details may be made without departing from the scope and / or spirit of the invention as claimed in the claims below, defined in the constituent statements and / or as described in the specific examples above, for example, the bacterial composition is characterized in that it improves the aeration of the substrate (e.g., soil) after soil inoculation, improves the aeration of anaerobic soil and / or absorbs carbon dioxide from the atmosphere, thereby reducing carbon dioxide in the atmosphere, and for example, the substrate includes any substrate suitable for supporting the growth of the intended plant species, including solid-based root growth media, such as soil, sand, gravel, volcanic ash and calcined clay; liquid-based root growth media, such as media used in hydroponic systems; and air-based (i.e., gaseous) root growth media, such as media used in aeroponic systems.

Claims

1. A bacterial composition for substrate inoculation to improve substrate fertility, the composition include: at least one autotrophic bacterial strain at a predetermined concentration, wherein the autotrophic bacteria is characterized by the ability to produce an organic compound in a predetermined substrate; as well as a predetermined concentration of at least one mineral nutrient producing bacterial strain selected from the group consisting of nitrogen fixing bacteria, potassium solubilizing bacteria, and phosphate solubilizing bacteria, wherein the bacteria is characterized by having the ability to produce mineral nutrients in such a predetermined substrate; The autotrophic bacteria part of the composition is inoculated into the substrate to increase the yield of metabolizable organic compounds, and the bacteria part of the composition that produces mineral nutrients is inoculated into the substrate to increase the metabolism of organic compounds, thereby increasing the yield of metabolizable mineral nutrients in the predetermined substrate.

2. The bacterial composition according to claim 1, wherein the predetermined substrate is pre-selected based on its suitability for supporting the growth of a predetermined plant or animal.

3. The bacterial composition according to claim 1, wherein the predetermined matrix is ​​selected from the group consisting of: Plant substrate, include: Solid-based root growing media, including earth, sand, gravel, volcanic ash, and calcined clay, Liquid-based root growing medium, such as that used in hydroponic systems, and Air-based root growing media, including air and other gas mixtures, such as media used in aeroponic systems; and Animal substrates, which include large bodies of water, such as oceans or seawater, natural and artificial freshwater lakes, rivers, dams, etc.

4. The bacterial composition according to claim 1, wherein the composition comprises autotrophic bacteria at a concentration of 30% to 70% of the total weight of the composition.

5. The bacterial composition according to claim 1, wherein the composition comprises mineral nutrient producing bacteria at a concentration of 30% to 70% of the total weight of the composition.

6. The bacterial composition according to claim 1, wherein the composition is characterized in that it improves the pH balance of the substrate.

7. The bacterial composition according to claim 1, wherein the composition is characterized in that it absorbs carbon dioxide in the atmosphere, thereby reducing carbon dioxide in the atmosphere.

8. The bacterial composition according to claim 1, wherein the autotrophic bacteria comprises photoautotrophic bacteria capable of producing organic compounds by photosynthesis.

9. The bacterial composition according to claim 1, wherein the autotrophic bacteria include chemoautotrophic bacteria capable of producing organic compounds through chemical reactions.

10. The bacterial composition according to claim 1, wherein the autotrophic bacteria comprises a bacterial culture of one or more strains selected from the group consisting of Rhodopseudomonas and Rhodospirilla.

11. The bacterial composition according to claim 1, wherein the composition is characterized in that it improves the aeration of the predetermined substrate after inoculation.

12. The bacterial composition according to claim 11, wherein the composition is characterized in that it improves the aeration of anaerobic substrates.

13. The bacterial composition according to claim 1, wherein the composition is inoculated into a substrate to improve nitrogen fixation in the substrate, wherein the composition include: at least one strain of nitrogen-fixing bacteria at a predetermined concentration, wherein the nitrogen-fixing bacteria is characterized by the ability to biologically fix atmospheric nitrogen in such predetermined substrate; The nitrogen-fixing bacteria portion of the composition is inoculated into a substrate to improve the metabolism of organic compounds, thereby increasing the yield of nitrogen in metabolizable form in the predetermined substrate.

14. The bacterial composition according to claim 13, wherein the nitrogen-fixing bacteria comprises a bacterial culture of one or more strains selected from the genera Azotobacter, Azospirillum, Bacillus brevis and Paenibacillus polymyxa.

15. The bacterial composition according to claim 1, wherein the composition is inoculated into a substrate to increase the content of metabolizable potassium in the substrate, wherein the composition include: a predetermined concentration of at least one strain of potassium-solubilizing bacteria characterized by the ability to solubilize potassium-containing minerals, convert insoluble potassium into soluble potassium, and produce metabolizable potassium in such predetermined matrix; The potassium-solubilizing bacteria portion of the composition is inoculated into a substrate to improve the metabolism of organic compounds, thereby increasing the yield of potassium in a metabolizable form in the predetermined substrate.

16. The bacterial composition according to claim 15, wherein the potassium-solubilizing bacteria comprises a bacterial culture of one or more strains selected from the genera Pseudomonas, Burkholderia, Acidithiobacillus ferrooxidans, Bacillus and Paenibacillus.

17. The bacterial composition according to claim 1, wherein the composition is inoculated into a substrate to increase the content of metabolizable phosphate in the substrate, wherein the composition include: a predetermined concentration of at least one strain of phosphate-solubilizing bacteria characterized by the ability to solubilize phosphate-containing minerals, convert insoluble phosphates into soluble phosphates, and produce metabolizable phosphates in such predetermined substrate; The phosphate-solubilizing bacteria portion of the composition is inoculated into a substrate to improve the metabolism of organic compounds, thereby increasing the yield of metabolizable phosphate in the predetermined substrate.

18. The bacterial composition according to claim 17, wherein the phosphate-solubilizing bacteria comprises a bacterial culture of one or more strains selected from the genera Bacillus coagulans, Bacillus megaterium, Pseudomonas fluorescens, Rhodococcus, Arthrobacter, Serratia, Chryseobacterium, Gordonia, Leaf bacterium, Delftia, Azotobacter, Xanthomonas, Enterobacter, Pantoea and Flavobacterium agility.

19. The bacterial composition according to claim 1, wherein the composition is selected from the group consisting of: Fermented liquid preparations, with or without a culture period; freeze-dried formulations that are rehydrated before application to the matrix, with or without a brief incubation period; Spray-dried formulations that are rehydrated prior to application to a substrate, with or without a brief incubation period; and Refracted dried formulations were rehydrated before application to the matrix, with or without a brief incubation period.

20. The bacterial composition of claim 3, wherein the composition is applied to a solid-based root growth medium at a concentration of 1 billion to 1 trillion colony forming units (CFU) per hectare of medium.

21. The bacterial composition according to claim 3, wherein the predetermined substrate is a solid-based root growth medium selected from the group consisting of sandy soil, clay, loam, low-carbon soil, and high-carbon soil.

22. A method for inoculating a predetermined substrate to improve its fertility by bacteria, said method The following steps are involved: inoculating at least one autotrophic bacterial strain at a predetermined concentration, wherein the autotrophic bacteria is characterized by the ability to produce an organic compound in a predetermined substrate; and inoculating a predetermined concentration of at least one bacterial strain producing mineral nutrients selected from the group consisting of nitrogen-fixing bacteria, potassium-solubilizing bacteria, and phosphate-solubilizing bacteria, wherein the bacteria is characterized by having the ability to produce mineral nutrients in such a predetermined substrate; The production of metabolizable organic compounds is increased by inoculating the autotrophic bacteria portion of the composition, and the metabolism of organic compounds is increased by inoculating the mineral nutrient producing bacteria portion of the composition, thereby increasing the production of metabolizable mineral nutrients in the predetermined substrate to a predetermined level.

23. The method of claim 22, wherein the method improves nitrogen fixation in a preselected substrate by bacteria, the method The following steps are involved: inoculating with a predetermined concentration of at least one strain of nitrogen-fixing bacteria, wherein the nitrogen-fixing bacteria is characterized by the ability to biologically fix atmospheric nitrogen in such predetermined substrate; Wherein, by inoculating the nitrogen-fixing bacteria portion of the composition, the metabolism of organic compounds is improved, thereby increasing the yield of metabolizable nitrogen in the predetermined substrate to a predetermined level.

24. The method according to claim 22, wherein the method increases the metabolizable potassium content in a predetermined substrate by bacteria, the method The following steps are involved: inoculating a predetermined concentration of at least one strain of potassium-solubilizing bacteria, the potassium-solubilizing bacteria being characterized by the ability to dissolve potassium-containing minerals, convert insoluble potassium into soluble potassium, and produce metabolizable potassium in such a predetermined matrix; Wherein, by inoculating the potassium-solubilizing bacteria portion of the composition, the metabolism of organic compounds is increased, thereby increasing the yield of metabolizable potassium in the predetermined substrate to a predetermined level.

25. The method according to claim 22, wherein the method increases the content of metabolizable phosphate in a predetermined substrate by bacteria, the method The following steps are involved: inoculating a predetermined concentration of at least one strain of phosphate-solubilizing bacteria characterized by the ability to solubilize phosphate-containing minerals, convert insoluble phosphates into soluble phosphates, and produce metabolizable phosphates in such a predetermined substrate; The inoculation of the phosphate-solubilizing bacteria portion of the composition improves the metabolism of organic compounds, thereby increasing the yield of metabolizable phosphate in the predetermined substrate to a predetermined level.

26. The method of claim 22, wherein the step of inoculating a predetermined substrate in the form of a solid-based root growth medium include: applying the bacterial composition to the planting trench; applying the bacterial composition to a planting furrow and watering the furrow; as well as The existing vegetation or its surroundings are sprayed with a liquid solution containing the composition in a predetermined volume and at predetermined intervals.