Biogas slurry composite biological agent for improving grape quality and application method thereof

Abstract

The invention relates to a biogas slurry composite biological agent for improving grape quality and an application method thereof, and belongs to the technical field of grape planting. Aiming at the problems that the direct application effect of the existing biogas slurry is unstable, the nutrients are unbalanced, and the traditional chemical fertilizer is easy to cause soil hardening and fruit quality reduction, the invention provides a technical scheme of taking the biogas slurry as a basic solution and scientifically compounding functional additives. Each liter of basic liquid of the preparation comprises 1.0-3.0 g of potassium humate, 0.5-2.0 ml of sugar alcohol calcium and 0.5-2.0 g of carboxymethyl chitosan, and at least one of 0.3-1.0 g of bacillus subtilis powder with the effective viable count being greater than or equal to 10 billion / g or 0.1-0.3 ml of a brassinolide aqueous solution with the concentration being 0.01% is selectively added. The preparation is mainly used for improving the fruit quality of grapes, and high-yield and high-quality cultivation of the grapes is realized through multiple effects of synergistically providing nutrients, promoting absorption, enhancing stress resistance and the like.

Description

Technical Field

[0001] This invention relates to the field of grape cultivation technology. More specifically, this invention relates to a biogas slurry compound biological agent for improving grape quality and its application method. Background Technology

[0002] Biogas slurry is a byproduct of biogas projects. It is rich in nitrogen, phosphorus, potassium and organic matter, but its nutrients are usually unbalanced, its direct application has unstable effects, and it may have problems such as excessive salt content and heavy metal risks, which limit its high-value utilization.

[0003] High-end grape varieties such as Shine Muscat have high requirements for fruit size, sugar content, and flavor. Traditional fertilizer application can easily lead to soil compaction and a decline in fruit quality. Although biostimulants (such as humic acid and seaweed extracts) are effective, they are expensive and their synergistic effect with the soil substrate is poor when applied alone.

[0004] Currently, there is a lack of systematic solutions for scientifically combining inexpensive biogas slurry with various functional additives and precisely controlling its growth during key stages of grape development. Most studies focus only on the effects of a single additive or a single period. Summary of the Invention

[0005] One object of the present invention is to solve at least the above-mentioned problems and to provide at least the advantages that will be described later.

[0006] To achieve these objectives and other advantages according to the present invention, a biogas slurry compound biological agent for improving grape quality is provided, with biogas slurry as the base liquid, wherein each liter of the base liquid contains the following active ingredients: potassium humate: 1.0~3.0 g; calcium sugar alcohol: 0.5~2.0 ml; carboxymethyl chitosan: 0.5~2.0 g; at least one of Bacillus subtilis powder and brassinolide aqueous solution, wherein the effective viable count of Bacillus subtilis powder is ≥10 billion / g, and its addition amount is 0.3~1.0 g per liter of base liquid; the concentration of brassinolide aqueous solution is 0.01%, and its addition amount is 0.1~0.3 ml per liter of base liquid.

[0007] Preferably, it also includes seaweed extract, which is added at a concentration of 0 to 1.0 g / L of base solution.

[0008] Preferably, the amount of potassium humate used is 2.0 g, based on a purity of 50%; the amount of sugar alcohol calcium used is 1.0 ml; and the amount of carboxymethyl chitosan used is 1.0 g, based on a purity of 90%. The amount of Bacillus subtilis powder added is 0.6 g per liter of base solution; the amount of brassinolide aqueous solution added is 0.2 ml per liter of base solution. The seaweed extract is added at a rate of 1.0 g / L of base solution during the grape veraison stage.

[0009] Preferably, the potassium humate, calcium sugar alcohol, and carboxymethyl chitosan are pre-formed into solid micronuclei with a particle size of 0.5~1.5mm using a fluidized bed granulation process, wherein the mass ratio of potassium humate to carboxymethyl chitosan is (1.8~2.2):1, and the granulation binder is a solution of brassinolide aqueous solution diluted 10-20 times. The Bacillus subtilis powder and seaweed extract are co-loaded on a porous silica nanocarrier to form a functional composite. The porous silica nanocarrier has a pore size of 8-15 nm and a specific surface area ≥500 m². 2 / g; The base liquid undergoes two stages of treatment before use: first, it is treated for 3-5 minutes in a pulsed magnetic field with a frequency of 15-25 kHz and a field strength of 0.8-1.2 T, and then treated with a wavelength of 365 nm and a power density of 50-80 mW / cm². 2 The solution was subjected to circulatory treatment under ultraviolet light for 20-30 minutes. The base liquid after two-stage treatment is injected into the reactor. At a stirring speed of 200-300 rpm, the solid micronuclei are added first and dispersed for 10 minutes. Then the functional complex is added and stirring is continued for 15-20 minutes to form a stable suspension system.

[0010] Preferably, while adding the functional complex and continuing to stir, an aqueous solution of a coating stabilizer composed of phospholipids and soluble soybean protein is sprayed into the suspension system in an atomized form at a dosage of 0.1-0.3 g per liter of suspension system; the aqueous solution of the coating stabilizer is an aqueous solution containing 5% coating stabilizer by mass, and the mass ratio of phospholipids to soluble soybean protein in the coating stabilizer is 1:(1.5-2.5); after spraying, the temperature of the suspension system is uniformly reduced to 15-20°C within 5-8 minutes, and stirred at 100-150 rpm for 10-15 minutes at this temperature, finally obtaining the biogas slurry compound biological agent.

[0011] Preferably, the coating stabilizer further includes 8-15% by mass of succinylated pectin, which is prepared by the following method: high ester pectin is dispersed in anhydrous pyridine, and a solution containing succinic anhydride is slowly added under nitrogen protection and stirring at 60°C. The reaction is carried out for 6 hours. After the reaction is completed, the mixture is poured into ice-cold ethanol to precipitate, filtered, washed several times with ethanol, and dried under vacuum to obtain succinylated pectin.

[0012] Preferably, the base liquid originates from an anaerobic fermentation process using grape winemaking waste as fermentation raw material, and must meet the following specific indicators before use: a) Chemical oxygen demand is 15,000~25,000 mg / L; b) The ammonium nitrogen content is 250~350 mg / L; c) Electrical conductivity ≤ 4.0 mS / cm; d) The pH value at the end of anaerobic fermentation is stable at 7.0~7.5, and the mixture has been left to stand for aging for no less than 15 days.

[0013] This invention also provides a method for fertilizing grapes using the above-mentioned biogas slurry compound biological agent, comprising the following steps: The total annual amount of biogas slurry is distributed among the five key growth stages: 25% during the germination stage, 20% during the flowering stage, 35% during the fruit enlargement stage, 10% during the color change stage, and 10% during the post-harvest recovery stage. Specifically, the germination stage is applied 3 times, the flowering stage is applied 2 times, the fruit enlargement stage is applied 3 times, the color change stage is applied once, and the post-harvest recovery stage is applied once. Before each fertilization, the biogas slurry compound biological agent is prepared according to the single application amount of biogas slurry corresponding to the growth stage, and then applied to the grape root zone; When applying the biogas slurry compound biological agent during the color-changing period, seaweed extract is added. At the same time as applying the biogas slurry compound biological agent, potassium sulfate is applied as a basic fertilizer at a rate of 8 kg per mu.

[0014] This invention includes at least the following beneficial effects: The biogas slurry compound biological agent is based on biogas slurry and is formulated by adding potassium humate, sugar alcohol calcium, carboxymethyl chitosan, Bacillus subtilis powder, and / or brassinolide aqueous solution. The biogas slurry provides basic nutrients and organic matter; potassium humate promotes root growth and water retention; sugar alcohol calcium supplements calcium and prevents cracking; carboxymethyl chitosan induces resistance and improves quality; and seaweed extract promotes color change and increases sugar content. The addition of Bacillus subtilis or brassinolide further enhances the system's stress resistance and nutrient absorption efficiency from the perspective of microbial regulation or endogenous hormones.

[0015] Regarding the protection of active ingredients, by embedding bioactive substances such as Bacillus subtilis and brassinolide into solid micronuclei or anchoring them within nanocarriers, they are effectively isolated from potential inhibitors or harmful substances in biogas slurry. This design significantly improves the retention rate of bioactivity in complex biogas slurry environments. It has been proven that the survival rate of Bacillus subtilis and the active half-life of brassinolide are both increased several times, thus ensuring the stable and reliable bioefficacy of the formulation when applied in the field and solving the core problem of easy inactivation of active ingredients.

[0016] Simultaneously, by subjecting the biogas slurry base liquid to two-stage synergistic treatment using pulsed magnetic field and ultraviolet light, its colloidal state and redox environment were effectively optimized, transforming it into a homogeneous and stable carrier medium. Combining the solid micronuclei formed by fluidized bed granulation with the functional composite loaded with a nanoporous carrier, a structured formulation system was constructed. This system maintains a uniform suspension state during long-term storage, is not prone to stratification or sedimentation, and greatly improves the product's shelf life and ease of application.

[0017] By introducing environmentally responsive materials such as succinylated pectin to construct a smart coating, functional complexes loaded with active ingredients can respond to pectinase secreted by roots in the rhizosphere region, achieving targeted and timely release. This root-to-root release characteristic greatly improves the utilization efficiency of nutrients and biostimulants, reduces loss, and achieves a precise balance between protection and release.

[0018] Field application results show that this invention can systematically improve grape yield and fruit quality. Combined with a precise fertilization program designed according to the nutrient requirements of grapes during their key growth stages, this formulation effectively promotes vine growth, enhances leaf photosynthetic capacity, improves the sugar-acid ratio, color, and firmness of the fruit, and increases the content of nutrients such as vitamin C. Simultaneously, its effects on promoting root development and improving the rhizosphere microecology enhance the grapevine's resistance to stress and disease, achieving a synergistic improvement in high yield, quality, and health.

[0019] Furthermore, this invention uses biogas slurry produced from the fermentation of grape winemaking waste as the main raw material, achieving efficient resource recycling of agricultural waste and reducing production costs. The entire technical solution deeply integrates materials science, biotechnology, and agronomic practices, forming a stable, efficient, and environmentally friendly green fertilizer product and technology model, providing strong technical support for improving the efficiency and sustainable development of the grape industry.

[0020] Other advantages, objectives and features of the present invention will become apparent in part from the following description, and in part from those skilled in the art through study and practice of the invention. Attached Figure Description

[0021] Figure 1 This is a schematic diagram of the release characteristics described in Test Example 2 of the present invention. Detailed Implementation

[0022] The present invention will be further described in detail below with reference to embodiments, so that those skilled in the art can implement it based on the description.

[0023] It should be noted that, unless otherwise specified, the experimental methods described in the following implementation plan are all conventional methods, and the reagents and materials described are all commercially available unless otherwise specified.

[0024] Example 1: Preparation of biogas slurry compound biological agent Take the biogas slurry after it has been fully fermented in the biogas project and filter it to remove large particulate impurities. Add the following per liter of biogas slurry: 2.0g potassium humate (50% purity), 1.0ml sugar alcohol calcium, and 1.0g carboxymethyl chitosan (90% purity). Stir mechanically for 10 minutes until well mixed. Before fertilization, add 0.6g of Bacillus subtilis powder (effective viable count ≥10 billion / g) per liter of biogas slurry to the above mixture and stir gently; For formulations used during the color-changing period, an additional 1.0g of seaweed extract per liter of biogas slurry needs to be added.

[0025] The biogas slurry originates from an anaerobic fermentation process using grape winemaking waste as fermentation raw material, and must meet the following specific indicators before use: a) Chemical oxygen demand (COD) is 15,000~25,000 mg / L; b) Ammonium nitrogen (NH4) + The content of -N) is 250~350 mg / L; c) Electrical conductivity (EC value) ≤ 4.0 mS / cm; d) The pH value at the end of anaerobic fermentation is stable at 7.0~7.5, and the mixture has been left to stand for aging for no less than 15 days.

[0026] Example 2: Preparation of biogas slurry compound biological agent The biogas slurry, obtained after full fermentation in the biogas project, is filtered to remove large particulate impurities. This biogas slurry originates from an anaerobic fermentation process using grape winemaking waste as fermentation material and must meet the following specific indicators before use: a) Chemical oxygen demand (COD) is 15,000~25,000 mg / L; b) Ammonium nitrogen (NH4) + The content of -N) is 250~350 mg / L; c) Electrical conductivity (EC value) ≤ 4.0 mS / cm; d) The pH value at the end of anaerobic fermentation is stable at 7.0~7.5, and the mixture has been left to stand for aging for no less than 15 days.

[0027] Based on the dosage of biogas slurry, measure the following per liter: 2.0g potassium humate (50% purity), 1.0ml sugar alcohol calcium, 1.0g carboxymethyl chitosan (90% purity), 0.6g Bacillus subtilis powder (effective viable count ≥10 billion / g), and 0.2ml brassinolide aqueous solution (concentration 0.01%). For formulations used during the color-changing period, an additional 1.0g of seaweed extract per liter of biogas slurry is required.

[0028] The potassium humate, calcium sugar alcohol, and carboxymethyl chitosan are pre-formed into solid micronuclei with a particle size of 1 mm using a fluidized bed granulation process. The granulation binder is a solution of brassinolide aqueous solution diluted 10 times.

[0029] The Bacillus subtilis powder and seaweed extract were co-loaded onto a porous silica nanocarrier to form a functional composite. The preparation process of the functional composite is as follows: First, the porous silica nanocarrier (pore size 8~15 nm, specific surface area ≥500 m²) was loaded onto the porous silica nanocarrier. 2 The silica nanocarrier ( / g) was calcined at 350℃ for 2 hours for activation pretreatment. After cooling, it was mixed with a pre-prepared 10 mg / mL seaweed extract PBS solution at a mass ratio of 10:1 (the porous silica nanocarrier to the seaweed extract in the PBS solution). The mixture was shaken and adsorbed at 25℃ and 150 rpm for 4 hours. The solid was collected by centrifugation and washed to obtain the seaweed extract pre-loaded carrier. Subsequently, Bacillus subtilis powder and an appropriate amount of PBS buffer were added to the above moistened carrier. The mixture was gently stirred into a uniform paste under sterile conditions. After pre-freezing at -40℃ for 2 hours, it was freeze-dried at -40℃ and a vacuum degree ≤10 Pa for 24 hours. Finally, the dried solid was lightly ground and passed through a 200-mesh sieve to obtain the porous silica nanocarrier functional composite co-loaded with Bacillus subtilis and seaweed extract.

[0030] The biogas slurry undergoes two stages of treatment before use: first, it is treated for 4 minutes in a pulsed magnetic field with a frequency of 20kHz and a field strength of 1T, and then treated with a pulsed magnetic field with a wavelength of 365nm and a power density of 70mW / cm³. 2 The solution was subjected to circulatory treatment under ultraviolet light for 20 minutes. The biogas slurry after two-stage treatment is injected into the reactor. At a stirring speed of 250 rpm, the solid micronuclei are added first and dispersed for 10 minutes. Then, the functional complex is added and stirring is continued for 20 minutes to form a stable suspension system.

[0031] Example 3: Preparation of biogas slurry compound biological agent Based on Example 2, while adding the functional complex to the suspension system and continuing to stir, an aqueous solution of a coating stabilizer composed of phospholipids and soluble soybean protein was sprayed into the suspension system in an atomized form at a dosage of 0.2 g per liter of suspension system. The aqueous solution of the coating stabilizer was an aqueous solution containing 5% coating stabilizer by mass, and the mass ratio of phospholipids to soluble soybean protein in the coating stabilizer was 1:2. After spraying, the temperature of the suspension system was uniformly reduced to 15°C within 6 minutes, and stirred at 150 rpm for 10 minutes at this temperature to finally obtain the biogas slurry compound biological agent.

[0032] Example 4: Preparation of biogas slurry compound biological agent Based on Example 3, the coating stabilizer further includes 10% by mass of succinylated pectin, which is prepared by the following method: 10g of high-ester pectin is dispersed in 100mL of anhydrous pyridine, and under nitrogen protection and stirring at 60°C, a solution containing 5g of succinic anhydride is slowly added. The reaction is carried out for 6 hours. After the reaction is completed, the mixture is poured into ice-cold ethanol to precipitate, filtered, washed several times with ethanol, and vacuum dried to obtain succinylated pectin.

[0033] Test Example 1: Verifying the effect of different preparation processes on the stability and retention rate of active ingredients in biogas slurry compound biological agents. Compound biogas slurry biological agents were prepared according to Example 1 (T1) and Example 2 (T2) respectively, and the original biogas slurry (CK) without any compounding treatment was taken as a control.

[0034] Detection indicators and methods: All samples were stored in a constant temperature incubator at 25±1℃ in the dark. Samples were taken for testing on day 0 and day 30 of storage.

[0035] Physical stability: ① Visually observe the precipitation and stratification; ② Use a laser particle size and zeta potential analyzer (equipment: Malvern Zetasizer Nano ZS90, Malvern, UK) to determine the zeta potential and particle size distribution of the sample (characterized by D90 value).

[0036] Bioactivity retention: ① Bacillus subtilis viable cell count: The dilution plating method (culture medium: nutrient agar) was used, and the cells were counted after incubation at 30℃ for 48 hours to calculate the viability. ② Brassinolide content determination: High performance liquid chromatography (HPLC) was used (equipment: Agilent 1260 Infinity II; column: C18 column; mobile phase: methanol-water; detection wavelength: 210 nm), and the retention rate was calculated using the external standard method.

[0037] Results and Analysis: The physical stability results are shown in Table 1. After 30 days of storage, obvious precipitation occurred in the CK and T1 treatments, while the T2 treatment remained in a uniform suspension. The zeta potential and particle size distribution results (Table 1) show that the T2 formulation, which has undergone two-stage treatment and structural design, has a significantly higher absolute value of zeta potential, finer particles, and a more uniform distribution, confirming that its colloidal stability is far superior to that of the simply mixed T1.

[0038] Table 1. Physical stability indicators of each treatment formulation after 30 days of storage Note: Different letters after the data in the same column indicate that the differences are significant at the P<0.05 level according to Duncan's new multiple range test, and the same applies below.

[0039] The results of bioactivity retention are shown in Table 2. After 30 days of storage, the viable bacterial survival rate of T2 (structured preparation) was significantly higher than that of T1 (simple mixing). This demonstrates that the structured process, including two-stage treatment and loading with micronuclei and nanocarriers, has an excellent effect on protecting microbial activity and preventing rapid inactivation in complex biogas slurry environments. The brassinolide retention rate of T2, as high as 82.6%, indicates that this structured process can also effectively protect the activity of plant growth regulators.

[0040] Test Example 2: Verifying the effect of coating stabilization process on formulation storage stability and smart release characteristics Biogas slurry compound biological agents were prepared according to Example 3 (T3) and Example 4 (T4) respectively, and the biogas slurry compound biological agent prepared in Example 2 (T2) was used as a control.

[0041] Detection indicators and methods: Storage stability: Samples T2 (uncoated), T3, and T4 were stored for 15 days in a constant temperature incubator at 30±1℃. The polydispersity index before and after storage was determined using a particle size and zeta potential analyzer.

[0042] Intelligent Response Release Characteristics: To visually verify the release behavior, a water-soluble fluorescent tracer (sodium fluorescein) was pre-loaded onto the functional complex. Equal volumes of the T3 and T4 finished formulations were placed in: A. deionized water; B. phosphate buffer (pH 5.5) containing pectinase (1.0 U / mL, Sigma Pectinase). The mixture was shaken at 25°C and 100 rpm, and samples were taken at 0, 2, 6, 12, 24, and 48 hours. The fluorescence intensity of the supernatant was measured using a fluorescence spectrophotometer (equipment: Hitachi F-7000) to calculate the cumulative release rate.

[0043] Results and Analysis: The storage stability results are shown in Table 3. After accelerated storage, the polydispersity index of T3 and T4 was significantly lower than that of the uncoated T2, indicating that the coating process effectively inhibited particle aggregation. Among them, the T4 system containing succinylated pectin showed the best homogeneity.

[0044] Table 3. Polydispersity index of each treatment formulation after accelerated storage (30°C, 15 days) Release characteristics results as follows Figure 1As shown, in deionized water (environment A), the release curves of T3 and T4 were similar, with cumulative release rates of less than 25% after 48 hours, exhibiting good sustained-release characteristics. In the pectinase-containing solution (environment B, simulating rhizosphere), T4 showed significant enzyme-triggered release behavior: a release inflection point appeared at 12-24 hours, and the cumulative release rate reached 78.5% after 48 hours, significantly higher than the release rate of T3 (45.2%) under the same environment. This demonstrates that the introduction of succinylated pectin endows the coated rhizosphere with intelligent release function in response to enzymes.

[0045] Example 5: Field application effect verification (taking "Sunshine Rose" grape as an example) Vineyards with consistent soil conditions were selected, and a randomized block design was adopted with 6 treatments: Example 1 (T1), Example 2 (T2), Example 3 (T3), Example 4 (T4), single biogas slurry (T5), and clean water (CK).

[0046] Field management: Plant spacing 2m × 3m; Prepare 5 tons of biogas slurry per mu (approximately 45 kg / plant) throughout the year. Calculate the dosage per plant at each stage according to the following proportions: 25% during germination, 20% during flowering, 35% during fruit enlargement, 10% during color change, and 10% during post-harvest recovery. The dosages are: germination stage: 45 kg / plant × 25% ÷ 3 times = 3.75 kg / plant / time; flowering stage: 45 kg / plant × 20% ÷ 2 times = 4.5 kg / plant / time; fruit enlargement stage: 45 kg / plant × 35% ÷ 3 times = 5.25 kg / plant / time; color change stage: 45 kg / plant × 10% ÷ 1 time = 4.5 kg / plant / time; post-harvest recovery stage: 45 kg / plant × 10% ÷ 1 time = 4.5 kg / plant / time. Before each fertilization, the biogas slurry compound biological agent is prepared according to the single application amount of biogas slurry corresponding to the growth stage, and then applied to the grape root zone; When applying the biogas slurry compound biological agent during the color-changing period, seaweed extract is added. Each time the biogas slurry compound biological agent is applied as a top dressing, potassium sulfate is also applied as a base fertilizer at a rate of 8 kg per acre (approximately 70 g / plant).

[0047] Test items and methods: At the fruit ripening stage (110 days after full bloom), 10 clusters of fruit were randomly collected from each replicate for each treatment and their quality was determined.

[0048] Single fruit weight and bunch weight: Weighed using an electronic balance (accuracy 0.01g).

[0049] Soluble solids content: determined using a handheld digital refractometer (equipment: PAL-1, Atago, Japan).

[0050] Fruit firmness: Measured using a fruit firmness tester (equipment: GY-4, Zhejiang Top).

[0051] Vitamin C content: determined by 2,6-dichlorophenolindophenol titration.

[0052] New shoot growth and diseases: Measure the thickness of the internodes in the middle of spring shoots; investigate the incidence of downy mildew.

[0053] Results and Analysis: The field application effects are shown in Table 4. The results show that the treatments (T1-T4) using the biogas slurry compound biological agent of this invention are significantly better than the single biogas slurry treatment (T5) and the water control (CK) in terms of grape fruit quality and tree growth indicators. Furthermore, the effects show an increasing trend with the upgrading of the preparation process and the addition of key technologies (from T1 to T4).

[0054] Table 4. Effects of different treatments on fruit quality and vine growth of Shine Muscat grapes The T4 treatment showed the most outstanding effect, with the soluble solids content of the fruit increasing by 3.9°Brix and 2.7°Brix compared to CK and T5, respectively, and the incidence of downy mildew decreasing by about 60% and 50%, respectively, demonstrating significant overall advantages.

[0055] Field trials fully validated the practical application value of the complete technical solution of this invention (from specific biogas slurry raw materials and multi-level structured formulation preparation to precision fertilization methods). Results showed that this invention not only significantly and stably increases grape yield and core quality indicators but also enhances vine robustness and disease resistance. The progressive improvement in effect from T1 to T4 clearly demonstrates a substantial synergistic effect, ultimately achieving optimal agronomic results through the application method.

[0056] Although embodiments of the present invention have been disclosed above, they are not limited to the applications listed in the specification and embodiments. They can be applied to various fields suitable for the present invention. For those skilled in the art, other modifications can be easily made. Therefore, without departing from the general concept defined by the claims and their equivalents, the present invention is not limited to the specific details and embodiments shown and described herein.

Claims

1. A biogas slurry compound biological agent for improving grape quality, characterized in that, Using biogas slurry as the base liquid, each liter of the base liquid contains the following effective ingredients: potassium humate: 1.0~3.0 g; sugar alcohol calcium: 0.5~2.0 ml; carboxymethyl chitosan: 0.5~2.0 g; at least one of Bacillus subtilis powder and brassinolide aqueous solution, wherein the effective viable count of Bacillus subtilis powder is ≥10 billion / g, and its addition amount is 0.3~1.0 g per liter of base liquid; the concentration of brassinolide aqueous solution is 0.01%, and its addition amount is 0.1~0.3 ml per liter of base liquid.

2. The biogas slurry compound biological agent as described in claim 1, characterized in that, It also includes seaweed extract, which is added at a rate of 0 to 1.0 g / L of base solution.

3. The biogas slurry compound biological agent as described in claim 2, characterized in that, The amount of potassium humate used is 2.0 g, based on a purity of 50%; the amount of sugar alcohol calcium used is 1.0 ml; the amount of carboxymethyl chitosan used is 1.0 g, based on a purity of 90%. The amount of Bacillus subtilis powder added is 0.6 g per liter of base solution; the amount of brassinolide aqueous solution added is 0.2 ml per liter of base solution. The seaweed extract is added at a rate of 1.0 g / L of base solution during the grape veraison stage.

4. The biogas slurry compound biological agent as described in claim 3, characterized in that, The potassium humate, calcium sugar alcohol, and carboxymethyl chitosan are pre-formed into solid micronuclei with a particle size of 0.5~1.5mm using a fluidized bed granulation process, wherein the mass ratio of potassium humate to carboxymethyl chitosan is (1.8~2.2):1, and the granulation binder is a solution of brassinolide aqueous solution diluted 10-20 times. The Bacillus subtilis powder and seaweed extract are co-loaded on a porous silica nanocarrier to form a functional composite. The porous silica nanocarrier has a pore size of 8-15 nm and a specific surface area ≥500 m². 2 / g; The base liquid undergoes two stages of treatment before use: first, it is treated for 3-5 minutes in a pulsed magnetic field with a frequency of 15-25 kHz and a field strength of 0.8-1.2 T, and then treated with a wavelength of 365 nm and a power density of 50-80 mW / cm². 2 The solution was subjected to circulatory treatment under ultraviolet light for 20-30 minutes. The base liquid after two-stage treatment is injected into the reactor. At a stirring speed of 200-300 rpm, the solid micronuclei are added first and dispersed for 10 minutes. Then the functional complex is added and stirring is continued for 15-20 minutes to form a stable suspension system.

5. The biogas slurry compound biological agent as described in claim 4, characterized in that, While adding the functional complex and continuing to stir, an aqueous solution of a coating stabilizer composed of phospholipids and soluble soybean protein is sprayed into the suspension system in an atomized form at a dosage of 0.1-0.3 g per liter of suspension system. The aqueous solution of the coating stabilizer is an aqueous solution containing 5% coating stabilizer by mass, and the mass ratio of phospholipids to soluble soybean protein in the coating stabilizer is 1:(1.5-2.5). After spraying, the temperature of the suspension system is uniformly reduced to 15-20°C within 5-8 minutes, and stirred at 100-150 rpm for 10-15 minutes at this temperature to finally obtain the biogas slurry compound biological agent.

6. The biogas slurry compound biological agent as described in claim 4, characterized in that, The coating stabilizer also includes 8-15% by mass of succinylated pectin, which is prepared by the following method: high ester pectin is dispersed in anhydrous pyridine, and a solution containing succinic anhydride is slowly added under nitrogen protection and stirring at 60°C. The reaction is carried out for 6 hours. After the reaction is completed, the mixture is poured into ice-cold ethanol to precipitate, filtered, washed several times with ethanol, and dried under vacuum to obtain succinylated pectin.

7. The biogas slurry compound biological agent as described in claim 1, characterized in that, The base liquid is derived from an anaerobic fermentation process using grape winemaking waste as fermentation raw material, and must meet the following specific indicators before use: a) Chemical oxygen demand is 15,000~25,000 mg / L; b) The ammonium nitrogen content is 250~350 mg / L; c) Electrical conductivity ≤ 4.0 mS / cm; d) The pH value at the end of anaerobic fermentation is stable at 7.0~7.5, and the mixture has been left to stand for aging for no less than 15 days.

8. A method for fertilizing grapes using a biogas slurry compound biological agent as described in any one of claims 1 to 7, characterized in that, Includes the following steps: The total annual amount of biogas slurry is distributed among the five key growth stages: 25% during the germination stage, 20% during the flowering stage, 35% during the fruit enlargement stage, 10% during the color change stage, and 10% during the post-harvest recovery stage. Specifically, the germination stage is applied 3 times, the flowering stage is applied 2 times, the fruit enlargement stage is applied 3 times, the color change stage is applied once, and the post-harvest recovery stage is applied once. Before each fertilization, the biogas slurry compound biological agent is prepared according to the single application amount of biogas slurry corresponding to the growth stage, and then applied to the grape root zone; When applying the biogas slurry compound biological agent during the color-changing period, seaweed extract is added. At the same time as applying the biogas slurry compound biological agent, potassium sulfate is applied as a basic fertilizer at a rate of 8 kg per mu.

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