Phosphorus-solubilizing and potassium-promoting bacterium SWYP-1 and application thereof

Abstract

The invention belongs to the technical field of microorganisms, and particularly relates to a phosphorus-solubilizing and potassium-promoting bacterium SWYP-1 and application thereof. The phosphorus-solubilizing and potassium-promoting bacterium is SWYP-1, the taxonomic name of the phosphorus-solubilizing and potassium-promoting bacterium is Phyllobacterium trunii, the preservation number of the phosphorus-solubilizing and potassium-promoting bacterium is CGMCC (China General Microbiological Culture Collection Center) No.28728, and the phosphorus-solubilizing and potassium-promoting bacterium is preserved in the China General Microbiological Culture Collection Center on October 24, 2023. The strain can decompose solidified phosphorus resources in soil into phosphorus which can be absorbed and utilized by plants, promote phosphorus absorption of the plants, effectively utilize coal gangue resources and relieve the pressure of coal gangue on the environment, and has a wide application prospect. The invention also provides a biological agent, a soil conditioner and a microbial fertilizer containing the phosphorus-solubilizing and potassium-promoting bacterium, which can be conveniently applied to soil or rhizosphere environment around plants, decompose phosphorus and potassium in the soil into quick-acting phosphorus and quick-acting potassium, decompose mineral elements, improve the fertilizer quality of the soil, increase both production and income and improve the environment.

Description

Technical Field

[0001] The present application relates to the field of microbial technology, and in particular to a phosphate-solubilizing and potassium-promoting bacterium SWYP-1 and its application. Background Art

[0002] Phosphorus is one of the three essential nutrients for crop growth and development, and plays an important role in agriculture. Phosphorus mainly exists in the form of insoluble phosphates in the soil, and crops have difficulty absorbing and utilizing this insoluble phosphorus, which cannot meet their growth needs. More than 95% of the phosphorus in the soil is in an ineffective form, and plants have difficulty absorbing and utilizing this insoluble phosphorus for their growth and development. At present, in agricultural production, the problem of soil deficiency is generally solved by applying phosphorus fertilizers to meet the plant's demand for phosphorus. However, more and more unfavorable conditions have restricted the use of phosphorus fertilizers. On the one hand, the continuous depletion of high-quality phosphate resources and the increasing shortage of new energy sources have led to an increase in the production and transportation costs of phosphorus fertilizers, increasing the burden on agricultural production. On the other hand, despite the application of a large amount of phosphorus fertilizers, the utilization rate of phosphorus fertilizers by crops is very low.

[0003] The main treatment methods for coal gangue are physical, chemical and biological methods. The biological method mainly uses the metabolic activities of organisms or microorganisms to improve the physical and chemical properties of the soil and enhance the biological activity of the soil, thereby partially or completely restoring the soil ecological environment to its original state. Among them, some microorganisms with phosphorus and potassium solubilizing functions can dissolve the insoluble nitrogen, phosphorus and potassium in coal gangue, thereby accelerating the weathering of coal gangue and increasing the nutrient content of coal gangue available for plant absorption and utilization.

[0004] Therefore, screening microbial strains that can solubilize phosphorus and promote potassium can effectively solve the problems of severe phosphorus deficiency in soil and the comprehensive utilization of coal gangue. Summary of the invention

[0005] In order to effectively solve the problems of severe phosphorus deficiency in the soil and the comprehensive utilization of coal gangue, the present application provides a microbial strain that can dissolve phosphorus and promote potassium. The strain can decompose the solidified phosphorus resources in the soil into phosphorus that can be absorbed and utilized by plants, thereby promoting plant phosphorus absorption. At the same time, it can also effectively decompose the mineral elements in coal gangue, especially the decomposition of mineral elements in coal gangue in a solid-state reaction, and has broad application prospects.

[0006] On the one hand, the present application provides a phosphate-solubilizing and potassium-promoting bacterium SWYP-1, which has been identified as Phyllobacterium ifriqiyense and has been deposited. The deposit information is as follows: Strain: SWYP-1 Deposit number: CGMCC No.28728 Taxonomic name: Phyllobacterium ifriqiyense Collection date: October 24, 2023 Depository: China National Microbiological Culture Collection Administration General Microbiology Center Deposit address: Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.

[0007] Using the above technical scheme, the phosphate-solubilizing and potassium-promoting bacteria SWYP-1 is Phyllobacteriumifriqiyense, which can decompose the solidified phosphorus resources in the soil into phosphorus that can be absorbed and utilized by plants, promote plant phosphorus absorption, and can also effectively decompose the mineral elements in coal gangue, especially the decomposition of mineral elements in coal gangue in solid-state reactions, and has broad application prospects.

[0008] On the other hand, a biological bacterial agent for solubilizing phosphate and promoting potassium is provided, wherein the active ingredient of the biological bacterial agent is the phosphate-solubilizing and potassium-promoting bacteria SWYP-1.

[0009] Furthermore, the active ingredients of the biological bacterial agent include the bacterial body, bacterial liquid or fermentation liquid of the phosphate-solubilizing and potassium-promoting bacteria SWYP-1.

[0010] Furthermore, the biological agent is in the form of a wettable powder, a water dispersant, a water suspension or a dispersible oil suspension.

[0011] By adopting the above technical scheme, the phosphate-solubilizing and potassium-promoting bacteria SWYP-1 is prepared into a biological bacterial agent. The bacterial body or bacterial liquid can be obtained by centrifuging the phosphate-solubilizing and potassium-promoting bacteria SWYP-1 through cultivation, or the phosphate-solubilizing and potassium-promoting bacteria SWYP-1 can be fermented in a suitable culture environment to obtain a fermentation liquid, which is mixed with other components to prepare a biological bacterial agent. The dosage form of the biological bacterial agent can be a wettable powder, a water dispersant, a water suspension or a dispersible oil suspension, which is convenient to use.

[0012] On the other hand, the present application also provides a soil conditioner for solubilizing phosphorus and promoting potassium, wherein the active ingredient of the soil conditioner includes the phosphorus solubilizing potassium promoting bacteria SWYP-1.

[0013] By adopting the above technical scheme, the phosphate-solubilizing and potassium-promoting bacteria SWYP-1 is prepared into a soil conditioner. The soil conditioner is prepared by preparing the bacteria of the phosphate-solubilizing and potassium-promoting bacteria SWYP-1 alone or mixing it with commonly used components with soil improvement properties. When applied to the soil, it can effectively improve the phosphorus and potassium content in the soil, increase the phosphorus absorption or potassium absorption coefficient of the soil, improve the soil's ability to absorb and fix phosphorus / potassium, improve soil fertility, increase production and income, and improve the environment.

[0014] On the other hand, the present application also provides a microbial fertilizer for solubilizing phosphorus and promoting potassium, wherein the active ingredient of the microbial fertilizer includes the phosphorus solubilizing potassium promoting bacteria SWYP-1.

[0015] By adopting the above technical scheme, the phosphate-solubilizing and potassium-promoting bacteria SWYP-1 is prepared into a microbial fertilizer. By applying the microbial fertilizer to the rhizosphere surrounding environment of plant growth, such as soil, soilless culture medium or plant growth nutrient solution, the plant's absorption of phosphorus and potassium elements can be effectively promoted, thereby promoting plant growth.

[0016] On the other hand, the present application also provides the use of the phosphate-solubilizing and potassium-promoting bacteria SWYP-1, the biological bacterial agent, the soil conditioner or the microbial fertilizer in solubilizing phosphate and potassium, adjusting soil pH, decomposing mineral elements or promoting plant growth.

[0017] Furthermore, the application of the mineral element decomposition is to decompose the mineral elements in calcium phosphate or coal gangue.

[0018] Furthermore, the decomposition is to decompose the coal gangue in a solid state reaction.

[0019] Furthermore, the application of promoting plant growth is specifically to apply the phosphate-solubilizing and potassium-promoting bacteria SWYP-1, the biological agent, the soil conditioner or the microbial fertilizer to the rhizosphere environment where the plants grow.

[0020] Compared with the prior art, this application has the following technical effects: 1) The present application conducts extensive soil sampling from the habitat of the strain, and obtains a phosphate-solubilizing and potassium-promoting bacterium SWYP-1 through separation, purification and screening. The bacterium is biologically identified as Phyllobacterium ifriqiyense. The strain can decompose the solidified phosphorus resources in the soil into phosphorus that can be absorbed and utilized by plants, thereby promoting plant phosphorus absorption. At the same time, it can also effectively utilize coal gangue resources and alleviate the pressure caused by coal gangue on the environment. It has broad application prospects.

[0021] 2) The phosphate-solubilizing and potassium-promoting bacteria SWYP-1 provided in the present application is used to prepare biological agents, soil conditioners and biological fertilizers, which can be conveniently applied to the soil or the rhizosphere environment around plants to decompose phosphorus and potassium in the soil into fast-acting phosphorus and fast-acting potassium, thereby improving soil fertility, increasing production and income, and improving the environment.

[0022] 3) The phosphate-solubilizing and potassium-promoting bacteria SWYP-1 provided in the present application can decompose the mineral elements in the coal gangue by carrying out a solid-state reaction with the coal gangue under the condition of controlling the water content of the bacterial sludge at 10%. The reaction completely simulates the industrial production process, indicating that the strain has a strong decomposition ability and industrial application value. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] For better understanding and implementation, the present invention is described in detail below with reference to the accompanying drawings.

[0024] Figure 1 This is the colony appearance and Gram staining results of the phosphate-solubilizing and potassium-promoting bacteria SWYP-1.

[0025] Figure 2 is the standard curve of phosphate ion. DETAILED DESCRIPTION

[0026] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention. The experimental methods in the following examples without specifying specific conditions are usually carried out according to conventional conditions, conditions described in laboratory manuals or conditions recommended by manufacturers.

[0027] The composition of the culture medium used is as follows.

[0028] PVK plate culture medium: glucose 10.0 g / L, ammonium sulfate 0.5 g / L, yeast extract powder 0.5 g / L, sodium chloride 0.3 g / L, potassium chloride 0.3 g / L, magnesium sulfate 0.3 g / L, ferrous sulfate 0.03 g / L, manganese sulfate 0.03 g / L, calcium phosphate 5.0 g / L, agar 15.0 g / L.

[0029] LB medium: sodium chloride 10 g / L, peptone 10 g / L, yeast powder 5 g / L.

[0030] Medium B (inorganic phosphorus liquid medium with calcium phosphate as the phosphorus source): glucose 10 g / L, ammonium sulfate 0.5 g / L, sodium chloride 0.3 g / L, magnesium sulfate heptahydrate 0.3 g / L, manganese sulfate tetrahydrate 0.03 g / L, potassium chloride 0.3 g / L, ferrous sulfate heptahydrate 0.03 g / L, calcium phosphate 5 g / L, yeast extract 0.5 g / L.

[0031] Culture medium C (inorganic phosphorus liquid culture medium with coal gangue as phosphorus source): glucose 10 g / L, ammonium sulfate 0.5 g / L, sodium chloride 0.3 g / L, magnesium sulfate heptahydrate 0.3 g / L, manganese sulfate tetrahydrate 0.03 g / L, potassium chloride 0.3 g / L, ferrous sulfate heptahydrate 0.03 g / L, coal gangue 50 g / L, yeast extract 0.5 g / L.

[0032] The main components of the coal gangue used are shown in Table 1.

[0033] Table 1 Content of each component of coal gangue (ω(B) / %) Silicon dioxide Aluminum oxide Iron (III) oxide Titanium Dioxide Calcium Oxide Magnesium Oxide <![CDATA[SiO2]]> <![CDATA[Al2O3]]> <![CDATA[Fe2O3]]> <![CDATA[TiO2]]> CaO MgO 49.2 22.36 1.44 0.752 0.296 1.81 Potassium Oxide Sodium oxide Manganese oxide Phosphorus pentoxide Loss on ignition sulfur <![CDATA[K2O]]> <![CDATA[Na2O]]> MnO <![CDATA[P2O5]]> LOI S 1.84 1.05 0.031 0.081 20.9 0.11 Example 1 Isolation and screening of strains 1. Sample collection and pretreatment Sample collection: Conduct extensive sample collection in the likely habitat of the strain.

[0034] Sample pretreatment: Select 7-8 plant roots from the sample and place them in a sample treatment tube. Add 20 ml of sterilized distilled water, add glass beads, and vortex for 10 minutes.

[0035] 2. Initial screening Pipette 1 ml of the pretreated sample solution and dilute to 10 -3 , 10 -4 , 10 -5 , pipette 100ul onto the PVK plate, spread it evenly with a coating rod, make three parallel plates, and culture in a 37℃ incubator for 2-3 days.

[0036] 3. Rescreening Pick a single colony with a transparent circle on the plate and culture it in a 5 ml LB test tube at 37°C with shaking for 24 h.

[0037] 4. Strain Isolation Take 1 ml of the above bacterial solution and dilute it to 10 -5 , 10 -6 , 10 -7 , pipette 100ul onto the PVK plate, spread it evenly with a coating rod, make three parallel plates, and culture in a 37℃ incubator for 2-3 days.

[0038] 5. Enrichment culture Single colonies with obvious transparent zones and vigorous growth were selected for enrichment and preservation.

[0039] Example 2 Identification of strains 1) Appearance of colonies like Figure 1 As shown in the figure, the colony of the bacteria is white, round in shape, with smooth edges and a convex and moist surface. After Gram staining, the bacteria appear purple, which is a Gram-positive bacteria.

[0040] 2) Molecular biological identification The genomic DNA of the strain obtained by screening was extracted and 16S rRNA was amplified by PCR. The product was recovered by electrophoresis gel and then sequenced to obtain the 16S rRNA nucleotide sequence of the strain, as shown in SEQ ID NO.1.

[0041] After comparison in the bioinformatics database, phylogenetic analysis was performed based on the neighbor joining method using software such as ClustalW (1.6) and MEGA5.05 to construct a phylogenetic tree. The strain was identified as Phyllobacterium ifriqiyense and was named phosphate-solubilizing and potassium-promoting bacteria SWYP-1.

[0042] Example 3 The colonies of phosphate-solubilizing and potassium-promoting bacteria SWYP-1 were activated and expanded in LB medium, and cultured at 37°C, 200r / min in a shaking incubator for 24h. Take 5ml of the culture fluid of the expanded strain and add it to 300ml of B medium and C medium, and culture at 37°C, 200r / min in a shaking incubator. Take 1ml of samples on the 0th day, 1st day, 2nd day, and 3rd day, respectively, and after measuring the pH value, dilute the B medium to 250 times with deionized water, and dilute the C medium to 500 times with deionized water, and determine the absorbance using the molybdenum antimony colorimetric method. At the same time, use the phosphorus standard solution to prepare a standard curve using the molybdenum antimony colorimetric method (such as Figure 2 As shown), the concentration of phosphate ions was calculated according to the standard curve. The concentration of phosphate ions was C = 1.3881 × absorbance A + 0.0019, R 2 =0.9988, the test results are shown in Table 2.

[0043] Table 2 Phosphate ion concentration As can be seen from Table 2, in the inorganic phosphorus liquid culture medium with calcium phosphate as the phosphorus source and coal gangue as the phosphorus source, the concentration of phosphate increases continuously with the extension of the culture time, indicating that the phosphate-solubilizing and potassium-promoting bacteria SWYP-1 can decompose both calcium phosphate and coal gangue.

[0044] Example 4 Take 50 ml of the culture solution of the selected bacteria after 16 hours of shaking culture at 37°C, 200 r / min, centrifuge, remove the supernatant, resuspend with 30 ml of deionized water, and divide it into 3 parts for standby use. Take 600 g of coal gangue that has passed through a 10-mesh sieve and divide it into 6 parts, add one part of the standby solution to each of 3 parts, which is the experimental group, and add 10 ml of deionized water to each of the other 3 parts, which is the blank group, and culture in a constant temperature incubator at 37°C.

[0045] One week later, 5 g of samples were taken, vortexed and extracted with 20 ml of deionized water for 1 min, allowed to stand for 30 min, centrifuged at 7000 r / min for 5 min, the supernatant was taken, filtered with a 2-micron filter membrane, and the contents of potassium, sodium, calcium, magnesium and iron were determined by atomic absorption spectrometry. The results are shown in Table 3.

[0046] Table 3 Content of each element element Experimental group (mg / L) Blank group (mg / L) Potassium 12.16 3.44 sodium 14.54 4.86 calcium 12.46 3.24 magnesium 3.20 0.86 iron 5.26 1.38 It can be seen from Table 3 that after adding the phosphate-degrading and potassium-promoting bacteria SWYP-1, the contents of potassium, sodium, calcium, magnesium and iron in the culture medium are significantly improved compared with the blank group, indicating that the Tunisian Leaf Bacillus (Phyllobacteriumifriqiyense) SWYP-1 provided in the present application can react with coal gangue in a solid state at a water content of 10%, decompose the chemical elements in the solid coal gangue, and has good industrial application value.

[0047] This specific embodiment is merely an explanation of the present application and is not a limitation of the present application. After reading this specification, those skilled in the art may make modifications to the present embodiment without any creative contribution as needed, but such modifications are protected by the patent law as long as they are within the scope of the claims of the present application.

Claims

1. A phosphate-solubilizing and potassium-promoting bacterium SWYP-1, characterized in that: The phosphate-solubilizing and potassium-promoting bacteria SWYP-1 is taxonomically named Phyllobacterium ifriqiyense, with a preservation number of CGMCC No. 28728. It was deposited in the General Microbiology Center of the China National Microbiological Culture Collection Administration on October 24, 2023.

2. A biological agent for dissolving phosphorus and promoting potassium, characterized in that: The active ingredients of the biological bacterial agent include the phosphate-solubilizing and potassium-promoting bacteria SWYP-1 described in claim 1.

3. The biological agent according to claim 2, characterized in that: The active ingredients of the biological bacterial agent include the bacterial body, bacterial liquid or fermentation liquid of the phosphate-solubilizing and potassium-promoting bacteria SWYP-1 described in claim 1.

4. The biological agent according to claim 3, characterized in that: The biological bacterial agent is in the form of a wettable powder, a water dispersant, a water suspension or a dispersible oil suspension.

5. A soil conditioner for dissolving phosphorus and promoting potassium, characterized in that: The active ingredient of the soil conditioner includes the phosphate-solubilizing and potassium-promoting bacteria SWYP-1 described in claim 1.

6. A microbial fertilizer for dissolving phosphorus and promoting potassium, characterized in that: The active ingredient of the microbial fertilizer includes the phosphate-solubilizing and potassium-promoting bacteria SWYP-1 described in claim 1.

7. Use of the phosphate-solubilizing and potassium-promoting bacteria SWYP-1 described in claim 1, the biological agent described in any one of claims 2 to 4, the soil conditioner described in claim 5 or the microbial fertilizer described in claim 6 in solubilizing phosphate and potassium, adjusting soil pH, decomposing mineral elements or promoting plant growth.

8. The use according to claim 7, characterized in that: The application of the mineral element decomposition is to decompose the mineral elements in calcium phosphate or coal gangue.

9. The use according to claim 8, characterized in that: The decomposition is to decompose the coal gangue in a solid state reaction.

10. The use according to claim 8, characterized in that: The application for promoting plant growth is to apply the phosphate-solubilizing and potassium-promoting bacteria SWYP-1 described in claim 1, the biological agent described in any one of claims 2-4, the soil conditioner described in claim 5 or the microbial fertilizer described in claim 6 to the rhizosphere surrounding environment where the plants grow.

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