CN111925971A - Environment-responsive bacillus emulsion and preparation method thereof - Google Patents

Abstract

The invention relates to an environment-responsive bacillus emulsion, belonging to the technical field of agricultural microbial agent development. The emulsion comprises an inner oil phase and an outer water phase, wherein the oil phase is added with bacterial powder, grease and surfactant to form the emulsion type inner oil phase, the outer water phase is added with a polysaccharide compound formed by porous starch and chitosan to form an electrostatic response type emulsifier, and the oil phase and the water phase are mixed to form a gel type emulsion. In the land with higher salinization degree, the polysaccharide compound in the emulsion is rapidly disintegrated due to overhigh pH value or electrostatic shielding effect of salt ions, and the microporous structure of the porous starch can increase the contact between the external environment and the oil phase and improve the sensitivity of the emulsion. The dissipated oil drops are in a small drop state due to the oil phase emulsifier contained in the oil drops, so that the release of the embedded strains is further promoted, and the soil environment is improved in a short time. In meta-acid or neutral soil, the emulsion can effectively slow down the release speed of strains, prolong the action time of the strains and improve the utilization efficiency and time of agricultural microorganisms.

Description

Environment-responsive bacillus emulsion and preparation method thereof

The technical field is as follows:

the invention relates to an environment-responsive bacillus emulsion and a preparation method thereof, belonging to the technical field of agricultural microbial agent development.

Background art:

the bacillus subtilis is an aerobic gram-positive bacterium, has stronger stress resistance compared with general strains, can resist heat and drought, and related documents also prove that the bacillus subtilis can effectively prevent and treat diseases caused by plant fungi when being applied to an agricultural field. Bacillus subtilis is a typical widely accepted bacterium for promoting and maintaining the normal growth of plants, widely distributed in all natural environments, in soil, on crop surfaces and on animal surfaces, and even in parts of the interior of plants.

In recent years, due to the abuse of artificially synthesized fertilizers and pesticides, partial agricultural land in China has the problems of over-high content of acid, alkali or salt ions in soil and the like. The physical and chemical properties of the soil have great influence on the growth of crops and the fertilizer efficiency of the soil. For example, salinized farmlands are not suitable for normal growth of crops, and a large amount of free hydrogen ions exist in acid soil, which can cause loss of fertilizer and strain activity, and greatly influence the utilization rate of the used agricultural microbial agent.

In the existing bacterial manure technology, the treatment of bacillus subtilis is mainly focused on spray drying, fluidized bed granulation and the like, and even a mode of directly diluting a strain stock solution and applying the diluted strain stock solution to a farmland is adopted. The agricultural microbial agent produced by the processes can only play a role in a short time in farmlands, and as most strains are inactivated or lost in severe conditions and the action time is relatively short, the agricultural microbial agent cannot effectively improve the growth conditions of crops, has an antagonistic effect with pathogenic bacteria and inhibits the amplification and the propagation of the pathogenic bacteria.

The invention content is as follows:

the invention aims to overcome the defects of the prior art and provides an environment-responsive bacillus emulsion, and the emulsion stabilized by a porous starch/chitosan electrostatic compound provided by the invention can adjust the number of released viable bacteria according to the pH value of the soil environment. In agricultural land with high salinization degree, the polysaccharide complex in the emulsion is rapidly disintegrated due to overhigh pH value or electrostatic shielding effect of salt ions, so that the emulsion is rapidly broken and the oil phase is released. The porous structure of the porous starch can further accelerate the contact between the external environment and the oil phase, and increase the sensitivity of the emulsion to the external environment, thereby releasing the thalli in a short time and improving the soil environment. Meanwhile, oil drops in the soil can concentrate on the roots of the plants and release thalli in a concentrated manner along with the absorption effect of the roots of the plants on water, so that the loss of strains is reduced, and the pesticide effect is enhanced. In meta-acid or neutral soil, the emulsion can effectively slow down the release speed of the strains, prolong the action duration of the strains and improve the utilization efficiency and the action time of the agricultural microbial agent.

In order to achieve the purpose, the invention provides an environment-responsive bacillus emulsion, which comprises an inner oil phase and an outer water phase, wherein bacterial powder, grease and a surfactant are added into the oil phase to form an emulsion-type inner oil phase, a polysaccharide compound formed by porous starch and chitosan is added into the outer water phase to be dissolved in a polyethylene glycol aqueous solution to form an electrostatic responsive emulsifier, the oil phase and the water phase are dispersed or uniformly mixed to form a gel-type emulsion, and the porous starch adsorbed on the interface of the emulsion can realize the responsive contact of the emulsion-type oil phase and the environment outside a saline-alkali soil;

further, the bacterial powder can be at least one of bacillus subtilis bacterial powder, bacillus licheniformis bacterial powder, bacillus thuringiensis bacterial powder or bacillus amyloliquefaciens bacterial powder;

furthermore, the addition amount of the bacterial powder accounts for 20-30% of the mass of the oil phase;

furthermore, the addition amount of the surfactant accounts for 1-2% of the mass of the oil phase;

furthermore, the adding amount of the porous starch is 2-7% of the mass of the external water phase;

the diameter of the small hole of the porous starch is 0.5-1.5 μm, preferably 1 μm;

the porous starch can be rice porous starch, corn porous starch, cassava porous starch, potato porous starch or wheat porous starch and the like;

preferably, the porous starch is corn porous starch;

further, the adding amount of the chitosan is 1 to 2 percent of the mass of the external water phase;

further, the solution in the water phase is 40-60% (m: m) of polyethylene glycol aqueous solution, and the electrostatic response type external water phase is obtained after a polysaccharide compound formed by porous starch and chitosan is dissolved;

preferably, the solution in the aqueous phase is a 50% aqueous solution of polyethylene glycol.

The preparation method of the environment-responsive bacillus emulsion comprises the following specific steps:

(1) preparation of oil phase: adding the bacterial powder and the grease in proportion, stirring at room temperature for 25-30min, and then adding the surfactant in proportion to obtain an oil phase;

further, the oil and fat is corn oil, rapeseed oil, palm oil or soybean oil and the like;

further, the surfactant is an anionic emulsifier or a nonionic emulsifier;

preferably, the surfactant is a mixture of alkylphenol polyoxyethylene, fatty alcohol polyoxyethylene and sodium dodecyl sulfate, and the mass ratio is 2:2: 1;

(2) preparation of the aqueous phase: mixing the following components in a mass ratio of 2-7: 1-2, adding porous starch and chitosan, dissolving in 40-60% polyethylene glycol solution according to a material-liquid ratio of 6-9% (m: v), adding acetic acid accounting for 1-2% (v: v) of the water phase, stirring for 20-30min, and then adjusting pH to 6.5-6.0 with alkali liquor;

preferably, the alkali liquor is 0.5mol/L sodium hydroxide aqueous solution;

(3) preparing an emulsion: mixing the oil phase and the water phase, wherein the inner oil phase accounts for 75-90% (m: m) of the emulsion, emulsifying, and packaging to obtain the product;

preferably, the inner oil phase is 90% by weight.

The invention also provides application of the bacillus emulsion in the field of agricultural microbial agents.

Has the advantages that:

1. the invention adopts pH and salt ion response type polysaccharide electrostatic compound formed by electrostatic complexation of porous starch/chitosan as an emulsifier for the first time, and the compound can be quickly adsorbed on the interface of oil drops in the shearing process of the oil drops and can be simultaneously adsorbed on the interfaces of two or more oil drops to form a stable emulsion oil drop network, so that the emulsion is changed into gel from fluid state (shown in figure 1). The contact of the strains with oxygen is reduced, the strains are prevented from recovering in advance, the physiological activity of the strains is protected to the maximum extent in the storage and transportation processes, and the bacterial activity of the strains is ensured;

2. the bacillus agent with acid-base salt sensitivity is obtained through the difference of charges of the emulsifier in different acid-base environments, and the bacillus agent can adjust the release rate of a strain according to the acid-base degree and the salt ion content of a soil environment: in an acidic or neutral environment, the emulsion exists in the form of emulsion gel with anti-dilution capacity, so that the release rate of the strain is reduced; in an alkaline environment, the charge of the chitosan is extremely low, so that the electrostatic interaction with the porous starch is reduced, and the embedded strains are rapidly released; in an environment with higher salt ion concentration, the salt ions shield the charges of the emulsifier, reduce the electrostatic effect and increase the release of strains;

3. in the release process of the emulsion, the microporous structure of the porous starch can increase the contact between the external environment and the oil phase, and improve the environmental sensitivity of the emulsion. Meanwhile, the oil phase continues to use and optimizes oil drop stabilizing process and components, on the premise of reducing environmental pollution, the oil drops can be combined with water in soil quickly from micron-sized large oil drops to nanometer-sized small oil drops after being released from an emulsion system and maintain the state of small liquid drops, when the grain size of the oil drops is smaller than the size of the strain, the strain can be leaked from the oil drops quickly, and the technology and the mechanism can accelerate the release of internal strains;

4. the external water phase adopts 40-60% polyethylene glycol aqueous solution as solution, and can construct micron-sized emulsion with the internal phase of the missible oil type to successfully construct the internal environment of the oil phase, load the strain and improve the stress resistance and storage stability of the strain;

5. the water content of the invention is relatively low in the emulsion, the emulsion is pasty, the harm of dust flying to users is reduced compared with common powdery products, and the storage and the transportation are easier compared with liquid products;

6. the bacillus emulsion provided by the invention is simple in preparation process, environment-friendly and suitable for industrial mass production and application.

Description of the drawings:

FIG. 1 is a schematic diagram of the product structure;

FIG. 2 is a diagram of a field trial;

FIG. 3 shows the microstructure under a cryo-scanning electron microscope;

FIG. 4 is a graph of storage modulus and loss modulus;

FIG. 5 is a schematic diagram of product release;

FIG. 6 product instability coefficient.

The specific implementation scheme is as follows:

in order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the present patent and are not intended to limit the present invention.

The structure of the environment-responsive microbial inoculum provided by the invention is shown in fig. 1, and the invention will be further explained with reference to specific embodiments.

Example 1: environment-responsive bacillus subtilis emulsion

(1) Preparation of oil phase: stirring 150g of bacillus subtilis powder and 450g of rapeseed oil at room temperature for 30min, and then adding 10g of a mixture (mass ratio is 2:2: 1) of surfactant alkylphenol ethoxylate, fatty alcohol polyoxyethylene ether and lauryl sodium sulfate to obtain an oil phase;

(2) preparation of the aqueous phase: dissolving 2g of corn porous starch (the diameter of a pore of the porous starch is 0.5-1.5 mu m) and 1g of chitosan in 50ml of 50% polyethylene glycol solution (polyethylene glycol 400 is selected in the embodiment of the invention), adding 1ml of acetic acid, stirring for 30min, and adjusting the pH to 6.5 by using 0.5mol/L sodium hydroxide;

(3) preparing an emulsion: mixing the oil phase and the water phase, wherein the internal oil phase accounts for 90%, adding the mixture into a dispersion machine, carrying out 12000-15000 rotation, carrying out dispersion for 3-5 minutes for emulsification, and packaging the emulsified mixture to obtain the product.

Example 2: environment-responsive bacillus subtilis emulsion

(1) Preparation of oil phase: stirring 150g of bacillus subtilis powder and 450g of rapeseed oil at room temperature for 30min, and then adding 10g of a mixture (mass ratio is 2:2: 1) of surfactant alkylphenol ethoxylate, fatty alcohol polyoxyethylene ether and lauryl sodium sulfate to obtain an oil phase;

(2) preparation of the aqueous phase: dissolving 2g corn porous starch (porous starch pore diameter 0.5-1.5 μm) in 50ml 50% polyethylene glycol solution, adding 1ml acetic acid, and adjusting pH to 6.5 with 0.5mol/l sodium hydroxide;

(3) preparing an emulsion: mixing the oil phase and the water phase, wherein the internal oil phase accounts for 90%, adding the mixture into a dispersion machine, carrying out 12000-15000 rotation, carrying out dispersion for 3-5 minutes for emulsification, and packaging the emulsified mixture to obtain the product.

Example 3: environment-responsive bacillus subtilis emulsion

(1) Preparation of oil phase: stirring 150g of bacillus subtilis powder and 450g of rapeseed oil for 30min at room temperature to obtain an oil phase; (2) preparation of the aqueous phase: dissolving 2g corn porous starch (with pore diameter of 0.5-1.5 μm) and 1g chitosan in 50ml 50% polyethylene glycol solution, adding 1ml acetic acid, stirring for 30min, and adjusting pH to 6.5 with 0.5mol/l sodium hydroxide;

(3) preparing an emulsion: mixing the oil phase and the water phase, wherein the internal oil phase accounts for 90%, adding the mixture into a dispersion machine, carrying out 12000-15000 rotation, carrying out dispersion for 3-5 minutes for emulsification, and packaging the emulsified mixture to obtain the product.

Case effect detection:

(1) detecting the release amount of viable bacteria under different pH environments:

15g of samples prepared by the three schemes of the embodiment 1 to 3 are respectively taken, evenly divided into three parts, respectively placed in 100ml of sterile water with pH values of 5, 7 and 9 for 1 day, and then the number of strains in the sterile water under different pH value environments is detected.

The detection results are shown in table 1, the release speed of the live bacteria wrapped by the environment-responsive bacillus subtilis emulsion prepared by the method in embodiment 1 can be adjusted according to different pH values, when the pH value is alkaline, the release amount of the live bacteria is obviously higher than that of the live bacteria in other environments, and in an acidic environment, the release of the strains is relatively minimum. The result shows that the environmental response type bacillus subtilis emulsion provided by the invention can adjust the release amount of the live bacteria wrapped by the bacillus subtilis emulsion according to the pH value of the environment so as to prolong the fertilizer effect and improve the saline-alkali soil environment.

(2) Detecting the release amount of the living bacteria under the salt ion environment:

15g of each sample prepared by the three schemes of the examples 1 to 3 are equally divided into three parts, the three parts are respectively placed in 100ml of sterile water with the salt ion concentration of 0 to 120mM for 1 day, and then the number of the strains in the sterile water under the environment with different salt ion concentrations is detected.

The detection results are shown in table 2, the release rate of the live bacteria wrapped by the environmental response type bacillus subtilis emulsion prepared by the method in embodiment 1 can be adjusted according to different salt ion concentrations, when the salt ion concentration is low, the release amount of the live bacteria is obviously lower than that of other embodiments, and the release of the strains is obviously increased in the environment with high salt ion concentration. The result shows that the environmental response type bacillus subtilis emulsion provided by the invention can adjust the release amount of live bacteria wrapped by the emulsion according to the salt ion concentration of the environment, thereby prolonging the fertilizer effect and improving the saline-alkali soil environment.

(3) The effect of storage time on strain activity was examined:

the samples prepared by the three schemes of examples 1-3 are stored in a sealed mode at room temperature for 0-180 days, 15g of the samples are diluted in 100g of physiological saline after storage, 0.5 wt% of Tween 80 is added, and the samples are placed in a conical flask and shaken for 30min-1h in a shaking table (200rpm) by using glass beads (flatly paved at the bottom of the flask). Then diluted with physiological saline according to the activity and number of the strain, the diluted solution is spread on an LB culture medium, and cultured in an incubator at 37 ℃ for 12-16h, and counted. The results, as shown in Table 3, show that the inactivation of the bacterial species during storage of the product prepared in example 1 is significantly lower than that of the other samples, indicating that the bacterial powder prepared by the method can better protect the bacterial cells during storage.

(4) Field test

And (3) carrying out a fertilizer efficiency experiment on celery seedlings (with the plant height of 15 cm), and carrying out data acquisition and analysis on an experiment sample. The experiment was carried out in a sunlight greenhouse, seeding was carried out 7, 15 days in 2019, transplanting was carried out 8 days in 9 months, and prepared samples were added into soil pits before transplanting, about 5g per plant. Then carrying out conventional crop irrigation fertilization culture, and collecting and processing experimental results. The results are shown in table 4, and it is obvious that the example 1 can promote the growth of celery, the yield is relatively increased, fig. 2 shows that the plant height is obviously higher than that of other experimental groups, the root system is more developed, and the product release principle is shown in fig. 5.

TABLE 1

TABLE 2

TABLE 3

TABLE 4

Example 4 physical Properties

(1) Micro and macro structure

2g of each of the samples prepared in the three protocols of examples 1 to 3 was taken, and the samples were placed on a flat plate and covered with a film. Rapid freezing was performed with liquid nitrogen, followed by observation under a scanning electron microscope and image recording.

The detection results are shown in fig. 3, and the samples prepared according to the methods of embodiments 1 and 3 have relatively uniform oil droplet size, no obvious oil droplet aggregation phenomenon, and a polysaccharide network structure is formed in the water phase, and the oil droplets are embedded therein to form a gel structure. In comparison, the sample prepared in example 2 has uneven oil drop size, obvious large oil drops and no obvious network structure, and indicates that the emulsion structure is unstable.

(2) Rheological Properties

Samples prepared by the three schemes of the embodiment 1 to 3 are respectively taken for 5g, the samples are placed on a rheometer to be subjected to frequency scanning, the storage modulus and the loss modulus of the samples are obtained, the frequency range is 0.1 to 100rad/s, and the testing temperature is constant at 25 ℃. As shown in FIG. 4, the samples prepared by the methods described in examples 1 and 3 have significantly higher storage modulus (G ') and loss modulus (G') than other samples because the gel network formed therein imparts stronger gel characteristics.

EXAMPLE 5 emulsion instability coefficient determination

The emulsion was prepared in the same manner as in example 1, except that the aqueous solution used was replaced, as follows:

group A: 30% polyethylene glycol aqueous solution; group B: 50% aqueous polyethylene glycol solution; group C: 70% polyethylene glycol aqueous solution; group D: glycerol; group E: propylene glycol;

the stability measurements were performed on 5 groups of emulsion samples obtained from the preparation using a LUMiSizer. After 0.4 g of sample is added into the sample cell, centrifugation is carried out for 30 minutes at 3000 r/min, near infrared light is used for irradiation in the process, the light intensity is evaluated to calculate the instability coefficient of the emulsion, and the obtained instability index is analyzed and compared.

The results are shown in FIG. 6, and the instability index of the emulsion with 50% polyethylene glycol aqueous solution as the external aqueous phase solution is significantly lower than that of other proportions of polyethylene glycol solutions, as well as glycerol and propylene glycol, indicating that the emulsion is preferred to have 50% polyethylene glycol as the external aqueous phase when the emulsion-based oil phase is the internal phase during production and preparation. This result is simultaneously reflected in the stability of the emulsion during the storage and transport phases.

Example 6: environment-responsive bacillus emulsion

(1) Preparation of oil phase: stirring 150g of bacillus licheniformis powder and 340g of rapeseed oil for 25min at room temperature, and then adding 10g of surfactant calcium dodecyl sulfonate to obtain an oil phase;

(2) preparation of the aqueous phase: dissolving 3g rice porous starch (porous starch pore diameter 0.5-1.5 μm) and 1g chitosan in 50ml 40% polyethylene glycol solution, adding 1ml acetic acid, stirring for 20min, and adjusting pH to 6.5 with 0.5mol/L sodium hydroxide;

(3) preparing an emulsion: mixing the oil phase and the water phase, adding the oil phase accounting for 88 percent into a dispersion machine, carrying out 12000-15000 rotation, carrying out emulsification after dispersion for 3-5 minutes, and packaging after emulsification to obtain the product.

Example 7: environment-responsive bacillus emulsion

(1) Preparation of oil phase: stirring 122.5g of bacillus thuringiensis powder and 350g of soybean oil for 25min at room temperature, and then adding 7.5g (mass ratio is 2:2: 1) of a mixture of surfactant alkylphenol ethoxylate, fatty alcohol-polyoxyethylene ether and lauryl sodium sulfate to obtain an oil phase;

(2) preparation of the aqueous phase: dissolving 3g porous potato starch (with pore diameter of 0.5-1.5 μm) and 1g chitosan in 50ml 60% polyethylene glycol solution, adding 1ml acetic acid, stirring for 20min, and adjusting pH to 6.5 with 0.5mol/L sodium hydroxide;

(3) preparing an emulsion: mixing the oil phase and the water phase, adding the oil phase accounting for 87.5 percent into a dispersion machine, carrying out 12000-15000 turns, carrying out dispersion for 3-5 minutes for emulsification, and packaging after emulsification to obtain the product.

The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the patent. It should be noted that, for those skilled in the art, various changes, combinations and improvements can be made in the above embodiments without departing from the patent concept, and all of them belong to the protection scope of the patent. Therefore, the protection scope of this patent shall be subject to the claims.

Claims (10)

1. An environment response type bacillus emulsion is characterized in that the emulsion comprises an inner oil phase and an outer water phase, wherein bacterial powder, grease and a surfactant are added into the oil phase to form an emulsion type inner oil phase, a polysaccharide compound formed by porous starch and chitosan is added into the outer water phase and dissolved in a polyethylene glycol aqueous solution to form an electrostatic response type emulsifier, and the oil phase and the water phase are dispersed or uniformly mixed to form a gel type emulsion.

2. The environment-responsive bacillus emulsion as recited in claim 1, wherein in the oil phase, the addition amount of the bacterial powder accounts for 20-30% of the mass of the oil phase, and the addition amount of the surfactant accounts for 1-2% of the mass of the oil phase;

in the water phase, the adding amount of the porous starch is 2-7% of the mass of the external water phase, and the adding amount of the chitosan is 1-2% of the mass of the external water phase.

3. The environmentally responsive bacillus emulsion of claim 1, wherein the oil is corn oil, rapeseed oil, palm oil, or soybean oil; the surfactant is an anionic emulsifier or a nonionic emulsifier.

4. The environmentally responsive bacillus emulsion of claim 1, wherein the aqueous solution of polyethylene glycol is 40-60% by mass.

5. The environmentally responsive bacillus emulsion of claim 1, wherein the porous starch is rice porous starch, corn porous starch, tapioca porous starch, potato porous starch, or wheat porous starch; the diameter of the small hole of the porous starch is 0.5-1.5 μm.

6. The environmentally responsive bacillus emulsion of claim 1, wherein the bacterial powder is at least one of bacillus subtilis bacterial powder, bacillus licheniformis bacterial powder, bacillus thuringiensis bacterial powder, or bacillus amyloliquefaciens bacterial powder.

7. The environmentally responsive bacillus emulsion of any one of claims 1-6, prepared by a process comprising:

(1) preparation of oil phase: adding the bacterial powder and the grease in proportion, stirring at room temperature for 25-30min, and then adding the surfactant in proportion to obtain an oil phase;

(2) preparation of the aqueous phase: mixing the following components in a mass ratio of 2-7: 1-2, adding porous starch and chitosan, dissolving in 40-60% polyethylene glycol solution according to the material-liquid ratio of 6-9%, adding acetic acid accounting for 1-2% of the volume ratio of the water phase, stirring for 20-30min, and then adjusting the pH to 6.5-6.0 with alkali liquor;

(3) preparing an emulsion: mixing the oil phase and the water phase, wherein the oil phase accounts for 75-90% of the weight of the emulsion, emulsifying and packaging to obtain the product.

8. The environmentally responsive bacillus emulsion of claim 7, prepared by the following method:

(1) preparation of oil phase: adding the bacterial powder and the grease in proportion, stirring at room temperature for 25-30min, and then adding the surfactant in proportion to obtain an oil phase;

the surfactant is a mixture of alkylphenol polyoxyethylene, fatty alcohol polyoxyethylene ether and lauryl sodium sulfate, and the mass ratio is 2:2: 1;

(2) preparation of the aqueous phase: mixing the following components in a mass ratio of 2-7: 1-2, adding corn porous starch and chitosan, dissolving the corn porous starch and the chitosan into 50% polyethylene glycol solution according to the material-liquid ratio of 6-9%, adding acetic acid accounting for 1-2% of the volume ratio of the water phase, stirring for 20-30min, and then adjusting the pH value to 6.5-6.0 by using 0.5mol/L sodium hydroxide solution;

(3) preparing an emulsion: mixing the oil phase and the water phase, wherein the internal oil phase accounts for 90%, emulsifying and packaging to obtain the product.

9. The environmentally responsive bacillus emulsion of claim 7, prepared by the following method:

(1) preparation of oil phase: stirring 150g of bacillus subtilis powder and 450g of rapeseed oil at room temperature for 30min, and then adding a mixture of bacillus subtilis powder and rapeseed oil in a mass ratio of 2:2:1, 10g of a mixture of surfactants of alkylphenol polyoxyethylene ether, fatty alcohol polyoxyethylene ether and lauryl sodium sulfate to obtain an oil phase;

(2) preparation of the aqueous phase: dissolving 2g of corn porous starch and 1g of chitosan in 50ml of 50% polyethylene glycol solution, adding 1ml of acetic acid, stirring for 30min, and adjusting the pH to 6.5 by using 0.5mol/L sodium hydroxide;

(3) preparing an emulsion: mixing the oil phase and the water phase, wherein the internal oil phase accounts for 90%, adding the mixture into a dispersion machine, carrying out 12000-15000 rotation, carrying out dispersion for 3-5 minutes for emulsification, and packaging the emulsified mixture to obtain the product.

10. Use of an environmentally responsive bacillus emulsion according to any of claims 1-9 in the field of agricultural microbial agents.

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