CN113151088A

CN113151088A - Multi-bacterium fusion symbiotic fermentation process for resisting diseases and promoting growth

Info

Publication number: CN113151088A
Application number: CN202110448885.1A
Authority: CN
Inventors: 杨建国; 赵猛; 屈峰
Original assignee: Shandong Lihaoyuan Bioengineering Co ltd
Current assignee: Li Haoyuan Bioengineering (Shandong) Co.,Ltd.
Priority date: 2021-04-25
Filing date: 2021-04-25
Publication date: 2021-07-23
Anticipated expiration: 2041-04-25
Also published as: CN113151088B

Abstract

The application relates to the field of microbial fermentation, in particular to a multi-bacterium fusion symbiotic fermentation process for resisting diseases and promoting growth, which comprises the following steps: step (1), preparing bacterial liquid: respectively preparing paenibacillus polymyxa liquid, yeast liquid and streptococcus thermophilus liquid; step (2), fusing symbiotic fermentation: adding a fermentation medium into the fermentation equipment, controlling the pH value and the temperature of the fermentation medium, adding paenibacillus polymyxa liquid while stirring the fermentation medium, and continuing stirring and fermenting; then controlling the pH value and the temperature of the system, adding yeast liquid while stirring, and continuously stirring and fermenting; and then controlling the pH value and the temperature of the system, adding the streptococcus thermophilus liquid while stirring, and continuously stirring and fermenting to obtain the multi-bacterium fusion symbiotic fermentation liquid. The multi-bacterium fusion symbiotic fermentation process has the advantages that the growth-promoting rhizosphere probiotics and the disease-resistant rhizosphere probiotics are mixed and fermented, the production cost of the microbial inoculum is reduced, the microbial community is various, the environmental adaptability is high, and the application range is wide.

Description

Multi-bacterium fusion symbiotic fermentation process for resisting diseases and promoting growth

Technical Field

The application relates to the field of microbial fermentation, in particular to a multi-bacterium fusion symbiotic fermentation process for resisting diseases and promoting growth.

Background

In a natural ecosystem, different microorganisms are mutually restricted and balanced in terms of living space and nutrient requirements. In the soil environment, the manual fertilization and pesticide intervention destroys the original microorganism types and quantity, breaks the balance, increases the pathogenic bacteria biomass of crops, causes the occurrence of soil-borne diseases and bacterial fungal diseases, and further influences the growth, yield and quality of plants.

In the prior art, beneficial microorganisms at plant rhizosphere are artificially added, a community system of the microorganisms is changed, pathogenic bacteria are inhibited, and the plant growth promoting and stress resisting effects are achieved.

At present, batch fermentation or fed-batch fermentation is adopted in the conventional fermentation process of beneficial microorganisms of plant rhizosphere, strains in the microbial agent prepared by the process are single, the single strain is weak in environmental adaptability due to single metabolic type, single respiratory type and single action function, and is difficult to form dominant flora, and the strain is usually required to be reasonably compounded when used as the microbial agent in the later stage.

Aiming at the related technologies, the inventor considers that the strains prepared by batch fermentation or fed-batch fermentation at present are single, and the strains are required to be added in a compounding manner when being used as a microbial agent in the later period, so that the fermentation process has high cost and complex use procedures.

Disclosure of Invention

In order to solve the problems of single bacterial colony and high process cost of the existing fermentation process, the application provides a multi-bacterium fusion symbiotic fermentation process for resisting diseases and promoting growth.

The application provides a multi-bacterium fusion symbiotic fermentation process for disease resistance and growth promotion, which adopts the following technical scheme: a multi-bacterium fusion symbiotic fermentation process for resisting diseases and promoting growth comprises the following steps:

step (1), preparing bacterial liquid: inoculating the activated paenibacillus polymyxa into the culture medium A, shaking up, and culturing to obtain paenibacillus polymyxa liquid; inoculating the activated yeast into a culture medium B, shaking up, and culturing to obtain a yeast liquid; inoculating the activated streptococcus thermophilus into a culture medium C, shaking up, and culturing to obtain streptococcus thermophilus liquid;

step (2), fusing symbiotic fermentation: adding a fermentation medium into a fermentation device, controlling the pH value of the fermentation medium to be 7.0-7.2 and the temperature to be 35-37 ℃, adding a Paenibacillus polymyxa liquid while stirring the fermentation medium, continuously stirring, and fermenting for 24-36 hours to obtain a fermentation liquid A; then controlling the pH value of the system to be 6.0-6.5 and the temperature to be 35-37 ℃, adding yeast liquid while stirring the fermentation liquid A, continuously stirring, and fermenting for 45-50h to obtain fermentation liquid B; and controlling the pH value of the system to be 5.0-5.5 and the temperature to be 30-32 ℃, adding the streptococcus thermophilus liquid while stirring the fermentation liquid B, continuously stirring, and fermenting for 3-4 days to obtain the multi-bacterium fusion symbiotic fermentation liquid.

This application adopts above-mentioned many fungus to fuse symbiotic fermentation process, the mixed fermentation of rhizosphere probiotic and the disease-resistant rhizosphere probiotic of promoting growth, disposable input, disposable output, change traditional batch fermentation process, reduce the manufacturing cost of microbial inoculum, and the fungus crowd that mixed production made is various, it is strong to environmental suitability, the problem that traditional batch fermentation process bacterial strain and effect are comparatively single has been solved, the range of application is extensive, shorten the process when using as the microbial inoculum, play disease-resistant effect of promoting growth to the plant.

The paenibacillus polymyxa exists in a spore form, has good temperature resistance, stable quality, durable pesticide effect and long pesticide and fertilizer effect, is an ideal medicament for preventing and treating soil-borne diseases such as bacterial wilt, fusarium wilt, root rot, soft rot and the like, has obvious effects of promoting growth, improving yield and improving quality, has no pesticide injury and residue, and is a preferred microbial preparation for producing pollution-free, green and organic vegetables.

The saccharomycete and the streptococcus thermophilus can promote crop growth, secrete metabolic active matter, stimulate crop growth and development, regulate crop balance, absorb nutrients, induce the expression of resistance gene, raise the disease and stress resisting capacity of crop, promote the synthesis of effective matter, improve the quality of agricultural product and other functions.

Through the multi-bacterium fusion symbiotic fermentation process, paenibacillus polymyxa, saccharomycetes and streptococcus thermophilus are mixed and fermented, and system condition parameters before each colony is added are controlled, so that the balance of soil flora can be improved by the prepared mixed fermentation microbial inoculum, and three strains are symbiotically fermented through co-culture, so that a feasible fermentation culture process is provided for large-scale production of composite strains, the synergistic effect of each strain is improved, the plant disease and insect pest resistance effect is improved, the root growth can be promoted, and the crop stress resistance is improved. And finally, performing mixed fermentation on the multiple strains to obtain a multi-strain fusion symbiotic fermentation liquid, wherein the viable count of paenibacillus polymyxa is more than 20 hundred million/mL, the viable count of yeast liquid is more than 20 hundred million/mL, and the viable count of streptococcus thermophilus liquid is more than 20 hundred million/mL, stopping the tank, and stopping fermentation.

Preferably, in the step (1), the culture medium A is prepared by the following steps: weighing 1-5g of beef extract, 8-12g of peptone, 3-8g of sodium chloride and 15-25g of agar, adding into distilled water, uniformly mixing, supplementing the distilled water to 1000mL, adjusting the pH value to 7.0-7.3, and then sterilizing at the temperature of 118-.

The culture medium A is prepared by mixing the materials, culture nutrients are provided for the Paenibacillus polymyxa, the Paenibacillus polymyxa is inoculated to the culture medium A and cultured in a shaking flask, the OD600 of the growth curve is detected every two hours after inoculation, the growth curve is drawn, the middle period or the later period of logarithmic phase of growth is reached, and the Paenibacillus polymyxa liquid reaches the standard of inoculation to a fermentation tank. And before inoculating the paenibacillus polymyxa, the culture medium A is sterilized at high temperature and under pressure, so that the influence of other strains or microorganisms on the cultivation of the paenibacillus polymyxa is reduced, and the cleanness and the singleness of the cultured strains are ensured.

Preferably, in the step (1), the culture medium B is prepared by the following steps: weighing 180-220g of potato filtrate, 18-22g of glucose and 15-25g of agar, adding into distilled water, mixing uniformly, supplementing the distilled water to a system of 1000mL, and sterilizing at the temperature of 118-123 ℃ and the pressure of 0.08-0.12MPa for 28-32min to obtain the culture medium B.

The culture medium B is prepared by mixing the materials, culture nutrients are provided for the saccharomycetes, after the saccharomycetes are inoculated to the culture medium B, shake-flask culture is carried out, the OD600 of the growth curve is detected every two hours after inoculation, the growth curve is drawn, and the middle period or the later period of logarithmic phase growth is reached, namely the saccharomycetes liquid reaches the standard of being inoculated to a fermentation tank. And before the culture medium B is inoculated with the saccharomycetes, the culture medium B is sterilized at high temperature and under pressure, so that the influence of other strains or microorganisms on the culture of the saccharomycetes is reduced, and the cleanness and the singleness of the cultured strains are ensured.

Wherein the potato filtrate is obtained by steaming potato and filtering.

Preferably, in the step (1), the culture medium C is prepared by the following steps: weighing 8-12g of peptone, 8-12g of beef extract, 3-8g of yeast extract, 18-22g of glucose, 3-8g of sodium acetate, 1-3g of citric diamine, 0.8-1.2g of tween-80, 1.5-2.5g of dipotassium hydrogen phosphate, 0.5-0.6g of magnesium sulfate, 0.20-0.30g of manganese sulfate, 18-22g of calcium carbonate and 15-25g of agar, adding into distilled water, uniformly mixing, supplementing distilled water to a system volume of 1000mL, and sterilizing at a temperature of 118-.

The culture medium C is prepared by mixing the materials, culture nutrients are provided for the streptococcus thermophilus, the streptococcus thermophilus is inoculated to the culture medium C, shake flask culture is carried out, the OD600 of the growth curve is detected every two hours after inoculation, the growth curve is drawn, the middle period or the later period of logarithmic phase growth is achieved, and the streptococcus thermophilus liquid reaches the standard of being inoculated to a fermentation tank. And before inoculating the streptococcus thermophilus, the culture medium C is sterilized at high temperature and under high pressure, so that the influence of other strains or microorganisms on the culture of the streptococcus thermophilus is reduced, and the cleanness and the singleness of the cultured strains are ensured.

Preferably, in the step (2), the fermentation medium is prepared by the following steps: weighing 1-3g of brown sugar, 1-3g of corn starch, 1.5-2.5g of monopotassium phosphate, 8-12g of peptone, 3-8g of yeast powder, 1-5g of beef extract, 18-22g of glucose, 1-2g of trace elements, 1-2g of growth factors and 1-2g of vitamins, adding the components into distilled water, uniformly mixing, supplementing the distilled water to a system of 1000mL, and then performing steam sterilization for 30-40min under the conditions of temperature of 115 and 125 ℃ and pressure of 0.10-0.15MPa to prepare the fermentation culture medium.

The fermentation culture medium is prepared by mixing the materials, so that sufficient fermentation nutrients are provided for paenibacillus polymyxa, saccharomycetes and streptococcus thermophilus, and the fermentation culture medium is subjected to sterilization pretreatment at high temperature and under pressure, so that the influence of other strains or microorganisms on a multi-bacterium symbiotic fermentation process is reduced, and the normal fermentation is ensured.

Wherein the vitamins need to be filtered and sterilized before being added, and the microelements are one or more of ferric sulfate, zinc sulfate, calcium chloride, manganese sulfate, sodium acetate, triammonium citrate, ferric trichloride, calcium sulfate and magnesium chloride; the growth factor comprises 0.95-1.9g choline and 0.1-0.5mg betaine.

Preferably, in the step (2), when the paenibacillus polymyxa liquid is added, after the fermentation medium is added into the fermentation equipment, oxygen is supplied to the fermentation equipment, the dissolved oxygen of the fermentation medium is controlled to account for 8-12% of the volume of the fermentation medium, then the paenibacillus polymyxa liquid is added while stirring at the rotation speed of 150-;

when adding yeast liquid, supplying oxygen to the fermentation equipment, controlling the dissolved oxygen of the system to account for 5-10% of the volume of the fermentation liquid A, then adding the yeast liquid while stirring at the rotation speed of 150-180rpm, and continuing stirring and fermenting at the rotation speed of 150-180 rpm;

adding streptococcus thermophilus liquid, supplying oxygen to the fermentation equipment, controlling the dissolved oxygen amount of the system to be 5-10% of the volume of the fermentation liquid B, then adding the streptococcus thermophilus liquid while stirring at the rotating speed of 50-80rpm, and continuously stirring and fermenting at the rotating speed of 50-80 rpm; .

Before each bacterial colony is added into the fermentation tank, the dissolved oxygen amount, the pH value, the temperature and the rotating speed of a system in the fermentation tank are regulated, and then the bacterial colony is added, so that the bacterial colony is easy to symbiotic after being added, and the bacterial colony is fermented in a synergistic manner without producing antagonistic action, so that the multi-bacterial fusion symbiotic fermentation broth is prepared. After each addition of each bacterium, the pH value of the fermentation process system is reduced, the pH value is adjusted after the fermentation is stable, and then the next bacterium colony is added. For example, after adding Paenibacillus polymyxa liquid, the pH value of the fermentation process system is reduced, after the fermentation is carried out until the pH value is stable, the pH value of the fermentation liquid A is adjusted to 6.0-6.5, and then yeast liquid is added; similarly, after adding yeast liquid, the pH value of the fermentation process system is reduced, after the fermentation is carried out until the pH value is stable, the pH value of the fermentation liquid B is adjusted to 5.0-5.5, and then the streptococcus thermophilus liquid is added.

Preferably, the inoculation amount of the paenibacillus polymyxa liquid in the fermentation medium is 2-5% of the volume of the fermentation medium, the inoculation amount of the yeast liquid in the fermentation liquid A is 2-5% of the volume of the fermentation liquid A, and the inoculation amount of the streptococcus thermophilus liquid in the fermentation liquid B is 2-5% of the volume of the fermentation liquid B.

By controlling the inoculation amount of each strain, a plurality of strains are symbiotic and are fermented in a synergistic manner without producing antagonistic action, so that the prepared multi-strain fusion symbiotic fermentation broth can act on plants to resist diseases and insect pests, promote root growth and improve the stress resistance of crops.

Preferably, the fermentation equipment comprises a fermentation tank body and a stirring mechanism arranged in the fermentation tank body, wherein the stirring mechanism comprises a main stirring rod, a first driving motor for driving the main stirring rod to rotate and side stirring assemblies symmetrically arranged on two sides of the main stirring rod; the side stirring assembly comprises a side stirring rod and a second driving motor for driving the side stirring rod to rotate, a connecting piece is sleeved on the outer wall of the main stirring rod through a bearing, and the second driving motor is arranged on the connecting piece; the top of the fermentation tank body is provided with a feed inlet for feeding, an air inlet for supplying oxygen into the fermentation tank body and an exhaust outlet for discharging fermentation waste gas; the outer wall of the fermentation tank body is provided with a controller, and the first driving motor and the second driving motor are electrically connected with the controller.

Through adopting above-mentioned fermentation equipment, in the intergrowth fermentation technology of many fungus fusions, main puddler can realize rotating under first driving motor's drive, and the puddler that inclines can realize rotating under second driving motor's drive for main puddler and the equal accessible rotation of puddler that inclines improve the stirring homogeneity in the fermentation cylinder, and a plurality of intervals stir in the zymotic fluid, can improve the dissolved oxygen degree of system.

Preferably, the connecting piece comprises a sleeve and connecting rods symmetrically arranged at the bottom of the sleeve, the outer wall of the bearing is connected with the inner wall of the sleeve, and the second driving motor is arranged on the connecting rods;

the telescopic outer wall ring is equipped with first flange, the outer wall of main puddler in the top ring of first flange is equipped with the second flange, first flange is provided with the fastener, the second flange corresponds the fastener is concave to be equipped with the storage tank, the one end of fastener is articulated in first flange, the other end of fastener inlay to establish to the storage tank.

Through adopting above-mentioned technical scheme, the fastener inlays to establish to the storage tank for the looks block of first flange and second flange is connected, and then realizes being connected of sleeve and main puddler, and when making main puddler rotate, drive the side puddler and rotate, stirs the zymotic fluid system. Preferably, when oxygen is supplied to the system, the clamping connection of the fastener to the accommodating groove is released, the first driving motor and the second driving motor both operate simultaneously, so that the main rotating rod and the two side connecting rods rotate simultaneously, three vortex-like flow fields are generated in the fermentation liquor, more oxygen can be carried into the fermentation liquor in the rotating process, and the dissolved oxygen degree is improved; and when adding the bacterial colony in toward the system, the fastener inlays to establish and carries out looks lock to the storage tank for two side stirring subassemblies are connected with main dwang, and first driving motor functions, and second driving motor does not function, and first driving motor drive main dwang rotates, drives two side stirring subassemblies and rotates, stirs the zymotic fluid, improves the mixing homogeneity.

Preferably, the fermentation tank body comprises an inner tank body and an outer tank body, a water flow interlayer is arranged between the inner tank body and the outer tank body, the outer tank body is provided with a water inlet and a water outlet which are communicated with the water flow interlayer, and the controller is arranged on the outer wall of the outer tank body;

the lateral wall of inner tank body is provided with pH sensor, dissolved oxygen sensor and temperature sensor, pH sensor, dissolved oxygen sensor and temperature sensor all are connected with the controller electricity.

The warm water or the cooling water flows into the water flow interlayer through the water inlet and the water outlet to regulate and control the temperature of the fermentation liquor in the inner tank body; the pH value of the fermentation liquor can be monitored in real time through the pH sensor, the dissolved oxygen of the fermentation liquor can be monitored in real time through the dissolved oxygen sensor, and the temperature of the fermentation liquor can be monitored in real time through the temperature sensor, so that the conditions of the dissolved oxygen, the pH value and the temperature of the fermentation liquor can be regulated and controlled in real time.

In summary, the present application has the following beneficial effects:

1. the utility model provides a many fungus fusion symbiosis fermentation process, the mixed fermentation of rhizosphere probiotic and the disease-resistant rhizosphere probiotic of promoting growth, realize disposable input among the bacterial fermentation process, disposable output, change traditional batch fermentation technology, reduce the manufacturing cost of microbial inoculum, and the fungus crowd that mixed production made is various, it is strong to environmental suitability, the problem that traditional batch fermentation process bacterial strain and effect are comparatively single has been solved, the range of application is extensive, shorten the process when using as the microbial inoculum, play disease-resistant effect of promoting growth to the plant.

2. Before each strain is added, the dissolved oxygen amount, the pH value, the temperature and the rotating speed of a system in the fermentation tank are regulated and controlled, and then the bacterial colony is added, so that the bacterial colony is easy to symbiotic after being added, and the bacterial colony is fermented in a synergistic manner without producing an antagonistic effect, so that the multi-bacterium fusion symbiotic fermentation broth with the disease-resistant growth-promoting effect is prepared.

3. According to the fermentation equipment, in the multi-bacterium fusion symbiotic fermentation process, the main stirring rod can rotate under the drive of the first drive motor, and the side stirring rod can rotate under the drive of the second drive motor, so that the main stirring rod and the side stirring rod can rotate; when oxygen is supplied to the system, the first driving motor and the second driving motor operate simultaneously, so that multiple parts of the fermentation liquor can be stirred, and the dissolved oxygen of the fermentation liquor is increased; and when adding the bacterial colony in the past system, two side stirring subassemblies are connected with main dwang, and first driving motor functions, and second driving motor does not function, and first driving motor drive main dwang rotates, drives two side stirring subassemblies and rotates, stirs the zymotic fluid, improves the homogeneity of mixing.

Drawings

FIG. 1 is a schematic view of the structure of a fermentation apparatus according to the present application;

FIG. 2 is a sectional view of a fermentation apparatus of the present application;

FIG. 3 is an enlarged view of portion A of FIG. 2;

FIG. 4 is a schematic structural diagram of a stirring mechanism in the fermentation apparatus of the present application;

description of the drawings: 1. a fermentation tank body; 11. a feed inlet; 12. an air inlet; 13. an exhaust port; 14. an inner tank body; 15. an outer tank body; 16. a water flow interlayer; 17. a water inlet; 18. a water outlet; 19. a tank cover; 191. a cover plate; 2. a stirring mechanism; 21. a main stirring rod; 211. a second flange; 2111. a containing groove; 22. a first drive motor; 23. a side stirring assembly; 231. a side stirring rod; 232. a second drive motor; 24. a bearing; 251. a sleeve; 252. a connecting rod; 253. a first flange; 2531. a fastener; 3. a controller; 4. a pH sensor; 5. a dissolved oxygen sensor; 6. a temperature sensor.

Detailed Description

The present application is described in further detail below with reference to figures 1-4 and examples.

The following examples and comparative examples of Paenibacillus polymyxa were purchased from the institute of Biotechnology, Innovation and Biotechnology, Beijing, with resource number BNCC185335 and were purified, activated and cultured using NA solid medium before inoculation.

The yeast is purchased from Beijing Beinan Chuanglian union of Industrial science and Biotechnology with the resource number of BNCC164204, and before inoculation, the yeast is purified, activated and cultured by adopting a PDA solid culture medium.

The streptococcus thermophilus is purchased from Beijing Beinanna institute of Biotechnology with resource number BNCC162965, and is purified, activated and cultured by adopting an MRS solid culture medium before inoculation.

The lactococcus lactis is the lactococcus lactis purchased from Beijing Beinanna institute of Biotechnology with resource number BNCC 195305.

Examples

Example 1

A multi-bacterium fusion symbiotic fermentation process for resisting diseases and promoting growth comprises the following steps:

step (1-1), preparing a culture medium:

preparing a culture medium A: weighing 3g of beef extract, 10g of peptone, 5g of sodium chloride and 20g of agar, adding into distilled water, mixing uniformly, supplementing distilled water until the system is 1000mL, adjusting the pH value to 7.2, and sterilizing at 121 ℃ and 0.10MPa for 30min to obtain the culture medium A.

Preparing a culture medium B: weighing 200g of potato filtrate, 20g of glucose and 20g of agar, adding into distilled water, mixing uniformly, supplementing distilled water to a system of 1000mL, and sterilizing at 121 ℃ and 0.10MPa for 30min to obtain a culture medium B.

Preparing a culture medium C: weighing 10g of peptone, 12g of beef extract, 5g of yeast extract, 20g of glucose, 5g of sodium acetate, 2g of diamine citrate, 1.0g of tween-80, 2.0g of dipotassium hydrogen phosphate, 0.58g of magnesium sulfate, 0.25g of manganese sulfate, 20g of calcium carbonate and 20g of agar, adding into distilled water, uniformly mixing, supplementing distilled water until the system is 1000mL, and then sterilizing at the temperature of 121 ℃ and under the pressure of 0.10MPa for 30min to obtain the culture medium C.

Preparing a fermentation medium: weighing 2g of brown sugar, 2g of corn starch, 2.0g of monopotassium phosphate, 10g of peptone, 5g of yeast powder, 3g of beef extract, 20g of glucose, 1.5g of trace elements, 1.5g of choline, 0.3mg of betaine, 0.7g of manganese sulfate and 0.8g of ferric chloride, adding into distilled water, uniformly mixing, supplementing distilled water to a system of 1000mL, and then performing steam sterilization for 35min under the conditions of temperature of 120 ℃ and pressure of 0.15MPa to obtain the fermentation medium.

Step (1-2), preparing bacterial liquid: inoculating the activated paenibacillus polymyxa into the culture medium A, wherein the inoculation amount is 10%, shaking up, and culturing at 36 +/-1 ℃ for 36h to obtain paenibacillus polymyxa liquid; inoculating the activated yeast into a culture medium B, wherein the inoculation amount is 10%, shaking up, and culturing at 36 +/-1 ℃ for 36h to obtain a yeast liquid; inoculating the activated streptococcus thermophilus into a culture medium C, wherein the inoculation amount is 10%, shaking up, and culturing at 31 +/-1 ℃ for 36h to obtain streptococcus thermophilus liquid.

Step (2), fusing symbiotic fermentation: adding a fermentation medium into fermentation equipment, controlling the pH value of the fermentation medium to be 7.0 and the temperature to be 35 ℃, supplying oxygen into the fermentation equipment, controlling the dissolved oxygen amount of the fermentation medium to account for 8% of the volume of the fermentation medium, then adding paenibacillus polymyxa liquid under the condition of rotating speed of 150rpm while stirring, continuously stirring, and fermenting for 24 hours to obtain fermentation liquid A;

then controlling the pH value of the fermentation liquor A system to be 6.0 and the temperature to be 35 ℃, supplying oxygen to fermentation equipment, controlling the dissolved oxygen of the system to account for 5% of the volume of the fermentation liquor A, then adding yeast liquid while stirring under the condition that the rotating speed is 150rpm, continuously stirring the yeast liquid with the inoculum size of 2% of the volume of the fermentation liquor A, and fermenting for 45 hours to obtain fermentation liquor B;

and controlling the pH value of the fermentation liquid B system to be 5.0 and the temperature to be 30 ℃, supplying oxygen to fermentation equipment, controlling the dissolved oxygen of the system to account for 5% of the volume of the fermentation liquid B, then adding the streptococcus thermophilus liquid under the condition of 50rpm while stirring, wherein the inoculation amount of the streptococcus thermophilus liquid is 2% of the volume of the fermentation liquid B, continuously stirring, and after fermenting for 3 days, the viable count of the paenibacillus polymyxa, the saccharomycetes and the streptococcus thermophilus is more than 20 hundred million/mL, so that the multi-bacterium fusion symbiotic fermentation liquid is obtained.

Example 2

This embodiment differs from embodiment 1 described above in that: and (3) combining the condition parameter change in the symbiotic fermentation process in the step (2).

Example 3

The condition parameters in the above examples 1 to 3 are shown in the following table 1:

TABLE 1 Condition parameter Table for multiple bacteria fusion symbiotic fermentation in examples 1-3

Example 4

This embodiment differs from embodiment 2 described above in that: the condition parameters in the preparation of the culture medium A in the step (1) are changed, and the details are shown in the following table 2.

Example 5

This embodiment differs from embodiment 2 described above in that: step (1) preparation of the changes in the condition parameters in medium A, see in particular Table 2 below.

TABLE 2 Condition parameter Table for Medium A preparation in examples 2, 4 and 5

Example 6

This embodiment differs from embodiment 2 described above in that: step (1) preparation of the changes in the condition parameters in medium B, see in particular Table 3 below.

Example 7

TABLE 3 Condition parameter Table for Medium B preparation in examples 2, 6 and 7

Example 8

This embodiment differs from embodiment 2 described above in that: step (1) preparation of the medium C in the condition parameters change, see table 4 below.

Example 9

TABLE 4 Condition parameter Table for Medium C preparation in examples 2, 8 and 9

Example 10

This embodiment differs from embodiment 2 described above in that: the changes in the condition parameters in the configured fermentation medium of step (1) are specifically seen in table 5 below.

Example 11

TABLE 5 Condition parameter Table for fermentation Medium preparation in examples 2, 10 and 11

Example 12

This embodiment differs from embodiment 2 described above in that:

the fermentation equipment adopted in the fermentation process in the step (2) is shown in fig. 1 and fig. 2 and comprises a fermentation tank body 1 and a stirring mechanism 2, wherein the fermentation tank body 1 comprises an inner tank body 14, an outer tank body 15 and a tank body cover 19, a sealed water interlayer 16 is arranged between the inner tank body 14 and the outer tank body 15 in a surrounding manner, the tank body cover 19 covers the inner tank body 14, a water inlet 17 and a water outlet 18 are formed in the side wall of the outer tank body 15, the water inlet 17 and the water outlet 18 are communicated with the water interlayer 16 so as to introduce warm water or cooling water into the water interlayer 16, and the fermentation liquor loaded in the inner tank body 14 is subjected to water bath temperature control; the top of the tank cover 19 is provided with a feed inlet 11, an air inlet 12 and an air outlet, the feed inlet 11 is used for adding a fermentation medium, a paenibacillus polymyxa liquid and a yeast liquid streptococcus thermophilus liquid, the air inlet 12 is used for connecting an external ventilation pipeline and supplying oxygen into the inner tank 14, carbon dioxide and other gases can be generated in the fermentation process, and the air outlet is used for discharging the carbon dioxide in the inner tank 14 outwards so as to avoid the influence on the normal fermentation process caused by high concentration of the carbon dioxide in the inner tank 14. Preferably, the feeding hole 11 can be used as a discharging hole, and the fermentation liquid is pumped out of the inner tank body 14 to realize discharging through the pipeline passing through the feeding hole 11 and the tail end of the pipeline extending below the liquid level of the fermentation liquid.

Referring to fig. 2, the stirring mechanism 2 includes a main stirring rod 21, a first driving motor 22, a connecting member and two side stirring assemblies 23, the first driving motor 22 drives the main stirring rod 21 to rotate, wherein the first driving motor 22 is arranged at the outer side of the top of the outer tank body 15, and a triangular stirring member is arranged at the bottom end of the main stirring rod 21 to stir the fermentation liquor and reduce the shearing damage effect of the outer edge of the blade on the thallus of the fermentation liquor. Referring to fig. 2 and 3, the connecting member includes a sleeve 251 and a connecting rod 252, a through hole for the main stirring rod 21 to pass through is formed in the middle of the connecting rod 252, a bearing 24 is sleeved on the outer wall of the main stirring rod 21, and the outer wall of the bearing 24 is connected with the inner wall of the sleeve 251, so that the main stirring rod 21 passes through the sleeve 251 and the through hole and can rotate in the through hole without driving the connecting rod 252 to rotate. Two side stirring subassemblies 23 are in the axial symmetry setting of main puddler 21, and every side stirring subassembly 23 includes side puddler 231 and second driving motor 232, and second driving motor 232 sets up in connecting rod 252, and the rotation of second driving motor 232 drive side puddler 231, and the bottom of side puddler 231 is provided with triangle form stirring piece equally to stir the zymotic fluid, reduce the shearing destruction effect of blade outer edge to the zymotic fluid thalli. When oxygen is supplied to the inner tank 14, the first driving motor 22 drives the main stirring rod 21 to rotate, the second driving motor 232 drives the side stirring rod 231 to rotate, the main stirring rod 21 and the side stirring rod 231 rotate simultaneously, three flow fields similar to vortexes are generated in the fermentation broth, more oxygen can be carried into the fermentation broth in the rotating process, the dissolved oxygen degree is improved, in addition, in the rotating process of the main stirring rod 21 and the side stirring rod 231, slight movement and deviation can occur in the fermentation broth, the slight stirring of the fermentation broth can also be driven, and the dissolved oxygen degree is improved.

In addition, referring to fig. 3 and 4, the outer wall of the sleeve 251 is provided with a first flange 253, the outer wall of the main stirring rod 21 is provided with a second flange 211, the second flange 211 is disposed above the first flange 253, the side of the first flange 253 is hinged with a plurality of fasteners 2531, the second flange 211 is correspondingly provided with a containing groove 2111, the tail end of the fastener 2531 far away from the hinged end is embedded into the containing groove 2111, the fastening connection between the fastener 2531 and the containing groove 2111 is realized, the connection between the sleeve 251 and the main stirring rod 21 is realized, and when the main stirring rod 21 rotates, the side stirring rod 231 is driven to rotate. Preferably, the fastening end of the fastening member 2531 is a magnet, the inner wall of the accommodating groove 2111 is provided with a magnetic layer, and the fastening end of the fastening member 2531 and the accommodating groove 2111 are magnetically attracted to each other, so that the fastening member 2531 and the second flange 211 are stably fastened. When oxygen is required to be supplied to the fermentation liquor system, the fastener 2531 is released from clamping the accommodating groove 2111, so that the main stirring rod 21 and the two side stirring rods 231 rotate to improve the dissolved oxygen degree of the fermentation liquor; when adding the bacterial colony in the zymotic fluid, fastener 2531 block to storage tank 2111 for first flange 253 is connected with second flange 211, and then main puddler 21 drives connecting rod 252 and rotates, and then drives two root side puddlers 231 and rotates, second driving motor 232 does not operate this moment, two side puddlers 231 do not have rotation motion, make main puddler 21 and two side puddlers 231 carry out the stirring of great degree to the zymotic fluid, improve the mixing homogeneity of zymotic fluid system in fungus liquid and the inner tank body 14. Referring back to fig. 1, the top of the tank cover 19 is opened with an opening and is covered by a cover plate 191 to ensure the sealing state of the inner tank 14 and the normal fermentation; the cover plate 191 can be connected with the tank body cover 19 in a hinged mode, the cover plate 191 is turned over, and a hand is stretched into the opening, so that the clamping connection or the releasing connection of the fastener 2531 is realized.

Referring back to fig. 1, a controller 3 for controlling the operation of the fermentation apparatus is disposed on the side wall of the outer tank 15, preferably, a PLC controller is selected, and the first driving motor 22, the second driving motor 232, the pH sensor 4, the dissolved oxygen sensor 5 and the temperature sensor 6 are all electrically connected to the controller 3.

The working principle of the fermentation equipment of the embodiment is as follows: adding a fermentation medium into the inner tank body 14 through the feeding hole 11, enabling the fastener 2531 to be not buckled with the accommodating groove 2111, starting the first driving motor 22 and the second driving motor 232, enabling the main stirring rod 21 and the side stirring rod 231 to rotate simultaneously, communicating the air inlet 12 with external oxygen pumping equipment through a pipeline, supplying oxygen into the inner tank body 14, and promoting oxygen dissolution and the fermentation medium through rotation of the main stirring rod 21 and the side stirring rod 231; after reaching the target dissolved oxygen volume, close first driving motor 22 and second driving motor 232, open apron 191, be connected fastener 2531 and storage tank 2111 looks block, close apron 191, restart first driving motor 22, second driving motor 232 does not start, and then drive main puddler 21 and rotate, and drive connecting rod 252, the rotation of side puddler 231, two side puddlers 231 use the axis of main puddler 21 to rotate as the center, stir the fermentation culture medium, then add the paenibacillus polymyxa fungus liquid while stirring, continue stirring, fermentation after finishing adding.

After the fermentation is carried out for a certain period of time, closing the first driving motor 22 and the second driving motor 232, turning over the cover plate 191, opening the exhaust hole, discharging the gases such as carbon dioxide and the like generated by the fermentation in the inner tank body 14 to the outside of the tank body, simultaneously releasing the clamping connection between the fastener 2531 and the accommodating groove 2111, closing the cover plate 191, starting the first driving motor 22 and the second driving motor 232 again, supplying oxygen to the inner tank body 14, and promoting the dissolution of oxygen into the fermentation liquid A by the rotation of the main stirring rod 21 and the side stirring rod 231; after the target dissolved oxygen amount is reached, the first driving motor 22 and the second driving motor 232 are closed, the cover plate 191 is opened, the fastener 2531 is connected with the accommodating groove 2111 in a clamping mode, the cover plate 191 is closed, the first driving motor 22 is started again, the second driving motor 232 is not started, the stirring action is repeated, the fermentation liquid A is stirred, the yeast liquid is added while stirring, and stirring and fermentation are continued after the addition is finished.

The addition of the Streptococcus thermophilus solution is similarly performed by the oxygen supply operation and the Streptococcus thermophilus solution addition operation, and the description thereof will not be repeated.

Comparative example

Comparative example 1

This comparative example differs from example 2 above in that: the symbiotic fermentation system for the multi-bacterium fusion is the symbiotic fermentation of paenibacillus polymyxa and yeast, does not contain the configuration of a culture medium C, the preparation of streptococcus thermophilus liquid, the inoculation of the streptococcus thermophilus liquid and the control of related condition parameters, and the step (2) is as follows:

step (2), fusing symbiotic fermentation: adding a fermentation medium into fermentation equipment, controlling the pH value of the fermentation medium to be 7.1 and the temperature to be 36 ℃, supplying oxygen into the fermentation equipment, controlling the dissolved oxygen of the fermentation medium to account for 10% of the volume of the fermentation medium, then adding paenibacillus polymyxa liquid under the condition of a rotating speed of 165rpm while stirring, continuously stirring, and fermenting for 30 hours to obtain fermentation liquid A;

and then controlling the pH value of the fermentation liquor A system to be 6.3 and the temperature to be 36 ℃, supplying oxygen to fermentation equipment, controlling the dissolved oxygen of the system to account for 8% of the volume of the fermentation liquor A, then adding yeast liquid while stirring at the rotating speed of 165rpm, continuously stirring, and fermenting for 48 hours to obtain the multi-bacteria fusion symbiotic fermentation liquor.

Comparative example 2

This comparative example differs from example 2 above in that: the symbiotic fermentation system of the paenibacillus polymyxa and the streptococcus thermophilus is symbiotic fermentation in the multi-bacterium fusion symbiotic fermentation system, and does not contain the configuration of a culture medium B, the preparation of saccharomycetes, the inoculation of yeast liquid and the control of related condition parameters, and the step (2) is specifically as follows:

and controlling the pH value of the fermentation liquor A system to be 5.3, controlling the temperature to be 31 ℃, supplying oxygen to fermentation equipment, controlling the dissolved oxygen of the system to account for 8% of the volume of the fermentation liquor A, then adding the streptococcus thermophilus liquid under the condition of a rotating speed of 65rpm while stirring, continuously stirring, and fermenting for 3.5 days to obtain the multi-bacterium fusion symbiotic fermentation liquor.

Comparative example 3

This comparative example differs from example 2 above in that: the symbiotic fermentation system for the yeast and the streptococcus thermophilus in the multi-bacterium fusion symbiotic fermentation system does not contain the configuration of a culture medium A, the preparation of paenibacillus polymyxa, the inoculation of paenibacillus polymyxa liquid and the control of related condition parameters, and the step (2) is as follows:

step (2), fusing symbiotic fermentation: adding a fermentation medium into fermentation equipment, controlling the pH value of the fermentation medium to be 6.3 and the temperature to be 36 ℃, supplying oxygen into the fermentation equipment, controlling the dissolved oxygen of a system to account for 8% of the volume of the fermentation medium, then adding yeast liquid under the condition of a rotating speed of 165rpm while stirring, continuously stirring, and fermenting for 48 hours to obtain fermentation liquid A;

Comparative example 4

This comparative example differs from example 2 above in that: lactococcus lactis was used instead of streptococcus thermophilus of example 2.

The lactococcus lactis culture medium comprises 10.0g of beef extract, 5.0g of peptone, 5.0g of NaCl, 5g of yeast extract, 5g of glucose, 15.0g of agar and 1.0L of distilled water, and is sterilized for 30min at the temperature of 121 ℃ and the pressure of 0.10MPa to prepare the culture medium.

And (3) in the process of inoculating the fermentation liquor B in the step (2), controlling the pH value of a fermentation liquor B system to be 7.0 and the temperature to be 30 ℃, supplying oxygen to fermentation equipment, controlling the dissolved oxygen of the system to account for 5% of the volume of the fermentation liquor B, then adding lactococcus lactis under the condition of 50rpm while stirring, wherein the inoculation amount of the lactococcus lactis is 2% of the volume of the fermentation liquor B, continuously stirring, and after fermenting for 3 days, the number of viable bacteria of paenibacillus polymyxa, saccharomycetes and lactococcus lactis is more than 20 hundred million/mL to obtain the multi-bacteria fusion symbiotic fermentation liquor.

Comparative example 5

This comparative example differs from example 2 above in that: in the comparative example, paenibacillus polymyxa, saccharomycetes and streptococcus thermophilus are respectively inoculated to a fermentation medium for independent fermentation, and then are mixed according to the proportion of 1:1:1 before use to prepare the compound microbial agent; the fermentation of Paenibacillus polymyxa alone is as follows: adding a fermentation medium into fermentation equipment, controlling the pH value of the fermentation medium to be 7.1 and the temperature to be 36 ℃, supplying oxygen into the fermentation equipment, controlling the dissolved oxygen of the fermentation medium to account for 10% of the volume of the fermentation medium, then adding paenibacillus polymyxa liquid under the condition of a rotating speed of 165rpm while stirring, continuously stirring, and fermenting for 30 hours to obtain paenibacillus polymyxa fermentation liquid;

the individual fermentations of yeast are as follows: adding a fermentation medium into fermentation equipment, controlling the pH value of the fermentation medium to be 6.3 and the temperature to be 36 ℃, supplying oxygen into the fermentation equipment, controlling the dissolved oxygen of a system to account for 8% of the volume of the fermentation medium, then adding yeast liquid under the condition of a rotating speed of 165rpm while stirring, continuously stirring, and fermenting for 48 hours to obtain yeast fermentation liquid;

the fermentation of S.thermophilus alone is as follows: adding a fermentation medium into a fermentation device, controlling the pH value of the fermentation medium to be 5.3 and the temperature to be 31 ℃, supplying oxygen into the fermentation device, controlling the dissolved oxygen of a system to account for 8% of the volume of the fermentation medium, then adding streptococcus thermophilus liquid while stirring under the condition of a rotating speed of 65rpm, continuously stirring, and fermenting for 3.5 days to obtain the streptococcus thermophilus fermentation liquid.

Performance test

The multi-bacterium fusion symbiotic fermentation broth prepared in the above examples 1-12, the multi-bacterium fusion symbiotic fermentation broth prepared in the comparative examples 1-3, and the compound microbial agent prepared in the comparative example 4 were subjected to soil microbial testing and cucumber disease-resistant growth-promoting tests.

The soil microorganism test comprises the following steps: specifically, after fully mixing a sample (the multi-bacterium fusion symbiotic fermentation broth prepared in examples 1 to 12, the multi-bacterium fusion symbiotic fermentation broth prepared in comparative examples 1 to 3 and the composite microbial agent prepared in comparative example 4) with water (the ratio is 1: 300), spraying and applying the mixture to test soil (the test soil is taken from the profit area of eastern province reclamation city in Shandong province), wherein the usage amount of each square meter is 20L of diluted bacterial liquid; meanwhile, a blank control group is set, fermentation liquor is replaced by clear water, and clear water is added into the soil. Each treatment sample is applied for three times in a total, once a week, and seven days after the third application, sampling and investigation are carried out, and the total number of the soil microorganisms is counted.

(II) cucumber growth promotion test: the cucumber variety is selected from Vinci needle, and purchased from Taixing Miao GmbH of Xintai city, Shandong; the planting soil is taken from a reclamation area in Dongying city of Shandong province, the planting soil is sterilized at 100 ℃ for 1h, the planting soil is filled into seedling trays with the length of 34.5cm, the width of 24cm and the height of 11cm, the cucumber seeds are sown after pregermination, 100 seeds are planted in each tray, thinning is carried out after seedling emergence, and 50 seedlings with uniform growth are reserved. And 5d after sowing, when the true leaves are initially spread, selecting plants with consistent growth vigor, and inoculating a fermentation liquid sample in a root irrigation mode along with water, wherein three plants are irrigated in each sample embodiment/comparative example. Meanwhile, a blank control group is set, fermentation liquor is replaced by clear water, and clear water is added into the soil.

After 7d of treatment, measuring the growth index and disease resistance index of the cucumber seedlings, and measuring the stem thickness at a position 1cm below the cotyledon nodes; measuring the distance between the stem base and the growing point to obtain the plant height; the plants were repeatedly washed with clean water, then with toilet paper to remove free water, and the fresh weight was measured with an electronic balance.

The stem thickness, plant height and fresh weight were averaged.

And (III) cucumber disease resistance test: according to the cucumber seedling in the above test (II), the grading standard of cucumber seedling blight is as follows:

level 0: no symptoms;

level 1: the yellowing or wilting area of the true leaves and the cotyledons does not exceed 50 percent of the total area;

and 2, stage: the yellowing or wilting area of the true leaves and the cotyledons exceeds 50 percent of the total area;

and 3, level: leaves wither or die, only growing points survive;

4, level: the whole plant withers severely to die.

The disease index [ Σ (number of diseased plants × representative number of stages)/(total number of plants × highest representative value) ] × 100%.

The results of the soil microorganism test and the cucumber disease resistance and growth promotion test are specifically shown in the following table 6:

TABLE 6 Total microbial count of soil and cucumber disease-resistant growth-promoting test results

According to the embodiment, the multi-bacterium fusion symbiotic fermentation broth prepared by the multi-bacterium fusion symbiotic fermentation process has strong adaptability to the environment, and can play a role in resisting diseases and promoting growth of plants.

Compared with the example 2, in the periodic soil spraying treatment, the number of soil microorganisms tends to increase, and the stem thickness, the plant height and the fresh weight of the cucumber also tend to increase in the growth promotion test of the cucumber in the periodic soil spraying treatment, which shows that the fermentation equipment can improve the dissolved oxygen of fermentation liquor, promote the mixing uniformity of materials, promote the growth of bacterial colonies, improve the number of bacterial colonies in the mixed fermentation liquor, and further improve the symbiosis in the soil and the growth promotion and disease resistance effects on plants when applied to a multi-bacteria fusion symbiotic fermentation process.

Wherein, the symbiotic fermentation broth of Paenibacillus polymyxa and yeast is prepared in a comparative example 1 by adopting a multi-bacterium fusion symbiotic fermentation process, the symbiotic fermentation broth of Paenibacillus polymyxa and Streptococcus thermophilus is prepared in a comparative example 2 by adopting a multi-bacterium fusion symbiotic fermentation process, and the symbiotic fermentation broth of yeast and Streptococcus thermophilus is prepared in a comparative example 3 by adopting a multi-bacterium fusion symbiotic fermentation process; in the comparative example 4, the symbiotic fermentation broth of Paenibacillus polymyxa, yeast and lactococcus lactis is prepared by adopting the multi-bacterium fusion symbiotic fermentation process. Compared with the embodiment 2, the four comparative examples have the advantages that after the soil is periodically sprayed and treated, the number of microorganisms in the soil is obviously lower, and in the cucumber growth promotion test, the stem thickness, the plant height and the fresh weight average of the cucumber are obviously reduced compared with the embodiment 2, and the disease index is increased; the application shows that the paenibacillus polymyxa, the saccharomycetes and the streptococcus thermophilus are subjected to symbiotic fermentation, and the prepared symbiotic fermentation liquid has strong adaptability to the environment and plays a role in resisting diseases and promoting growth of plants.

In the embodiment 5, the paenibacillus polymyxa, the saccharomycetes and the streptococcus thermophilus are fermented independently, then are mixed according to the ratio of 1:1:1 to prepare the compound microbial agent, and then are subjected to periodic spraying treatment on soil and a growth promotion test on cucumbers, compared with the embodiment 2, the soil microbial agent in the comparative example 5 is remarkably lower in number, the stem thickness, the plant height and the fresh weight of the cucumbers are remarkably reduced compared with the embodiment 2, and the disease index is increased; the method has the advantages that the symbiotic fermentation broth is prepared by adopting the multi-bacterium fusion symbiotic fermentation process, and the prepared symbiotic fermentation broth has strong adaptability to the environment, can improve the balance of soil flora and plays a role in resisting diseases and promoting growth of plants.

The present embodiment is only for explaining the present application, and it is not limited to the present application, and those skilled in the art can make modifications of the present embodiment without inventive contribution as needed after reading the present specification, but all of them are protected by patent law within the scope of the claims of the present application.

Claims

1. A multi-bacterium fusion symbiotic fermentation process for resisting diseases and promoting growth is characterized in that: the method comprises the following steps:

2. The multi-bacterium fusion symbiotic fermentation process for resisting diseases and promoting growth according to claim 1, characterized in that: in the step (1), the culture medium A is prepared by the following steps: weighing 1-5g of beef extract, 8-12g of peptone, 3-8g of sodium chloride and 15-25g of agar, adding into distilled water, uniformly mixing, supplementing the distilled water to 1000mL, adjusting the pH value to 7.0-7.3, and then sterilizing at the temperature of 118-.

3. The multi-bacterium fusion symbiotic fermentation process for resisting diseases and promoting growth according to claim 1, characterized in that: in the step (1), the culture medium B is prepared by the following steps: weighing 180-220g of potato filtrate, 18-22g of glucose and 15-25g of agar, adding into distilled water, mixing uniformly, supplementing the distilled water to a system of 1000mL, and sterilizing at the temperature of 118-123 ℃ and the pressure of 0.08-0.12MPa for 28-32min to obtain the culture medium B.

4. The multi-bacterium fusion symbiotic fermentation process for resisting diseases and promoting growth according to claim 1, characterized in that: in the step (1), the culture medium C is prepared by the following steps: weighing 8-12g of peptone, 8-12g of beef extract, 3-8g of yeast extract, 18-22g of glucose, 3-8g of sodium acetate, 1-3g of citric diamine, 0.8-1.2g of tween-80, 1.5-2.5g of dipotassium hydrogen phosphate, 0.5-0.6g of magnesium sulfate, 0.20-0.30 of manganese sulfate, 18-22g of calcium carbonate and 15-25g of agar, adding into distilled water, mixing uniformly, supplementing distilled water to a system volume of 1000mL, and sterilizing at a temperature of 118-.

5. The multi-bacterium fusion symbiotic fermentation process for resisting diseases and promoting growth according to claim 1, characterized in that: in the step (2), the fermentation medium is prepared by the following steps: weighing 1-3g of brown sugar, 1-3g of corn starch, 1.5-2.5g of monopotassium phosphate, 8-12g of peptone, 3-8g of yeast powder, 1-5g of beef extract, 18-22g of glucose, 1-2g of trace elements, 1-2g of growth factors and 1-2g of vitamins, adding the components into distilled water, uniformly mixing, supplementing the distilled water to a system of 1000mL, and then performing steam sterilization for 30-40min under the conditions of temperature of 115 and 125 ℃ and pressure of 0.10-0.15MPa to prepare the fermentation culture medium.

6. The multi-bacterium fusion symbiotic fermentation process for resisting diseases and promoting growth according to claim 1, characterized in that: in the step (2), when the paenibacillus polymyxa liquid is added, after a fermentation culture medium is added into the fermentation equipment, oxygen is supplied to the fermentation equipment, the dissolved oxygen of the fermentation culture medium is controlled to be 8-12% of the volume of the fermentation culture medium, and then the paenibacillus polymyxa liquid is added while stirring at the rotation speed of 150-180 rpm;

when adding yeast liquid, supplying oxygen to the fermentation equipment, controlling the dissolved oxygen of the system to account for 5-10% of the volume of the fermentation liquid A, and then adding the yeast liquid while stirring at the rotation speed of 150-;

adding the streptococcus thermophilus liquid, supplying oxygen to the fermentation equipment, controlling the dissolved oxygen amount of the system to be 5-10% of the volume of the fermentation liquid B, and then adding the streptococcus thermophilus liquid while stirring at the rotating speed of 50-80 rpm.

7. The multi-bacterium fusion symbiotic fermentation process for resisting diseases and promoting growth according to claim 1 or 6, characterized in that: the inoculation amount of the paenibacillus polymyxa liquid in the fermentation medium is 2-5% of the volume of the fermentation medium, the inoculation amount of the yeast liquid in the fermentation liquid A is 2-5% of the volume of the fermentation liquid A, and the inoculation amount of the streptococcus thermophilus liquid in the fermentation liquid B is 2-5% of the volume of the fermentation liquid B.

8. The multi-bacterium fusion symbiotic fermentation process for resisting diseases and promoting growth according to claim 1 or 6, characterized in that: the fermentation equipment comprises a fermentation tank body (1) and a stirring mechanism (2) arranged in the fermentation tank body (1), wherein the stirring mechanism (2) comprises a main stirring rod (21), a first driving motor (22) for driving the main stirring rod (21) to rotate and side stirring assemblies (23) symmetrically arranged on two sides of the main stirring rod (21); the side stirring assembly (23) comprises a side stirring rod (231) and a second driving motor (232) for driving the side stirring rod (231) to rotate, a connecting piece is sleeved on the outer wall of the main stirring rod (21) through a bearing (24), and the second driving motor (232) is arranged on the connecting piece;

the top of the fermentation tank body (1) is provided with a feed inlet (11) for feeding materials, an air inlet (12) for supplying oxygen into the fermentation tank body (1) and an exhaust port (13) for discharging fermentation waste gas; the outer wall of the fermentation tank body (1) is provided with a controller (3), and the first driving motor (22) and the second driving motor (232) are electrically connected with the controller (3).

9. The multi-bacterium fusion symbiotic fermentation process for resisting diseases and promoting growth according to claim 8, characterized in that: the connecting piece comprises a sleeve (251) and connecting rods (252) symmetrically arranged at the bottom of the sleeve (251), the outer wall of the bearing (24) is connected with the inner wall of the sleeve (251), and the second driving motor (232) is arranged on the connecting rods (252);

the outer wall ring of sleeve (251) is equipped with first flange (253), the outer wall of main puddler (21) in the top ring of first flange (253) is equipped with second flange (211), first flange (253) are provided with fastener (2531), second flange (211) correspond fastener (2531) are sunken to be equipped with the storage tank, the one end of fastener (2531) is articulated in first flange (253), the other end of fastener (2531) inlays to establish to the storage tank.

10. The multi-bacterium fusion symbiotic fermentation process for resisting diseases and promoting growth according to claim 8, characterized in that: the fermentation tank body (1) comprises an inner tank body (14) and an outer tank body (15), a water flow interlayer (16) is arranged between the inner tank body (14) and the outer tank body (15), the outer tank body (15) is provided with a water inlet (17) and a water outlet (18) which are communicated with the water flow interlayer (16), and the controller (3) is arranged on the outer wall of the outer tank body (15);

the lateral wall of inner tank body (14) is provided with pH sensor (4), dissolved oxygen sensor (5) and temperature sensor (6), pH sensor (4), dissolved oxygen sensor (5) and temperature sensor (6) all are connected with controller (3) electricity.