Microbial agent for promoting growth and development of orchid and preparation process of microbial agent

Abstract

The invention relates to the technical field of orchid seedling culture, in particular to a microbial agent for promoting growth and development of orchid and a preparation process of the microbial agent. The preparation process comprises the following steps: preparing a cottonseed hull biochar material; preparing a carbon-based microbial stationary liquid; and preparing a microbial agent for promoting the growth and development of orchid plants. According to the invention, the oyster mushroom strain is fixed in the cottonseed hull with a lignocellulose structure, a plant framework is broken by using an enzyme secreted by natural growth of fungi to obtain a biochar precursor with well-developed pores, and after the cottonseed hull is prepared into the biochar under the action of high temperature, the strain can be fixed in the pores of the biochar material to grow and propagate; and a layer of continuous and homologous biological membrane is formed, so that the reproduction of microorganisms in the same population is expanded, the concentration of dominant bacteria in the microbial liquid is increased, and the promotion effect of the microbial agent on plant growth is enhanced.

Description

Technical Field

[0001] The invention relates to the technical field of orchid seedling cultivation, and in particular to a microbial agent for promoting the growth and development of orchid plants and a preparation process thereof. Background Art

[0002] Microbial agents are a type of product that uses beneficial microorganisms such as bacteria, fungi, actinomycetes, etc., and their metabolites to improve plant growth, soil health, or prevent and control pests and diseases. According to their uses, they can be divided into soil conditioners, plant growth regulators, and biopesticides.

[0003] Orchids are a type of plant that has high requirements for environmental conditions. Due to their high requirements for nutrient absorption and soil environment, orchids usually form a symbiotic structure with mycorrhizal fungi, which greatly expands the root absorption range of orchids and enhances their ability to absorb water and nutrients. Therefore, during the growth and development of orchids, microbial agents can be used to increase the seedling formation rate of orchids.

[0004] However, the commonly used strains of existing microbial agents are generally nitrogen-fixing bacteria such as rhizobia and Frankia, and plant hormone-producing strains such as Bacillus subtilis and Pseudomonas fluorescens. These strains enrich the nutrients in the soil through their life activities, or produce plant hormones to promote plant growth. However, orchids absorb less nutrients in the process of forming seedlings, and their response to external hormone stimulation is also relatively weak. Therefore, the common microbial agent strains have limited promoting effect on the development of orchid seeds. Therefore, the present invention provides a microbial agent for promoting the growth and development of orchids and a preparation process thereof, and achieves the effect of promoting the growth and development of orchids by preparing a microbial agent using endophytic fungi of orchids. Summary of the invention

[0005] In view of the shortcomings of the prior art, the object of the present invention is to provide a microbial agent for promoting the growth and development of orchid plants and a preparation process thereof.

[0006] S1: Preparation of cottonseed hull biochar material Add cottonseed hulls, MgSO4, KH2PO4, urea, lime and sugar to deionized water, then add Pleurotus ostreatus strains and mix evenly, culture at a constant temperature, then steam inactivate to obtain microbial treated cottonseed hulls, pyrolyze the microbial treated cottonseed hulls to obtain cottonseed hull powder, grind and sieve, then add NaOH, heat and activate to obtain heat-treated powder, wash and dry the heat-treated powder to obtain cottonseed hull biochar material; S2: Preparation of carbon-based microbial fixative The rice husk, corn stalk and rice stalk are mixed, crushed and sieved, and then loaded into a quartz boat, flattened and compacted, wrapped with tin foil and sealed, and placed in a vacuum tube high temperature furnace, and pyrolyzed in a vacuum tube high temperature furnace using nitrogen as a carrier gas and a protective gas to obtain a mixed biochar material, and the mixed biochar material is mixed with a cottonseed hull biochar material to obtain a mixed powder, and the mixed powder is mixed with a PDA culture medium, and adsorbed and fixed by shaking to obtain a carbon-based microbial fixative; S3: Preparation of composite humic acid hydrogel powder Take a healthy Paphiopedilum sclerophyllum and rinse the surface matrix under running water, and use filter paper to absorb the water, cut out the roots of the Paphiopedilum sclerophyllum and disinfect them, first soak the roots of the Paphiopedilum sclerophyllum in alcohol, then disinfect them with mercuric chloride to obtain treated roots, place the treated roots on a culture dish, cut them into tissue segments, place them on a PDA culture medium for inoculation, and add a charcoal-based microbial fixative, and culture them in an incubator away from light. After colonies grow out of the tissue segments, pick up the hyphae at the edge of the colony, transfer them to a new culture dish containing a PDA culture medium for purification and culture, purify and obtain growth-promoting microorganisms, dilute the growth-promoting microorganisms with deionized water, and then add L-tryptophan to obtain a microbial agent that promotes the growth and development of orchids.

[0007] Furthermore, step S1 of preparing cottonseed hull biochar material comprises the following steps: S1.1: Add 4-5 parts by mass of cottonseed hulls, 0.2-0.5 parts by mass of MgSO4, 1-3 parts by mass of KH2PO4, 0.5-2 parts by mass of urea, 0.1-0.5 parts by mass of lime and 1-3 parts by mass of white sugar to 100 parts by mass of deionized water, then add 0.2-0.4 parts by mass of Pleurotus ostreatus spawn, mix well, and culture at a constant temperature of 24-26°C for 40 days, then steam inactivate at 120-125°C, then wash with deionized water 3 times, and dry at 100-105°C to constant weight to obtain microbial treated cottonseed hulls; S1.2: The cottonseed hull treated with microorganisms is heated to 500-550°C in an argon atmosphere at a flow rate of 100 mL / min, the heating rate is 3°C / min, and the pyrolysis is performed for 1-1.5 hours to obtain cottonseed hull powder, which is then ground through an 80-mesh sieve, and then 4-4.5 times the mass of NaOH is added, and argon is passed through the argon atmosphere at a flow rate of 100 mL / min, heated to 800-850°C, and activated for 3-3.5 hours to obtain heat-treated powder; S1.3: The heat-treated powder is washed with 0.1 mol / L HCl and dried at 60-80° C. to a constant weight to obtain a cottonseed hull biochar material.

[0008] Furthermore, step S2 prepares a carbon-based microbial fixative solution, comprising the following steps: S2.1: Mix rice husk, corn stalk and rice straw in a mass ratio of 1:1:(1-2), crush them through an 18-mesh sieve, put them into a quartz boat, flatten and compact them, wrap them with tin foil and seal them in a vacuum tube high-temperature furnace, use nitrogen as carrier gas and protective gas, pyrolyze them in a vacuum tube high-temperature furnace for 2-2.5h, and the pyrolysis temperature is 600℃ to obtain a mixed biochar material; S2.2: Mix the mixed biochar material with the cottonseed hull biochar material in a mass ratio of 1: (2-3) to obtain a mixed powder, mix the mixed powder with PDA culture medium in a ratio of 1 g: (10-12) mL, and fix by shaking adsorption at 35-36°C and 160-180 r / min for 24-25 hours to obtain a carbon-based microbial fixative.

[0009] Furthermore, step S3 prepares a microbial agent for promoting the growth and development of orchid plants, comprising the following steps: S3.1: Take a healthy Paphiopedilum sclerophyllum and rinse the surface substrate under running water and use filter paper to absorb the water. Cut out the roots of Paphiopedilum sclerophyllum and disinfect them. First, soak the roots of Paphiopedilum sclerophyllum in alcohol for 60-65 seconds, rinse them with sterile water 2-3 times, wipe the surface moisture with sterilized filter paper, then disinfect them with 0.1-0.15% mercuric chloride for 5-9 minutes, rinse them with sterile water 2-3 times, wipe the surface moisture with sterilized filter paper, and get the treated roots. S3.2: Place the treated roots on a culture dish and cut them into 4.8-5.2 mm tissue segments, inoculate them on PDA culture medium, and add 5-20% of the total mass of the system's carbon-based microbial fixative solution. Incubate them in an incubator at 25-26°C in the dark for 7 days. After colonies grow out of the tissue segments, pick up the hyphae at the edge of the colonies, transfer them to a new culture dish containing PDA culture medium for purification and culture, purify 2-3 times, and obtain growth-promoting microorganisms; S3.3: Dilute the growth-promoting microorganism with 10 times the volume of deionized water, and then add 0.5-0.6 g / L of L-tryptophan to obtain a microbial agent that promotes the growth and development of orchids.

[0010] Furthermore, the oyster mushroom species is a fungus with the scientific name of Pleurotus ostreatus.

[0011] Furthermore, the HCl concentration is 0.1 mol / L.

[0012] Furthermore, the flow rate of nitrogen was 100 mL / min, and the temperature heating rate was set at 18-20° C. / min.

[0013] Furthermore, the PDA culture medium is prepared by mixing 15 g of agar, 200 g of peeled potatoes, 20 g of glucose, and 1 L of pure water.

[0014] Further, the Paphiopedilum micranthum is a triennial plant with the scientific name Paphiopedilum micranthum Tang & FT Wang.

[0015] A microbial agent for promoting the growth and development of orchid plants is prepared by the above-mentioned preparation process of the microbial agent for promoting the growth and development of orchid plants.

[0016] Compared with the prior art, the present invention has at least the following beneficial effects: 1. The present invention fixes the Pleurotus ostreatus strains inside the cottonseed hulls with a lignocellulose structure, uses the enzymes secreted by the natural growth of the fungus to break the plant framework, obtains a biochar precursor with well-developed pores, and uses NaOH for secondary modification to make the pores larger and looser. After such cottonseed hulls are made into biochar through high temperature, the strains can be fixed in the gaps of the biochar material for growth and reproduction, and form a continuous, homogeneous biofilm, so that the microorganisms of the same population can multiply and expand, and the concentration of the dominant strains in the microbial liquid is increased, thereby enhancing the promoting effect of the microbial agent on plant growth.

[0017] 2. The present invention separates endophytic fungi from the roots of Paphiopedilum sclerophyllum. The endophytic fungi help the growth and development of orchid plants by producing growth hormones, promoting the absorption of mineral nutrients, and enhancing the stress resistance of plants. Since orchid seeds are tiny and lack endosperm, they rely heavily on fungi to provide nutrients required for seed germination. After the hyphae invade the seeds, the nutrients are transferred to the seeds through the embryo. Therefore, the microbial agent prepared from the endophytic fungi extracted from Paphiopedilum sclerophyllum can coexist with the seeds of orchid plants after spraying, and promote the germination of orchid seeds and the growth and development of plants by secreting hormones, vitamins, hydroxyproline-rich glycoproteins and other substances. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] The accompanying drawings, which are incorporated herein and constitute a part of the specification, illustrate embodiments of the present disclosure and, together with the description, further serve to explain the principles of the present disclosure and to enable those skilled in the relevant art to make and use the present disclosure.

[0019] Figure 1 The present invention is a flow chart of the preparation process of a microbial agent for promoting the growth and development of orchid plants used in an embodiment of the present invention. DETAILED DESCRIPTION

[0020] The following is a detailed description of a microbial agent for promoting the growth and development of orchid plants and its preparation process provided by the present invention in conjunction with the accompanying drawings and specific embodiments. At the same time, it is explained here that in order to make the embodiments more detailed, the following embodiments are the best and preferred embodiments, and those skilled in the art may also adopt other alternatives to implement some known technologies; and the accompanying drawings are only for a more specific description of the embodiments, and are not intended to specifically limit the present invention.

[0021] Example 1: A process for preparing a microbial agent for promoting the growth and development of orchid plants, such as Figure 1 As shown, the following steps are included: S1: Preparation of cottonseed hull biochar material S1.1: 4 parts by mass of cottonseed hulls, 0.2 parts by mass of MgSO4, 1 part by mass of KH2PO4, 0.5 parts by mass of urea, 0.1 parts by mass of lime and 1 part by mass of white sugar are added to 100 parts by mass of deionized water, and then 0.2 parts by mass of Pleurotus ostreatus are added, mixed evenly, and cultured at a constant temperature of 24°C for 40 days, then steam-inactivated at 120°C, then washed with deionized water for 3 times, and dried at 100°C to constant weight to obtain microbial-treated cottonseed hulls; S1.2: The cottonseed hull treated with microorganisms was heated to 500°C in an argon atmosphere with a flow rate of 100 mL / min, and the heating rate was 3°C / min. The cottonseed hull powder was obtained by pyrolysis for 1 hour, and then ground through an 80-mesh sieve, and then 4 times the mass of NaOH was added, and argon was passed through at a flow rate of 100 mL / min, heated to 800°C, and activated for 3 hours to obtain heat-treated powder; S1.3: The heat-treated powder was washed with 0.1 mol / L HCl and dried at 60°C to constant weight to obtain cottonseed hull biochar material.

[0022] S2: Preparation of carbon-based microbial fixative S2.1: Rice husk, corn stalk and rice stalk were mixed in a mass ratio of 1:1:1 and crushed through an 18-mesh sieve. The mixture was then placed in a quartz boat, flattened and compacted, wrapped with tin foil and sealed in a vacuum tube high-temperature furnace. Nitrogen was used as carrier gas and protective gas. The flow rate of nitrogen was 100 mL / min. The mixture was pyrolyzed in a vacuum tube high-temperature furnace for 2-2.5 h. The pyrolysis temperature was 600 °C and the temperature heating rate was set to 18 °C / min to obtain a mixed biochar material. S2.2: Mix the mixed biochar material and cottonseed hull biochar material in a mass ratio of 1:2 to obtain a mixed powder. Mix the mixed powder with PDA culture medium in a ratio of 1g:10mL. The PDA culture medium is prepared by mixing 15g agar, 200g peeled potato, 20g glucose and 1L pure water. Shake and adsorb and fix at 35°C and 160r / min for 24h to obtain a carbon-based microbial fixative.

[0023] S3: Preparation of microbial agents for promoting the growth and development of orchid plants S3.1: Take a healthy Paphiopedilum micranthum (scientific name: Tang & FT Wang), rinse the surface substrate under running water and dry it with filter paper, cut out the roots of Paphiopedilum micranthum and disinfect them. First, soak the roots of Paphiopedilum micranthum in 70% alcohol for 60 seconds, rinse them twice with sterile water, wipe the surface moisture with sterilized filter paper, then disinfect them with 0.1% mercuric chloride for 5 minutes, rinse them twice with sterile water, wipe the surface moisture with sterilized filter paper, and get the treated roots; S3.2: Place the treated roots on a culture dish and cut them into 4.8-5.2mm tissue segments, inoculate them on PDA culture medium, and add 5% carbon-based microbial fixative solution of the total mass of the system. Culture them in an incubator at 25°C in the dark for 7 days. After colonies grow out of the tissue segments, pick the hyphae at the edge of the colony and transfer them to a new culture dish containing PDA culture medium for purification and culture. Purify twice to obtain growth-promoting microorganisms. S3.3: Dilute the growth-promoting microorganism with 10 times the volume of deionized water, and then add 0.5 g / L of L-tryptophan to obtain a microbial agent that promotes the growth and development of orchids.

[0024] Example 2: A preparation process of a microbial agent for promoting the growth and development of orchid plants, such as Figure 1 As shown, the following steps are included: S1: Preparation of cottonseed hull biochar material S1.1: 5 parts by mass of cottonseed hulls, 0.5 parts by mass of MgSO4, 3 parts by mass of KH2PO4, 2 parts by mass of urea, 0.5 parts by mass of lime and 3 parts by mass of white sugar are added to 100 parts by mass of deionized water, and then 0.4 parts by mass of Pleurotus ostreatus is added, mixed evenly, and cultured at a constant temperature of 24°C for 40 days, then steam-inactivated at 120°C, then washed with deionized water for 3 times, and dried at 100°C to constant weight to obtain microbial-treated cottonseed hulls; S1.2: The cottonseed hull treated with microorganisms was heated to 500°C in an argon atmosphere with a flow rate of 100 mL / min, and the heating rate was 3°C / min. The cottonseed hull powder was obtained by pyrolysis for 1 hour, and then ground through an 80-mesh sieve, and then 4 times the mass of NaOH was added, and argon was passed through at a flow rate of 100 mL / min, heated to 800°C, and activated for 3 hours to obtain heat-treated powder; S1.3: The heat-treated powder was washed with 0.1 mol / L HCl and dried at 60°C to constant weight to obtain cottonseed hull biochar material.

[0025] S2: Preparation of carbon-based microbial fixative S2.1: Rice husk, corn stalk and rice stalk were mixed in a mass ratio of 1:1:2 and crushed through an 18-mesh sieve. The mixture was then placed in a quartz boat, flattened and compacted, wrapped with tin foil and sealed in a vacuum tube high-temperature furnace. Nitrogen was used as carrier gas and protective gas. The flow rate of nitrogen was 100 mL / min. The mixture was pyrolyzed in a vacuum tube high-temperature furnace for 2-2.5 h. The pyrolysis temperature was 600 °C and the temperature heating rate was set to 18 °C / min to obtain a mixed biochar material. S2.2: Mix the mixed biochar material and cottonseed hull biochar material in a mass ratio of 1:3 to obtain a mixed powder. Mix the mixed powder with PDA culture medium in a ratio of 1g:12mL. The PDA culture medium is prepared by mixing 15g agar, 200g peeled potato, 20g glucose and 1L pure water. Shake and adsorb and fix at 35°C and 160r / min for 24h to obtain a carbon-based microbial fixative.

[0026] S3: Preparation of microbial agents for promoting the growth and development of orchid plants S3.1: Take a healthy Paphiopedilum micranthum (scientific name: Tang & FT Wang), rinse the surface substrate under running water and dry it with filter paper, cut out the roots of Paphiopedilum micranthum and disinfect them. First, soak the roots of Paphiopedilum micranthum in 70% alcohol for 60 seconds, rinse them twice with sterile water, wipe the surface moisture with sterilized filter paper, then disinfect them with 0.1% mercuric chloride for 5 minutes, rinse them twice with sterile water, wipe the surface moisture with sterilized filter paper, and get the treated roots; S3.2: Place the treated roots on a culture dish and cut them into 4.8-5.2 mm tissue segments, inoculate them on PDA culture medium, and add 20% of the total mass of the system's carbon-based microbial fixative solution. Culture them in an incubator at 25°C in the dark for 7 days. After colonies grow out of the tissue segments, pick up the hyphae at the edge of the colony and transfer them to a new culture dish containing PDA culture medium for purification and culture. Purify twice to obtain growth-promoting microorganisms. S3.3: Dilute the growth-promoting microorganism with 10 times the volume of deionized water, and then add 0.6 g / L of L-tryptophan to obtain a microbial agent that promotes the growth and development of orchids.

[0027] Example 3: A process for preparing a microbial agent for promoting the growth and development of orchid plants, such as Figure 1 As shown, the following steps are included: S1: Preparation of cottonseed hull biochar material S1.1: 4 parts by mass of cottonseed hulls, 0.2 parts by mass of MgSO4, 1 part by mass of KH2PO4, 0.5 parts by mass of urea, 0.1 parts by mass of lime and 1 part by mass of white sugar are added to 100 parts by mass of deionized water, and then 0.2 parts by mass of Pleurotus ostreatus are added, mixed evenly, and cultured at a constant temperature of 26°C for 40 days, then steam-inactivated at 125°C, then washed with deionized water for 3 times, and dried at 105°C to constant weight to obtain microbial-treated cottonseed hulls; S1.2: The cottonseed hull treated with microorganisms was heated to 500°C in an argon atmosphere with a flow rate of 100 mL / min, the heating rate was 3°C / min, and the pyrolysis was performed for 1.5 h to obtain cottonseed hull powder, which was ground through an 80-mesh sieve, and then 4.5 times the mass of NaOH was added, and argon was passed through the argon atmosphere at a flow rate of 100 mL / min, heated to 850°C, and activated for 3.5 h to obtain heat-treated powder; S1.3: The heat-treated powder was washed with 0.1 mol / L HCl and dried at 80°C to constant weight to obtain cottonseed hull biochar material.

[0028] S2: Preparation of carbon-based microbial fixative S2.1: Rice husk, corn stalk and rice stalk were mixed in a mass ratio of 1:1:1, crushed and passed through an 18-mesh sieve, then loaded into a quartz boat, flattened and compacted, wrapped with tin foil and sealed in a vacuum tube high-temperature furnace, and nitrogen was used as carrier gas and protective gas. The flow rate of nitrogen was 100 mL / min, and pyrolysis was performed in a vacuum tube high-temperature furnace for 2-2.5 h. The pyrolysis temperature was 650 °C, and the temperature heating rate was set to 20 °C / min to obtain a mixed biochar material; S2.2: Mix the mixed biochar material and cottonseed hull biochar material in a mass ratio of 1:2 to obtain a mixed powder. Mix the mixed powder with PDA culture medium in a ratio of 1g:10mL. The PDA culture medium is prepared by mixing 15g agar, 200g peeled potato, 20g glucose and 1L pure water. Shake and adsorb and fix at 35°C and 180r / min for 25h to obtain a carbon-based microbial fixative.

[0029] S3: Preparation of microbial agents for promoting the growth and development of orchid plants S3.1: Take a healthy Paphiopedilum micranthum (scientific name: Tang & FT Wang), rinse the surface substrate under running water and dry the water with filter paper, cut out the roots of Paphiopedilum micranthum and disinfect them. First, soak the roots of Paphiopedilum micranthum in 75% alcohol for 65 seconds, rinse them with sterile water for 3 times, wipe the surface moisture with sterilized filter paper, then disinfect them with 0.1% mercuric chloride for 5 minutes, rinse them with sterile water for 3 times, wipe the surface moisture with sterilized filter paper, and get the treated roots; S3.2: Place the treated roots on a culture dish and cut them into 5.2 mm tissue segments, inoculate them on PDA medium, and add 5% carbon-based microbial fixative solution of the total mass of the system. Culture them in an incubator at 26°C in the dark for 7 days. After colonies grow out of the tissue segments, pick the hyphae at the edge of the colonies and transfer them to a new culture dish containing PDA medium for purification and culture. Purify 3 times to obtain growth-promoting microorganisms. S3.3: Dilute the growth-promoting microorganism with 10 times the volume of deionized water, and then add 0.5 g / L of L-tryptophan to obtain a microbial agent that promotes the growth and development of orchids.

[0030] Comparative Example 1: Compared with Example 1, Comparative Example 1 is a commercially available microbial agent for promoting plant growth and development.

[0031] Comparative Example 2: Compared with Example 1, the difference of Comparative Example 2 is that no cottonseed hull biochar material is added in step S2.2, specifically "S2.2: Mix the mixed biochar material with PDA culture medium in a ratio of 1 g:10 mL, the PDA culture medium is prepared by mixing 15 g agar, 200 g peeled potato, 20 g glucose, and 1 L of pure water, and fix by shaking adsorption at 35°C and 160 r / min for 24 hours to obtain a carbon-based microbial fixed solution". The prepared microbial agent for promoting the growth and development of orchids is recorded as Comparative Example 2.

[0032] Comparative Example 3: Compared with Example 1, the difference of Comparative Example 3 is that no mixed biochar material is added in step S2.2, specifically "S2.2: Mix the cottonseed hull biochar material with the PDA culture medium in a ratio of 1 g:10 mL, the PDA culture medium is prepared by mixing 15 g agar, 200 g peeled potato, 20 g glucose, and 1 L of pure water, and fix by shaking adsorption at 35°C and 160 r / min for 24 hours to obtain a carbon-based microbial fixed solution". The prepared microbial agent for promoting the growth and development of orchids is recorded as Comparative Example 3.

[0033] The examples 1-3 and the comparative examples 1-3 were divided into six groups, and symbiotically germinated with 25 orchid seeds in each group, and the seedling formation rate of the orchid seeds was observed, as shown in Table 1.

[0034] Table 1 Seedling formation rate (%) Example 1 16 Example 2 16 Example 3 16 Comparative Example 1 12 Comparative Example 2 8 Comparative Example 3 8 The seedling formation rates of Examples 1-3 are all 16%, while that of Comparative Example 1, i.e., the commercially available product, is 12%. It can be seen that the microbial agent of the present invention improves the seedling formation rate of orchid plants compared with commercially available products, and has the effect of promoting the growth and development of orchid plants.

[0035] The seedling formation rate of Comparative Examples 2 and 3 is 8%. It can be seen that the biochar material prepared from cottonseed hulls treated with Pleurotus ostreatus strains, after supporting the carbon-based microbial fixative, can enable the endophytic fungi of Paphiopedilum rigidum to have a better reproductive advantage during cultivation, and the concentration in the bacterial liquid is higher, thereby having a better effect in promoting plant growth.

[0036] The above embodiments are merely illustrative of the principles and effects of the present invention, and are not intended to limit the present invention. Anyone familiar with the art may modify or alter the above embodiments without departing from the spirit and scope of the present invention. Therefore, all equivalent modifications or alterations made by a person of ordinary skill in the art without departing from the spirit and technical concept disclosed by the present invention shall still be covered by the claims of the present invention.

Claims

1. A preparation process of a microbial agent for promoting the growth and development of orchid plants, characterized in that: The steps include: S1: Preparation of cottonseed hull biochar material Add cottonseed hulls, MgSO4, KH2PO4, urea, lime and sugar to deionized water, then add Pleurotus ostreatus strains and mix evenly, culture at a constant temperature, then steam inactivate to obtain microbial treated cottonseed hulls, pyrolyze the microbial treated cottonseed hulls to obtain cottonseed hull powder, grind and sieve, then add NaOH, heat and activate to obtain heat-treated powder, wash and dry the heat-treated powder to obtain cottonseed hull biochar material; S2: Preparation of carbon-based microbial fixative The rice husk, corn stalk and rice stalk are mixed, crushed and sieved, and then loaded into a quartz boat, flattened and compacted, wrapped with tin foil and sealed, and placed in a vacuum tube high temperature furnace, and pyrolyzed in a vacuum tube high temperature furnace using nitrogen as a carrier gas and a protective gas to obtain a mixed biochar material, and the mixed biochar material is mixed with a cottonseed hull biochar material to obtain a mixed powder, and the mixed powder is mixed with a PDA culture medium, and adsorbed and fixed by shaking to obtain a carbon-based microbial fixative; S3: Preparation of composite humic acid hydrogel powder Take a healthy Paphiopedilum sclerophyllum and rinse the surface matrix under running water, and use filter paper to absorb the water, cut out the roots of the Paphiopedilum sclerophyllum and disinfect them, first soak the roots of the Paphiopedilum sclerophyllum in alcohol, then disinfect them with mercuric chloride to obtain treated roots, place the treated roots on a culture dish, cut them into tissue segments, place them on a PDA culture medium for inoculation, and add a charcoal-based microbial fixative, and culture them in an incubator away from light. After colonies grow out of the tissue segments, pick up the hyphae at the edge of the colony, transfer them to a new culture dish containing a PDA culture medium for purification and culture, purify and obtain growth-promoting microorganisms, dilute the growth-promoting microorganisms with deionized water, and then add L-tryptophan to obtain a microbial agent that promotes the growth and development of orchids.

2. The preparation process of a microbial agent for promoting the growth and development of orchid plants according to claim 1, characterized in that: Step S1: preparing cottonseed hull biochar material, comprising the following steps: S1.1: Add 4-5 parts by mass of cottonseed hulls, 0.2-0.5 parts by mass of MgSO4, 1-3 parts by mass of KH2PO4, 0.5-2 parts by mass of urea, 0.1-0.5 parts by mass of lime and 1-3 parts by mass of white sugar to 100 parts by mass of deionized water, then add 0.2-0.4 parts by mass of Pleurotus ostreatus spawn, mix well, and culture at a constant temperature of 24-26°C for 40 days, then steam inactivate at 120-125°C, then wash with deionized water 3 times, and dry at 100-105°C to constant weight to obtain microbial treated cottonseed hulls; S1.2: The cottonseed hull treated with microorganisms is heated to 500-550°C in an argon atmosphere at a flow rate of 100 mL / min, the heating rate is 3°C / min, and the pyrolysis is performed for 1-1.5 hours to obtain cottonseed hull powder, which is then ground through an 80-mesh sieve, and then 4-4.5 times the mass of NaOH is added, and argon is passed through the argon atmosphere at a flow rate of 100 mL / min, heated to 800-850°C, and activated for 3-3.5 hours to obtain heat-treated powder; S1.3: The heat-treated powder is washed with 0.1 mol / L HCl and dried at 60-80° C. to a constant weight to obtain a cottonseed hull biochar material.

3. The preparation process of a microbial agent for promoting the growth and development of orchid plants according to claim 2, characterized in that: Step S2 is to prepare a carbon-based microbial fixative solution, comprising the following steps: S2.1: Mix rice husk, corn stalk and rice straw in a mass ratio of 1:1:(1-2), crush them through an 18-mesh sieve, put them into a quartz boat, flatten and compact them, wrap them with tin foil and seal them in a vacuum tube high-temperature furnace, use nitrogen as carrier gas and protective gas, pyrolyze them in a vacuum tube high-temperature furnace for 2-2.5h, and the pyrolysis temperature is 600℃ to obtain a mixed biochar material; S2.2: Mix the mixed biochar material with the cottonseed hull biochar material in a mass ratio of 1: (2-3) to obtain a mixed powder, mix the mixed powder with PDA culture medium in a ratio of 1 g: (10-12) mL, and fix by shaking adsorption at 35-36°C and 160-180 r / min for 24-25 hours to obtain a carbon-based microbial fixative.

4. The preparation process of a microbial agent for promoting the growth and development of orchid plants according to claim 3, characterized in that: Step S3 is to prepare a microbial agent for promoting the growth and development of orchid plants, comprising the following steps: S3.1: Take a healthy Paphiopedilum sclerophyllum and rinse the surface substrate under running water and use filter paper to absorb the water. Cut out the roots of Paphiopedilum sclerophyllum and disinfect them. First, soak the roots of Paphiopedilum sclerophyllum in alcohol for 60-65 seconds, rinse them with sterile water 2-3 times, wipe the surface moisture with sterilized filter paper, then disinfect them with 0.1-0.15% mercuric chloride for 5-9 minutes, rinse them with sterile water 2-3 times, wipe the surface moisture with sterilized filter paper, and get the treated roots. S3.2: Place the treated roots on a culture dish and cut them into 4.8-5.2 mm tissue segments, inoculate them on PDA culture medium, and add 5-20% of the total mass of the system's carbon-based microbial fixative solution. Incubate them in an incubator at 25-26°C in the dark for 7 days. After colonies grow out of the tissue segments, pick up the hyphae at the edge of the colonies, transfer them to a new culture dish containing PDA culture medium for purification and culture, purify 2-3 times, and obtain growth-promoting microorganisms; S3.3: Dilute the growth-promoting microorganism with 10 times the volume of deionized water, and then add 0.5-0.6 g / L of L-tryptophan to obtain a microbial agent that promotes the growth and development of orchids.

5. The preparation process of a microbial agent for promoting the growth and development of orchid plants according to claim 2, characterized in that: The oyster mushroom species is a fungus with the scientific name of Pleurotus ostreatus.

6. The preparation process of a microbial agent for promoting the growth and development of orchid plants according to claim 2, characterized in that: The HCl concentration is 0.1 mol / L.

7. The preparation process of a microbial agent for promoting the growth and development of orchid plants according to claim 3, characterized in that: The flow rate of nitrogen was 100 mL / min, and the temperature heating rate was set at 18-20 °C / min.

8. The preparation process of a microbial agent for promoting the growth and development of orchid plants according to claim 4, characterized in that: The PDA medium was prepared by mixing 15 g of agar, 200 g of peeled potatoes, 20 g of glucose, and 1 L of pure water.

9. The preparation process of a microbial agent for promoting the growth and development of orchid plants according to claim 4, characterized in that: Paphiopedilum micranthum is a three-year-old plant with the scientific name Paphiopedilum micranthum Tang & FT Wang.

10. A microbial agent for promoting the growth and development of orchid plants, which is prepared by the preparation process of the microbial agent for promoting the growth and development of orchid plants according to any one of claims 1 to 9.

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