Compound microbial agent capable of resisting root rot and hemp mouth disease of angelica sinensis and preparation method of compound microbial agent

Abstract

The invention discloses a compound microbial agent capable of resisting root rot and hemp mouth disease of angelica sinensis and a preparation method of the compound microbial agent, and belongs to the technical field of microorganisms. The compound microbial agent consists of 20-40 parts of compound microbial powder, 3-5 parts of an active agent, 10-20 parts of a carrier and 5-10 parts of a nutritional agent, the compound microorganism bacterium powder is prepared from bacillus clausii, streptomyces lilacinus and matsutake pemophila; the carrier is prepared from fly ash, bentonite and rice hull biochar; the active agent is prepared from a phyllostachys parviflora extract and cocamidopropyl betaine according to the mass ratio of 5: 1. The compound microbial agent can play a good role in preventing and treating root rot and hemp mouth disease of angelica sinensis at the same time, the yield increasing effect of angelica sinensis is remarkable, and the quality and yield of angelica sinensis are improved; the carrier used in the invention is a ceramic-based porous material, not only has abundant pore structures and good biocompatibility, but also has especially stable physical and chemical properties, ensures the long-term stability and activity of the microbial agent on the carrier, reduces the usage amount of the microbial agent, and has good application prospects.

Description

Technical Field

[0001] The invention belongs to the technical field of microorganisms, and particularly relates to a composite microbial agent capable of resisting angelica root rot and rashes and a preparation method thereof. Background Art

[0002] Angelica sinensis Angelica sinensis The dried root of (Oliv.) Diels has the effects of nourishing blood and activating blood circulation, moistening the intestines, regulating menstruation and relieving pain, and strengthening the body and replenishing the body. It is a traditional food and medicine substance and has been included in the list of dual-purpose medicine and food (National Health Commission Food Letter

[2019] No. 8), and is eaten as a spice and condiment. As a perennial herb, Angelica sinensis is a common medicinal material in traditional Chinese medicine treatment. In recent years, with the increase in market demand for Angelica sinensis, the planting area of ​​Angelica sinensis has gradually expanded. Cropping and continuous cropping have become the norm. The soil-borne diseases brought about by this have rapidly increased, becoming a major problem in production, seriously affecting the production and medicinal value of Angelica sinensis. Plant soil-borne diseases are a general term for diseases that occur in the roots or stems of plants and are transmitted through soil as a medium, including root rot, wilt, damping-off, damping-off, blight, and Verticillium wilt. The pathogens of this type of disease have part or most of their life cycle in the soil. When conditions are suitable, the pathogens germinate and infect the roots or stems of plants, causing plant diseases. Among them, root rot and numbness are two common diseases in angelica cultivation.

[0003] The main pathogens of root rot are Fusarium spp. Fusarium ) fungi. Fusarium has strong ecological adaptability and can live a parasitic or saprophytic life. It is widely distributed in soil, air, animals and plants. It is an important plant pathogenic fungus that can cause root rot, stem base rot, wilt and fruit rot in different plants. Currently, the known pathogens of Angelica root rot caused by Fusarium include Fusarium solani ( Fusarium solani (Mart.) Sacc), Fusarium oxysporum ( Fusarium oxysporum Schlecht.), Fusarium odoratum ( Fusarium redolens Wollenw), Fusarium acuminate ( Fusarium acuminatum Ellis & Everhart.) and Fusarium solani ( Fusarium avenaceum According to the survey, the incidence of this disease in the main angelica production areas is about 35%, and in severe cases it reaches 80% or more, which is one of the important reasons for the reduction or even complete failure of angelica production.

[0004] The cause of the paronychia of Angelica sinensis is quite complicated. The main pathogen is Fusarium solani, and there are also cross-infections and complex infections such as Chaetomium globosum Kunze., Alternaria olternatas pp. and actinomycetes. At the same time, the paronychia of Angelica sinensis is also called stem nematode disease, which can be caused by infection with the rot stem nematode. The damage of these stem nematodes and underground pests is also directly related to fungi. While the nematodes are causing damage, the epidermis of the Angelica sinensis roots is longitudinally cracked, and a large amount of yellow-brown powdery particles are contained in the cracks. The spongy lignification of the root tissue loses its oiliness, and the medicinal quality is reduced. The paronychia is dominated by pathogens, and the wounds caused by underground pests are conducive to the invasion of pathogens, which leads to the aggravation of the paronychia and is the cause of the disease.

[0005] At present, for the main diseases of Angelica sinensis, root rot and hemp-mouth disease, traditional prevention and control methods mostly rely on chemical pesticides, but the long-term and large-scale use of chemical pesticides will not only cause environmental pollution, but also may lead to increased resistance of pests and diseases. Biological control has become a research hotspot for replacing chemical pesticides because of its advantages such as less environmental pollution and effective prevention and control of plant diseases. Chinese patent CN18743398 A discloses a composite microbial agent for resisting Angelica sinensis hemp-mouth disease and its preparation method and application; Chinese patent CN18561643 A discloses a composite microbial fertilizer for inhibiting Angelica sinensis root rot and its preparation method; although the prior art has disclosed a variety of microbial agents for the prevention and control of different types of diseases of Angelica sinensis, there are few microbial agents that can simultaneously achieve excellent prevention and control effects on Angelica sinensis root rot and hemp-mouth disease. Therefore, it is particularly important to develop a safe, environmentally friendly and efficient microbial agent to simultaneously achieve effective prevention and control of Angelica sinensis root rot and hemp-mouth disease. Summary of the invention

[0006] The purpose of the present invention is to provide a composite microbial agent that can simultaneously prevent and treat angelica root rot and rashes. By using one microbial agent, the excellent effect of preventing and treating two common diseases of angelica can be achieved, and the yield and quality of angelica can be improved.

[0007] In order to achieve the above technical objectives, the technical solution adopted by the present invention is: A composite microbial agent capable of resisting angelica root rot and rashes is prepared from the following raw materials in parts by weight: 20-40 parts of composite microbial powder, 3-5 parts of an active agent, 10-20 parts of a carrier, and 5-10 parts of a nutrient agent; the composite microbial agent comprises Bacillus clausii, Streptomyces lilacinus, and Masseillaris timonei; the carrier is made of fly ash, bentonite, and rice husk biochar; the active agent is composed of golden bamboo grass extract and coconut oil amidopropyl betaine in a mass ratio of 5:1.

[0008] Preferably, the carrier is prepared by the following method: fly ash, bentonite and rice husk biochar are mixed in a mass ratio of 1:1:3 and mechanically ground for 30 minutes, then placed in a high-temperature muffle furnace and heated to 800°C in stages, kept warm for 1 hour, then heated to 1100°C, kept warm for 2 hours, and ground into powder after cooling to obtain the carrier.

[0009] Preferably, the temperature of the muffle furnace is raised to 800° C. at a rate of 5° C. / min, and then raised to 1100° C. at a rate of 3° C. / min.

[0010] Preferably, the preparation method of the golden bamboo grass extract is as follows: (1) Weigh 100 g of golden bamboo grass, clean it, dry it in an oven, crush it, and pass it through an 80-mesh sieve to obtain powdered golden bamboo grass; (2) Take 15 g of golden bamboo grass powder, add it to 150 ml of 2% wt oxalic acid solution, stir, then transfer it to a hydrothermal reactor, let it stand for 45 minutes, and then place the hydrothermal reactor in a forced air drying oven and keep it at 150°C for 3 hours. After the insulation is completed, filter and take filtrate A; (3) adding 2%wt calcium carbonate suspension to the filtrate A obtained in step (2) until no bubbles are generated, letting it stand for 30-40 minutes and then filtering to remove the precipitate to obtain filtrate B; (4) Add an appropriate amount of activated carbon to the filtrate B, set the temperature to 70°C in a constant temperature oscillator, and oscillate at 150 rpm for 2 h, then filter to obtain the golden bamboo grass extract.

[0011] Preferably, in step (4), the material-liquid ratio of activated carbon to filtrate B is 1 g:30 ml.

[0012] Preferably, the composite microbial agent is prepared by the following method: thawing and activating Bacillus clausii, Streptomyces lilacinus, and Massimoya timonei strains respectively, streaking the plates and culturing them in a 28°C incubator for 48 hours, taking a loop of the cultured strains in LB liquid culture medium, placing it in a shaker at 30°C and 180 r / min for 24 hours to obtain seed liquid; inoculating the seed liquids of the three strains into TSB liquid culture medium at an inoculation rate of 3%, bottling 200 mL, 30°C, 200 r / min shaking culture for 72 hours to obtain fermentation liquid; mixing the three fermentation liquids in a mass ratio of 1:1:1 and freeze-drying them to obtain bacterial powder, which is the composite microbial powder.

[0013] The LB liquid culture medium is composed of: 5 g yeast extract, 10 g tryptone, 10 g sodium chloride, 1000 mL distilled water, and the pH is adjusted to 7.0; and the medium is sterilized at a high temperature of 121° C. with steam for 20 min.

[0014] The TSB liquid culture medium is composed of: 15 g of tryptone, 5 g of soy peptone, 5 g of sodium chloride, and 1000 ml of distilled water. The pH is adjusted to 7.2±0.2 and sterilized by pressure steam at 121° C. for 20 min.

[0015] Preferably, the Bacillus clausii strain is numbered as CGMCC No.1.3672, purchased from China General Microbiological Culture Collection Administration Center, with a deposit date of June 11, 2004, and a deposit address of No. 3, No. 1 Beichen West Road, Chaoyang District, Beijing; the Streptomyces lilacinus strain is numbered as CGMCC No.4.6512, purchased from China General Microbiological Culture Collection Administration Center, with a deposit date of September 3, 2008, and a deposit address of No. 3, No. 1 Beichen West Road, Chaoyang District, Beijing; the timonene masseilli strain is numbered as CGMCC No.1.12742, purchased from China General Microbiological Culture Collection Administration Center, with a deposit date of December 26, 2013, and a deposit address of No. 3, No. 1 Beichen West Road, Chaoyang District, Beijing. The Bacillus clausii, Streptomyces lilacinus, and timonene masseilli strains used in the present invention can all be purchased through the strain catalog of the collection center without the need for additional deposit.

[0016] Preferably, the nutrient agent is composed of urea, potassium dihydrogen phosphate, amino acid chelated zinc, amino acid chelated magnesium, and amino acid chelated manganese in a mass ratio of 1:0.8:0.05:0.02:0.01.

[0017] The present invention also provides a method for preparing the composite microbial agent capable of resisting both angelica root rot and rashes, which comprises the following steps: A. preparing composite microbial powder; B. preparing the carrier; C. preparing a golden bamboo grass extract, and then combining the golden bamboo grass extract with cocoamidopropyl betaine in a mass ratio of 5:1 to form an active agent; D. uniformly mixing the composite microbial powder, carrier and active agent, and drying at 40° C. to obtain a mixture; E. Evenly mix the above mixture with the nutrient agent to obtain the final product.

[0018] All raw materials used in the present invention are commercially available.

[0019] The dosage and usage of the composite microbial agent for resisting angelica root rot and rashes are as follows: it is mixed with conventional fertilizers as base fertilizer during transplanting, and the dosage is 10 kg / mu.

[0020] The beneficial effects of the present invention are: (1) The composite microbial powder is prepared by screening and compounding three strains of Bacillus clausii, Streptomyces lilacinus and Massimodium timonei, which are activated and fermented and cultured respectively and then mixed in equal proportions. The three strains have a significant synergistic effect and can secrete a variety of antibacterial substances, such as phenols, flavonoids, alkaloids and other biologically active substances, which have good preventive and therapeutic effects on angelica root rot and numbness. The present invention also adds active substances of specific components, among which the golden bamboo grass extract contains appropriate concentrations of D-xylose, arabinose, galactose, glucose and other carbohydrate active substances, which can promote the growth and reproduction of microorganisms and improve the activity of the microorganisms of the present invention, and an appropriate amount of coconut oil amidopropyl betaine promotes the microorganisms to better absorb the active substances. The two work together to achieve the effect of improving the activity of the composite microbial agent.

[0021] (2) The present invention uses a product prepared by high-temperature calcination of fly ash, bentonite and rice husk biochar as a carrier, which is a ceramic-based porous material. Compared with traditional biochar carriers, the carrier of the present invention has a richer and more controllable pore structure (such as Figure 2 As shown in the figure, the ceramic-based porous material has a larger specific surface area, better biocompatibility, better stability, especially stability in extreme environments, which can adsorb more microorganisms and provide a good colonization environment for them, ensuring the long-term stability and activity of the microbial agent on the carrier. At the same time, the ceramic-based porous material also has unique physical surface properties, which is conducive to the diffusion of metabolites, thereby achieving better prevention and control of angelica root rot and numbness.

[0022] (3) The nutrient agent of the present invention contains a variety of nutrients that are easier for microorganisms and plants to absorb. On the one hand, it provides sufficient nutrients for the microorganisms to facilitate their growth and reproduction. On the other hand, it also provides sufficient nutrients for the plants, which is more conducive to the growth of angelica sinensis and improves the yield and quality of angelica sinensis. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 This is a diagram showing the antibacterial effect of the composite microbial fermentation liquid prepared in Example 3 and Comparative Examples 3-5 of the present invention on Fusarium ovata; Figure 2 The microscopic morphology images (SEM images) of the carrier prepared by the present invention and ordinary rice husk biochar, wherein a is the SEM of rice husk biochar, and b is the porous material carrier prepared by the present invention. DETAILED DESCRIPTION

[0024] The technical solution of the present invention is further described below in conjunction with specific embodiments, but is not limited thereto.

[0025] Example 1 A composite microbial agent capable of resisting angelica root rot and rashes is prepared from the following raw materials in parts by weight: 40 parts of composite microbial powder, 5 parts of an active agent, 20 parts of a carrier, and 10 parts of a nutrient agent; the composite microbial agent comprises Bacillus clausii, Streptomyces lilacinus, and Masseillaris timonei; the carrier is made of fly ash, bentonite, and rice husk biochar; the active agent is composed of golden bamboo grass extract and coconut oil amidopropyl betaine in a mass ratio of 5:1.

[0026] The carrier is prepared by the following method: fly ash, bentonite and rice husk biochar are mixed in a mass ratio of 1:1:3, mechanically ground for 30 minutes, then placed in a high-temperature muffle furnace and heated to 800°C at a speed of 5°C / min and kept for 1 hour, then heated to 1100°C at a speed of 3°C / min, kept for 2 hours, and ground into powder after cooling to obtain the carrier.

[0027] The preparation method of the golden bamboo grass extract is as follows: (1) Weigh 100 g of golden bamboo grass, clean it, dry it in an oven, crush it, and pass it through an 80-mesh sieve to obtain powdered golden bamboo grass; (2) Take 15 g of golden bamboo grass powder, add it to 150 ml of 2% wt oxalic acid solution, stir, then transfer it to a hydrothermal reactor, let it stand for 45 minutes, and then place the hydrothermal reactor in a forced air drying oven and keep it at 150°C for 3 hours. After the insulation is completed, filter and take filtrate A; (3) adding 2%wt calcium carbonate suspension to the filtrate A obtained in step (2) until no bubbles are generated, letting it stand for 30-40 minutes and then filtering to remove the precipitate to obtain filtrate B; (4) Add an appropriate amount of activated carbon to filtrate B at a material-liquid ratio of 1 g:30 ml, set the temperature to 70 °C in a constant temperature oscillator, oscillate at 150 rpm for 2 h, and then filter to obtain the golden bamboo grass extract.

[0028] The composite microbial agent is prepared by the following method: thawing and activating Bacillus clausii, Streptomyces lilacinus and Massimodium timonei strains respectively, culturing in a 28°C incubator for 48 hours after streaking on a plate, taking a loop of the cultured strains in an LB liquid culture medium, placing the culture medium in a shaking incubator at 30°C and 180 r / min for 24 hours to obtain a seed solution; inoculating the seed solutions of the three strains into a TSB liquid culture medium at an inoculation rate of 3% respectively, bottling the culture medium at a volume of 200 mL, 30°C, and 200 r / min for 72 hours to obtain a fermentation solution; and mixing the three fermentation solutions in a mass ratio of 1:1:1 and freeze-drying the mixture to obtain a bacterial powder, namely the composite microbial powder.

[0029] The LB liquid culture medium is composed of: 5 g yeast extract, 10 g tryptone, 10 g sodium chloride, 1000 mL distilled water, and the pH is adjusted to 7.0; and the medium is sterilized at a high temperature of 121° C. with steam for 20 min.

[0030] The TSB liquid culture medium is composed of: 15 g of tryptone, 5 g of soy peptone, 5 g of sodium chloride, and 1000 ml of distilled water. The pH is adjusted to 7.2±0.2 and sterilized by pressure steam at 121° C. for 20 min.

[0031] The Bacillus clausii strain is numbered as CGMCC No.1.3672, purchased from China General Microbiological Culture Collection Administration Center, with a preservation date of June 11, 2004, and a preservation address of No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing; the Streptomyces lilacinus strain is numbered as CGMCC No.4.6512, purchased from China General Microbiological Culture Collection Administration Center, with a preservation date of September 3, 2008, and a preservation address of No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing; the Marseilles timonei strain is numbered as CGMCC No.1.12742, purchased from China General Microbiological Culture Collection Administration Center, with a preservation date of December 26, 2013, and a preservation address of No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing.

[0032] The nutrient agent is composed of urea, potassium dihydrogen phosphate, amino acid chelated zinc, amino acid chelated magnesium, and amino acid chelated manganese in a mass ratio of 1:0.8:0.05:0.02:0.01.

[0033] A method for preparing the composite microbial agent for resisting both angelica root rot and rashes comprises the following steps: A. preparing composite microbial powder; B. preparing the carrier; C. preparing a golden bamboo grass extract, and then combining the golden bamboo grass extract with cocoamidopropyl betaine in a mass ratio of 5:1 to form an active agent; D. uniformly mixing the composite microbial powder, carrier and active agent, and drying at 40° C. to obtain a mixture; E. Evenly mix the above mixture with the nutrient agent to obtain the final product.

[0034] Example 2 A composite microbial agent capable of resisting angelica root rot and rashes is prepared from the following raw materials in parts by weight: 20 parts of composite microbial powder, 3 parts of an active agent, 10 parts of a carrier, and 5 parts of a nutrient agent; the composite microbial agent comprises Bacillus clausii, Streptomyces lilacinus, and Masseillaris timonei; the carrier is made of fly ash, bentonite, and rice husk biochar; the active agent is composed of golden bamboo grass extract and coconut oil amidopropyl betaine in a mass ratio of 5:1.

[0035] The carrier is prepared by the following method: fly ash, bentonite and rice husk biochar are mixed in a mass ratio of 1:1:3, mechanically ground for 30 minutes, then placed in a high-temperature muffle furnace and heated to 800°C at a speed of 5°C / min and kept for 1 hour, then heated to 1100°C at a speed of 3°C / min, kept for 2 hours, and ground into powder after cooling to obtain the carrier.

[0036] The preparation method of the golden bamboo grass extract is as follows: (1) Weigh 100 g of golden bamboo grass, clean it, dry it in an oven, crush it, and pass it through an 80-mesh sieve to obtain powdered golden bamboo grass; (2) Take 15 g of golden bamboo grass powder, add it to 150 ml of 2% wt oxalic acid solution, stir, then transfer it to a hydrothermal reactor, let it stand for 45 minutes, and then place the hydrothermal reactor in a forced air drying oven and keep it at 150°C for 3 hours. After the insulation is completed, filter and take filtrate A; (3) adding 2%wt calcium carbonate suspension to the filtrate A obtained in step (2) until no bubbles are generated, letting it stand for 30-40 minutes and then filtering to remove the precipitate to obtain filtrate B; (4) Add an appropriate amount of activated carbon to filtrate B at a material-liquid ratio of 1 g:30 ml, set the temperature to 70 °C in a constant temperature oscillator, oscillate at 150 rpm for 2 h, and then filter to obtain the golden bamboo grass extract.

[0037] The composite microbial agent is prepared by the following method: thawing and activating Bacillus clausii, Streptomyces lilacinus and Massimodium timonei strains respectively, culturing in a 28°C incubator for 48 hours after streaking on a plate, taking a loop of the cultured strains in an LB liquid culture medium, placing the culture medium in a shaking incubator at 30°C and 180 r / min for 24 hours to obtain a seed solution; inoculating the seed solutions of the three strains into a TSB liquid culture medium at an inoculation rate of 3% respectively, bottling the culture medium at a volume of 200 mL, 30°C, and 200 r / min for 72 hours to obtain a fermentation solution; and mixing the three fermentation solutions in a mass ratio of 1:1:1 and freeze-drying the mixture to obtain a bacterial powder, namely the composite microbial powder.

[0038] The LB liquid culture medium is composed of: 5 g yeast extract, 10 g tryptone, 10 g sodium chloride, 1000 mL distilled water, and the pH is adjusted to 7.0; and the medium is sterilized at a high temperature of 121° C. with steam for 20 min.

[0039] The TSB liquid culture medium is composed of: 15 g of tryptone, 5 g of soy peptone, 5 g of sodium chloride, and 1000 ml of distilled water. The pH is adjusted to 7.2±0.2 and sterilized by pressure steam at 121° C. for 20 min.

[0040] The Bacillus clausii strain is numbered as CGMCC No.1.3672, purchased from China General Microbiological Culture Collection Administration Center, with a preservation date of June 11, 2004, and a preservation address of No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing; the Streptomyces lilacinus strain is numbered as CGMCC No.4.6512, purchased from China General Microbiological Culture Collection Administration Center, with a preservation date of September 3, 2008, and a preservation address of No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing; the Marseilles timonei strain is numbered as CGMCC No.1.12742, purchased from China General Microbiological Culture Collection Administration Center, with a preservation date of December 26, 2013, and a preservation address of No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing.

[0041] The nutrient agent is composed of urea, potassium dihydrogen phosphate, amino acid chelated zinc, amino acid chelated magnesium, and amino acid chelated manganese in a mass ratio of 1:0.8:0.05:0.02:0.01.

[0042] A method for preparing the composite microbial agent for resisting both angelica root rot and rashes comprises the following steps: A. preparing composite microbial powder; B. preparing the carrier; C. preparing a golden bamboo grass extract, and then combining the golden bamboo grass extract with cocoamidopropyl betaine in a mass ratio of 5:1 to form an active agent; D. uniformly mixing the composite microbial powder, carrier and active agent, and drying at 40° C. to obtain a mixture; E. Evenly mix the above mixture with the nutrient agent to obtain the final product.

[0043] Example 3 A composite microbial agent capable of resisting angelica root rot and rashes is prepared from the following raw materials in parts by weight: 30 parts of composite microbial powder, 4 parts of an active agent, 15 parts of a carrier, and 8 parts of a nutrient agent; the composite microbial agent comprises Bacillus clausii, Streptomyces lilacinus, and Masseillaris timonei; the carrier is made of fly ash, bentonite, and rice husk biochar; the active agent is composed of golden bamboo grass extract and coconut oil amidopropyl betaine in a mass ratio of 5:1.

[0044] The carrier is prepared by the following method: fly ash, bentonite and rice husk biochar are mixed in a mass ratio of 1:1:3, mechanically ground for 30 minutes, then placed in a high-temperature muffle furnace and heated to 800°C at a speed of 5°C / min and kept for 1 hour, then heated to 1100°C at a speed of 3°C / min, kept for 2 hours, and ground into powder after cooling to obtain the carrier.

[0045] The preparation method of the golden bamboo grass extract is as follows: (1) Weigh 100 g of golden bamboo grass, clean it, dry it in an oven, crush it, and pass it through an 80-mesh sieve to obtain powdered golden bamboo grass; (2) Take 15 g of golden bamboo grass powder, add it to 150 ml of 2% wt oxalic acid solution, stir, then transfer it to a hydrothermal reactor, let it stand for 45 minutes, and then place the hydrothermal reactor in a forced air drying oven and keep it at 150°C for 3 hours. After the insulation is completed, filter and take filtrate A; (3) adding 2%wt calcium carbonate suspension to the filtrate A obtained in step (2) until no bubbles are generated, letting it stand for 30-40 minutes and then filtering to remove the precipitate to obtain filtrate B; (4) Add an appropriate amount of activated carbon to filtrate B at a material-liquid ratio of 1 g:30 ml, set the temperature to 70 °C in a constant temperature oscillator, oscillate at 150 rpm for 2 h, and then filter to obtain the golden bamboo grass extract.

[0046] The composite microbial agent is prepared by the following method: thawing and activating Bacillus clausii, Streptomyces lilacinus and Massimodium timonei strains respectively, culturing in a 28°C incubator for 48 hours after streaking on a plate, taking a loop of the cultured strains in an LB liquid culture medium, placing the culture medium in a shaking incubator at 30°C and 180 r / min for 24 hours to obtain a seed solution; inoculating the seed solutions of the three strains into a TSB liquid culture medium at an inoculation rate of 3% respectively, bottling the culture medium at a volume of 200 mL, 30°C, and 200 r / min for 72 hours to obtain a fermentation solution; and mixing the three fermentation solutions in a mass ratio of 1:1:1 and freeze-drying the mixture to obtain a bacterial powder, namely the composite microbial powder.

[0047] The LB liquid culture medium is composed of: 5 g yeast extract, 10 g tryptone, 10 g sodium chloride, 1000 mL distilled water, and the pH is adjusted to 7.0; and the medium is sterilized at a high temperature of 121° C. with steam for 20 min.

[0048] The TSB liquid culture medium is composed of: 15 g of tryptone, 5 g of soy peptone, 5 g of sodium chloride, and 1000 ml of distilled water. The pH is adjusted to 7.2±0.2 and sterilized by pressure steam at 121° C. for 20 min.

[0049] The Bacillus clausii strain is numbered as CGMCC No.1.3672, purchased from China General Microbiological Culture Collection Administration Center, with a preservation date of June 11, 2004, and a preservation address of No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing; the Streptomyces lilacinus strain is numbered as CGMCC No.4.6512, purchased from China General Microbiological Culture Collection Administration Center, with a preservation date of September 3, 2008, and a preservation address of No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing; the Marseilles timonei strain is numbered as CGMCC No.1.12742, purchased from China General Microbiological Culture Collection Administration Center, with a preservation date of December 26, 2013, and a preservation address of No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing.

[0050] The nutrient agent is composed of urea, potassium dihydrogen phosphate, amino acid chelated zinc, amino acid chelated magnesium, and amino acid chelated manganese in a mass ratio of 1:0.8:0.05:0.02:0.01.

[0051] A method for preparing the composite microbial agent for resisting both angelica root rot and rashes comprises the following steps: A. preparing composite microbial powder; B. preparing the carrier; C. preparing a golden bamboo grass extract, and then combining the golden bamboo grass extract with cocoamidopropyl betaine in a mass ratio of 5:1 to form an active agent; D. uniformly mixing the composite microbial powder, carrier and active agent, and drying at 40° C. to obtain a mixture; E. Evenly mix the above mixture with the nutrient agent to obtain the final product.

[0052] Comparative Example 1 A composite microbial agent capable of resisting angelica root rot and rashes, comprising the following raw materials in parts by weight: 30 parts of composite microbial powder, 4 parts of an active agent, 15 parts of a carrier, and 8 parts of a nutrient agent; the composite microbial agent comprises Bacillus clausii, Streptomyces lilacinus, and Masseillaris timonei; the carrier is rice husk biochar; the active agent is a composition of golden bamboo grass extract and coconut oil amidopropyl betaine in a mass ratio of 5:1.

[0053] The preparation method of the golden bamboo grass extract is as follows: (1) Weigh 100 g of golden bamboo grass, clean it, dry it in an oven, crush it, and pass it through an 80-mesh sieve to obtain powdered golden bamboo grass; (2) Take 15 g of golden bamboo grass powder, add it to 150 ml of 2% wt oxalic acid solution, stir, then transfer it to a hydrothermal reactor, let it stand for 45 minutes, and then place the hydrothermal reactor in a forced air drying oven and keep it at 150°C for 3 hours. After the insulation is completed, filter and take filtrate A; (3) adding 2%wt calcium carbonate suspension to the filtrate A obtained in step (2) until no bubbles are generated, letting it stand for 30-40 minutes and then filtering to remove the precipitate to obtain filtrate B; (4) Add an appropriate amount of activated carbon to filtrate B at a material-liquid ratio of 1 g:30 ml, set the temperature to 70 °C in a constant temperature oscillator, oscillate at 150 rpm for 2 h, and then filter to obtain the golden bamboo grass extract.

[0054] The composite microbial agent is prepared by the following method: thawing and activating Bacillus clausii, Streptomyces lilacinus and Massimodium timonei strains respectively, culturing in a 28°C incubator for 48 hours after streaking on a plate, taking a loop of the cultured strains in an LB liquid culture medium, placing the culture medium in a shaking incubator at 30°C and 180 r / min for 24 hours to obtain a seed solution; inoculating the seed solutions of the three strains into a TSB liquid culture medium at an inoculation rate of 3% respectively, bottling the culture medium at a volume of 200 mL, 30°C, and 200 r / min for 72 hours to obtain a fermentation solution; and mixing the three fermentation solutions in a mass ratio of 1:1:1 and freeze-drying the mixture to obtain a bacterial powder, namely the composite microbial powder.

[0055] The LB liquid culture medium is composed of: 5 g yeast extract, 10 g tryptone, 10 g sodium chloride, 1000 mL distilled water, and the pH is adjusted to 7.0; and the medium is sterilized at a high temperature of 121° C. with steam for 20 min.

[0056] The TSB liquid culture medium is composed of: 15 g of tryptone, 5 g of soy peptone, 5 g of sodium chloride, and 1000 ml of distilled water. The pH is adjusted to 7.2±0.2 and sterilized by pressure steam at 121° C. for 20 min.

[0057] The Bacillus clausii strain is numbered as CGMCC No.1.3672, purchased from China General Microbiological Culture Collection Administration Center, with a preservation date of June 11, 2004, and a preservation address of No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing; the Streptomyces lilacinus strain is numbered as CGMCC No.4.6512, purchased from China General Microbiological Culture Collection Administration Center, with a preservation date of September 3, 2008, and a preservation address of No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing; the Marseilles timonei strain is numbered as CGMCC No.1.12742, purchased from China General Microbiological Culture Collection Administration Center, with a preservation date of December 26, 2013, and a preservation address of No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing.

[0058] The nutrient agent is composed of urea, potassium dihydrogen phosphate, amino acid chelated zinc, amino acid chelated magnesium, and amino acid chelated manganese in a mass ratio of 1:0.8:0.05:0.02:0.01.

[0059] A method for preparing the composite microbial agent for resisting both angelica root rot and rashes comprises the following steps: A. preparing composite microbial powder; B. preparing a golden bamboo grass extract, and then combining the golden bamboo grass extract with cocoamidopropyl betaine in a mass ratio of 5:1 to form an active agent; C. uniformly mixing the composite microbial powder, carrier and active agent, and drying at 40° C. to obtain a mixture; D. Evenly mix the above mixture with the nutrient agent to obtain the final product.

[0060] This comparative example is basically the same as Example 3, the only difference being that the carrier is commercially available rice husk biochar.

[0061] Comparative Example 2 A composite microbial agent capable of resisting angelica root rot and rashes is prepared from the following raw materials in parts by weight: 30 parts of composite microbial powder, 15 parts of a carrier, and 8 parts of a nutrient agent; the composite microbial agent comprises Bacillus clausii, Streptomyces lilacinus, and Masseillaris timonei; and the carrier is made of fly ash, bentonite, and rice husk biochar.

[0062] The carrier is prepared by the following method: fly ash, bentonite and rice husk biochar are mixed in a mass ratio of 1:1:3, mechanically ground for 30 minutes, then placed in a high-temperature muffle furnace and heated to 800°C at a speed of 5°C / min and kept for 1 hour, then heated to 1100°C at a speed of 3°C / min, kept for 2 hours, and ground into powder after cooling to obtain the carrier.

[0063] The composite microbial agent is prepared by the following method: thawing and activating Bacillus clausii, Streptomyces lilacinus and Massimodium timonei strains respectively, culturing in a 28°C incubator for 48 hours after streaking on a plate, taking a loop of the cultured strains in an LB liquid culture medium, placing the culture medium in a shaking incubator at 30°C and 180 r / min for 24 hours to obtain a seed solution; inoculating the seed solutions of the three strains into a TSB liquid culture medium at an inoculation rate of 3% respectively, bottling the culture medium at a volume of 200 mL, 30°C, and 200 r / min for 72 hours to obtain a fermentation solution; and mixing the three fermentation solutions in a mass ratio of 1:1:1 and freeze-drying the mixture to obtain a bacterial powder, namely the composite microbial powder.

[0064] The LB liquid culture medium is composed of: 5 g yeast extract, 10 g tryptone, 10 g sodium chloride, 1000 mL distilled water, and the pH is adjusted to 7.0; and the medium is sterilized at a high temperature of 121° C. with steam for 20 min.

[0065] The TSB liquid culture medium is composed of: 15 g of tryptone, 5 g of soy peptone, 5 g of sodium chloride, and 1000 ml of distilled water. The pH is adjusted to 7.2±0.2 and sterilized by pressure steam at 121° C. for 20 min.

[0066] The Bacillus clausii strain is numbered as CGMCC No.1.3672, purchased from China General Microbiological Culture Collection Administration Center, with a preservation date of June 11, 2004, and a preservation address of No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing; the Streptomyces lilacinus strain is numbered as CGMCC No.4.6512, purchased from China General Microbiological Culture Collection Administration Center, with a preservation date of September 3, 2008, and a preservation address of No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing; the Marseilles timonei strain is numbered as CGMCC No.1.12742, purchased from China General Microbiological Culture Collection Administration Center, with a preservation date of December 26, 2013, and a preservation address of No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing.

[0067] The nutrient agent is composed of urea, potassium dihydrogen phosphate, amino acid chelated zinc, amino acid chelated magnesium, and amino acid chelated manganese in a mass ratio of 1:0.8:0.05:0.02:0.01.

[0068] A method for preparing the composite microbial agent for resisting both angelica root rot and rashes comprises the following steps: A. preparing composite microbial powder; B. preparing the carrier; C. after uniformly mixing the composite microbial powder and the carrier, drying at 40° C. to obtain a mixture; D. Evenly mix the above mixture with the nutrient agent to obtain the final product.

[0069] This comparative example is basically the same as Example 3, the only difference being that it does not contain the active agent and the preparation method thereof.

[0070] Comparative Example 3 A composite microbial agent capable of resisting angelica root rot and rashes, comprising the following raw materials in parts by weight: 30 parts of composite microbial powder, 4 parts of an active agent, 15 parts of a carrier, and 8 parts of a nutrient agent; the composite microbial agent comprises Bacillus clausii and Streptomyces lilacinus; the carrier is made of fly ash, bentonite, and rice husk biochar; the active agent is composed of golden bamboo grass extract and coconut oil amidopropyl betaine in a mass ratio of 5:1.

[0071] The carrier is prepared by the following method: fly ash, bentonite and rice husk biochar are mixed in a mass ratio of 1:1:3, mechanically ground for 30 minutes, then placed in a high-temperature muffle furnace and heated to 800°C at a speed of 5°C / min and kept for 1 hour, then heated to 1100°C at a speed of 3°C / min, kept for 2 hours, and ground into powder after cooling to obtain the carrier.

[0072] The preparation method of the golden bamboo grass extract is as follows: (1) Weigh 100 g of golden bamboo grass, clean it, dry it in an oven, crush it, and pass it through an 80-mesh sieve to obtain powdered golden bamboo grass; (2) Take 15 g of golden bamboo grass powder, add it to 150 ml of 2% wt oxalic acid solution, stir, then transfer it to a hydrothermal reactor, let it stand for 45 minutes, and then place the hydrothermal reactor in a forced air drying oven and keep it at 150°C for 3 hours. After the insulation is completed, filter and take filtrate A; (3) adding 2%wt calcium carbonate suspension to the filtrate A obtained in step (2) until no bubbles are generated, letting it stand for 30-40 minutes and then filtering to remove the precipitate to obtain filtrate B; (4) Add an appropriate amount of activated carbon to filtrate B at a material-liquid ratio of 1 g:30 ml, set the temperature to 70 °C in a constant temperature oscillator, oscillate at 150 rpm for 2 h, and then filter to obtain the golden bamboo grass extract.

[0073] The composite microbial agent is prepared by the following method: thawing and activating Bacillus clausii and Streptomyces lilacinus strains respectively, culturing in a 28°C incubator for 48 hours after streaking on a plate, taking a loop of the cultured strains into an LB liquid culture medium, placing the culture medium in a shaking incubator at 30°C and 180 r / min for 24 hours to obtain a seed solution; inoculating the seed solutions of the two strains into a TSB liquid culture medium at an inoculation rate of 3%, bottling 200 mL, 30°C, 200 r / min shaking culture for 72 hours to obtain a fermentation solution; mixing the two fermentation solutions in a mass ratio of 1:1 and freeze-drying the mixture to obtain a bacterial powder, namely the composite microbial powder.

[0074] The LB liquid culture medium is composed of: 5 g yeast extract, 10 g tryptone, 10 g sodium chloride, 1000 mL distilled water, and the pH is adjusted to 7.0; and the medium is sterilized at a high temperature of 121° C. with steam for 20 min.

[0075] The TSB liquid culture medium is composed of: 15 g of tryptone, 5 g of soy peptone, 5 g of sodium chloride, and 1000 ml of distilled water. The pH is adjusted to 7.2±0.2 and sterilized by pressure steam at 121° C. for 20 min.

[0076] The Bacillus clausii strain is numbered CGMCC No.1.3672, purchased from the China General Microbiological Culture Collection Center, the preservation date is June 11, 2004, and the preservation address is: No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing; the Streptomyces lilacinus strain is numbered CGMCC No.4.6512, purchased from the China General Microbiological Culture Collection Center, the preservation date is September 3, 2008, and the preservation address is: No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing.

[0077] The nutrient agent is composed of urea, potassium dihydrogen phosphate, amino acid chelated zinc, amino acid chelated magnesium, and amino acid chelated manganese in a mass ratio of 1:0.8:0.05:0.02:0.01.

[0078] A method for preparing the composite microbial agent for resisting both angelica root rot and rashes comprises the following steps: A. preparing composite microbial powder; B. preparing the carrier; C. preparing a golden bamboo grass extract, and then combining the golden bamboo grass extract with cocoamidopropyl betaine in a mass ratio of 5:1 to form an active agent; D. uniformly mixing the composite microbial powder, carrier and active agent, and drying at 40° C. to obtain a mixture; E. Evenly mix the above mixture with the nutrient agent to obtain the final product.

[0079] This comparative example is basically the same as Example 3, the only difference being that the composite microbial powder does not contain Masseille timonei and the corresponding culture method.

[0080] Comparative Example 4 A composite microbial agent capable of resisting angelica root rot and dermatophyte disease is prepared from the following raw materials in parts by weight: 30 parts of composite microbial powder, 4 parts of an active agent, 15 parts of a carrier, and 8 parts of a nutrient agent; the composite microbial agent comprises Bacillus clausii and Masseillaris timonei; the carrier is made of fly ash, bentonite, and rice husk biochar; the active agent is composed of golden bamboo grass extract and coconut oil amidopropyl betaine in a mass ratio of 5:1.

[0081] The carrier is prepared by the following method: fly ash, bentonite and rice husk biochar are mixed in a mass ratio of 1:1:3, mechanically ground for 30 minutes, then placed in a high-temperature muffle furnace and heated to 800°C at a speed of 5°C / min and kept for 1 hour, then heated to 1100°C at a speed of 3°C / min, kept for 2 hours, and ground into powder after cooling to obtain the carrier.

[0082] The preparation method of the golden bamboo grass extract is as follows: (1) Weigh 100 g of golden bamboo grass, clean it, dry it in an oven, crush it, and pass it through an 80-mesh sieve to obtain powdered golden bamboo grass; (2) Take 15 g of golden bamboo grass powder, add it to 150 ml of 2% wt oxalic acid solution, stir, then transfer it to a hydrothermal reactor, let it stand for 45 minutes, and then place the hydrothermal reactor in a forced air drying oven and keep it at 150°C for 3 hours. After the insulation is completed, filter and take filtrate A; (3) adding 2%wt calcium carbonate suspension to the filtrate A obtained in step (2) until no bubbles are generated, letting it stand for 30-40 minutes and then filtering to remove the precipitate to obtain filtrate B; (4) Add an appropriate amount of activated carbon to filtrate B at a material-liquid ratio of 1 g:30 ml, set the temperature to 70 °C in a constant temperature oscillator, oscillate at 150 rpm for 2 h, and then filter to obtain the golden bamboo grass extract.

[0083] The composite microbial agent is prepared by the following method: thawing and activating Bacillus clausii and Massimo timonei strains respectively, culturing in a 28°C incubator for 48 hours after streaking on a plate, taking a loop of the cultured strains into an LB liquid culture medium, placing the culture medium in a shaking incubator at 30°C and 180 r / min for 24 hours to obtain a seed solution; inoculating the seed solutions of the two strains into a TSB liquid culture medium at an inoculation rate of 3%, bottling 200 mL, 30°C, 200 r / min shaking culture for 72 hours to obtain a fermentation solution; mixing the two fermentation solutions in a mass ratio of 1:1 and freeze-drying the mixture to obtain a bacterial powder, namely the composite microbial powder.

[0084] The LB liquid culture medium is composed of: 5 g yeast extract, 10 g tryptone, 10 g sodium chloride, 1000 mL distilled water, and the pH is adjusted to 7.0; and the medium is sterilized at a high temperature of 121° C. with steam for 20 min.

[0085] The TSB liquid culture medium is composed of: 15 g of tryptone, 5 g of soy peptone, 5 g of sodium chloride, and 1000 ml of distilled water. The pH is adjusted to 7.2±0.2 and sterilized by pressure steam at 121° C. for 20 min.

[0086] The Bacillus clausii strain is numbered CGMCC No.1.3672, purchased from the China General Microbiological Culture Collection Center, the preservation date is June 11, 2004, and the preservation address is: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing; the Marseille timonei strain is numbered CGMCC No.1.12742, purchased from the China General Microbiological Culture Collection Center, the preservation date is December 26, 2013, and the preservation address is: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.

[0087] The nutrient agent is composed of urea, potassium dihydrogen phosphate, amino acid chelated zinc, amino acid chelated magnesium, and amino acid chelated manganese in a mass ratio of 1:0.8:0.05:0.02:0.01.

[0088] A method for preparing the composite microbial agent for resisting both angelica root rot and rashes comprises the following steps: A. preparing composite microbial powder; B. preparing the carrier; C. preparing a golden bamboo grass extract, and then combining the golden bamboo grass extract with cocoamidopropyl betaine in a mass ratio of 5:1 to form an active agent; D. uniformly mixing the composite microbial powder, carrier and active agent, and drying at 40° C. to obtain a mixture; E. Evenly mix the above mixture with the nutrient agent to obtain the final product.

[0089] This comparative example is basically the same as Example 3, the only difference being that the composite microbial powder does not contain Streptomyces lilacinus and the corresponding culture method.

[0090] Comparative Example 5 A composite microbial agent capable of resisting angelica root rot and rashes, comprising the following raw materials in parts by weight: 30 parts of composite microbial powder, 4 parts of an active agent, 15 parts of a carrier, and 8 parts of a nutrient agent; the composite microbial agent comprises Streptomyces lilacinus and Masseillaris timonei; the carrier is made of fly ash, bentonite, and rice husk biochar; the active agent is composed of golden bamboo grass extract and coconut oil amidopropyl betaine in a mass ratio of 5:1.

[0091] The carrier is prepared by the following method: fly ash, bentonite and rice husk biochar are mixed in a mass ratio of 1:1:3, mechanically ground for 30 minutes, then placed in a high-temperature muffle furnace and heated to 800°C at a speed of 5°C / min and kept for 1 hour, then heated to 1100°C at a speed of 3°C / min, kept for 2 hours, and ground into powder after cooling to obtain the carrier.

[0092] The preparation method of the golden bamboo grass extract is as follows: (1) Weigh 100 g of golden bamboo grass, clean it, dry it in an oven, crush it, and pass it through an 80-mesh sieve to obtain powdered golden bamboo grass; (2) Take 15 g of golden bamboo grass powder, add it to 150 ml of 2% wt oxalic acid solution, stir, then transfer it to a hydrothermal reactor, let it stand for 45 minutes, and then place the hydrothermal reactor in a forced air drying oven and keep it at 150°C for 3 hours. After the insulation is completed, filter and take filtrate A; (3) adding 2%wt calcium carbonate suspension to the filtrate A obtained in step (2) until no bubbles are generated, letting it stand for 30-40 minutes and then filtering to remove the precipitate to obtain filtrate B; (4) Add an appropriate amount of activated carbon to filtrate B at a material-liquid ratio of 1 g:30 ml, set the temperature to 70 °C in a constant temperature oscillator, oscillate at 150 rpm for 2 h, and then filter to obtain the golden bamboo grass extract.

[0093] The composite microbial agent is prepared by the following method: thawing and activating Streptomyces lilacinus and Massimodium timonei strains respectively, culturing in a 28°C incubator for 48 hours after streaking on a plate, taking a loop of the cultured strains in an LB liquid culture medium, placing the culture medium in a shaker at 30°C and 180 r / min for 24 hours to obtain a seed solution; inoculating the seed solutions of the two strains into a TSB liquid culture medium at an inoculation rate of 3%, bottling 200 mL, 30°C, 200 r / min shaking culture for 72 hours to obtain a fermentation solution; mixing the two fermentation solutions in a mass ratio of 1:1 and freeze-drying the mixture to obtain a bacterial powder, namely the composite microbial powder.

[0094] The LB liquid culture medium is composed of: 5 g yeast extract, 10 g tryptone, 10 g sodium chloride, 1000 mL distilled water, and the pH is adjusted to 7.0; and the medium is sterilized at a high temperature of 121° C. with steam for 20 min.

[0095] The TSB liquid culture medium is composed of: 15 g of tryptone, 5 g of soy peptone, 5 g of sodium chloride, and 1000 ml of distilled water. The pH is adjusted to 7.2±0.2 and sterilized by pressure steam at 121° C. for 20 min.

[0096] The Streptomyces lilacinus strain is numbered CGMCC No.4.6512, purchased from the China General Microbiological Culture Collection Center, with a preservation date of September 3, 2008, and a preservation address of No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing; the Marseilles timonei strain is numbered CGMCC No.1.12742, purchased from the China General Microbiological Culture Collection Center, with a preservation date of December 26, 2013, and a preservation address of No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing.

[0097] The nutrient agent is composed of urea, potassium dihydrogen phosphate, amino acid chelated zinc, amino acid chelated magnesium, and amino acid chelated manganese in a mass ratio of 1:0.8:0.05:0.02:0.01.

[0098] A method for preparing the composite microbial agent for resisting both angelica root rot and rashes comprises the following steps: A. preparing composite microbial powder; B. preparing the carrier; C. preparing a golden bamboo grass extract, and then combining the golden bamboo grass extract with cocoamidopropyl betaine in a mass ratio of 5:1 to form an active agent; D. uniformly mixing the composite microbial powder, carrier and active agent, and drying at 40° C. to obtain a mixture; E. Evenly mix the above mixture with the nutrient agent to obtain the final product.

[0099] This comparative example is basically the same as Example 3, the only difference being that the composite microbial powder does not contain Bacillus Clausii and the corresponding culture method.

[0100] Test verification Antibacterial effect verification: Pathogen: Fusarium solani strain of Angelica sinensis, isolated from Angelica sinensis root rot samples, identified and preserved by the applicant.

[0101] The plate confrontation test was used to verify the antibacterial effect of different composite bacterial liquids. The specific operation method was as follows: the pathogen was inoculated into the PDA culture medium and cultured at 28°C. After the colony covered the plate, a bacterial cake (6 mm in diameter) was punched out with a hole puncher. The bacterial cake was inverted in the center of a 9 cm PDA plate. Four equidistant points were taken on the vertical intersection line centered on the bacterial plate, and 5 μL of different mixed fermentation bacterial liquids of the composite microbial powder described in Example 3, Comparative Example 3, Comparative Example 4, and Comparative Example 5 before freeze-drying were inoculated respectively. The control group was inoculated with an equal volume of LB liquid culture medium, which was placed in an incubator at 28°C for culture. The inhibition zone was observed after 5 days. The results are as follows Figure 1 As shown. For Fusarium spp. in Angelica sinensis, the bacterial solution obtained by fermentation and culture of the composite strain in the embodiment of the present invention showed good antibacterial effect. However, after any one of the strains was missing in Comparative Examples 3-5, the antibacterial effect on Fusarium spp. in Angelica sinensis became smaller, which shows that the three preferred strains of the present invention have good synergistic effects and have good preventive effects on Fusarium spp., the main pathogen of Angelica sinensis root rot and hemp-mouth disease.

[0102] Planting test A plot of land with severe angelica root rot year after year was selected as the planting site. The last crop was angelica, and the planting variety was Mingui No. 2. The test plot was divided into 7 treatment groups, each with an area of ​​1 mu, a row spacing of 40 cm, a plant spacing of 30 cm, and an aisle of 0.5 m between cells. Artificial hole transplanting was adopted, with 2 plants per hole. When transplanting, the composite microbial agent prepared by Example 3 of the present invention and Comparative Examples 1-5 was mixed and applied as a base fertilizer with conventional chemical fertilizers, and the dosage was 10 kg / mu. At the same time, a blank control group was set up, and only conventional chemical fertilizers were applied. All others were managed according to conventional field planting. Until the angelica was harvested, the yield (in fresh weight), the first-class return rate, the number of root rot diseased plants and the incidence rate were statistically counted, and the results are shown in Table 1.

[0103] Table 1 Statistics of angelica yield and root rot incidence in each treatment group It can be seen from the data in Table 1 above that the yield and quality of Angelica sinensis are significantly improved after use of the composite microbial agent prepared by the present invention, which is significantly better than comparative examples 1-5, and the rate of Angelica sinensis root rot diseased plants is also significantly lower than that of the blank control group. This is mainly due to the interaction between the raw materials of the present invention, which can have a good preventive and therapeutic effect on Angelica sinensis root rot, and has a significant effect on improving the yield and quality of Angelica sinensis.

[0104] Another plot of land with severe Angelica sinensis paroxysmal disease year after year was selected as the planting site. The last crop was Angelica sinensis, and the planting variety was Mingui No. 2. The test plot was divided into 7 treatment groups, each with an area of ​​1 mu, a row spacing of 40 cm, a plant spacing of 30 cm, and a corridor of 0.5 m between cells. Artificial hole transplanting was adopted, with 2 plants per hole. When transplanting, the composite microbial inoculant prepared by Example 3 of the present invention and Comparative Examples 1-5 was mixed and applied as a base fertilizer with conventional chemical fertilizers, and the dosage was 10 kg / mu. At the same time, a blank control group was set up, and only conventional chemical fertilizers were applied. All others were managed according to conventional field planting. Until the Angelica sinensis was harvested, the yield (in fresh weight), the first-class return rate, the number of plants with paroxysmal disease, and the incidence rate were statistically analyzed, and the results are shown in Table 2.

[0105] Table 2 Statistics of angelica yield and the incidence of leprosy in each treatment group From the results in Table 2 above, it can be seen that the composite microbial agent prepared by the present invention also has a good preventive and therapeutic effect on Angelica sinensis paronychia, and compared with the blank control, the yield and quality of Angelica sinensis are also significantly improved.

[0106] It can be seen from the results of the above embodiments and test examples that the composite microbial agent prepared by the present invention has a good prevention and control effect on common root rot and hemp mouth disease of Angelica sinensis. At the same time, the prepared composite microbial agent contains rich active microorganisms. While effectively inhibiting the growth of pathogens, its various metabolites can also promote the growth of Angelica sinensis, effectively improve the yield and quality of Angelica sinensis, and have good application prospects.

[0107] It should be noted that the above embodiments are only partial embodiments of the preferred methods of implementing the present invention, rather than all embodiments. Obviously, based on the above embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work should fall within the scope of protection of the present invention.

Claims

1. A composite microbial agent for resisting angelica root rot and rashes, characterized in that: The invention is made of the following raw materials in parts by weight: 20-40 parts of composite microbial powder, 3-5 parts of an active agent, 10-20 parts of a carrier, and 5-10 parts of a nutrient agent; the composite microbial agent comprises Bacillus clausii, Streptomyces lilacinus, and Massimodium timonei; the carrier is made of fly ash, bentonite, and rice husk biochar; the active agent is composed of golden bamboo grass extract and coconut oil amide propyl betaine in a mass ratio of 5:

1.

2. The composite microbial agent for resisting angelica root rot and rashes according to claim 1, characterized in that: The carrier is prepared by the following method: fly ash, bentonite and rice husk biochar are mixed in a mass ratio of 1:1:3, mechanically ground for 30 minutes, then placed in a high-temperature muffle furnace, heated to 800°C in stages, kept warm for 1 hour, then heated to 1100°C, kept warm for 2 hours, and ground into powder after cooling to obtain the carrier.

3. The composite microbial agent for resisting angelica root rot and rashes according to claim 2, characterized in that: The temperature of the muffle furnace was raised to 800° C. at a rate of 5° C. / min, and then raised to 1100° C. at a rate of 3° C. / min.

4. The composite microbial agent for resisting angelica root rot and rashes according to claim 1, characterized in that: The preparation method of the golden bamboo grass extract is as follows: (1) Weigh 100 g of golden bamboo grass, clean it, dry it in an oven, crush it, and pass it through an 80-mesh sieve to obtain powdered golden bamboo grass; (2) Take 15 g of golden bamboo grass powder, add it to 150 ml of 2% wt oxalic acid solution, stir, then transfer it to a hydrothermal reactor, let it stand for 45 minutes, and then place the hydrothermal reactor in a forced air drying oven and keep it at 150°C for 3 hours. After the insulation is completed, filter and take filtrate A; (3) adding 2%wt calcium carbonate suspension to the filtrate A obtained in step (2) until no bubbles are generated, letting it stand for 30-40 minutes and then filtering to remove the precipitate to obtain filtrate B; (4) Add an appropriate amount of activated carbon to the filtrate B, set the temperature to 70°C in a constant temperature oscillator, and oscillate at 150 rpm for 2 h, then filter to obtain the golden bamboo grass extract.

5. The composite microbial agent for resisting angelica root rot and rashes according to claim 4, characterized in that: In the step (4), the material-liquid ratio of activated carbon to filtrate B is 1 g:30 ml.

6. The composite microbial agent for resisting angelica root rot and rashes according to claim 1, characterized in that: The composite microbial agent is prepared by the following method: thawing and activating Bacillus clausii, Streptomyces lilacinus, and Massimodium timonei strains respectively, streaking the plates and culturing them in a 28°C incubator for 48 hours, taking a loop of the cultured strains in LB liquid culture medium, placing the culture medium in a shaking incubator at 30°C and 180 r / min for 24 hours, and obtaining a seed solution; The seed liquid of the three strains was inoculated into TSB liquid culture medium at an inoculation rate of 3%, the bottling volume was 200 mL, and the culture was shaken at 30°C and 200 r / min for 72 h to obtain fermentation liquid; the three fermentation liquids were mixed in a mass ratio of 1:1:1 and freeze-dried to obtain bacterial powder, which is the composite microbial bacterial powder.

7. The composite microbial agent for resisting angelica root rot and rashes according to claim 1 or 6, characterized in that: The strain number of the Bacillus clausii is CGMCC No.1.3672, the strain number of Streptomyces lilacinus is CGMCC No.4.6512, and the strain number of Masseillione timonei is CGMCC No.1.12742.

8. The composite microbial agent for resisting angelica root rot and rashes according to claim 1, characterized in that: The nutrient agent is composed of urea, potassium dihydrogen phosphate, amino acid chelated zinc, amino acid chelated magnesium, and amino acid chelated manganese in a mass ratio of 1:0.8:0.05:0.02:0.

01.

9. A method for preparing the composite microbial agent for resisting angelica root rot and dermatophyte according to any one of claims 1 to 9, characterized in that: It includes the following steps: A. preparing composite microbial powder; B. preparing the carrier; C. preparing a golden bamboo grass extract, and then combining the golden bamboo grass extract with cocoamidopropyl betaine in a mass ratio of 5:1 to form an active agent; D. uniformly mixing the composite microbial powder, carrier and active agent, and drying at 40° C. to obtain a mixture; E. Evenly mix the above mixture with the nutrient agent to obtain the final product.

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