CN115161247A - Bacillus subtilis 201015 and application thereof - Google Patents
Bacillus subtilis 201015 and application thereof Download PDFInfo
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- CN115161247A CN115161247A CN202211007174.1A CN202211007174A CN115161247A CN 115161247 A CN115161247 A CN 115161247A CN 202211007174 A CN202211007174 A CN 202211007174A CN 115161247 A CN115161247 A CN 115161247A
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- C—CHEMISTRY; METALLURGY
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- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
- A01N63/20—Bacteria; Substances produced thereby or obtained therefrom
- A01N63/22—Bacillus
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01P—BIOCIDAL, PEST REPELLANT, PEST ATTRACTANT OR PLANT GROWTH REGULATORY ACTIVITY OF CHEMICAL COMPOUNDS OR PREPARATIONS
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- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/07—Bacillus
- C12R2001/125—Bacillus subtilis ; Hay bacillus; Grass bacillus
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- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
- Y02W30/00—Technologies for solid waste management
- Y02W30/40—Bio-organic fraction processing; Production of fertilisers from the organic fraction of waste or refuse
Abstract
The bacillus subtilis 201015 disclosed by the invention not only can effectively release phosphorus and potassium which are not dissolved in soil or fertilizer, but also has a certain inhibition effect on the growth of plant pathogenic fungi including sclerotinia sclerotiorum, rhizoctonia solani and fusarium. Practice proves that the microbial agent prepared by the strain can effectively promote the growth of crops, and has better control effect on potato rhizoctonia solani caused by rhizoctonia solani.
Description
Technical Field
The invention belongs to the technical field of plant growth-promoting bacteria, and particularly relates to a bacillus subtilis strain 201015 with a phosphate solubilizing effect, a plant growth promoting effect, fusarium, rhizoctonia solani and sclerotinia sclerotiorum inhibiting effect, and an application of the bacillus subtilis strain 201015 in bacterial fertilizer development and prevention and control of potato rhizoctonia solani.
Background
Chemical fertilizers have long played a very important role in increasing the yield per unit area of crops, but the application of fertilizers in one taste has currently had some negative effects. Some farmlands, especially for greenhouse soil, have a series of problems, such as soil hardening, function deterioration, pH imbalance, soil microflora imbalance and environmental pollution.
The microbial fertilizer is also called microbial fertilizer, and is a fertilizer which is prepared by fermenting and culturing plant growth-promoting bacteria beneficial to soil and a culture medium together to fully exert the interaction among floras so as to achieve the optimal application effect. Researches show that plant growth-promoting bacteria (azotobacter, phosphate-solubilizing bacteria, potassium-solubilizing bacteria, plant growth-promoting bacteria and biological control bacteria) in the microbial agent can release organic acid in soil and dissolve compounds such as phosphorus and potassium which are not dissolved in the soil, so that nutrients in the soil are converted into soluble state to be absorbed and utilized by plants again, and the nutritional status of crops and the nutritional conditions of the soil are improved. On the other hand, some microbial agents can improve the community structure of soil microorganisms after being applied, and simultaneously prevent and treat various diseases caused by harmful fungi, bacteria and the like. The microbial agents of different manufacturers have different strains and functions and different effects, so that the selection of effective strains is of great importance to the research of the microbial agents.
One of the problems in the prior art is that the effect of strains contained in some microbial fertilizers is limited, a plurality of strains are required to be added for improving the efficacy of the microbial fertilizers, but when the plurality of strains are mixed and cultured, the growth antagonism often exists, so that the strains are required to be separately cultured and then mixed together, which undoubtedly increases the procedure and cost for preparing the microbial fertilizers, and therefore, the search for plant growth promoting bacteria integrating multiple functions is very important.
The bacillus subtilis is a mesophilic and aerobic gram-positive rod-shaped bacterium, can generate spores with strong resistance, and has broad-spectrum activity and strong stress resistance. The bacillus subtilis is added into part of biological bacterial manure in the market at present, has the advantages of inhibiting the growth of plant pathogenic fungi, promoting the growth of plants, inducing the resistance of plants and the like, and has the characteristics of no harm to people and livestock, high propagation speed, no pollution to environment, strong stress resistance and the like. In addition, the strain has long survival time, transportation resistance, storage resistance, difficult pollution and convenient and safe use.
The rhizoctonia solani of the potato is also called as the stem rot of the potato and the black nevus of the potato, is a common disease of the potato, often causes seedling and ridge shortage and influences agricultural planting and the yield of the potato. The disease occurs in connection with cold spring and humid conditions. The disease is serious when the soil temperature is low after the sowing. The seedbed or the greenhouse has high temperature, much soil moisture, application of non-decomposed fertilizer, poor ventilation and insufficient light, and is most prone to diseases. The pathogenic bacteria of the disease is a fungus, the asexual stage of the disease is rhizoctonia solani, and many wild plants and crops in the world are hosts of the fungus and have wide distribution range. Its sclerotium can overwinter on plant residues, tubers or in soil. When the temperature and the humidity are proper in the spring of the next year, sclerotia germinates, seedlings, buds, stolons, underground stems, tubers and roots of potatoes are infected, sclerotia is formed on new potato blocks, sclerotia also remains in soil, and the sclerotia becomes a bacteria source of the next year after winter. Therefore, the survival amount of rhizoctonia in the non-rotation or less rotation land is increased, and if the infected seed potato is planted again, the bacteria source becomes more and the harm is more serious.
Research and development of novel microbial fertilizers and exploration of new products capable of replacing chemical fertilizers are development trends of green agriculture in China and meet requirements of people on food safety which are more and more emphasized. The microbial agent has good application value, and becomes an important product for developing ecological agriculture in China.
Disclosure of Invention
The invention discloses a Bacillus subtilis 201015 capable of promoting plant growth, which is classified and named as Bacillus subtilis, and is preserved in China general microbiological culture Collection center in 1 month and 19 days in 2021, wherein the preservation unit address is No. 3 of No. 1 Xilu Beijing of the Chaoyang district in Beijing, and the preservation number is CGMCC NO:21665.
The bacillus subtilis has a high-efficiency phosphate solubilizing function. The culture medium has initial pH of 7.0, inoculum size of 6.0% (V/V), and culture conditions of 26 deg.C suitable for the growth of the strain, and phosphorus dissolving amount can reach 567.3 + -1.7 mg/L. Is far higher than the phosphate solubilizing amount of phosphate solubilizing bacteria reported in the prior art. The phosphorus dissolving effect is the key point of the research of phosphorus dissolving microorganisms, and the research finds that the phosphorus dissolving capacity of the microorganisms is greatly different. In the current research, the phosphorus content of the supernatant of the phosphorus-dissolving strain PSB3 screened by Yuxianmei et al (2008) is gradually increased along with the increase of the culture days, reaches 218.6Lg/mL on the 4 th day, and starts to decrease after the peak appears. Li Yue E et al (2010) obtained LM18 with the strongest phosphorus-dissolving capacity from 5 strains of phosphorus-dissolving bacteria of alfalfa and red bean grass, and the phosphorus-dissolving amount was 300.3mg/mL. The effect of different culture conditions on the phosphorus dissolving capacity of the bacillus subtilis is researched by Chenling and the like, and the result shows that the phosphorus dissolving capacity of the strain and the pH value of a culture solution are in extremely obvious negative correlation, and the phosphorus amount reaches the highest peak and is 105.82 mg/L.
The bacillus subtilis has a high-efficiency potassium-dissolving function. The culture medium has initial pH of 7.0, inoculation amount of 6.0% (V/V), and culture condition at 26 deg.C suitable for the growth of the strain, and potassium-dissolving amount of 0.50 + -1.21 mg/L.
The bacillus subtilis has the function of inhibiting plant pathogenic fungi in a plate confronting experiment, and field biological control experiments prove that the microbial inoculum has better control effect on potato rhizoctonia solani caused by rhizoctonia solani.
Activating the strain preserved on the inclined plane, inoculating the strain on a beef extract peptone culture medium, and culturing for 18-24 h; picking up on an activation mediumInoculating the strain into a triangular flask containing 200mL of liquid fermentation medium, culturing at 30-35 ℃ for 12-24 h to obtain seed fermentation liquid, inoculating the seed liquid into a 7-liter fermentation tank according to the inoculation amount of 3-10% for fermentation culture, then inoculating the seed liquid into a 30-liter fermentation tank according to the inoculation amount of 3-10% for fermentation culture to obtain culture liquid, wherein the number of thalli can reach 1.31 multiplied by 10, and the number of thalli can reach 1.31 multiplied by 10 10 ～9.56×10 10 CFU/mL。
The preparation method of the strain solid microbial inoculum comprises the steps of crushing corncobs by using a micro powder sample machine, mixing the crushed corncobs with turfy soil according to the proportion of 1. The total viable bacteria number of the strain is not less than 2.62 × 10 8 CFU/mL. The bacterial fertilizer contains various secondary metabolites generated by microbial fermentation, can promote the formation of soil granular structure, can effectively convert phosphorus insoluble in water in a solution into phosphorus soluble in water, increases the absorption of phosphorus elements by crops, inhibits the growth of some plant pathogenic fungi in soil, and is beneficial to the water retention, fertilizer retention, ventilation and root system development promotion of soil. Meanwhile, the bacterial manure provided by the invention can promote the growth of various crops.
Has the beneficial effects that:
drawings
The accompanying drawings, which are included to provide a further understanding of the invention and are incorporated in and constitute a part of this specification, illustrate embodiments of the invention and together with the description serve to explain the principles of the invention and not to limit the invention. In the drawings:
FIG. 1 shows the amount of potassium released by Bacillus subtilis.
FIG. 2 shows the inhibition of Bacillus subtilis on the growth of sclerotinia sclerotiorum, rhizoctonia solani and Fusarium hyphae;
FIG. 3 is a gram stain of the strain;
FIG. 4 is the phosphate solubilizing circle produced by the strain;
FIG. 5 is a homology alignment of Bacillus subtilis 16S rDNA sequences;
FIG. 6 shows the effect of microbial inoculum on the height of Chinese cabbage;
FIG. 7 is the effect of microbial inoculum on the dry weight and fresh weight of pakchoi;
FIG. 8 shows the effect of microbial inoculum on growth of pakchoi;
FIG. 9 is a picture of the control effect of a microbial inoculum on Rhizoctonia solani, wherein a is not sprayed in advance with the microbial inoculum; picture, b: spraying the microbial inoculum in advance;
FIG. 10 is a symptom of potato root caused by Rhizoctonia solani;
FIG. 11 is a potato which is not infected with pathogenic bacteria after being treated with the microbial inoculum;
FIG. 12 is a graph showing the effect of diluting 10 times the inoculum on the number and size of potato tubers.
Detailed Description
The preferred embodiments of the present invention will be described in conjunction with the accompanying drawings, and it will be understood that they are described herein for the purpose of illustration and explanation and not limitation.
Examples
The bacillus subtilis 201015 is obtained by screening in farmland soil of Yangtze township Chongcun in lake of Tang river in 2019 for 4 months.
(1) Morphological characteristics of colonies: culturing the beef extract peptone agar medium at 25-35 deg.c for 2-3 days, and observing the nutritive cell in the medium, which is unicellular, rod-shaped, gram-positive bacterium 0.5-3 times 0.5-3 microns and has elliptic spore inside the bacterium. The cells were cultured in the above medium at 30 ℃ for 12 hours to allow mass growth. The colony is milky white, the edge is relatively regular, the colony is nearly circular, the surface is rough and irregular, a transparent ring is obviously formed when the colony grows on an inorganic phosphorus PKO culture medium (as shown in figure 4), and the colony can grow within the pH range of 5-10.
(2) Physiological and biochemical properties: see Table 1
TABLE 1 physiological and biochemical Properties of Bacillus subtilis strains
Remarking: +: positive; -: negative; v.p. reaction: voges-Proskauer
(3) Molecular biological characterization of strains
And (3) carrying out 16S rDNA amplification and sequencing on the pure culture strain obtained by separation and purification. The experimental process is as follows: PCR reaction 20 μ L:10 XTaqTM Buffer 2. Mu.L, dNTP mix (2.5 mmol/L each) 1.8. Mu.L, primer C1.4. Mu.L, primer C2.4. Mu.L, template DNA 0.5. Mu.L, taKaRa TaqTM 0.12. Mu.L, and sterilized deionized water 14.78. Mu.L. The reaction conditions are as follows: 94 ℃,5min;94 ℃ for 30s, and the annealing temperature is 49.5 ℃, and the cycle time is 25; after the reaction was complete, extension was carried out for 7min at 72 ℃. Gel recovery was performed using a small gel recovery kit (Shunhua Bioengineering, inc., shanghai), and the purified DNA fragment was stored at-20 ℃ and used for sequencing. After PCR amplification, the obtained sequences are compared in a Genbank database, and the result shows that the homology of the 16S rRNA gene sequence of the strain and the bacillus subtilis reaches 100 percent, and the strain is identified as the bacillus subtilis by combining with physiological and biochemical characteristic determination. As shown in fig. 5.
Determination of inorganic phosphorus dissolving capacity of the strain: the culture solution is put into a centrifuge tube and centrifuged (10000 r/min) for 15mim at 4 ℃, and the effective phosphorus content is measured by molybdenum blue colorimetry after the supernatant is absorbed.
Measurement of potassium-releasing function of the strain: culturing with Behcet's culture medium at 28 deg.C for four days, picking colony after bacteria growth, and inoculating to fermentation culture medium.
Fermentation medium: 10.0g of glucose, 0.2HO4.2g of Na2HPO4, 0.2g of MgSO4.7H2O, 0.2g of NaCL, 0.2g of CaSO 4.2H2O, 0.0g of CaCO35.0g, 2.5g of potassium feldspar powder (150 meshes), 1000mL of deionized water and pH7.2.
Inoculating the bacterial strain obtained by screening into 50mL of fermentation medium, carrying out shake culture at 28 ℃ under the condition of 160r/min, taking 5mL of fermentation liquor every 12h, centrifuging at 4 000r/min for 10min, collecting supernatant, measuring the content of potassium in the fermentation liquor by using an inductively coupled plasma emission spectrometer (ICP-OES), comparing with blank which is not inoculated, and calculating the decomposition rate of the bacterial strain on potassium feldspar ore powder.
Potassium dissolution rate (%) = ((potassium content in bacteria liquid-potassium content in control liquid) x bacteria liquid volume)/potassium feldspar powder amount x total potassium content x100%
The bacillus subtilis has the following inhibition effect on the growth of sclerotinia sclerotiorum, rhizoctonia solani and fusarium hyphae: adopting a plate opposing culture method to preliminarily screen antagonistic strains. Preparing PDA culture medium, inoculating the grown strain on a flat plate, culturing at constant temperature of 25 deg.C with the distance between two inoculating points being 30mm, and observing the growth condition of the flat plate strain with the non-inoculated test strain as control.
The phosphate solubilizing bactericide is prepared by the following steps:
activating bacillus subtilis stored in an ultra-low temperature refrigerator, inoculating the bacillus subtilis to a beef extract peptone culture medium, culturing for 18 hours, selecting strains on the activated culture medium, inoculating the strains into a triangular flask containing 250mL of liquid fermentation culture medium, culturing for 12 hours at 37 ℃ and 200rpm/min to obtain seed fermentation liquid, inoculating the seed liquid into a 7-liter fermentation tank according to 3% of inoculum size for amplification culture, then inoculating the seed liquid into a 30-liter fermentation tank according to 4% of inoculum size for fermentation culture to obtain culture liquid, wherein the number of the strains can reach 2.3 x10 10 CFU/mL。
Crushing corncobs by using a miniature powder sample machine, mixing the crushed corncobs with turfy soil according to the proportion of 1:1, finally adding bacillus subtilis fermentation liquor, uniformly mixing, and drying at the temperature of below 35 ℃ until the total moisture of the bacterial fertilizer is less than or equal to 12 percent to obtain the microbial fertilizer. The total viable bacteria number of the bacterial strains in the bacterial manure is not less than 2.52 multiplied by 10 9 CFU/g；
Example 2:
selecting the strain stored on the inclined plane at 4 ℃, inoculating the strain on a beef extract peptone culture medium, and culturing for 16h; selecting thallus on the activated culture medium, carrying out streak separation and purification culture, inoculating the purified single colony into a triangular flask containing 250mL beef extract peptone culture medium, culturing at 37 ℃ for 10h at 200rpm/min to obtain seed fermentation liquor, inoculating the seed liquor into a 7-liter fermentation tank according to the inoculation amount of 5% for fermentation culture, then inoculating the seed liquor into a 30-liter fermentation tank according to the inoculation amount of 6% for fermentation culture to obtain culture liquor, wherein the thallus number can reach 2.51 multiplied by 10 9 CFU/mL。
Pulverizing corn cob with a miniature powder sample machine, mixing with turfy soil, and adding Bacillus subtilisAnd (3) uniformly mixing the bacillus fermentation liquid, and drying at the temperature of below 35 ℃ until the total moisture of the bacterial manure is less than or equal to 15%, thus obtaining the microbial manure. The total viable bacteria number of the strain is not less than 1.65 multiplied by 10 9 CFU/mL。
Application example 1
The microbial agent (microbial agent for short) prepared in the embodiment 1 is applied to planting of the pakchoi and used as a base fertilizer, and the influence of the microbial agent on the growth of the pakchoi is observed; the microbial agent is applied to potato planting and used as a base fertilizer to observe the influence of the microbial agent on the growth of potatoes and the effect of the microbial agent on the control of rhizoctonia solani of the potatoes.
Application 1: in the effect of the microbial inoculum on the growth of the pakchoi, a single 10 represents that the fermentation liquid of the bacillus subtilis 201015 strain is diluted by 10 times, a single 30 represents that the fermentation liquid of the bacillus subtilis 201015 strain is diluted by 30 times, and ck is a control treatment, wherein the pakchoi is applied with a culture solution which is diluted by 10 times and is not inoculated with bacteria in the treatment. When the Chinese cabbages are planted, the liquid which is correspondingly treated is sprayed in the soil, and the using amount is 5 kg/mu.
As can be seen from FIG. 6, the heights of the pakchoi plants in the experimental group applied with the microbial inoculum are increased by 33% and 28% in different degrees compared with the control group, respectively, wherein the growth promoting effect is the best after the fermentation broth is diluted by 10 times.
From fig. 7, it can be seen that the two processed microbial agents have an effect on both the dry weight and the fresh weight of the pakchoi, wherein the dry weight and the fresh weight of the pakchoi after a single 10-time treatment (dilution of the fermentation broth by 10 times) are significantly different from those of the control group. The fresh weight of the pakchoi with different microbial inoculum increases 267.1% and 61.8% respectively, and the fresh weight of the pakchoi with different microbial inoculum increases 312.5% and 50.0% respectively (figure 8) compared with the control group.
Application example 2
The microbial agent prepared in example 2 has an effect on the growth of potatoes and a control effect on rhizoctonia solani
(1) The potato is sprayed with the microbial inoculum by a sprayer, the dosage is 5 kg/mu, and then rhizoctonia solani is inoculated to observe the biological control effect of the microbial inoculum.
After potato tubers of early white varieties are subjected to quantitative spray treatment, the potato tubers are immediately inoculated with the sclerotium rolfsii mycelium blocks subjected to plate culture, and each potato tuber is inoculated with about 1 square centimeter of mycelium blocks. The results of the experiment were recorded after 3d incubation. The results show that the disease incidence of the potato treated by the bacillus subtilis microbial inoculum is obviously lower than that of the control (only pathogenic bacteria are inoculated, and no microbial inoculum is used for any treatment), the control effect can reach 59.7 percent, and the biocontrol microbial inoculum sprayed before inoculation has a certain inhibition effect on the infection of the pathogenic bacteria (figure 4).
(2) Observing the biological control effect of the inoculant after the inoculant is sprayed 2 days after the inoculation of the rhizoctonia solani
The method comprises the steps of firstly inoculating rhizoctonia solani mycelium blocks on potato tubers of early white varieties, culturing for 2d at 28 ℃, then carrying out spray treatment by using a bacillus subtilis microbial inoculum, repeating each treatment for 3 times, taking potato slices inoculated with pathogenic bacteria without spraying a biocontrol microbial inoculum as a reference, spraying the biocontrol microbial inoculum without spraying the pathogenic bacteria, and culturing for 2d and then observing an experimental result (figure 9).
The experimental result shows that after the quantitative spraying treatment of the microbial inoculum, the pathogenic rate of pathogenic bacteria can be reduced, but the disease inhibiting effect is obviously weaker than the effect of spraying the microbial inoculum in advance. After the pathogenic bacteria infect the plants, the bacterial agent is used for treatment, and the bacterial inhibition control effect is not good. (Table 2).
TABLE 2 bacteriostatic test by inoculating pathogenic bacteria first and then using bacterial agent to prevent and treat
3) Effect of bacterial agent in preventing and controlling rhizoctonia solani of potato in field
After the seed potatoes are sown, the fermented microbial inoculum is diluted and then is subjected to spraying treatment, and the specific dosage is shown in the table. The application effect experiment of the bacillus subtilis strain in the field shows that the bacillus subtilis strain has certain control effect on diseases caused by rhizoctonia solani, and the inhibition rate of the bacillus subtilis strain on the diseases can reach more than 60% no matter the application amount of the microbial inoculum. FIG. 10 shows the root symptoms of potato caused by Rhizoctonia solani, and FIG. 11 shows potato treated with the microbial inoculum without infecting pathogenic bacteria (random block design of cell, row length of 5m,6 ridge area, row spacing of 24cm, ridge width of 0.65cm, cell area of 19.5m2, three times of repetition, after harvest, sampling for photography and index measurement, and the same steps as those in experiments (4) and (5)).
TABLE 3 field test of biological control effect of microbial inoculum
(4) Influence of microbial inoculum on number of potatoes in single plant
As can be seen from Table 4, the four seed dressings increased the number of potatoes grown per plant, the ratio of medium to small potatoes, compared to the control, with 3.5 kg/acre of the inoculant treatment having the highest number of potatoes per plant, and the inoculant treated potatoes compared to the compound fertilizer treatment.
TABLE 4 influence of inoculum on number and type of potatoes per plant
Remarking: small potatoes: the weight of each potato is less than 100g, and the weight of each potato is as follows: the weight of each potato is 100g-300 g; big potato: individual potatoes weigh more than 300g.
(5) Influence of microbial inoculum on potato yield
As can be seen from Table 5, compared with the control, the treatment with the microbial inoculum of different concentrations increases the potato yield, the microbial inoculum treatment with the application amount of 6 kg/mu increases the potato yield to the maximum, but compared with the treatment with the phosphate fertilizer, the effect of increasing the yield is not as good, which indicates that the microbial inoculum has a certain phosphate fertilizer effect (as shown in FIG. 12).
TABLE 5 Effect of microbial inoculum on Potato yield
Finally, it should be noted that: although the present invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that changes may be made in the embodiments and/or equivalents thereof without departing from the spirit and scope of the invention. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (7)
1. The Bacillus subtilis 201015 is classified and named as Bacillus subtilis, and is preserved in China general microbiological culture Collection center (CGMCC) at 19 th 1 month 2021, with the preservation number of CGMCC NO:21665.
2. The use of the bacillus subtilis 201015 of claim 1 for improving the nutritional status of crops and the nutritional conditions of soil by increasing the dissolved phosphorus and/or the dissolved potassium of soil species through a phosphorus and/or potassium solubilizing effect.
3. The use of bacillus subtilis 201015 of claim 1 for inhibiting fusarium, rhizoctonia solani and sclerotinia and controlling rhizoctonia solani.
4. The application of the bacillus subtilis 201015 of claim 1 in preparing a microbial fertilizer.
5. A microbial fertilizer, characterized in that the active ingredient is the Bacillus subtilis 201015 of claim 1.
6. The microbial fertilizer according to claim 5, characterized in that it is implemented as a root irrigation or a hole application.
7. The preparation method of the microbial fertilizer as claimed in claim 5, wherein the preparation method comprises the steps of crushing corncobs, mixing the crushed corncobs with turfy soil according to a ratio of 1.
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