Multifunctional microbial agent composite stabilizer as well as preparation method and application thereof

Abstract

The invention discloses a multifunctional microbial agent compound stabilizer as well as a preparation method and application thereof, and belongs to the technical field of microorganisms. The compound microorganism bacterial liquid disclosed by the invention is prepared from the following strains: Pseudomonas E sp017968885, Bacillus amyloliquefaciens XN-3 and Arthrobacter sp., and the number ratio of viable bacteria is 2 to 3 to 1. The invention further discloses a preparation method of the compound microorganism bacterial liquid, which is characterized in that the compound microorganism bacterial liquid is prepared from the following components: Pseudomonas E sp017968885, Bacillus amyloliquefaciens XN-3 and Arthrobacter sp. The formula of the compound stabilizer consists of the following substances: 3% w / v of starch as a nutritional ingredient; the protective agent is 2% w / v ascorbic acid; and a stabilizer is 0.2% w / v xanthan gum. After the microbial agent adopting the formula is stored for 287 days under a room-temperature dark storage condition, the viable count is 1.42 * 10 < 8 > CFU / mL, and the survival rate is 35.95%.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of microbial technology, in particular to a multifunctional microbial agent composite stabilizer and its preparation method and application. BACKGROUND

[0002] Microbial agents are widely used in agriculture, environmental protection and other fields, and can effectively improve soil quality, enhance plant disease resistance and repair polluted environment. However, microbial agents have poor stability, single function and are easily affected by the environment, which limits their large-scale promotion.

[0003] In the field of agriculture, with the development of agricultural modernization, the demand for soil improvement and healthy growth of crops is increasingly urgent. The extensive use of chemical fertilizers and pesticides has led to soil structure destruction, microbial community imbalance and enhanced resistance to pests and diseases. Although microbial agents can regulate soil microecology, promote nutrient transformation and inhibit pathogenic bacteria, they are easily affected by temperature, humidity, pH and other factors in the soil due to their poor stability, making it difficult to maintain activity for a long time and affecting application effect. For example, in the southern region with high temperature and humidity, the activity of some microbial agents decreases rapidly, and they cannot fully play the role of improving soil and promoting crop growth.

[0004] In the field of environmental protection, microbial agents are used for wastewater treatment, waste treatment and soil pollution remediation. In wastewater treatment, stable and efficient microbial agents are needed to decompose organic pollutants and remove nitrogen and phosphorus nutrients. However, due to poor stability, existing microbial agents are easily inhibited by toxic and harmful substances in complex wastewater environments, affecting their activity and function, and the treatment effect is unstable. For example, in the treatment of industrial wastewater containing heavy metals and organic pollutants, the activity of ordinary microbial agents is inhibited by heavy metals, making it difficult to effectively degrade organic pollutants.

[0005] Currently, there are limited methods to improve the stability of microbial agents. Adding conventional protective agents such as sugars and proteins has little effect on improving stability. Optimizing the preparation process can improve it to some extent, but it still cannot meet the needs in complex application environments. It is crucial to develop new and efficient microbial agent composite stabilizers. SUMMARY

[0006] The purpose of the present application is to provide a multifunctional microbial agent composite stabilizer and its preparation method and application to solve the problems existing in the prior art. The present application solves the problem of easy loss of activity of liquid microbial agents during processing, transportation and storage by improving the microbial agent composite stabilizer.

[0007] To achieve the above purpose, the present application provides the following solutions:

[0008] The application provides a multifunctional microbial agent composite stabilizer, which comprises three components of starch, ascorbic acid and xanthan gum; the mass ratio of the starch, the ascorbic acid and the xanthan gum is 15:10:1; and the effective use concentration of the three components is 3% w / v of the starch, 2% w / v of the ascorbic acid and 0.2% w / v of the xanthan gum.

[0009] The application further provides a multifunctional microbial agent, which comprises a composite microbial agent and the composite stabilizer; the composite microbial agent comprises Pseudomonas E sp017968885, Bacillus amyloliquefaciens XN-3 and Arthrobacter sp.

[0010] Preferably, the viable cell quantity ratio of the Pseudomonas, the Bacillus amyloliquefaciens and the Arthrobacter in the composite microbial agent is 2:3:1.

[0011] The application further provides a preparation method of the multifunctional microbial agent, which comprises the following steps:

[0012] After the Pseudomonas, the Bacillus amyloliquefaciens and the Arthrobacter are respectively cultured, the three are mixed in equal volume to obtain a composite microbial suspension seed solution; the composite microbial suspension seed solution is inoculated into LB culture medium for composite culture to obtain a composite microbial agent; and the composite stabilizer is added into the composite microbial agent to obtain the multifunctional microbial agent.

[0013] Preferably, the mass-volume fraction of the composite stabilizer and the composite microbial agent is 5.2%; the culture medium for culturing the Arthrobacter is R2A culture medium; and the culture medium for culturing the Pseudomonas or the Bacillus amyloliquefaciens is LB or NA culture medium.

[0014] Preferably, the initial inoculation total cell amount of the composite culture is 4x10 6 CFU / mL; the composite culture condition is 30 DEG C and 150 rpm; and the composite culture time is 48 h.

[0015] The application further provides application of the composite stabilizer in preparation of the multifunctional microbial agent.

[0016] The application further provides application of the composite stabilizer in improving the room temperature stability of the microbial agent.

[0017] The application further provides application of the multifunctional microbial agent in preparation of a fertilizer additive.

[0018] The application also provides application of the multifunctional microbial agent in sewage treatment, garbage treatment and soil pollution remediation.

[0019] The application discloses the following technical effects:

[0020] (1) The application significantly improves the stability of the microbial agent. Long-term survival rate: under the room temperature and light-free storage condition, the microbial liquid agent adopting the composite stabilizer (3% starch, 2% ascorbic acid and 0.2% xanthan gum) still maintains 1.42*10 8 CFU / mL of viable bacteria after 287 days, and the survival rate reaches 35.95%. The viable bacteria number is equivalent to the initial value (the survival rate is 100%) when the storage time is 167 days, which is much higher than the effect of the conventional protective agent. The synergistic effect of the composite stabilizer: the starch provides carbon source as a nutrient matrix, the ascorbic acid protects the cells from oxidation, and the xanthan gum enhances the solution viscosity to reduce the settlement and physical damage, and the three components are combined to significantly prolong the shelf life of the microbial agent.

[0021] (2) The viable bacteria ratio and compatibility of the composite microbial agent are optimized. The strain combination is scientific, the strain combination (Pseudomonas E sp017968885, Bacillus amyloliquefaciens XN-3 and Arthrobacter sp.) with good compatibility is selected through the single-strain compatibility test, the viable bacteria number ratio is fixed as 2:3:1, and the function complementation (such as growth promotion, disease resistance and pollutant degradation) is ensured. The culture medium adaptability is selected. The optimal culture medium is selected for different strains to culture the single-strain as the seed liquid, and then the composite microbial agent is prepared through the composite culture, the microbial population grows synergistically, and the importance of the culture medium adaptability is embodied.

[0022] (3) Wide application scenarios. Agricultural field: as a fertilizer additive, the soil micro-ecology can be improved, and the crop growth can be promoted, and the composite microbial agent is especially suitable for the areas with high stability requirements in high-temperature and high-humidity environments. Environmental protection field: the composite microbial agent performs well in sewage treatment, garbage treatment and soil pollution remediation, and the composite microbial agent has high tolerance to heavy metals and organic pollutants, and the treatment effect is stable.

[0023] The application directly aims at the problems of poor stability and single function of the existing microbial agent, and significantly improves the adaptability of the microbial agent in complex environments through the composite stabilizer and strain matching innovation, and promotes the large-scale application of the microbial technology.

[0024] The application realizes the technical breakthroughs of long-term stability and diversified functions of the microbial agent through strain optimization, stabilizer compounding and process optimization, and has the characteristics of innovation and practicability, and has a wide market prospect in the fields of agricultural sustainable development and environmental governance. BRIEF DESCRIPTION OF DRAWINGS

[0025] In order to more clearly illustrate the technical solutions in the embodiments of the present application or the prior art, the following will briefly introduce the drawings needed in the embodiments. Obviously, the drawings in the following description only constitute some embodiments of the present application, and for those skilled in the art, other drawings can also be obtained from these drawings without creative effort.

[0026] Figure 1 Single bacterial compatibility test; 1: Bacillus mobilis; 2: Pseudomonas E sp017968885; 3: Bacillus amyloliquefaciens XN-3, 4: Arthrobacter sp.;

[0027] Figure 2 Colony morphology of each strain on different culture media; 1: Bacillus mobilis; 2: Pseudomonas E sp017968885; 3: Bacillus amyloliquefaciens XN-3, 4: Arthrobacter sp.

[0028] Figure 3 Microscopic morphology of each strain; 1: Bacillus mobilis; 2: Pseudomonas E sp017968885; 3: Bacillus amyloliquefaciens XN-3, 4: Arthrobacter sp.

[0029] Figure 4 Viable cell count change during the culture process of the compound microbial inoculant; 1-18: 18 kinds of compound microbial inoculants prepared by the stabilizer formulations. DETAILED DESCRIPTION

[0030] The various illustrative embodiments of the present application will now be described in detail below, which should not be considered as limiting the present application, but should be understood as a more detailed description of some aspects, characteristics and embodiments of the present application.

[0031] It should be understood that the terms described in the present application are only for describing the specific embodiments, and are not used to limit the present application. In addition, for the numerical range in the present application, it should be understood that each intermediate value between the upper limit and the lower limit of the range is also specifically disclosed. Each smaller range between the intermediate values in any stated value or stated range, and any other stated value or intermediate value in the range is also included in the present application. The upper limit and the lower limit of these smaller ranges can be independently included or excluded from the range.

[0032] Unless otherwise indicated, all technical and scientific terms used herein have the same meaning as those commonly understood by one of ordinary skill in the art to which this application pertains. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present application, preferred methods and materials are described. All publications mentioned in this specification are herein incorporated by reference to disclose and describe the methods and / or materials in connection with which the publications are cited. In case of conflict between the content of the specification and that of any incorporated literature, the content of the specification controls.

[0033] Many modifications and variations of the present application described in the specification are possible without departing from the scope or spirit of the application. Other embodiments of the application will be apparent to those skilled in the art from consideration of the specification and practice of the application disclosed herein. The specification and examples are illustrative only.

[0034] As used herein, the terms “comprises,” “comprising,” “includes,” “including,” “has,” “having,” and the like are open-ended terms that are intended to mean including, but not limited to.

[0035] Strain 1 Bacillus mobilis has been deposited with China General Microbiological Culture Collection Center on October 24, 2024, at the address of China Beijing, with the deposit number of CGMCC No. 32327.

[0036] Strain 2 Pseudomonas E sp017968885 was isolated from the rhizosphere soil of Picris japonica Thunb. and purified, and is now preserved in the Microbial Engineering Research Room of Liaoning Provincial Microbial Science Research Institute (Strain 2 public literature: Hu Q, Wu H, Yu M, Song L, Feng J, Ma Y, Ding F, Guo L and Wang Z (2025) Optimizing the application strategy of phosphorus fertilizer by Pseudomonas Esp017968885 to increase phosphorus yield of pepper. Front. Agron. 7:1628083. doi: 10.3389 / fagro.2025.1628083), i.e. Pseudomonas E sp017968885.

[0037] Strain 3 Bacillus amyloliquefaciens XN-3 was isolated from the soil sample of the cucumber greenhouse in Liuguanyingzi Village, Kazuo County, Chaoyang City, Liaoning Province (North Latitude 41.16° East Longitude 119.68°) (Strain 3 published literature: Aojing, Li Yang, Liu Xiaohui, et al. Screening and identification of a cucumber fusarium wilt antagonistic bacterium and its preliminary study on inhibition effect [J]. Journal of Yunnan Agricultural University (Natural Science), 2022, 37(03): 429-434.), i.e. Bacillus amyloliquefaciens XN-3.

[0038] Strain 4 Arthrobacter sp. was preserved in the China General Microbiological Culture Collection Center on March 17, 2020, and the address of the preservation unit is Beijing, China, with the preservation number CGMCC No. 19486, i.e. Arthrobacter sp.

[0039] NA medium formula: beef extract proteose peptone 10.0 g; sodium chloride (NaCl) 5.0 g; agar 16.0 g (solid medium is necessary, liquid medium does not contain agar); purified water 1000 mL, final pH 7.2±0.2 (25℃).

[0040] LB medium formula: tryptone 10.0 g; yeast extract 5.0 g; sodium chloride (NaCl) 10.0 g; purified water 1000 mL, final pH 7.0±0.2 (25℃).

[0041] R2A medium formula: yeast extract powder 0.5 g; proteose peptone 0.5 g; casein hydrolysate 0.5 g; glucose 0.5 g; soluble starch 0.5 g; potassium hydrogen phosphate (K2HPO4) 0.3 g; anhydrous magnesium sulfate (MgSO4) 0.024 g; sodium pyruvate 0.3 g; agar 15.0 g (liquid medium does not need to add agar); purified water 1000 mL, final pH 7.2±0.2 (25℃).

[0042] Example 1

[0043] 1. Single bacterial compatibility test

[0044] V-shaped streaking was performed on NA, LB and R2A plates respectively in the form of two-by-two combination of the four strains to determine the compatibility of the four strains. The results are shown in Figure 1 .

[0045] As Figure 1As shown, the single bacterial compatibility test results showed that strain 1 Bacillus mobilis was not compatible with strain 2 Pseudomonas E sp017968885, strain 3 Bacillus amyloliquefaciens XN-3, but was compatible with strain 4 Arthrobacter sp., and strain 2 Pseudomonas E sp017968885, strain 3 Bacillus amyloliquefaciens XN-3, and strain 4 Arthrobacter sp. were compatible with each other, so the subsequent complex bacterial liquid test will use strains 2, 3, and 4 as test strains.

[0046] 2. Single bacterial optimal medium screening

[0047] Each strain was cultured in LB, NA, and R2A media at 37°C and 180 rpm for 24 h; OD 600 and plate count were determined, and the medium with the highest growth of each strain was selected (referred to as "optimal medium"). (4 strains x 3 media = 12 treatments, 3 replicates), the results are shown in Tables 1, 2, and 3.

[0048] Table 1 OD 600 and plate count results (LB medium)

[0049]

[0050] Table 2 OD 600 and plate count results (NA medium)

[0051]

[0052] Table 3 OD 600 and plate count results (R2A medium)

[0053]

[0054] The single bacterial optimal medium screening results showed that the optimal medium for 4. Arthrobacter sp. was R2A; 1. Bacillus mobilis, 2. Pseudomonas E sp017968885, and 3. Bacillus amyloliquefaciens XN-3 could all grow well in LB and NA media, with OD 600 values of 1.21 x 10 8 ~ 1.54 x 10 8 CFU / mL after 24 h of culture, with no significant difference (p>0.05).

[0055] 3. Complex bacterial liquid enrichment test

[0056] ① Preparation of composite inoculum: single bacteria were respectively cultured with their respective "optimal base" (the culture medium of single bacteria Arthrobacter sp. was R2A medium; the culture medium of single bacteria Bacillus mobilis, Pseudomonas E sp017968885 and Bacillus amyloliquefaciens XN-3 was LB medium, cultured for 18 h → 5000 rpm, 5 min to collect bacterial bodies → 0.85% NaCl sterile resuspension → adjusted to 1x10 8 CFU / mL → mixed in equal volume to obtain a composite bacterial suspension (Total 4x10 8 CFU / mL). The morphology of various bacteria on the selective medium is shown in Figure 2 , and the microscopic morphology of each colony is shown in Figure 3 .

[0057] ② Composite culture: 1 mL of the composite suspension was inoculated into a 250 mL flask containing 100 mL of sterilized LB, NB, R2B (initial total bacteria 4x10 6 CFU / mL).

[0058] ③ Culture conditions: 30°C, 150 rpm (reducing the rotation speed to reduce flagella shedding, and taking into account Arthrobacter growth), 48 h.

[0059] ④ Sampling and determination: sampling at 0 h, 6 h, 12 h, 24 h, 48 h. Determination: OD 600 ; total viable bacteria count (PCA count); proportion of each bacteria (selective medium + colony microscopic morphology), and the results are shown in Table 2.

[0060] Table 4 Composite bacterial liquid culture results corresponding to different culture media during composite culture

[0061]

[0062] As shown in Figure 2 , plate colony morphology observation showed that strain 4 presented consistent yellowish colonies on NA, LB and R2A media, with obvious oil gloss on the surface, which was significantly different from the color and texture of the other strains. Strains 2 and 3 were white colonies in the above three media, among which strain 3 colonies had radial wrinkles on the surface, while strain 2 surface was flat, which could be used to preliminarily distinguish between the two under most conditions.

[0063] As shown in Figure 3As shown, the microscopic morphological identification further verified that strain 2 was a bacillus with spores, and strain 3 was a common bacillus without spores, which were easily distinguished under an oil microscope. By combining the macroscopic characteristics of the colonies and the microscopic morphology of the cells, strains 2, 3, and 4 can be accurately identified at one time, and the proportion of each bacterium in the complex bacterial solution can be quickly determined.

[0064] As shown in Table 4, the most suitable culture medium for complex culture is LB.

[0065] The results of the enrichment test of the complex bacterial solution showed that after 48 h of culture, the average OD 600 value of the complex bacterial solution was 0.9, and the total number of viable bacteria was 3.5 x 10 8 CFU / mL. The ratio of strain 2: strain 3: strain 4 was 2:3:1.

[0066] 4. Determination of stabilizer formula

[0067] The types of protective agents in the complex stabilizer include A (nutritional components), B (protective agents), and C (stabilizers). A has 3 levels, B has 2 levels, and C has 3 levels, which belong to a mixed level system. The types and amounts of components in each stabilizer formula are shown in Table 5. The experimental design combinations of each stabilizer formula are shown in Table 6.

[0068] Stabilizer test results: the complex microbial bacterial solution was fermented to a viable bacterial count of 1 x 10 8 CFU / mL or more, the pH was adjusted to between 4.0 and 6.5, 1-18 types of complex stabilizers were added according to the formula in Table 6, the proportion of each component in the complex microbial bacterial solution is shown in Table 5, and the mixture was thoroughly stirred until completely dissolved. Store at room temperature away from light, detect the change in viable bacterial count at regular intervals, and store for 7 months. The viable bacterial count detection results of the 18 types of complex microbial bacterial agents prepared by different complex stabilizers are shown in Table 7 and Figure 4 . After 167 days of storage, the viable bacterial count of the complex microbial bacterial agent prepared by adding stabilizer according to formula 11 was 3 x 10 8 , which was comparable to the initial viable bacterial count, with a survival rate of 100%. After 287 days of storage, the viable bacterial count of formula 11 was 1.42 x 10 8 CFU / mL, with a survival rate of 35.95%.

[0069] Table 5: Types of protective agent components

[0070]

[0071] Table 6: Experimental design combinations

[0072]

[0073] Table 7: Viable bacterial count detection results of 18 types of complex microbial bacterial agents prepared by different complex stabilizers

[0074]

[0075] Experimental results: the protection effect of combination 11 (formula 11) is the best (formula 11) Figure 4 ), that is, the combination of formula: nutrient component 3% starch (w / v); protective agent 2% ascorbic acid (w / v); stabilizer 0.2% xanthan gum (w / v) has the best microbial agent protection effect. Therefore, the mass ratio of starch, ascorbic acid and xanthan gum in the optimal composite stabilizer is 15:10:1, and the addition amount of the composite stabilizer in the composite microbial agent is 5.2% (w / v).

[0076] The following application example uses the composite microbial agent prepared by the stabilizer formula 11 as a multifunctional microbial agent to verify the application effect.

[0077] Application example 1 Application of multifunctional microbial agent in preparation of fertilizer additive

[0078] Purpose of implementation

[0079] The multifunctional microbial agent of the application is used as a fertilizer additive to improve the soil microecological structure, promote the nutrient absorption of crops, inhibit the reproduction of soil-borne pathogenic bacteria, solve the problems of soil compaction and microbial community imbalance caused by traditional fertilizers, and improve the yield and quality of crops. It is especially suitable for continuous planting, high temperature and high humidity, and other agricultural scenes with high requirements for microbial agent stability.

[0080] Materials and methods for implementation

[0081] Preparation of microbial agent: the multifunctional microbial agent is obtained by the preparation method described in example 1, and the viable bacterial count is ≥3.5×10 8 CFU / mL, the composition of the composite stabilizer is 3% w / v starch, 2% w / v ascorbic acid, and 0.2% w / v xanthan gum, and the viable bacterial count ratio of Pseudomonas E sp017968885, Bacillus amyloliquefaciens XN-3 and Arthrobacter sp. is 2:3:1.

[0082] Fertilizer adaptation: select conventional compound fertilizer (N-P-K ratio 15-15-15) as the base fertilizer, add 5% multifunctional microbial agent according to the mass of the fertilizer, mix thoroughly, prepare the compound microbial fertilizer, and store it in the dark at room temperature for standby.

[0083] Application object and condition: Greenhouse watermelons (variety: 8424) were used as test crops, and S2 type soil with a continuous planting history of 3 years and a fusarium wilt incidence of 80% was selected as the test field. The test field area was 0.4 mu, and three groups of repetitions were set up. The control group was applied with an equal amount of conventional compound fertilizer, and the test group was applied with compound microbial fertilizer.

[0084] Application method: Before sowing, the compound microbial fertilizer was uniformly applied on the soil surface, and the tillage depth was 20 cm to mix the fertilizer with the soil fully; during the growth period, 1 time of root irrigation treatment was carried out at the seedling stage and fruit setting stage respectively, and 500 mL of diluted 100 times microbial agent stock solution was irrigated per plant.

[0085] Effect determination: After the planting period ended, the soil urease, invertase, and alkaline phosphatase activities, watermelon plant height, single fruit weight, total yield, and fusarium wilt incidence were determined.

[0086] Implementation results

[0087] Significant improvement of soil enzyme activity: The soil urease activity of the test group was increased by 38.6% compared with the control group, the invertase activity was increased by 35.1%, and the alkaline phosphatase activity was increased by 9.9%, and the soil nutrient conversion efficiency was enhanced.

[0088] Optimization of crop growth indicators: The watermelon plant height of the test group was increased by 23.5% compared with the control group, the single fruit weight was increased by 7.7%, the total yield was increased by 5.3%, and the soluble solids content of the fruit was increased by 0.8 percentage points.

[0089] Significant disease prevention and control effect: The fusarium wilt incidence of watermelons in the test group was reduced to 15.2%, which was reduced by 64.8% compared with the control group (80%).

[0090] Application example 2 Application of multifunctional microbial agent in garbage treatment

[0091] Implementation purpose

[0092] The multifunctional microbial agent is applied to the co-composting of household garbage and kitchen garbage to accelerate the degradation of organic garbage, shorten the composting cycle, reduce the emission of foul gas during composting, increase the nutrient content of compost products, and realize the resource utilization of garbage.

[0093] Implementation materials and methods

[0094] Agent preparation: The viable count of the multifunctional microbial agent is 3.5×10 8 CFU / mL, and the compound stabilizer is added to ensure that the agent remains active during the high-temperature stage (55-65℃) of composting.

[0095] Garbage raw materials: mixed household garbage and kitchen garbage (mass ratio 3:2), garbage moisture content 60%, C / N ratio 25:1, main components including kitchen organic matter (vegetable residues, rice), paper, a small amount of plastic (already sorted out), total mass 50 kg.

[0096] Composting treatment: mix garbage raw materials with straw (conditioner) at a mass ratio of 10:1, the test group adds a multifunctional microbial agent at 3% of the total garbage mass, the control group does not add the agent, both groups use strip pile composting, regular turning (once every 3 days), and the moisture content of the pile is controlled at 55-65%.

[0097] Operation monitoring: composting period 45 days, regularly detect pile temperature, pH value, C / N ratio, and concentration of foul-smelling gases (ammonia, hydrogen sulfide), after composting, detect the contents of organic matter, total nitrogen, total phosphorus, and total potassium in the product, and the number of pathogenic bacteria (E. coli).

[0098] Implementation results

[0099] Shortening of the composting period: the test group reached the high-temperature stage (≥55°C) on the 3rd day, lasting for 15 days, which is 5 days longer than the control group (high-temperature lasting for 10 days); the composting maturity period was shortened to 38 days, which is 10 days shorter than the control group (48 days).

[0100] Reduction of foul-smelling gas emissions: the average emission concentration of ammonia in the test group was 8.5 mg / m 3 , and the average emission concentration of hydrogen sulfide was 0.3 mg / m 3 , which were 62.2% and 73.9% lower than the control group, respectively, and the strains inhibited the growth of odor-producing microorganisms (such as Desulfovibrio) and reduced the generation of foul-smelling substances.

[0101] Improvement of compost quality: after maturity, the organic matter content of the test group compost was 38.6%, the total nitrogen was 2.3%, the total phosphorus was 1.2%, and the total potassium was 1.8%, which were 12.5%, 18.4%, 20.0%, and 16.1% higher than the control group, respectively; the number of E. coli was <10 3 CFU / g, which met the health standards for organic fertilizers.

[0102] The above-described embodiments are only to describe the preferred modes of the present application, and do not limit the scope of the present application, and various modifications and improvements to the technical solutions of the present application made by those skilled in the art without departing from the design spirit of the present application shall fall within the protection scope determined by the claims of the present application.

Claims

1. A multifunctional microbial inoculant composite stabilizer, characterized by, The composite stabilizer comprises three components of starch, ascorbic acid and xanthan gum; the mass ratio of the starch, ascorbic acid and xanthan gum is 15:10:1; the effective use concentration of the three components is: 3% w / v of starch, 2% w / v of ascorbic acid and 0.2% w / v of xanthan gum.

2. A multi-functional microbial inoculant, characterized by, The microbial agent comprises: a composite microbial liquid and the composite stabilizer of claim 1; the composite microorganism comprises Pseudomonas E sp017968885, Bacillus amyloliquefaciens XN-3 and Arthrobacter sp.

3. The multifunctional microbial agent of claim 2, wherein the Bacillus sp. is Bacillus subtilis. The viable cell number ratio of Pseudomonas, Bacillus amyloliquefaciens and Arthrobacter in the composite microbial agent is 2:3:

1.

4. A method for preparing a multifunctional microbial agent as described in claim 2, characterized in that, The preparation method comprises the following steps: After culturing Pseudomonas, Bacillus amyloliquefaciens and Arthrobacter respectively, the equal volume is mixed to obtain a composite bacterial suspension seed liquid; the composite bacterial suspension seed liquid is inoculated into LB culture medium for composite culture to obtain a composite microbial liquid; the composite stabilizer of claim 1 is added into the composite microbial liquid to obtain a multifunctional microbial agent.

5. The production method according to claim 4, wherein The mass-volume fraction of the composite stabilizer and the composite microbial liquid is 5.2%; the culture medium for culturing Arthrobacter is R2A culture medium; the culture medium for culturing Pseudomonas or Bacillus amyloliquefaciens is LB or NA culture medium.

6. The production method according to claim 4, wherein The initial inoculation total bacteria amount of the complex culture is 4×10 6 CFU / mL; the condition of the complex culture is 30℃, 150 rpm; and the time of the complex culture is 48 h.

7. Application of the composite stabilizer of claim 1 in preparation of a multifunctional microbial agent.

8. Application of the composite stabilizer of claim 1 in improving the room temperature stability of a microbial agent.

9. Application of the multifunctional microbial agent of claim 2 or 3 in preparation of a fertilizer additive.

10. Application of the multifunctional microbial agent of claim 2 or 3 in sewage treatment, garbage treatment and soil pollution remediation.

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