Growth conditioner for tricholoma matsutake planting and preparation method thereof

Abstract

The invention discloses a growth conditioner for planting tricholoma matsutake and a preparation method thereof, and belongs to the technical field of tricholoma matsutake planting.The growth conditioner for planting tricholoma matsutake is prepared from, by weight, 5-8 parts of penicillium citrinum metabolite, 2 parts of arbuscular mycorrhizal fungi with the spore content being 20-30 / g, 5 parts of bacillus subtilis 108-1010 CFU / g and 10-12 parts of matrix. The pH value of the saline-alkali soil can be reduced, meanwhile, the organic matter content of the soil is increased, and the method is suitable for planting tricholoma matsutake; the beta-glucosidase activity of the tricholoma matsutake is promoted, the biomass of cell walls of pine trees serving as host trees is easily utilized, biological energy is obtained, and the growth of the tricholoma matsutake is promoted.

Description

Technical Field

[0001] The present invention relates to the technical field of Tricholoma matsutake cultivation, and particularly relates to a growth regulator for Tricholoma matsutake cultivation and a preparation method thereof. Background Art

[0002] Tricholoma matsutake, also known as pine mushroom, is one of the members of ectomycorrhizal fungi. Due to its unique flavor and taste, it has long been regarded as a highly regarded food in Asia and is widely popular because of its unique flavor and nutritional value. Tricholoma matsutake has special requirements for the growth environment. The ideal growth temperature is 15°C - 25°C, the humidity is maintained above 80%, it requires appropriate scattered light but cannot be directly exposed to the sun, and at the same time, good air circulation should be ensured. In the process of artificial cultivation of Tricholoma matsutake, using growth regulators is a common practice, mainly to improve the yield and quality of Tricholoma matsutake by improving the soil environment, providing nutrients, and promoting the growth of hyphae. Summary of the Invention

[0003] As described in the above prior art, one of the purposes of the present invention is to provide a growth regulator for Tricholoma matsutake cultivation. This growth regulator for Tricholoma matsutake cultivation can reduce the pH value of saline-alkali soil, increase the organic matter content of the soil at the same time, and is suitable for the cultivation of Tricholoma matsutake; it can promote the β-glucosidase activity of Tricholoma matsutake, easily utilize the biomass of the cell wall of the host tree pine, obtain bioenergy, and promote the growth of Tricholoma matsutake.

[0004] Another purpose of the present invention is to provide a manufacturing method of a growth regulator for Tricholoma matsutake cultivation. This method has simple steps and can industrially produce a growth regulator for Tricholoma matsutake cultivation.

[0005] One of the purposes of the present invention is achieved by adopting the following technical solutions:

[0006] A growth regulator for Tricholoma matsutake cultivation, calculated by weight, includes 5 - 8 parts of Penicillium digitatum metabolites, 2 parts of arbuscular mycorrhizal fungi with a spore content of 20 - 30 spores / g, 5 parts of Bacillus subtilis 10 8 -10 10 CFU / g, and 10 - 12 parts of matrix.

[0007] In this embodiment, calculated by weight, it includes 6 parts of Penicillium digitatum metabolites, 2 parts of arbuscular mycorrhizal fungi with a spore content of 30 spores / g, 5 parts of Bacillus subtilis 10 10 CFU / g, and 10 parts of matrix.

[0008] In this embodiment, the matrix is one of bagasse, corn stalks, wood chips, wheat straw or rice straw.

[0009] In this embodiment, the preparation method of the Penicillium digitatum metabolite comprises the following steps:

[0010] S1. Enrich Penicillium digitatum: S11. Collect citrus fruits with diseased spots from the orchard, and sample from the surface of the citrus fruits, especially the diseased spots;

[0011] S12. Perform 10-fold serial dilutions in sequence to prepare the sample into a serial dilution solution of 10 -1 to 10 -6 ;

[0012] S13. Take 0.1 mL of each serial dilution solution from the serial dilution solution of 10 -1 to 10 -6 , spread it on a PDA plate, add 300 ppm streptomycin to inhibit the growth of bacteria, isolate microfungi, culture at 25 °C for 3 - 5 days, select colonies with typical Penicillium colonies characteristics for pure culture, transfer them to a new PDA plate, and subculture at 25 °C for 5 - 7 days to obtain pure culture;

[0013] S2. Culture Penicillium: S21. Inoculate the pure-cultured Penicillium into Czapek broth medium, culture at 25 °C with a rotation speed of 120 r / min for 96 h;

[0014] S22. When used for fermentation culture, inoculate the seed medium into the fermentation medium according to an inoculation amount of 10%, and continue to culture for 120 h;

[0015] S3. Extract the metabolite: S31. Filter: After fermentation is completed, filter the culture solution using a syringe filter with a pore size of 0.45 mm to remove fungal spores and obtain a metabolite without spores;

[0016] S32. Concentrate: Concentrate the filtered metabolite 50 times using a freeze dryer to obtain the Penicillium digitatum metabolite.

[0017] In this embodiment, the composition of the seed medium for making the PDA plate is: 200 g / L of potato, 20 g / L of glucose, 15 g / L of agar, 1 L of distilled water, and the pH is 7.2.

[0018] In this embodiment, the composition of the fermentation medium is: 35.0 g / L of corn flour, 15.0 g / L of yeast peptone, 1.0 g / L of potassium dihydrogen phosphate, 5.0 g / L of calcium carbonate, 1 L of distilled water, and the pH is 7.2.

[0019] The second object of the present invention is achieved by the following technical solution:

[0020] A preparation method of a growth regulator for Tricholoma matsutake cultivation, comprising the following steps:

[0021] S1. Preparation of substrate: Weigh 10 - 12 parts of the substrate, crush it and sieve it through an 8 - mesh sieve to obtain the substrate;

[0022] S2. Preparation of microorganisms: Weigh 2 parts of the arbuscular mycorrhizal fungi, ensuring that its spore content is 20 - 30 spores / g; weigh 5 parts of the Bacillus subtilis, ensuring that its total viable count is 10^8 - 10^10 CFU / g;

[0023] S3. Mix the arbuscular mycorrhizal fungi and the Bacillus subtilis together, add them to sterile water to make a uniform bacterial suspension;

[0024] S4. Preparation of Penicillium digitatum metabolites: Weigh 5 - 8 parts of the Penicillium digitatum metabolites, dissolve them in sterile water to obtain the Penicillium digitatum metabolite solution;

[0025] S5. Add the uniform bacterial suspension from step S3 to the Penicillium digitatum metabolite solution, stir evenly, then add the substrate to this solution and stir evenly again to ensure that all components are fully mixed to obtain the growth regulator for Tricholoma matsutake cultivation.

[0026] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0027] The growth regulator for Tricholoma matsutake cultivation provided by the present invention selects a substrate which not only provides necessary nutrients and physical support for Tricholoma matsutake, but also can improve the water retention and air permeability of the soil, improve the physical properties of the soil, and create good conditions for the growth of Tricholoma matsutake; the combination of arbuscular mycorrhizal fungi and Bacillus subtilis can significantly reduce the pH value of saline - alkali soil and increase the organic matter content of the soil, which is suitable for the cultivation of Tricholoma matsutake; sampling from the surface of citrus fruits, especially the lesion parts, can enrich Penicillium digitatum from citrus and extract its metabolites, and use the metabolites of Penicillium digitatum to promote the growth of Tricholoma matsutake, promote the β - glucosidase activity of Tricholoma matsutake, easily utilize the biomass of the cell wall of the host tree pine, obtain bioenergy, and promote the growth of Tricholoma matsutake; collecting citrus fruits with lesion parts from the orchard to prepare the metabolites of Penicillium digitatum, the raw materials are easily available and the economic cost is low, enabling the resource utilization of citrus fruits with lesion parts. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] In order to more clearly illustrate the embodiments of the present invention, the accompanying drawings required for the embodiments will be briefly introduced below. Obviously, the accompanying drawings in the following description are only the embodiments of the present invention. For those of ordinary skill in the art, without creative efforts, other drawings can also be obtained based on the provided drawings.

[0029] Figure 1It is a result graph of the growth area measurement of matsutake by the growth regulators for matsutake cultivation in Examples 1-4 and Comparative Examples. Detailed implementation mode

[0030] Next, in combination with specific embodiments, the present invention will be further described. It should be noted that on the premise of no conflict, any combination of the following-described embodiments or technical features can form a new embodiment.

[0031] The syringe filter with a diameter of 0.45 mm is an Acrodisc syringe filter produced by Pall Corporation.

[0032] Example 1

[0033] This example provides a growth regulator for matsutake cultivation, which includes 6 parts of Penicillium digitatum metabolites, 2 parts of arbuscular mycorrhizal fungi with a spore content of 30 spores / g, 5 parts of Bacillus subtilis 10 CFU / g, and 10 parts of substrate bagasse according to weight.

[0034] In this example, the functions of arbuscular mycorrhizal fungi and Bacillus subtilis are introduced as follows:

[0035] Function of arbuscular mycorrhizal fungi: Arbuscular mycorrhizal fungi can form a symbiotic relationship with plant roots, expand the absorption area of plant roots, and promote the absorption and transformation of nutrients in the soil; arbuscular mycorrhizal fungi can also secrete organic acids to lower the soil pH value and improve the soil structure.

[0036] Secretion of organic acids: Both arbuscular mycorrhizal fungi and Bacillus subtilis can secrete organic acids. The organic acids secreted by arbuscular mycorrhizal fungi can neutralize the alkaline components in the soil and lower the soil pH value. Bacillus subtilis can also secrete a variety of organic acids, which can dissolve the minerals in the soil and release hydrogen ions, thereby lowering the soil pH value.

[0037] Increase soil organic matter content: The combination of arbuscular mycorrhizal fungi and Bacillus subtilis can significantly improve the growth indexes of plants, such as plant height, root length, etc. When plants grow vigorously, their root exudates and residues increase. After these organic substances enter the soil, they can increase the soil organic matter content.

[0038] Decompose organic matter: After Bacillus subtilis decomposes organic matter, some small-molecule organic substances and nutrient elements will be released. These small-molecule organic substances are more easily absorbed and utilized by plants, thus promoting plant growth. At the same time, arbuscular mycorrhizal fungi can transfer the nutrients absorbed by plant roots to the soil, promote the activities of soil microorganisms, and further increase the accumulation of soil organic matter.

[0039] In this embodiment, the preparation method of Penicillium digitatum metabolites comprises the following steps:

[0040] S1. Enrichment of Penicillium digitatum: S11. Collect citrus fruits with diseased spots from the orchard, and sample from the surface of the citrus fruits, especially the diseased spots;

[0041] S12. Perform 10-fold serial dilutions in sequence to make the sample into a serial dilution solution from 10 -1 to 10 -6 ;

[0042] S13. Take 0.1 mL of each serial dilution solution from the serial dilution solution from 10 -1 to 10 -6 , coat it on a PDA plate, add 300 ppm streptomycin to inhibit the growth of bacteria, isolate microfungi, culture at 25 °C for 4 days, select the colonies with typical Penicillium colonies characteristics for pure culture, transfer them to a new PDA plate, and subculture at 25 °C for 6 days to obtain pure culture;

[0043] S2. Culture of Penicillium: S21. Inoculate the pure cultured Penicillium into Czapek broth medium, culture at 25 °C, rotation speed 120 r / min, for 96 h;

[0044] S22. When used for fermentation culture, inoculate the seed medium into the fermentation medium according to the inoculation amount of 10%, and continue to culture for 120 h;

[0045] S3. Extraction of metabolites: S31. Filtration: After fermentation is completed, filter the culture solution with a syringe filter with a pore size of 0.45 mm to remove fungal spores and obtain spore-free metabolites;

[0046] S32. Concentration: Concentrate the filtered metabolites 50 times with a freeze dryer to obtain Penicillium digitatum metabolites.

[0047] In this embodiment, the composition of the seed medium required for making the PDA plate is: 200 g / L of potato, 20 g / L of glucose, 15 g / L of agar, 1 L of distilled water, and pH 7.2.

[0048] In this embodiment, the composition of the fermentation medium is: 35.0 g / L of corn flour, 15.0 g / L of yeast peptone, 1.0 g / L of potassium dihydrogen phosphate, 5.0 g / L of calcium carbonate, 1 L of distilled water, and pH 7.2.

[0049] This embodiment also provides a preparation method of a growth regulator for Tricholoma matsutake cultivation, comprising the following steps:

[0050] S1. Preparation of substrate: Weigh 10 parts of the substrate, crush it and sieve it through an 8-mesh sieve to obtain the substrate;

[0051] S2. Preparation of microorganisms: Weigh 2 parts of arbuscular mycorrhizal fungi to ensure that the spore content is 30 spores / g; weigh 5 parts of Bacillus subtilis to ensure that the total viable count is 10 10 CFU / g;

[0052] S3. Mix the arbuscular mycorrhizal fungi and Bacillus subtilis together and add them to sterile water to make a uniform bacterial suspension;

[0053] S4. Preparation of Penicillium digitatum metabolites: Weigh 6 parts of Penicillium digitatum metabolites and dissolve them in sterile water to obtain a Penicillium digitatum metabolite solution;

[0054] S5. Add the uniform bacterial suspension from step S3 to the Penicillium digitatum metabolite solution, stir evenly, then add the substrate to this solution and stir evenly again to ensure that all components are fully mixed to obtain a growth regulator for Tricholoma matsutake cultivation.

[0055] Example 2

[0056] This example provides a growth regulator for Tricholoma matsutake cultivation, which, by weight, includes 5 parts of Penicillium digitatum metabolites, 2 parts of arbuscular mycorrhizal fungi with a spore content of 25 spores / g, 5 parts of Bacillus subtilis with 10 9 CFU / g, and 10 parts of the substrate corn straw.

[0057] In this example, the preparation method of Penicillium digitatum metabolites includes the following steps:

[0058] S1. Enrich Penicillium digitatum: S11. Collect citrus fruits with diseased spots from the orchard and take samples from the surface of the citrus fruits, especially the diseased spots;

[0059] S12. Perform 10-fold serial dilutions in sequence to make the samples into serial dilutions of 10 -1 to 10 -6 ;

[0060] S13. Take 0.1 mL of each serial dilution from the serial dilutions of 10 -1 to 10 -6 and spread them on PDA plates, add 300 ppm streptomycin to inhibit bacterial growth, isolate microfungi, culture them at 25°C for 3 days, select colonies with typical Penicillium colonies characteristics for pure culture, transfer them to a new PDA plate, and subculture them at 25°C for 5 days to obtain pure culture;

[0061] S2. Culture Penicillium: S21. Inoculate the pure-cultured Penicillium into Czapek broth medium, culture it at 25°C with a rotation speed of 120 r / min for 96 h;

[0062] S22. For fermentation culture, inoculate the seed medium into the fermentation medium at an inoculation amount of 10%, and continue culturing for 120 h.

[0063] S3. Extract the metabolite: S31. Filtration: After fermentation is completed, filter the culture solution using a syringe filter with a pore size of 0.45 mm to remove fungal spores and obtain a metabolite without spores.

[0064] S32. Concentration: Concentrate the filtered metabolite 50 times using a freeze dryer to obtain the metabolite of Penicillium italicum.

[0065] In this example, the composition of the seed medium required for making the PDA plate is: 200 g / L of potato, 20 g / L of glucose, 15 g / L of agar, 1 L of distilled water, and the pH is 7.2.

[0066] In this example, the composition of the fermentation medium is: 35.0 g / L of corn flour, 15.0 g / L of yeast peptone, 1.0 g / L of potassium dihydrogen phosphate, 5.0 g / L of calcium carbonate, 1 L of distilled water, and the pH is 7.2.

[0067] This example also provides a preparation method of a growth regulator for Tricholoma matsutake cultivation, including the following steps:

[0068] S1. Preparation of the substrate: Weigh 10 parts of the substrate, crush it and pass it through an 8-mesh sieve to obtain the substrate.

[0069] S2. Preparation of microorganisms: Weigh 2 parts of arbuscular mycorrhizal fungi to ensure that the spore content is 25 per gram; weigh 5 parts of Bacillus subtilis to ensure that the total viable count is 10 9 CFU / g.

[0070] S3. Mix the arbuscular mycorrhizal fungi and Bacillus subtilis together, add them to sterile water, and make a uniform bacterial suspension.

[0071] S4. Preparation of the metabolite of Penicillium italicum: Weigh 5 parts of the metabolite of Penicillium italicum, dissolve it in sterile water, and prepare a metabolite solution of Penicillium italicum.

[0072] S5. Add the uniform bacterial suspension in step S3 to the metabolite solution of Penicillium italicum, stir evenly, then add the substrate to this solution, and stir evenly again to ensure that all components are fully mixed to obtain a growth regulator for Tricholoma matsutake cultivation.

[0073] Example 3

[0074] This embodiment provides a growth regulator for matsutake cultivation, which, by weight, includes 8 parts of Penicillium digitatum metabolites, 2 parts of arbuscular mycorrhizal fungi with a spore content of 20 spores / g, 5 parts of Bacillus subtilis 10 8 CFU / g, and 10 parts of substrate sawdust.

[0075] In this embodiment, the preparation method of Penicillium digitatum metabolites includes the following steps:

[0076] S1. Enrich Penicillium digitatum: S11. Collect citrus fruits with diseased spots from the orchard, and take samples from the surface of citrus fruits, especially the diseased spots;

[0077] S12. Perform 10-fold serial dilutions in sequence to make the samples into serial dilution solutions of 10 -1 to 10 -6 ;

[0078] S13. Take 0.1 mL of each serial dilution solution from the serial dilution solutions of 10 -1 to 10 -6 , coat them on PDA plates, add 300 ppm streptomycin to inhibit bacterial growth, isolate microfungi, culture them at 25°C for 5 days, select colonies with typical Penicillium colonies characteristics for pure culture, transfer them to new PDA plates, and subculture them at 25°C for 6 days to obtain pure culture;

[0079] S2. Culture Penicillium: S21. Inoculate the pure-cultured Penicillium into Czapek broth medium, culture it at 25°C with a rotation speed of 120 r / min for 96 h;

[0080] S22. When used for fermentation culture, inoculate the seed medium into the fermentation medium according to an inoculation amount of 10%, and continue to culture for 120 h;

[0081] S3. Extract metabolites: S31. Filtration: After fermentation is completed, filter the culture solution using a syringe filter with a pore size of 0.45 mm to remove fungal spores and obtain spore-free metabolites;

[0082] S32. Concentration: Concentrate the filtered metabolites 50 times using a freeze dryer to obtain Penicillium digitatum metabolites.

[0083] In this embodiment, the composition of the seed medium required for making PDA plates is: 200 g / L of potato, 20 g / L of glucose, 15 g / L of agar, 1 L of distilled water, and the pH is 7.2.

[0084] In this embodiment, the composition of the fermentation medium is: 35.0 g / L of corn flour, 15.0 g / L of yeast peptone, 1.0 g / L of potassium dihydrogen phosphate, 5.0 g / L of calcium carbonate, 1 L of distilled water, and the pH is 7.2.

[0085] This embodiment also provides a preparation method of a growth regulator for matsutake cultivation, including the following steps:

[0086] S1. Preparation of substrate: Weigh 10 parts of the substrate, crush it and sieve it through an 8-mesh sieve to obtain the substrate;

[0087] S2. Preparation of microorganisms: Weigh 2 parts of arbuscular mycorrhizal fungi, ensuring that its spore content is 20 spores / g; weigh 5 parts of Bacillus subtilis, ensuring that its total viable count is 10 8 CFU / g;

[0088] S3. Mix the arbuscular mycorrhizal fungi and Bacillus subtilis together, add them to sterile water, and make a uniform bacterial suspension;

[0089] S4. Preparation of Penicillium digitatum metabolites: Weigh 8 parts of Penicillium digitatum metabolites, dissolve them in sterile water, and prepare a Penicillium digitatum metabolite solution;

[0090] S5. Add the uniform bacterial suspension in step S3 to the Penicillium digitatum metabolite solution, stir evenly, then add the substrate to this solution, and stir evenly again to ensure that all components are fully mixed, obtaining a growth regulator for matsutake cultivation.

[0091] Example 4

[0092] This embodiment provides a growth regulator for matsutake cultivation. By weight, it includes 6 parts of Penicillium digitatum metabolites, 2 parts of arbuscular mycorrhizal fungi with a spore content of 30 spores / g, 5 parts of Bacillus subtilis with 10 10 CFU / g, and 10 parts of rice straw as the substrate.

[0093] In this embodiment, the preparation method of Penicillium digitatum metabolites includes the following steps:

[0094] S1. Enrichment of Penicillium digitatum: S11. Collect citrus fruits with diseased parts from the orchard, take samples from the surface of citrus fruits, especially the diseased parts;

[0095] S12. Perform 10-fold serial dilutions in sequence, and make the samples into serial dilution solutions from 10 -1 to 10 -6 ;

[0096] S13. Take 10 -1 to 10 -6For each gradient dilution in the gradient dilution solution, take 0.1 mL and spread it on a PDA plate. Add 300 ppm streptomycin to inhibit bacterial growth, isolate microfungi, culture at 25 °C for 4 days, select colonies with typical Penicillium colony characteristics for pure culture, transfer them to a new PDA plate, and subculture at 25 °C for 7 days to obtain pure culture;

[0097] S2. Cultivate Penicillium: S21. Inoculate the pure-cultured Penicillium into Czapek broth medium, culture at 25 °C with a rotation speed of 120 r / min for 96 h;

[0098] S22. When used for fermentation culture, inoculate the seed medium into the fermentation medium according to an inoculation amount of 10%, and continue to culture for 120 h;

[0099] S3. Extract metabolites: S31. Filtration: After fermentation is completed, filter the culture solution using a syringe filter with a pore size of 0.45 mm to remove fungal spores and obtain spore-free metabolites;

[0100] S32. Concentration: Concentrate the filtered metabolites 50 times using a freeze dryer to obtain Penicillium digitatum metabolites.

[0101] In this example, the composition of the seed medium required for making the PDA plate is: 200 g / L of potato, 20 g / L of glucose, 15 g / L of agar, 1 L of distilled water, and the pH is 7.2.

[0102] In this example, the composition of the fermentation medium is: 35.0 g / L of corn flour, 15.0 g / L of yeast peptone, 1.0 g / L of potassium dihydrogen phosphate, 5.0 g / L of calcium carbonate, 1 L of distilled water, and the pH is 7.2.

[0103] This example also provides a preparation method for a growth regulator for Tricholoma matsutake cultivation, including the following steps:

[0104] S1. Preparation of the substrate: Weigh 10 parts of the substrate, crush it and pass it through an 8-mesh sieve to obtain the substrate;

[0105] S2. Preparation of microorganisms: Weigh 2 parts of arbuscular mycorrhizal fungi to ensure that the spore content is 30 spores / g; weigh 5 parts of Bacillus subtilis to ensure that the total viable count is 10 10 CFU / g;

[0106] S3. Mix the arbuscular mycorrhizal fungi and Bacillus subtilis together, add them to sterile water, and make a uniform bacterial suspension;

[0107] S4. Preparation of Penicillium digitatum metabolites: Weigh 6 parts of Penicillium digitatum metabolites, dissolve them in sterile water, and prepare a Penicillium digitatum metabolite solution;

[0108] S5. Add the uniform bacterial suspension from step S3 into the Penicillium digitatum metabolite solution, stir evenly, then add the substrate into this solution and stir evenly again to ensure that all components are fully mixed, obtaining a growth regulator for Tricholoma matsutake cultivation.

[0109] Example 5

[0110] This example provides a method for using the growth regulator for Tricholoma matsutake cultivation. Specifically, dissolve the growth regulator for Tricholoma matsutake cultivation in an appropriate amount of water and spray it on the soil for Tricholoma matsutake cultivation once every 7 - 10 days to promote the growth of Tricholoma matsutake.

[0111] Comparative Example

[0112] Different from Example 1, this comparative example provides a growth regulator for Tricholoma matsutake cultivation. By weight, it includes 2 parts of arbuscular mycorrhizal fungi with a spore content of 30 spores / g, 5 parts of Bacillus subtilis 10 10 CFU / g, and 10 parts of the substrate bagasse.

[0113] Experimental Example

[0114] Dissolve the growth regulators for Tricholoma matsutake cultivation in Examples 1 - 4 and the comparative example in sterile water, take 10 μL, inject it onto the Tricholoma matsutake mycelium, and culture it on a Tricholoma matsutake seed medium (200 g / L of potato, 20 g / L of glucose, 15 g / L of agar, 1 L of distilled water, pH 7.2) at a temperature of 25°C for 1 month. After the cultivation ends, compare the growth area of Tricholoma matsutake in Example 1 with the control group treated with distilled water. In contrast, use imageJ software to measure the growth area of Tricholoma matsutake three times, and the measurement results are as Figure 1 shown.

[0115] It can be seen from the figure that the growth regulators for Tricholoma matsutake cultivation in Examples 1 - 4 have a significantly larger growth area of Tricholoma matsutake than the comparative example and the control group.

[0116] The above embodiments are only the preferred embodiments of the present invention and cannot be used to limit the scope of protection of the present invention. Any non - substantial changes and substitutions made by those skilled in the art based on the present invention fall within the scope of protection required by the present invention.

Claims

1. A growth conditioner for matsutake mushroom cultivation, characterized in that, By weight, it includes 5-8 parts of Penicillium digitatum metabolites, 2 parts of arbuscular mycorrhizal fungi with a spore content of 20-30 spores / g, 5 parts of Bacillus subtilis 8 -10 10 CFU / g, and 10-12 parts of matrix.

2. The growth regulator for matsutake mushroom cultivation according to claim 1, characterized in that, By weight, it includes 6 parts of Penicillium digitatum metabolites, 2 parts of arbuscular mycorrhizal fungi with a spore content of 30 spores / g, 5 parts of Bacillus subtilis 10 10 at 5 × 10⁶ CFU / g, and 10 parts of substrate.

3. A growth conditioner for matsutake cultivation according to claim 1, characterized in that, The substrate is one of bagasse, corn stalks, wood chips, wheat straw or rice straw.

4. A growth regulator for matsutake mushroom cultivation according to claim 1, characterized in that, The preparation method of the Penicillium digitatum metabolite comprises the following steps: S1. Enrich Penicillium digitatum: S11. Collect citrus fruits with diseased parts from the orchard, and take samples from the surface of the citrus fruits, especially the diseased parts; S12. Perform 10-fold serial dilutions in sequence to prepare serial dilution solutions of the sample from 10 -1 to 10 -6 . S13. Take 10 -1 to 10 -6 For each gradient dilution in the gradient dilution solution of 0.1 mL, spread it on a PDA plate, add 300 ppm streptomycin to inhibit bacterial growth, isolate microfungi, culture at 25 °C for 3 - 5 days, select colonies with typical Penicillium colony characteristics for pure culture, transfer them to a new PDA plate, and subculture at 25 °C for 5 - 7 days to obtain pure culture; S2. Culture Penicillium: S21. Inoculate the pure cultured Penicillium into the Czapek broth medium, culture at 25 °C with a rotation speed of 120 r / min for 96 h; S22. When used for fermentation culture, inoculate the seed medium into the fermentation medium according to the inoculation amount of 10%, and continue to culture for 120 h; S3. Extract the metabolite: S31. Filter: After fermentation is completed, filter the culture solution with a syringe filter with a pore size of 0.45 mm to remove fungal spores and obtain a spore-free metabolite; S32. Concentrate: Concentrate the filtered metabolite 50 times with a freeze dryer to obtain the Penicillium digitatum metabolite.

5. A growth regulator for matsutake mushroom cultivation according to claim 4, characterized in that, The composition of the seed medium for making the PDA plate is: 200 g / L of potato, 20 g / L of glucose, 15 g / L of agar, 1 L of distilled water, and the pH is 7.

2.

6. The growth regulator for matsutake mushroom cultivation according to claim 4, wherein, The composition of the fermentation medium is: 35.0 g / L of corn flour, 15.0 g / L of yeast peptone, 1.0 g / L of potassium dihydrogen phosphate, 5.0 g / L of calcium carbonate, 1 L of distilled water, and the pH is 7.

2.

7. The preparation method of a growth regulator for matsutake mushroom cultivation according to any one of claims 1 to 6, characterized in that, It comprises the following steps: S1. Preparation of the substrate: Weigh 10-12 parts of the substrate, crush it and pass it through an 8-mesh sieve to obtain the substrate; S2. Preparation of microorganisms: Weigh 2 parts of the arbuscular mycorrhizal fungi to ensure that the spore content is 20 - 30 spores / g; weigh 5 parts of the Bacillus subtilis to ensure that the total viable count is 10 8 -10 10 CFU / g; S3. Mix the arbuscular mycorrhizal fungi and the Bacillus subtilis together, and add them to sterile water to make a uniform bacterial suspension; S4. Preparation of the Penicillium digitatum metabolite: Weigh 5-8 parts of the Penicillium digitatum metabolite, dissolve it in sterile water to obtain the Penicillium digitatum metabolite solution; S5. Add the uniform bacterial suspension in step S3 to the Penicillium digitatum metabolite solution, stir evenly, then add the substrate to the solution, and stir evenly again to ensure that all components are fully mixed to obtain the growth regulator for Tricholoma matsutake cultivation.

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