A special bio-fertilizer for traditional Chinese medicinal materials based on rhizosphere microbiome remodeling

Abstract

The application discloses a special bio-fertilizer for Chinese medicinal materials based on rhizosphere microbiome remodeling, and belongs to the technical field of bio-fertilizers.The special bio-fertilizer for Chinese medicinal materials comprises the following raw materials in parts by weight: 0.2-3 parts of a fermented product of an aboveground part of Astragalus membranaceus and 2-8 parts of an enzymatic hydrolysate; the fermented product of the aboveground part of Astragalus membranaceus is obtained by fermenting the aboveground part of Astragalus membranaceus with Saccharomyces cerevisiae and Bacillus pumilus; and the enzymatic hydrolysate is obtained by enzymatically hydrolyzing soybean meal, bone meal and seaweed powder with a composite enzyme. It is verified through experiments that the special bio-fertilizer for Chinese medicinal materials has the effects of promoting rhizosphere microbiome remodeling of Astragalus membranaceus, promoting the growth of Astragalus membranaceus and improving the quality of Astragalus membranaceus.

Description

Technical Field

[0001] This invention belongs to the field of bio-fertilizer technology, and in particular relates to a bio-fertilizer for Chinese medicinal herbs based on rhizosphere microbiome remodeling. Background Technology

[0002] Medicinal herbs are the core raw materials of the traditional Chinese medicine industry, and their quality directly determines the efficacy and clinical treatment of Chinese medicines. The authenticity of medicinal herbs is the essence of their quality, and its formation is closely related to specific ecological environments, cultivation practices, and rhizosphere microecology. Currently, the cultivation of medicinal herbs is plagued by problems such as the overuse of chemical fertilizers and continuous cropping, leading to the deterioration of soil physicochemical properties, an imbalance in the rhizosphere microbial community structure, a decrease in the abundance of beneficial microorganisms, and a large accumulation of pathogenic microorganisms, resulting in frequent soil-borne diseases. This also leads to a decline in the content of effective components in medicinal herbs and a distortion of their authenticity, severely restricting the development of the medicinal herb industry.

[0003] The rhizosphere is a crucial region for the interaction between the roots and soil of medicinal herbs. The rhizosphere microbiome, acting as an invisible companion to the growth of these herbs, participates in multiple processes, including soil nutrient transformation, pest and disease control, plant growth regulation, and the synthesis of secondary metabolites, thus having a decisive impact on the growth and quality of medicinal herbs. Existing research indicates that the rhizosphere microbial communities of medicinal herbs from authentic producing areas possess unique structural characteristics, enriched with a large number of functional microorganisms related to the synthesis of active ingredients. Continuous cropping and overuse of chemical fertilizers can disrupt this unique microbial community structure, leading to the degradation of the quality of medicinal herbs.

[0004] Currently, most existing fertilizers specifically for Chinese medicinal herbs are based on single nutrient supplements or simple microbial agents, lacking the ability to target and regulate the rhizosphere microbiome, and thus failing to fundamentally solve the problem of rhizosphere microecological imbalance. While some bio-fertilizers contain functional bacteria, they suffer from drawbacks such as low bacterial survival rates, poor compatibility with the rhizosphere of Chinese medicinal herbs, and unclear remodeling effects. Therefore, developing a specialized bio-fertilizer capable of precisely remodeling the rhizosphere microbiome of Chinese medicinal herbs and improving their growth performance and quality has significant practical implications and application value. Summary of the Invention

[0005] In view of this, the purpose of this invention is to provide a special bio-fertilizer for Chinese medicinal materials based on rhizosphere microbiome remodeling, which promotes rhizosphere microbiome remodeling of Astragalus membranaceus, promotes Astragalus membranaceus growth and improves Astragalus membranaceus quality.

[0006] To achieve the above-mentioned objectives, the present invention provides the following technical solution: This invention provides a bio-fertilizer for Chinese medicinal herbs based on rhizosphere microbiome remodeling, comprising the following raw materials in parts by weight: 0.2-3 parts of fermented astragalus aerial parts and 2-8 parts of enzymatic hydrolysate; the fermented astragalus aerial parts are obtained by fermenting the astragalus aerial parts with brewer's yeast and Bacillus pumilus; the enzymatic hydrolysate is obtained by enzymatic hydrolysis of soybean meal, bone meal and seaweed powder with a compound enzyme.

[0007] Preferably, the mass ratio of the brewer's yeast to Bacillus pumilus is 1:0.5-2.5.

[0008] Preferably, the mass ratio of soybean meal, bone meal and seaweed powder is (1-3):(0.5-1.5):(0.5-1.5).

[0009] Preferably, the complex enzyme comprises neutral protease, cellulase and pectinase, wherein the mass ratio of neutral protease:cellulase:pectinase is (1-3):(0.5-1.5):(0.5-1.5).

[0010] This invention provides a method for preparing the bio-fertilizer specifically for Chinese medicinal materials, comprising the following steps: mixing the above-ground parts of Astragalus membranaceus with water, adding Saccharomyces cerevisiae and Bacillus pumilus for fermentation, sterilizing, and drying to obtain the fermented product of the above-ground parts of Astragalus membranaceus; mixing soybean meal, bone meal, and seaweed powder with water, adding a compound enzyme for enzymatic hydrolysis, inactivating, and drying to obtain the enzymatic hydrolysate; mixing the fermented product of the above-ground parts of Astragalus membranaceus with the enzymatic hydrolysate to obtain the bio-fertilizer specifically for Chinese medicinal materials.

[0011] Preferably, the mass-to-volume ratio of the aerial parts of Astragalus membranaceus to water is 1g:8-12mL.

[0012] Preferably, the inoculation amount of the brewing yeast and Bacillus pumilus is 2.5-5.5% of the mass of the aerial parts of Astragalus membranaceus; the fermentation temperature is 30-35℃ and the time is 2-6 days.

[0013] Preferably, the amount of the compound enzyme added is 1.5-3.5% of the total mass of soybean meal, bone meal, and seaweed powder; the enzymatic hydrolysis temperature is 45-50℃ and the time is 3-7h.

[0014] This invention provides the application of the bio-fertilizer specifically for Chinese medicinal materials or the bio-fertilizer specifically for Chinese medicinal materials prepared by the preparation method in reshaping the rhizosphere microbial community of plants.

[0015] This invention provides the application of the bio-fertilizer specifically for Chinese medicinal materials or the bio-fertilizer specifically for Chinese medicinal materials prepared by the preparation method in promoting the growth of Chinese medicinal materials and improving the quality of Chinese medicinal materials.

[0016] Compared with the prior art, the beneficial effects of the present invention are as follows: This invention uses the above-ground waste of Astragalus membranaceus after harvest as the core raw material. Through a process of directional fermentation with compound microbial agents, synergistic enzymatic hydrolysis with multiple enzymes, and precise compounding ratios, it achieves three core advantages: First, it completes the high-value resource utilization of Astragalus membranaceus waste, reducing both raw material costs and environmental pollution; second, through fermentation and enzymatic hydrolysis, it transforms macromolecular substances in the raw materials into small-molecule bioactive substances, significantly improving the fertilizer's speed and utilization rate, while simultaneously promoting the proliferation of beneficial bacteria in the Astragalus membranaceus rhizosphere and inhibiting pathogenic bacteria, thus reshaping the rhizosphere microecology; third, the appropriate compounding of fermented and hydrolyzed products achieves synergistic effects in microecological regulation and rapid nutrient supply, specifically addressing the obstacles of continuous cropping of Astragalus membranaceus, significantly increasing its yield and content of effective components, and combining economic, ecological, and production benefits. Attached Figure Description

[0017] Figure 1 This image shows different stages of Astragalus membranaceus cultivation. From top to bottom, the images represent the sowing stage, seedling stage, and flowering stage. Detailed Implementation

[0018] This invention provides a bio-fertilizer specifically for traditional Chinese medicine based on rhizosphere microbiome remodeling, comprising the following raw materials in parts by weight: 0.2-3 parts of Astragalus membranaceus aerial part fermentation product and 2-8 parts of enzymatic hydrolysate; the Astragalus membranaceus aerial part fermentation product is obtained by fermenting the aerial parts of Astragalus membranaceus with Saccharomyces cerevisiae and Bacillus pumilus; the enzymatic hydrolysate is obtained by enzymatic hydrolysis of soybean meal, bone meal, and seaweed powder with a compound enzyme. Preferably, this invention uses 0.5-2.5 parts of Astragalus membranaceus aerial part fermentation product and 3-7 parts of enzymatic hydrolysate, and more preferably 1 part of Astragalus membranaceus aerial part fermentation product and 5 parts of enzymatic hydrolysate. Unless otherwise specified, the Saccharomyces cerevisiae and Bacillus pumilus mentioned in this invention can be obtained from commercial channels. Preferably, the Saccharomyces cerevisiae has a viable count of 10 billion CFU / g and is purchased from Shandong Yihao Biotechnology Co., Ltd., and preferably, the Bacillus pumilus has a viable count of 50 billion CFU / g and is purchased from Hubei Yihao Biotechnology Co., Ltd.

[0019] In this invention, the mass ratio of *Saccharomyces cerevisiae* to *Bacillus pumilus* is 1:0.5-2.5, preferably 1:1. The mass ratio of soybean meal, bone meal, and seaweed powder is (1-3):(0.5-1.5):(0.5-1.5), preferably 2:1:1. The compound enzyme in this invention includes neutral protease, cellulase, and pectinase, and the mass ratio of neutral protease:cellulase:pectinase is (1-3):(0.5-1.5):(0.5-1.5), preferably 2:1:1. Unless otherwise specified, the soybean meal, bone meal, seaweed powder, neutral protease, cellulase, and pectinase in this invention are all commercially available. The enzyme activity of the neutral protease, cellulase, and pectinase in this invention is 30,000-70,000 U / g, the enzyme activity of cellulase is 30,000-70,000 U / g, and the enzyme activity of pectinase is 30,000-70,000 U / g, all preferably 60,000 U / g.

[0020] This invention also provides a method for preparing the special bio-fertilizer for traditional Chinese medicine, comprising the following steps: mixing the above-ground parts of Astragalus membranaceus with water, adding Saccharomyces cerevisiae and Bacillus pumilus for fermentation, sterilizing, and drying to obtain the fermented product of the above-ground parts of Astragalus membranaceus; mixing soybean meal, bone meal, and seaweed powder with water, adding a compound enzyme for enzymatic hydrolysis, inactivating, and drying to obtain the enzymatic hydrolysate; mixing the fermented product of the above-ground parts of Astragalus membranaceus with the enzymatic hydrolysate to obtain the special bio-fertilizer for traditional Chinese medicine. The above-ground parts of Astragalus membranaceus mentioned in this invention refer to the above-ground parts of Astragalus membranaceus after mature harvest, including stems and leaves. This invention, through the directional fermentation of the above-ground parts of Astragalus membranaceus and the compound enzymatic hydrolysis of soybean meal, bone meal, and seaweed powder, not only achieves the resource utilization of the above-ground parts of Astragalus membranaceus but also efficiently prepares a dual-functional raw material with both homologous active substances and small-molecule fast-acting nutrients; after the fermented product of the above-ground parts of Astragalus membranaceus is combined with the enzymatic hydrolysate of soybean meal, bone meal, and seaweed powder, a special bio-fertilizer is formed that simultaneously regulates the rhizosphere microecology of Astragalus membranaceus, alleviates continuous cropping obstacles, and provides sufficient nutrients for plant growth, achieving a synergistic effect of microecological reshaping and nutrient supply.

[0021] In this invention, the mass-to-volume ratio of the aerial parts of Astragalus membranaceus to water is 1g:8-12mL, preferably 1g:10mL. The inoculation amount of Saccharomyces cerevisiae and Bacillus pumilus is 2.5-5.5% of the mass of the aerial parts of Astragalus membranaceus, preferably 3%, 4%, and 5%. The fermentation temperature is 30-35℃, and the time is 2-6 days, preferably 31℃, 32℃, or 33℃, and the time is preferably 3 days, 4 days, or 5 days. The Saccharomyces cerevisiae of this invention can degrade the cellulose and polysaccharides of the aerial parts of Astragalus membranaceus, producing easily utilized nutrients such as small-molecule sugars and amino acids; Bacillus pumilus lowers the pH of the system by producing acid, inhibiting the growth of other bacteria. Together, they transform the aerial parts of Astragalus membranaceus into a fermentation substrate rich in active substances and odorless. The fermentation products not only retain and enrich the original flavonoids, saponins, and other bioactive components of Astragalus membranaceus, but also form functional substances such as organic acids and probiotic secondary metabolites, which can directionally promote the proliferation of beneficial microorganisms in the rhizosphere of Astragalus membranaceus, providing a dedicated nutrient source for the reshaping of the rhizosphere microecology.

[0022] In this invention, the compound enzyme comprises neutral protease, cellulase, and pectinase, wherein the mass ratio of neutral protease:cellulase:pectinase is (1-3):(0.5-1.5):(0.5-1.5), preferably 2:1:1. The amount of the compound enzyme added in this invention is 1.5-3.5% of the total mass of soybean meal, bone meal, and seaweed powder, preferably 2.5%. The enzymatic hydrolysis temperature in this invention is 45-50℃, and the time is 3-7 hours, preferably 46℃, 47℃, or 48℃, and the time is 4 hours, 5 hours, or 6 hours. The neutral protease of this invention can degrade large molecular proteins in soybean meal into free amino acids and small peptides, while cellulase and pectinase can decompose polysaccharides such as cellulose and pectin in soybean meal and seaweed powder. At the same time, calcium and phosphorus elements in bone meal are also partially activated, ultimately yielding a small molecular mixture with comprehensive nutritional components that is easily absorbed and utilized. This complex enzymatic hydrolysis product is rich in free amino acids, oligosaccharides, active peptides, and soluble mineral elements. It can not only provide Astragalus membranaceus and rhizosphere microorganisms with fast-acting nitrogen, carbon, and mineral nutrients, but also avoid the problems of slow decomposition and low utilization of large molecular raw materials, thus ensuring the speed and efficiency of fertilizer.

[0023] This invention provides the application of the bio-fertilizer specifically for Chinese medicinal herbs, or the bio-fertilizer specifically for Chinese medicinal herbs prepared by the aforementioned method, in reshaping the rhizosphere microbial community of plants. Unless otherwise specified, the bio-fertilizer specifically for Chinese medicinal herbs is applicable to any type of Chinese medicinal herb. Astragalus membranaceus is a preferred Chinese medicinal herb in this invention.

[0024] This invention provides the application of the bio-fertilizer specifically for Chinese medicinal herbs, or the bio-fertilizer specifically for Chinese medicinal herbs prepared by the aforementioned method, in promoting the growth and improving the quality of Chinese medicinal herbs. Unless otherwise specified, the bio-fertilizer is applicable to any type of Chinese medicinal herb. Astragalus membranaceus is a preferred Chinese medicinal herb in this invention.

[0025] The technical solutions provided by the present invention will be described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0026] Unless otherwise specified, the following embodiments are all conventional methods.

[0027] Unless otherwise specified, all materials and reagents used in the following examples are commercially available.

[0028] In the following examples, the viable count of *Saccharomyces cerevisiae* was 10 billion CFU / g, purchased from Shandong Yihao Biotechnology Co., Ltd.; the viable count of *Bacillus pumilus* was 50 billion CFU / g, purchased from Hubei Yihao Biotechnology Co., Ltd.; the viable count of *Bacillus subtilis* was ≥20 billion CFU / g, purchased from Shandong Huayunda New Materials Co., Ltd.; the viable count of *Bacillus amyloliquefaciens* was ≥20 billion CFU / g, purchased from Shandong Yihao Biotechnology Co., Ltd.; the neutral protease was purchased from Xiasheng (Beijing) Biotechnology Development Co., Ltd.; and the cellulase and pectinase were purchased from Taian Xindeli Bioengineering Co., Ltd., with an enzyme activity of 60,000 U / g.

[0029] Example 1 1. After harvesting mature Astragalus membranaceus, remove impurities from the above-ground parts (stems, leaves, and pods, sourced from the Astragalus membranaceus planting base in Yuanzhou District, Guyuan City, Ningxia), wash them, cut them into 2-3cm sections, dry them at 60℃ until the moisture content is ≤12%, and pulverize them through an 80-mesh sieve to obtain Astragalus membranaceus above-ground part powder.

[0030] 2. Add the powdered aerial parts of Astragalus membranaceus to water, adjust the material-to-liquid ratio to 1:10 (g / mL), mix well, and adjust the pH to 6.5; add 4% of the Astragalus membranaceus aerial part powder as a fermentation agent (Saccharomyces cerevisiae:Bacillus pumilus mass ratio of 1:1); ferment at a constant temperature of 32℃ for 4 days, stirring twice a day for 30 minutes each time, until the pH of the liquid drops to 4.5 and there is no off-odor; sterilize, dry until the moisture content is ≤10%, pulverize and pass through an 80-mesh sieve to obtain the fermented product of the aerial parts of Astragalus membranaceus.

[0031] 3. Mix soybean meal, bone meal, and seaweed powder in a mass ratio of 2:1:1, grind them through an 80-mesh sieve, add water to adjust the material-to-liquid ratio to 1:10 (g / mL), adjust the pH to 6.8, add 2.5% of the total mass of soybean meal, bone meal, and seaweed powder of a compound enzyme (neutral protease: cellulase: pectinase in a mass ratio of 2:1:1), and hydrolyze at a constant temperature of 48℃ for 5 hours. After hydrolysis, inactivate the enzyme at 92℃ for 18 minutes, dry it until the moisture content is ≤10%, grind it through an 80-mesh sieve, and obtain the hydrolysate.

[0032] 4. Mix the fermented above-ground parts of Astragalus membranaceus with the enzymatic hydrolysate at a mass ratio of 1:5 to obtain a special bio-fertilizer for Chinese medicinal materials.

[0033] Example 2 1. After harvesting mature Astragalus membranaceus, remove impurities from the above-ground parts (stems, leaves, and pods, sourced from the Astragalus membranaceus planting base in Yuanzhou District, Guyuan City, Ningxia), wash them, cut them into 2-3cm sections, dry them at 60℃ until the moisture content is ≤12%, and pulverize them through a 60-mesh sieve to obtain Astragalus membranaceus powder.

[0034] 2. Add the powdered aerial parts of Astragalus membranaceus to water, adjust the material-to-liquid ratio to 1:8 (g / mL), mix well, and adjust the pH to 6.8; add 3% of the Astragalus membranaceus aerial part powder as a fermentation agent (Saccharomyces cerevisiae:Bacillus pumilus mass ratio of 1:2); ferment at a constant temperature of 30℃ for 6 days, stirring twice a day for 30 minutes each time, until the pH of the liquid drops to 5.0 and there is no off-odor; sterilize, dry until the moisture content is ≤10%, pulverize and pass through an 80-mesh sieve to obtain the fermented product of the aerial parts of Astragalus membranaceus.

[0035] 3. Mix soybean meal, bone meal, and seaweed powder in a mass ratio of 1:0.5:0.5, grind them through a 60-mesh sieve, add deionized water to adjust the material-to-liquid ratio to 1:5 (g / mL), adjust the pH to 6.5, add 2.5% of the total mass of raw materials of a compound enzyme (neutral protease: cellulase: pectinase in a mass ratio of 1:1:1), and enzymatically hydrolyze at 50℃ for 6 hours. After enzymatic hydrolysis, inactivate the enzyme at 92℃ for 18 minutes, dry it until the moisture content is ≤10%, grind it through an 80-mesh sieve, and obtain the enzymatic hydrolysate.

[0036] 4. Mix the fermented above-ground parts of Astragalus membranaceus with the enzymatic hydrolysate at a mass ratio of 1:3 to obtain a special bio-fertilizer for Chinese medicinal materials.

[0037] Example 3 1. After harvesting mature Astragalus membranaceus, remove impurities from the above-ground parts (stems, leaves, and pods, sourced from the Astragalus membranaceus planting base in Yuanzhou District, Guyuan City, Ningxia), wash them, cut them into 2-3cm sections, dry them at 60℃ until the moisture content is ≤12%, and pulverize them through a 90-mesh sieve to obtain Astragalus membranaceus above-ground part powder.

[0038] 2. Add the powdered aerial parts of Astragalus membranaceus to deionized water, adjust the material-to-liquid ratio to 1:12 (g / mL), mix well, and adjust the pH to 6.3; add 5% of the mass of Astragalus membranaceus aerial parts powder as a fermentation agent (Saccharomyces cerevisiae:Bacillus pumilus mass ratio of 1:1); ferment at a constant temperature of 35℃ for 6 days, stirring twice a day for 30 minutes each time, until the pH of the liquid drops to 4.8 and there is no off-odor; sterilize, dry to a moisture content of ≤10%, pulverize and pass through a 90-mesh sieve to obtain the fermented product of Astragalus membranaceus aerial parts.

[0039] 3. Mix soybean meal, bone meal, and seaweed powder in a mass ratio of 2:1.5:1.5, grind them through a 90-mesh sieve, add deionized water to adjust the material-to-liquid ratio to 1:10 (g / mL), adjust the pH to 6.9, add 3.5% of the total mass of raw materials of a compound enzyme (neutral protease: cellulase: pectinase in a mass ratio of 2:1.5:1.5), and enzymatically hydrolyze at 45℃ for 7 hours. After enzymatic hydrolysis, inactivate the enzyme at 92℃ for 18 minutes, dry it until the moisture content is ≤10%, grind it through an 80-mesh sieve, and obtain the enzymatic hydrolysate.

[0040] 4. Mix the fermented above-ground parts of Astragalus membranaceus with the enzymatic hydrolysate at a mass ratio of 1:8 to obtain a special bio-fertilizer for Chinese medicinal materials.

[0041] Comparative Example 1 The above-ground parts of Astragalus membranaceus harvested in Example 1 were replaced with the dregs of Astragalus membranaceus decoction, and the remaining steps were the same as in Example 1.

[0042] Comparative Example 2 Replace Saccharomyces cerevisiae and Bacillus pumilus with Bacillus subtilis and Bacillus amyloliquefaciens, and follow the same steps as in Example 1.

[0043] Comparative Example 3 After harvesting, the aerial parts of Astragalus membranaceus were powdered and decocted in water to obtain an aqueous extract mixture. This mixture was then concentrated and dried to obtain the aqueous extract of the aerial parts of Astragalus membranaceus. The remaining steps were the same as in Example 1.

[0044] Comparative Example 4 1. After harvesting Astragalus membranaceus, remove impurities from the above-ground parts (stems and leaves), wash them, cut them into sections 2-3 cm long, dry them at 60℃ until the moisture content is ≤12%, pulverize them through an 80-mesh sieve to obtain Astragalus membranaceus powder; mix Astragalus membranaceus powder, soybean meal, bone meal and seaweed powder in a mass ratio of 10:5:2.5:2.5 to obtain a mixture.

[0045] 2. Add water to the mixture, adjust the material-to-liquid ratio to 1:10 (g / mL), mix well, and adjust the pH to 6.5; add 4% of the weight of Astragalus membranaceus aerial part powder as a fermentation agent (Saccharomyces cerevisiae:Bacillus pumilus mass ratio of 1:1); ferment at a constant temperature of 32℃ for 4 days, stirring twice a day for 30 minutes each time, until the pH of the liquid drops to 4.5 and there is no off-odor; sterilize, dry to ≤10% moisture, pulverize and pass through an 80-mesh sieve to obtain a special bio-fertilizer for Chinese medicinal materials.

[0046] Experimental Example 1 1. The experimental site was the Astragalus membranaceus (Huang Qi) planting base in Yuanzhou District, Guyuan City, Ningxia. Grouping: The experiment consisted of 7 treatment groups and 1 control group, with 3 replicates per group, using a randomized block design. The plot area was 20m². 2 (4m×5m), the treatments for each group are as follows: Examples 1-3 and Comparative Examples 1-4 were treated with the bio-fertilizer prepared in each group, while the blank control group was not treated with the bio-fertilizer prepared in this invention and was planted according to local planting methods. Fertilization was carried out 15 days after the Astragalus seedlings broke through the soil. Specifically, fertilization was carried out in trenches 8-10cm away from both sides of the seedlings, at a rate of 80g / m². 3 Fertilizer was applied in furrows, and the furrows were then leveled. All plots followed standard local planting methods in terms of sowing time, sowing density, and field management, with consistency maintained across groups. The experimental period was one year. Astragalus cultivation... Figure 1 As shown.

[0047] 2. Detection indicators and methods Rhizosphere microbiome analysis: After the experiment, a 5-point sampling method was used, and surface debris was removed before sampling. Rhizosphere soil (0-20cm around the roots) from 3 Astragalus plants in each plot was randomly collected and mixed as one sample. High-throughput sequencing technology was used to detect the rhizosphere microbial community structure (bacterial 16S rRNA gene and fungal ITS gene sequencing), and the microbial diversity indices (Shannon index and Simpson index), and the abundance changes of beneficial and pathogenic microorganisms were analyzed.

[0048] Astragalus yield testing: After harvest, all plants in each plot were harvested. Root diameter was measured using calipers at a point approximately 3mm from the base of the rhizome. Root length was measured using a ruler from the root tip to the point where the root connects to the rhizome. Stem length was measured from the root to the tip of the main stem. Fresh root weight was measured using an electronic balance with an accuracy of 0.01. Ten plants were randomly selected from each plot for testing, and the average value was calculated.

[0049] Quality indicators of Astragalus membranaceus: Astragaloside A in Astragalus membranaceus root was determined according to General Rule 0512 of the Chinese Pharmacopoeia by high performance liquid chromatography; total polysaccharide content in Astragalus membranaceus was determined according to General Rule 0832 of the Chinese Pharmacopoeia by phenol-sulfuric acid method.

[0050] Data processing: SPSS 26.0 software was used for statistical analysis, and P<0.05 was considered statistically significant.

[0051] 3. Results Analysis (1) Analysis of rhizosphere microbial diversity As shown in Table 1, the Shannon and Simpson indices of rhizosphere microorganisms in each example group were significantly higher than those in comparative groups 1-4 (P<0.05), with example group 1 showing the best effect, significantly higher than comparative groups 1-4 (P<0.05). The groups with Astragalus membranaceus residue replacing the aboveground parts and those with water decoction replacing fermentation had lower diversity indices, indicating that fermentation of the aboveground parts of Astragalus membranaceus by Saccharomyces cerevisiae and Bacillus pumilus can more effectively improve rhizosphere microbial diversity. The diversity indices of the groups with microbial agents replacing the aboveground parts and those with enzymatic hydrolysis replacing fermentation were lower than those in the example groups, demonstrating that the rationality of the compound fermentation agent and enzymatic hydrolysis process is crucial for improving microbial diversity.

[0052] Table 1 Results of rhizosphere microbial diversity analysis

[0053] Note: Different lowercase letters after the data in the same column indicate significant differences (P<0.05), and the same applies below.

[0054] (2) Abundance analysis of beneficial and pathogenic microorganisms in the rhizosphere As shown in Table 2, the relative abundance of beneficial microorganisms (Bacillus subtilis and Trichoderma) in each example group was significantly higher than that in each comparative group (P<0.05), while the relative abundance of pathogenic microorganisms (Fusarium oxysporum) was significantly lower than that in each comparative group (P<0.05). Example 1 group showed the best effect; in comparative group 1, the abundance of beneficial microorganisms decreased and the abundance of pathogenic bacteria increased due to the replacement of Astragalus membranaceus residue; in comparative group 2, the proliferation effect of beneficial microorganisms decreased after replacing the fermentation agent; in comparative group 3, water decoction could not effectively release active substances, resulting in weak microbial regulation; in comparative group 4, fermentation replaced enzymatic hydrolysis, resulting in insufficient small molecule nutrients and a lower abundance of beneficial microorganisms than in the example groups. This indicates that the fermentation + enzymatic hydrolysis process and raw material selection of the present invention can effectively and directionally enrich beneficial bacteria and inhibit pathogenic bacteria, thereby achieving rhizosphere microbiome remodeling.

[0055] Table 2. Abundance analysis results of beneficial and pathogenic microorganisms in the rhizosphere.

[0056] (3) Results of the influence of Astragalus membranaceus yield Table 3 shows that the root length, root diameter, stem length, and fresh weight of Astragalus membranaceus in each example group were significantly higher than those in each comparative group (P<0.05). In Comparative Group 1, due to raw material replacement, nutrient supply was insufficient, resulting in a decrease in growth indicators. In Comparative Group 2, after replacing the inoculant, the rhizosphere microecological regulation effect was poor, affecting Astragalus membranaceus growth. In Comparative Group 3, water decoction could not fully release nutrients, resulting in lower growth. In Comparative Group 4, enzymatic hydrolysis was replaced by fermentation, leading to insufficient supply of small-molecule nutrients, and growth indicators were lower than those in the example groups. This indicates that the preparation method of the present invention can significantly improve the growth of Astragalus membranaceus by optimizing the rhizosphere microecology and providing sufficient fast-acting nutrients.

[0057] Table 3 Results of the effects of each group on the growth indicators of Astragalus membranaceus

[0058] (4) Results of the influence of Astragalus membranaceus on quality As shown in Table 4, the contents of astragaloside A and total polysaccharides in each example group were significantly higher than those in each comparative group (P<0.05). In Comparative Example 1, the replacement of raw materials resulted in insufficient homologous active substances and a decline in quality indicators; in Comparative Example 2, the replacement of the microbial agent led to poor rhizosphere microecological regulation, affecting the synthesis of effective components; in Comparative Example 3, water decoction could not fully extract active substances, resulting in poor quality; and in Comparative Example 4, the replacement of enzymatic hydrolysis with fermentation resulted in unbalanced nutrient supply and lower content of effective components compared to the example groups. This indicates that the preparation method of the present invention can significantly improve the quality of Astragalus by reshaping the rhizosphere microecology and providing specific nutrients.

[0059] Table 4. Results of the effects of each group on the quality of Astragalus membranaceus.

[0060] In summary, the bio-fertilizers prepared in Examples 1-3, through fermentation of the aboveground parts of Astragalus membranaceus using Saccharomyces cerevisiae and Bacillus pumilus, and enzymatic hydrolysis of soybean meal, bone meal, and seaweed powder in a specific ratio, can significantly enhance the diversity of rhizosphere microorganisms in Astragalus membranaceus, directionally enrich beneficial microorganisms, inhibit pathogenic microorganisms, and achieve effective remodeling of the rhizosphere microbiome; at the same time, they significantly improve the yield and quality of Astragalus membranaceus, with Example 1 showing the best results. The effects of each comparative example significantly decreased due to changes in raw materials, inoculants, and processes (P<0.05), proving that the raw material selection, fermentation process, enzymatic hydrolysis process, and compounding ratio of this invention are irreplaceable, achieving synergistic effects of microecological remodeling and nutrient supply, effectively solving the problem of continuous cropping obstacles in Astragalus membranaceus, and improving the yield and quality of Astragalus membranaceus.

[0061] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A bio-fertilizer specifically for Chinese medicinal herbs based on rhizosphere microbiome remodeling, characterized in that, The raw materials include the following parts by weight: 0.2-3 parts of fermented astragalus aerial parts and 2-8 parts of enzymatic hydrolysate; The fermented product of the aerial parts of Astragalus membranaceus was obtained by fermenting the aerial parts of Astragalus membranaceus with Saccharomyces cerevisiae and Bacillus pumilus. The enzymatic hydrolysate is obtained by enzymatic hydrolysis of soybean meal, bone meal, and seaweed powder using a compound enzyme.

2. The bio-fertilizer specifically for Chinese medicinal materials as described in claim 1, characterized in that, The mass ratio of the brewer's yeast to Bacillus pumilus is 1:0.5-2.

5.

3. The bio-fertilizer specifically for Chinese medicinal materials as described in claim 1, characterized in that, The mass ratio of soybean meal, bone meal, and seaweed powder is (1-3):(0.5-1.5):(0.5-1.5).

4. The bio-fertilizer specifically for Chinese medicinal materials as described in claim 1, characterized in that, The complex enzyme comprises neutral protease, cellulase and pectinase, wherein the mass ratio of neutral protease:cellulase:pectinase is (1-3):(0.5-1.5):(0.5-1.5).

5. The method for preparing the bio-fertilizer specifically for Chinese medicinal materials as described in any one of claims 1-4, characterized in that, The process includes the following steps: the above-ground parts of Astragalus membranaceus are mixed with water, brewer's yeast and Bacillus pumilus are added for fermentation, sterilization and drying are performed to obtain fermented Astragalus membranaceus above-ground parts; soybean meal, bone meal and seaweed powder are mixed with water, compound enzymes are added for enzymatic hydrolysis, inactivation is performed, and drying is performed to obtain enzymatic hydrolysate; the fermented Astragalus membranaceus above-ground parts and enzymatic hydrolysate are mixed to obtain a special bio-fertilizer for Chinese medicinal materials.

6. The preparation method according to claim 5, characterized in that, The mass-to-volume ratio of the aerial parts of Astragalus membranaceus to water is 1g:8-12mL.

7. The preparation method according to claim 5, characterized in that, The inoculation amount of the brewing yeast and Bacillus pumilus is 2.5-5.5% of the weight of the aerial parts of Astragalus membranaceus; the fermentation temperature is 30-35℃ and the time is 2-6 days.

8. The preparation method according to claim 5, characterized in that, The amount of the compound enzyme added is 1.5-3.5% of the total mass of soybean meal, bone meal, and seaweed powder; the enzymatic hydrolysis temperature is 45-50℃ and the time is 3-7h.

9. The application of the bio-fertilizer for Chinese medicinal materials as described in any one of claims 1-4 or the bio-fertilizer for Chinese medicinal materials prepared by the preparation method described in any one of claims 5-8 in reshaping the rhizosphere microbial community of plants.

10. The application of the bio-fertilizer for Chinese medicinal materials as described in any one of claims 1-4 or the bio-fertilizer for Chinese medicinal materials prepared by the preparation method described in any one of claims 5-8 in promoting the growth of Chinese medicinal materials and improving the quality of Chinese medicinal materials.

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