CN114591851A - Agricultural microbial agent and preparation method thereof - Google Patents
Agricultural microbial agent and preparation method thereof Download PDFInfo
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- C—CHEMISTRY; METALLURGY
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- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- 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
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- A01N63/23—B. thuringiensis
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- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F11/00—Other organic fertilisers
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- C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
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Abstract
The invention belongs to the technical field of agricultural wastewater treatment, discloses an agricultural microbial agent and a preparation method thereof, and particularly discloses a culture medium, which comprises the following components: the tea leaf residue leachate, a carbon source, a nitrogen source and trace elements, and the preparation method of the tea leaf residue leachate comprises the following steps: and (4) carrying out filter pressing on the tea residues, collecting the percolate, and filtering to obtain the tea residue percolate. The invention provides a culture medium prepared from tea leaf residue percolate, which can be effectively used for culturing bacillus thuringiensis; the culture medium is prepared from the tea leaf residue percolate, so that the problem of treatment of high-concentration tea leaf residue wastewater can be effectively solved, loss of useful substances in the tea leaf residue wastewater is prevented, the utilization rate of the tea leaf residue wastewater is improved, and the pollution of the tea leaf residue wastewater to the environment is reduced.
Description
Technical Field
The invention belongs to the technical field of agricultural wastewater treatment, and particularly relates to an agricultural microbial agent and a preparation method thereof.
Background
With the rapid development of economy and improvement of living standard of people in China, tea drinks are more and more common in our lives. The tea residue leachate is high-concentration organic wastewater generated under physical and biological degradation such as compaction and fermentation of tea residues and seepage of precipitation and underground water in the tea beverage manufacturing process. The tea beverage manufacturing industry in China discharges more than ten million tons of high-concentration tea residue percolate every year, which is a main pollution source of tea beverage manufacturing enterprises and is the most difficult to treat, and the problem of wastewater treatment becomes a big problem in the tea beverage industry in China. The tea residue wastewater discharge amount is large, the nutrient content is large, the COD concentration is high, the tea residue wastewater is treated according to the traditional process, the treatment process is complex, the cost is high, the tea residue wastewater cannot be repeatedly used, and the enterprise burden is heavy.
Bacillus thuringiensis (Bt) is a Bacillus which parasitizes in insects to cause insect death and can produce parasporal crystals, Bt manufacturers registered in China by national pesticide administration departments currently have nearly 70 plants, the annual output is more than 3 multiplied by 10t, and the dosage forms of the product mainly comprise liquid and emulsion, and also comprise wettable powder and suspending agent. The preparation overcomes the defects of environmental pollution, harm to people and livestock, easy generation of resistance and the like of the traditional chemical pesticide, has the advantages of strong selectivity, safety, simple raw materials and the like, is a microbial pesticide which has the widest application and the largest yield in the world at present, accounts for 90-95 percent of the total amount of the microbial pesticide, and is widely applied to pest control in the aspects of agriculture, forestry, fruits, vegetables and urban gardens.
Disclosure of Invention
The first aspect of the present invention aims to provide a culture medium.
In a second aspect, the invention provides the use of a medium according to the first aspect of the invention for the cultivation of Bacillus thuringiensis.
The third aspect of the present invention is to provide a method for producing the agricultural microbial preparation of the first aspect of the present invention.
The fourth aspect of the invention aims to provide an agricultural microbial agent.
The fifth aspect of the present invention is directed to the use of the method of the third aspect of the present invention or the agricultural microbial preparation of the fourth aspect of the present invention for preparing a fertilizer.
The sixth aspect of the present invention is to provide the use of the agricultural microbial agent of the fourth aspect of the present invention for promoting the growth of crops.
In order to achieve the purpose, the technical scheme adopted by the invention is as follows:
in a first aspect of the invention, a culture medium is provided, which comprises the following components in parts by weight: the tea leaf residue leachate, a carbon source, a nitrogen source and trace elements, and the preparation method of the tea leaf residue leachate comprises the following steps: and (4) carrying out filter pressing on the tea residues, collecting the percolate, and filtering to obtain the tea residue percolate.
Preferably, the culture medium comprises the following components in parts by weight: 70-95 parts of tea leaf residue leachate, 1-15 parts of carbon source, 1-9 parts of nitrogen source and 0.1-5 parts of trace elements.
Further preferably, the culture medium comprises the following components in parts by weight: 80-93 parts of tea leaf residue leachate, 2-10 parts of carbon source, 3-7 parts of nitrogen source and 0.1-2 parts of trace elements.
Still more preferably, the culture medium comprises the following components in parts by weight: 80-90 parts of tea leaf residue leachate, 4-10 parts of carbon source, 4-7 parts of nitrogen source and 1-2 parts of trace elements.
Preferably, the trace elements include monopotassium phosphate, zinc sulfate and ammonium sulfate.
Preferably, the trace elements comprise the following components in parts by weight: 0.1-1 part of monopotassium phosphate, 0.1-0.8 part of zinc sulfate and 0.2-1.5 parts of ammonium sulfate.
Further preferably, the trace elements comprise the following components in parts by weight: 0.3-0.8 part of monopotassium phosphate, 0.3-0.5 part of zinc sulfate and 0.3-0.7 part of ammonium sulfate.
Still more preferably, the trace elements comprise the following components in parts by weight: 0.3-0.5 part of monopotassium phosphate, 0.3-0.4 part of zinc sulfate and 0.3-0.4 part of ammonium sulfate.
Preferably, the carbon source is one or more of glucose, lactose, sucrose, trehalose, fructose, mannitol and corn starch.
Preferably, the nitrogen source is one or more of corn steep liquor, casein, soybean meal, peptone, yeast extract, cotton seed powder and yeast powder.
Preferably, the filtration is filter paper filtration.
Preferably, the preparation method of the culture medium comprises the following steps: and sterilizing the tea residue percolate, and adding a carbon source, a nitrogen source and trace elements to obtain the culture medium.
Further preferably, the sterilization is carried out under the conditions of 115-150 ℃ and 0.1-0.3 MPa for 15-35 min.
In a second aspect of the invention, there is provided the use of a medium according to the first aspect of the invention in the cultivation of Bacillus thuringiensis.
In a third aspect of the present invention, a method for preparing an agricultural microbial agent is provided, which comprises the following steps: inoculating bacillus thuringiensis to the culture medium of the first aspect of the invention, and culturing to obtain the agricultural microbial agent.
Preferably, the inoculation amount of the bacillus thuringiensis is 0.1 w/w% -2 w/w%.
Further preferably, the inoculation amount of the bacillus thuringiensis is 0.1 w/w% -1.5 w/w%.
Still more preferably, the inoculation amount of the bacillus thuringiensis is 0.1 w/w% -1 w/w%.
Preferably, the culture condition is 25-37 ℃ and 100-200 r/min for 20-24 h.
Further preferably, the culture conditions are 30-37 ℃ and 150-200 r/min for 22-24 h.
Preferably, the agricultural microbial agent is a liquid formulation or a solid formulation.
Preferably, when the agricultural microbial agent is a solid preparation, the preparation method of the agricultural microbial agent further comprises the following steps: adding the carrier, mixing evenly and drying to obtain the agricultural microbial agent of the solid preparation.
Further preferably, the carrier is one or more of bran, rice bran, straw, peanut shell and wood shavings.
In a fourth aspect of the present invention, an agricultural microbial preparation is provided, wherein the agricultural microbial preparation is prepared by the preparation method of the agricultural microbial preparation of the third aspect of the present invention.
Preferably, the viable count of the bacillus thuringiensis in the agricultural microbial agent is (3-7) multiplied by 109CFU/mL。
Further preferably, the viable count of the bacillus thuringiensis in the agricultural microbial agent is (4-7) × 109CFU/mL。
More preferably, the viable count of the bacillus thuringiensis in the agricultural microbial agent is (4.5-6) × 109CFU/mL。
In a fifth aspect of the present invention, there is provided a method for preparing the agricultural microbial preparation of the third aspect of the present invention or use of the agricultural microbial preparation of the fourth aspect of the present invention in preparing a fertilizer.
In a sixth aspect of the present invention, there is provided the use of the agricultural microbial inoculant of the fourth aspect of the present invention for promoting the growth of crops.
Preferably, the crops are one or more of food crops, oil crops, vegetable crops, fruits, feed crops and medicinal crops.
Further preferably, the crop is a vegetable crop.
The invention has the beneficial effects that:
the invention provides a culture medium prepared from tea leaf residue percolate, which can be effectively used for culturing bacillus thuringiensis. By preparing the tea leaf residue percolate into the culture medium, the problem of treatment of high-concentration tea leaf residue wastewater can be effectively solved, loss of useful substances in the tea leaf residue wastewater is prevented, the utilization rate of the tea leaf residue wastewater is improved, and the pollution of the tea leaf residue wastewater to the environment is reduced; meanwhile, comprehensive utilization of resources is realized, the burden of environmental protection cost is reduced for enterprises, and new products can be developed to become new profit growth points of the enterprises, so that the effect of changing waste into valuable is achieved.
According to the invention, substances such as sugars, polyphenol, proteins and the like in the tea residue percolate are taken as nutrient components, a small amount of nitrogen source, carbon source and trace elements are supplemented, and bacillus thuringiensis can grow and proliferate at a proper temperature to further obtain the microbial agent.
The invention provides an agricultural microbial agent, all indexes of the agricultural microbial agent accord with national standard GB20287-2006 agricultural microbial agent, and effective viable bacteria bacillus thuringiensis is more than or equal to 2 multiplied by 109CFU/mL, safe and nontoxic. The agricultural microbial agent is used for planting crops, can effectively promote the growth of the crops, improves the yield of the crops, has no toxic or side effect on the crops, does not influence the quality of agricultural products, can effectively reduce the use of chemical fertilizers and pesticides, and does not cause pollution to the environment.
Drawings
FIG. 1 is a diagram showing the growth of Bacillus thuringiensis in the agricultural microbial preparation prepared in example 4.
FIG. 2 is a diagram showing the growth of Bacillus thuringiensis in the agricultural microbial preparation prepared in example 5.
FIG. 3 is a diagram showing the growth of Bacillus thuringiensis in the agricultural microbial preparation prepared in example 6.
FIG. 4 is a field diagram of a cabbage heart laboratory field in which one treatment is performed.
FIG. 5 is a field diagram of the cabbage heart experimental field of the second treatment.
FIG. 6 is a field diagram of the experimental vegetable heart field for the third treatment.
Detailed Description
The present invention will now be described in detail with reference to specific examples, but the scope of the present invention is not limited thereto.
The materials, reagents and the like used in the present examples are commercially available materials and reagents unless otherwise specified.
Example 1
A culture medium comprises the following components in parts by weight: 90 parts of tea leaf residue percolate, 4.5 parts of glucose, 4.4 parts of yeast powder and 1 part of trace elements;
the trace elements comprise 0.3 part of monopotassium phosphate, 0.3 part of zinc sulfate heptahydrate and 0.4 part of ammonium sulfate;
the preparation method of the tea residue percolate comprises the following steps: collecting the tea leftovers after soaking for 2 hours, performing filter pressing on the tea leftovers by using a plate-and-frame filter press to obtain percolate, and filtering the tea residue wastewater by using filter paper to obtain tea residue percolate;
the preparation method of the culture medium comprises the following steps:
sterilizing the tea residue percolate in a high temperature sterilization pot at high temperature and high pressure (125 deg.C, 0.15MPa) for 30min to obtain tea residue base solution, and adding glucose, yeast powder and microelements to obtain culture medium.
Example 2
A culture medium comprises the following components in parts by weight: 93 parts of tea leaf residue leachate, 2.5 parts of glucose, 3 parts of yeast powder and 1.2 parts of trace elements;
the trace elements comprise 0.5 part of monopotassium phosphate, 0.4 part of zinc sulfate heptahydrate and 0.3 part of ammonium sulfate;
the preparation method of the tea residue percolate comprises the following steps: collecting the tea leftovers after soaking for 2 hours, performing filter pressing on the tea leftovers by using a plate-and-frame filter press to obtain percolate, and filtering the tea residue wastewater by using filter paper to obtain tea residue percolate;
the preparation method of the culture medium comprises the following steps:
sterilizing the tea residue percolate in a high temperature sterilization pot at high temperature and high pressure (125 deg.C, 0.15MPa) for 30min to obtain tea residue base solution, and adding glucose, yeast powder and microelements to obtain culture medium.
Example 3
A culture medium comprises the following components in parts by weight: 80 parts of tea leaf residue percolate, 10 parts of glucose, 7 parts of yeast powder and 2 parts of trace elements;
the trace elements comprise 0.8 part of monopotassium phosphate, 0.5 part of zinc sulfate heptahydrate and 0.7 part of ammonium sulfate;
the preparation method of the tea residue percolate comprises the following steps: collecting the tea leftovers after soaking for 2 hours, performing filter pressing on the tea leftovers by using a plate-and-frame filter press to obtain percolate, and filtering the tea residue wastewater by using filter paper to obtain tea residue percolate;
the preparation method of the culture medium comprises the following steps:
sterilizing the tea residue percolate in a high temperature sterilization pot at high temperature and high pressure (125 deg.C, 0.15MPa) for 30min to obtain tea residue base solution, and adding glucose, yeast powder and microelements to obtain culture medium.
Comparative example 1.
A culture medium comprises the following components in parts by weight: 90 parts of deionized water, 4.5 parts of glucose, 4.4 parts of yeast powder and 1 part of trace elements;
the trace elements comprise 0.3 part of monopotassium phosphate, 0.3 part of zinc sulfate heptahydrate and 0.4 part of ammonium sulfate;
the preparation method of the culture medium comprises the following steps:
sterilizing the tea residue percolate at high temperature and high pressure (125 deg.C, 0.15MPa) for 30min to obtain tea residue base solution, and adding glucose, yeast powder and microelements to obtain culture medium.
Example 4
A preparation method of an agricultural microbial agent comprises the following steps:
(1) inoculating Bacillus thuringiensis (ATCC 10792) to NB medium, and shake culturing at 30 deg.C for 24 hr at 150 r/min; the number of viable bacteria was determined by plate count method after culturing (8.8X 10)9CFU/mL)；
(2) Inoculating the bacillus liquid obtained in the step (1) into the culture medium described in the embodiment 1, wherein the inoculation amount is 0.1% (w/w), the bacillus liquid is cultured in a constant temperature shaking table at 30 ℃ for 24 hours, and the rotating speed is set to be 150r/min, so that the agricultural microbial agent (the effective viable bacteria of bacillus thuringiensis is 5.8 multiplied by 10) is obtained9CFU/mL), the growth of Bacillus thuringiensis in the agricultural microbial agent is shown in figure 1.
Example 5
A preparation method of an agricultural microbial agent comprises the following steps:
(1) inoculating Bacillus thuringiensis to NB culture medium, shake culturing at 30 deg.C for 24 hr,the rotating speed is set to 150 r/min; the number of viable bacteria was determined by plate count method after culturing (8.2X 10)9CFU/mL)；
(2) Inoculating the bacillus liquid obtained in the step (1) into the culture medium described in the embodiment 2, wherein the inoculation amount is 0.3% (w/w), the bacillus liquid is cultured in a constant temperature shaking table at 30 ℃ for 24 hours, and the rotating speed is set to 150r/min, so that the agricultural microbial agent (the effective viable bacteria of bacillus thuringiensis is 5.2 multiplied by 10) is obtained9CFU/mL), the growth of Bacillus thuringiensis in the agricultural microbial agent is shown in figure 2.
Example 6
A preparation method of an agricultural microbial agent comprises the following steps:
(1) inoculating bacillus thuringiensis to an NB culture medium, and carrying out shake cultivation at a constant temperature of 30 ℃ for 24h at a rotation speed of 150 r/min; the number of viable bacteria was determined by plate count method after culturing (8.5X 10)9CFU/mL)；
(2) Inoculating the bacillus liquid obtained in the step (1) into the culture medium described in the embodiment 3, wherein the inoculation amount is 1% (w/w), carrying out shake culture at the constant temperature of 30 ℃ for 24 hours, and setting the rotating speed to be 150r/min, so as to obtain the agricultural microbial agent (the effective viable bacteria of bacillus thuringiensis is 4.9 multiplied by 10)9CFU/mL), the growth of Bacillus thuringiensis in the agricultural microbial agent is shown in figure 3.
Comparative example 2
A preparation method of an agricultural microbial agent comprises the following steps:
(1) inoculating bacillus thuringiensis to an NB culture medium, and carrying out shake cultivation at a constant temperature of 30 ℃ for 24h at a rotation speed of 150 r/min; the number of viable bacteria was determined by plate count method after culturing (8.8X 10)9CFU/mL)；
(2) Inoculating the bacillus liquid in the step (1) into the culture medium in the comparative example 1, wherein the inoculation amount is 0.1 percent (w/w), the bacillus liquid is cultured in a constant temperature shaking table at 30 ℃ for 24 hours, and the rotating speed is set to be 150r/min, so that the agricultural microbial agent (the effective viable bacteria of the bacillus thuringiensis are 5.4 multiplied by 10) is obtained9CFU/mL)。
Application examples
Own farm of Guangzhou Probence development agriculture company Limited (Tai located in the area of Guangzhou CityNo. 78 Chengxiang village army in Niuxin Ridge of Flat town) to plant cabbage (variety is '30 day cabbage'), wherein the soil of the farm is humid soil, the soil fertility is middle-grade, and the water source is river water. The experiment was divided into three treatments, treatment one: conventional fertilization + spraying of the agricultural microbial agent of example 4 every 7 days after planting of flowering cabbage (total spraying for 3 times in the whole period), wherein the concentration of the agricultural microbial agent is 1:500(v/v), and each spraying is 150 mL/mu; and (5) processing: conventional fertilization + spraying the agricultural microbial agent of the comparative example 2 every 7 days after planting the flowering cabbage (3 times of full-period total spraying), wherein the concentration of the agricultural microbial agent is 1:500(v/v), and each spraying is 150 mL/mu; and (3) treatment III: only conventional fertilization is carried out, and any microbial inoculum is not sprayed after the flowering cabbage is planted. Three replicates per treatment, randomized block arrangement, cell area 20m2And the flowering cabbage is uniformly planted, cultivated in open field and uniformly managed. The method is characterized in that field investigation is carried out on the cabbage heart in the growing period, main agronomic characters such as plant height, leaf width, leaf length, stem thickness and leaf number of the cabbage heart are regularly investigated at fixed points, and the yield of the cabbage heart is measured after 30 days of planting.
The experiment is carried out on the basis of conventional fertilization, wherein the conventional fertilization comprises the following steps: applying 250kg of commercial organic fertilizer (organic matter is more than or equal to 45 percent, and N + P + K is more than or equal to 5 percent) and 35kg of compound fertilizer (15-15-15) in the Bombay field as base fertilizer per mu; topdressing is carried out for 2 times after planting the flowering cabbage (200 kg/mu of dilute manure water and 8 kg/mu of compound fertilizer (15-15-15) for the Bombay field are respectively sprayed with water fertilizer once).
As can be seen from FIGS. 1 to 3, the flowering cabbage of each treatment group grew well. Further by investigating the influence of different treatments on the main agronomic characters of the flowering cabbage, the results are shown in table 1, and during harvesting, the plant height, the leaf width and the leaf length of the flowering cabbage obtained by spraying the agricultural microbial agent of example 4 are respectively improved by 294.3%, 145.7% and 235.5% compared with those before spraying; the plant height, the leaf width and the leaf length of the flowering cabbage obtained by spraying the agricultural microbial agent in the comparative example 2 are respectively improved by 268.6%, 134.3% and 222.5% compared with those before spraying; compared with the agronomic characters of the heart of the three treatment groups during harvesting, the plant height, the leaf width, the leaf length, the stem thickness and the leaf number of the heart of the first treatment group are respectively improved by 7.0 percent, 4.9 percent, 4.0 percent, 15.4 percent and 6.4 percent compared with the heart of the second treatment group, and are respectively improved by 8.2 percent, 7.5 percent, 5.1 percent, 25.0 percent and 3.8 percent compared with the heart of the third treatment group. It can be seen that spraying the agricultural microbial agent of example 4 3 times during the growth of the flowering cabbage can promote the growth of the flowering cabbage and obviously increase the plant height and stem thickness of the flowering cabbage.
As can be seen from Table 2, the average yield of flowering cabbage obtained by spraying the agricultural microbial preparation of example 4 was 61.2kg/20m2The yield per mu is 2040.0kg, which is 136.7kg more increased than that of the agricultural microbial agent sprayed in the comparative example 2, the yield is 7.2 percent, and the yield is increased obviously; compared with the method that other microbial agents are not additionally sprayed in the growth process of the flowering cabbage (treatment III), the yield of the agricultural microbial agent in the spraying embodiment 4 is improved by 163.3kg, the yield is increased by 8.7%, and the yield is remarkably increased. Compared with the second treatment and the third treatment, spraying the agricultural microbial agent containing the bacillus thuringiensis can improve the yield of the flowering cabbage by 26.6kg, and the yield increase by 1.4%. From the above results, it can be seen that spraying the agricultural microbial agent of example 4 3 times in total during the growth of the flowering cabbage can effectively improve the yield of the flowering cabbage, i.e., the microbial agent prepared from bacillus thuringiensis and tea leaf residue leachate has a better effect in improving the yield of the flowering cabbage.
TABLE 1 Effect of different treatments on the major agronomic traits of flowering cabbage
TABLE 2 Effect of different treatments on the yield of flowering cabbage
The embodiments of the present invention have been described in detail with reference to the drawings, but the present invention is not limited to the embodiments, and various changes can be made within the knowledge of those skilled in the art without departing from the gist of the present invention. Furthermore, the embodiments of the present invention and the features of the embodiments may be combined with each other without conflict.
Claims (10)
1. A culture medium comprising the following components: the tea leaf residue leachate, a carbon source, a nitrogen source and trace elements, and the preparation method of the tea leaf residue leachate comprises the following steps: and (4) carrying out filter pressing on the tea residues, collecting the percolate, and filtering to obtain the tea residue percolate.
2. The culture medium according to claim 1, comprising the following components in parts by weight: 70-95 parts of tea leaf residue leachate, 1-15 parts of carbon source, 1-9 parts of nitrogen source and 0.1-5 parts of trace elements.
3. The culture medium according to claim 2, comprising the following components in parts by weight: 80-93 parts of tea leaf residue leachate, 2-10 parts of carbon source, 3-7 parts of nitrogen source and 0.1-2 parts of trace elements.
4. A medium according to any one of claims 1 to 3, wherein the trace elements include potassium dihydrogen phosphate, zinc sulphate and ammonium sulphate.
5. Use of the medium according to any one of claims 1 to 4 in the cultivation of Bacillus thuringiensis.
6. A preparation method of an agricultural microbial agent comprises the following steps: inoculating bacillus thuringiensis into the culture medium of any one of claims 1-4, and culturing to obtain the agricultural microbial agent.
7. The method according to claim 6, wherein the amount of Bacillus thuringiensis inoculated is 0.1 to 2 w/w%; the culture conditions are preferably 25-37 ℃ and 100-200 r/min for 20-24 h.
8. An agricultural microbial agent prepared by the preparation method of claim 6 or 7, wherein the viable count of bacillus thuringiensis in the agricultural microbial agent is (3-7) x 109CFU/mL。
9. The method of claim 6 or 7 or the use of the agricultural microbial inoculant of claim 8 in the preparation of a fertilizer.
10. The use of the agricultural microbial inoculant of claim 8 for promoting the growth of crops.
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