CN108770593B - Lepista nuda strain and fruiting body cultivation method thereof - Google Patents

Abstract

The invention discloses a lilac mushroom strain and a fruiting body cultivation method thereof. The invention provides a Lepista nuda DCH560 with the preservation number of CGMCC No. 15376. The lepista nuda DCH560(CGMCC No.15376) provided by the invention is a strain separated from a specimen collected from autonomous county of the broad county of Dandelion, Liaoning, China, and the strain can be artificially domesticated to obtain a sporocarp. The pileus of the fruiting body obtained by the strain is purple, the flesh is thick, the fold is purple, and the pileus has strong fragrance and is consistent with the wild fruiting body. The invention domesticates to obtain the lilac mushroom fruiting body, develops a new rare edible mushroom variety, has important significance for protecting germplasm resources of the species, enriching edible mushroom variety resources, enriching vegetable baskets and the like, and has ecological significance for utilizing crop straws because the mushroom is a typical straw rotting fungus.

Description

Lepista nuda strain and fruiting body cultivation method thereof

Technical Field

The invention belongs to the technical field of microorganisms, and relates to a Lepista nuda strain and a fruiting body cultivation method thereof.

Background

Lepitta nuda (Bull.) Cooke is also called Tricholoma nudum, Lepista, and Lepista, belonging to Agaricales, Tricholomataceae, and Lentinus, and is called Lepista nuda because its stipe is in the color of lilac and has a very strong unique mushroom flavor. The lilac mushroom is pleasant in color and luster, aromatic in flavor, fresh, tender and sweet, and excellent in taste, is really reputed to eat the fragrance of one family and smell the fragrance of ten families, is popular in Europe, is named together with truffles and boletes, and is regarded as a top-grade food material in France.

Lepista nuda has high research and development values as a rare edible and medicinal fungus, but the main source is field collection so far, the natural resources are limited, and the quality cannot be guaranteed. Although some units develop domestication cultivation of the lepista nuda, no report on successful fruit body cultivation is found. The patent "a lilac mushroom culture medium and a lilac mushroom liquid spawn preparation method based on yellow wine lees" (CN201610552378.1) "provides a lilac mushroom culture medium and a lilac mushroom liquid spawn preparation method based on yellow wine lees, but does not obtain fruit bodies; "a Lepista nuda strain and its liquid strain and preparation method CN 201510877093.0" discloses a Lepista nuda strain and its liquid strain and preparation method, and no fruiting body is obtained. The culture characteristics of the mycelia of Lepista nuda and domestication studies thereof were studied and domesticated in the thesis of Lepista nuda Lepista academic thesis (Zhaotong, 2010) at Yanbian university, but fruiting bodies were not obtained as a result. The studies on the synthesis of the syringy mushroom primordium by the methods of Yaoshiwei et al revealed that no fruiting body was obtained (Yaoshiwei; Yuanyangbo; Gaojunwei, Leingiumo primordium induced synthesis medium initial probing chemistry and bioengineering, 2010, 27(1): 59-62.). 1994 reported domestication and cultivation of the self-growing edible fungi of the tea garden (Caiyun and Cuoming, 1994, 1: 24 tea leaves in Guizhou) reported that mushroom fruiting is carried out in the open field of the tea garden after 7 months.

Disclosure of Invention

It is an object of the present invention to provide Lepista nuda DCH560 strain.

The Lepista nuda (Lepista nuda) DCH560 strain provided by the invention has the preservation number of CGMCCNo.15376.

The application of the lepista nuda as a parent for crossbreeding is also within the protection scope of the invention.

The fruiting body obtained by cultivating the above-mentioned lepista nuda is also within the scope of the present invention.

The mycelium obtained by culturing the above-mentioned lepista nuda is also within the scope of the present invention.

Another object of the present invention is to provide a method for cultivating the fruit body.

The method provided by the invention comprises the following steps: and (3) sequentially carrying out stock seed preparation, cultivated seed preparation, inoculation and spawn running, soil covering, fruiting management and fruiting on the mycelia of the lepista nuda to obtain sporocarp.

In the above method, the method for preparing the stock comprises the following steps: inoculating the mycelium of the lepista nuda of claim 1 into a stock culture medium, and culturing at 15-25 ℃ in the dark, wherein the relative air humidity is 60% -70% until the mycelium overgrows the culture medium to obtain a stock;

or, the method for preparing the cultivar comprises the following steps: inoculating the stock seeds into a culture medium, and culturing at 15-25 ℃ in a dark place until hyphae overgrow the culture medium to obtain culture seeds, wherein the relative air humidity is 60% -70%;

or, the inoculation and spawn running method comprises the following steps: inoculating the cultivated species into a cultivation material, and culturing at 15-25 ℃ in the dark, wherein the relative humidity of air is 65% -75% until hypha overgrows the cultivation material;

or, the soil covering method comprises the following steps: in the inoculation and spawn running steps, after hyphae grow over the surface of the cultivation material, covering soil on the surface of the cultivation material, and culturing in a dark place at 15-25 ℃, wherein the relative humidity of air is 70-75%, and the relative concentration of carbon dioxide in the air is 0.1-0.5% until the hyphae grow to the surface of a soil layer;

or, the fruiting management method comprises the following steps: when the hypha obtained in the soil covering step grows to the surface of the soil layer, supplementing a layer of fine soil, maintaining the temperature at 15-23 ℃, the illumination intensity at 500-;

or, the fruiting method comprises the following steps: and transferring the mushroom primordium obtained in the fruiting management step to the condition that the ambient temperature is 18-20 ℃, the relative air humidity is 75-90% and the relative carbon dioxide concentration is 0.05-0.15% for culturing to obtain the fruiting body.

The method also comprises a step of harvesting the fruiting body, specifically, when the fruiting body grows to eight mature and the pileus is not completely unfolded, the mushroom can be harvested.

In the method, the stock culture medium and the cultivated species culture medium are both a wheat grain culture medium or a wheat grain cow dung culture medium; the stock culture medium and the cultivar culture medium are the same.

The wheat grain culture medium is prepared by mixing raw materials consisting of 98% of wheat grains, 1% of gypsum and 1% of lime by mass with water, wherein the water content of the raw materials is 65% by mass;

the wheat grain and cow dung culture medium is prepared by mixing raw materials consisting of 93% of wheat grains, 5% of cow dung, 1% of gypsum and 1% of lime with water, wherein the water content is 65% by weight.

In the above method, in the method for preparing the stock, the light-shielding culture at 15-25 ℃ is a light-shielding culture at 22 ℃. The cultivation material is a fermented manure and grass material and is prepared by fermenting crop straws, cow dung, soybean meal and gypsum.

The cultivation material can be obtained by performing secondary fermentation on the following cultivation raw materials: the cultivation raw material is prepared by mixing crop straws (rice straws and wheat straws), animal manure (such as dried cow manure and/or chicken manure), bean foil and gypsum powder. The mass (dry weight) ratio of the crop straws, the animal wastes, the bean foils and the gypsum powder is 55: 45: 1.2: 4.

in the present invention, the secondary fermentation is divided into pre-fermentation and post-fermentation. The pre-fermentation comprises the following steps: pre-wetting crop straws and cow dung 3-4 days before stacking, wherein the moisture content is 65% -70%, stacking and fermenting in the north and south directions, monitoring the temperature of a stack body, turning the stack body once when the internal temperature of the stack body begins to fall after reaching more than 70 ℃, and completing the pre-fermentation after 3-4 times of stack turning. The post-fermentation comprises the following steps: fermenting again the cultivation material after the pre-fermentation, raising the temperature inside the stack to 58-62 ℃ and keeping for 12-24h, then ventilating to lower the temperature of the stack to 50-52 ℃ and keeping for 4-6d, and then lowering the temperature of the stack to normal temperature (25-28 ℃). And finishing the post-fermentation, wherein the secondary fermentation is finished immediately. The cultivation raw material after the secondary fermentation is free of ammonia smell and excrement smell, a white actinomycete layer with an obvious material surface can be seen, the water content of the cultivation raw material after the secondary fermentation is adjusted to 65 percent (mass fraction), and the pH value is adjusted to 7.0-7.5, so that the cultivation material is obtained.

In the method, the cultivation mode can be shelf cultivation.

The layer frame cultivation comprises the following steps: the specification of the shelf is 3-5 layers, the height of the shelf is 0.6m, the width of the shelf is 1.5m, and the distance between the bottom layer and the ground is 0.3m (the length is determined according to the requirement); the cultivation material is spread on each layer of bed frame for cultivation and fruiting.

The application of the above sporocarp in food processing is also within the scope of the present invention.

The lepista nuda DCH560(CGMCC No.15376) provided by the invention is a strain separated from a specimen collected from autonomous county of the broad county of Dandelion, Liaoning, China, and the strain can be artificially domesticated to obtain a sporocarp. The pileus of the fruiting body obtained by the strain is purple, the flesh is thick, the fold is purple, and the pileus has strong fragrance and is consistent with the wild fruiting body. The invention domesticates to obtain the lilac mushroom fruiting body, develops a new rare edible mushroom variety, has important significance for protecting germplasm resources of the species, enriching edible mushroom variety resources, enriching vegetable baskets and the like, and has ecological significance for utilizing crop straws because the mushroom is a typical straw rotting fungus.

Deposit description

Suggested classification naming: lepista nuda (Levl.) Quel

Latin name: lepitta nuda

The referenced biological materials: DCH560

The preservation organization: china general microbiological culture Collection center

The preservation organization is abbreviated as: CGMCC (China general microbiological culture Collection center)

Address: xilu No.1 Hospital No. 3 of Beijing market facing Yang district

The preservation date is as follows: 17.4.2018

Registration number of the preservation center: CGMCC No.15376

Drawings

FIG. 1 shows a field collected Lepista nuda specimen.

FIG. 2 shows the Lepista nuda DCH560(CGMCC No.15376) strain tube strain (mother strain).

FIG. 3 shows the growth of mycelia of Lepista nuda DCH560(CGMCC No.15376) after 20 days of PDA plate culture.

FIG. 4 shows the growth of Lepista nuda DCH560(CGMCC No.15376) strain inoculated in 3 stock culture media with different formulations.

FIG. 5 shows the confluent fungus state after 30 days of seeding with Lepista nuda DCH560(CGMCC No.15376) strain.

FIG. 6 shows the primordial differentiation status of Lepista nuda DCH560(CGMCC No.15376) strain.

FIG. 7 shows fruiting status of Lepista nuda DCH560(CGMCC No.15376) strain.

FIG. 8 shows fruiting body basidiomycetes and basidiospores obtained by cultivating Lepista nuda DCH560(CGMCC No.15376) strain.

Detailed Description

The experimental procedures used in the following examples are all conventional procedures unless otherwise specified.

Materials, reagents and the like used in the following examples are commercially available unless otherwise specified.

Example 1 obtaining and identification of Lepista nuda DCH560(CGMCC No.15376)

The strain DCH560 is separated from natural lepista nuda specimens (the lepista nuda specimens are collected from the autonomous county of the broad county of Dandelion, Liaoning, China in 2016), and the strain is classified and identified by observing the morphological characteristics of colonies and hyphae and combining with sequencing of cell nucleus rDNA-ITS.

First, morphological characteristics

The strain DCH560 is cultured in a potato glucose agar (PDA) medium at 20 ℃ for 2 weeks, the diameter of the colony is about 5cm, the front of the colony is light purple and flocculent, the hyphae is loose, the back of the colony is slightly dark, and the young hyphae are darker than the old hyphae (see figure 1 and figure 2). The hyphae had septa, were transparent, had a locked association, had a diameter of 1.2 to 5 μm, and no asexual spore production was observed.

II, molecular biological characteristics

rDNA-ITS sequences are used extensively for fungal species identification as barcodes (Schoch, C.L. et al. nucleic acid ribosomal internal amplified spacer (ITS) region as a native DNA barcode marker for fungi. proceedings of the National Academy of Sciences,2012.109: 6241-. The invention carries out DNA extraction and rDNA-ITS amplification sequencing on the bacterial strain DCH560 (the used primer is a primer pair consisting of ITS4 and ITS 5), and the sequencing result is shown as a sequence 1 in a sequence table. BLAST online alignment of the sequence in the NCBI database revealed 99% homologous similarity to the corresponding sequence of Lepista nuda (GenBank: KJ 577474).

ITS4：5’-TCCTCCGCTTATTGATATGC-3’；

ITS5：5’-GGAAGTAAAAGTCGTAACAAGG-3’。

In view of the morphological characteristics and the classification and identification studies of rDNA-ITS sequences, the strain DCH560 of the present invention was determined to be a Lepista nuda (Lepista nuda). The strain is preserved in China general microbiological culture Collection center (CGMCC for short, the address is No. 3 of No.1 Xilu on North Chen of the south-rising area in Beijing), and the preservation number is CGMCC No. 15376.

Example 2 method for cultivating Lepista nuda DCH560(CGMCC No.15376) fruiting body

First, cultivation process

The cultivation process of the lilac mushroom DCH560(CGMCC No.15376) fruiting body is as follows: mother seed preparation → stock seed preparation → cultivar preparation → inoculation and spawn running → covering soil → fruiting management → fruiting → harvest.

(1) Preparing a mother seed: inoculating domesticated mycelia of Lepista nuda DCH560(CGMCC No.15376) to mother culture medium (test tube slant), culturing in dark at 15-25 deg.C (such as 20-23 deg.C), and culturing until mycelia overgrow the culture medium to obtain mycelia as mother culture (FIG. 3).

(2) Preparing stock seeds: transferring the mother seeds obtained in the step (1) to a stock seed culture medium, wherein 6-10 mother seed strain blocks with the diameter of 1.0cm are uniformly inoculated into the stock seed culture medium with the inoculation amount of every 750mL, the relative humidity of air is 60% -70%, and the mother seed strain blocks are cultured in a dark place at 15-25 ℃ (such as 20-23 ℃) until hyphae overgrows the culture medium, so that the stock seeds are obtained.

(3) Preparation of cultivars: transferring the stock seeds obtained in the step (2) to a culture medium of cultivated species, inoculating 30 bottles and 750mL of cultivated species into each 750mL stock seed bottle, culturing in a dark place at 15-25 ℃ (such as 20-23 ℃), and culturing until hyphae overgrow the culture medium to obtain the cultivated species, wherein the relative air humidity is 60% -70%.

(4) Inoculation and spawn running: inoculating the cultivation material obtained in the step (3) into cultivation material, wherein the inoculation ratio is that the cultivation material per square meter of cultivation area is inoculated into 2 bottles of cultivation seeds (750mL), the inoculation method is layer sowing, the cultivation seeds are uniformly scattered on the material surface, then a layer of material is paved, a film is covered for moisture preservation, the cultivation is carried out in the dark at 15-25 ℃ (such as 20-23 ℃), and the relative humidity of air is 65% -75% (figure 5).

(5) And (3) covering soil: covering soil on the surface of the cultivation material after hyphae grow over the surface of the material, culturing in a dark place at 15-25 ℃ (such as 20-23 ℃), controlling the relative humidity of air to be 70% -75%, and ventilating for 1 time every day to maintain the relative concentration of carbon dioxide in the air to be 0.1% -0.5% (volume fraction);

the soil covered with the soil is turfy soil, and the soil is required to have good air permeability and strong water holding capacity.

(6) And (3) fruiting management: and (5) when the hyphae cultured in the step (5) grow to the surface of the soil layer, supplementing a layer of fine soil, starting to cool, stimulating by using weak light with the illumination intensity of 500-800Lux, controlling the relative humidity of air to be 85-95%, ventilating for 2-3 times every day to reduce the relative concentration of carbon dioxide to 0.05-0.15% (volume fraction), and culturing for 5-10 days.

(7) Fruiting: after a large amount of mushroom primordia was found (FIG. 6), the mushroom primordia were transferred to the condition of the ambient temperature of 18 to 20 ℃, the relative humidity of air of 75 to 90%, and the relative concentration of carbon dioxide of 0.05 to 0.15% (volume fraction) for cultivation (FIG. 7).

(8) Harvesting: and (4) harvesting the first tide of mushrooms when the sporocarp grows to be eighty percent mature and the pileus is not completely unfolded. And cleaning residual mushroom roots and soil supplementing mushroom holes after picking, performing mushroom fruiting again after hypha is recovered, and harvesting the next tide of mushrooms.

The mother culture medium can be PDA culture medium, enriched PDA culture medium or bran extract culture medium;

each liter of PDA culture medium is prepared according to the following method: boiling potato 200g in 900mL water for 20min, adding agar 20g into the filtrate, heating to dissolve completely, adding water to 1000mL, and sterilizing at 121 deg.C for 30min to obtain 1LPDA culture medium.

The enriched PDA culture medium is prepared by the following method per liter: boiling 200g of potato in 900mL of water for 20min, adding 20g of agar, 20g of sucrose or glucose, 3g of monopotassium phosphate and 1.5g of magnesium sulfate into the filtrate, heating to completely dissolve the mixture, adding water to make the mixture fully dissolved, adding water to 1000mL, and sterilizing at 121 ℃ for 30min to obtain 1L of enriched PDA culture medium.

The bran extract culture medium is prepared by the following method per liter: boiling 100g of bran in 900mL of water for 20min, adding 20g of agar, 20g of sucrose or glucose, 3g of monopotassium phosphate and 1.5g of magnesium sulfate into the filtrate, heating to completely dissolve the mixture, and adding water to supplement the mixture to 1000mL to obtain 1L of the bran immersion culture medium;

the stock culture medium and the cultivated species culture medium are wheat grain culture medium or wheat grain cow dung culture medium.

The wheat grain culture medium formula comprises: 98% of wheat grains, 1% of gypsum and 1% of lime (mass fraction). The preparation method comprises soaking wheat grains for 12-24 hr, boiling for about 20min, and preventing wheat grains from swelling and cracking. Boiling, taking out, washing with clear water, draining off water, adding the rest components, dissolving, and stirring.

The wheat grain and cow dung culture medium comprises 93% of wheat grains, 5% of cow dung, 1% of gypsum and 1% of lime (mass fraction). The preparation method comprises soaking wheat grains for 12-24 hr, boiling for about 20min, and preventing wheat grains from swelling and cracking. Boiling, taking out, washing with clear water, draining off water, adding the rest components, and stirring.

The cultivation material is a fermented manure and grass material, is prepared by fermenting crop straws, cow dung, soybean meal and gypsum, and is specifically obtained by performing secondary fermentation on the following cultivation raw materials:

the cultivation raw materials are formed by mixing crop straws (rice straws and wheat straws), animal manure (such as dried cow manure and/or chicken manure), bean foils and gypsum powder, and the mass (dry weight) ratio of the crop straws, the animal manure, the bean foils and the gypsum powder is 55: 45: 1.2: 4.

the secondary fermentation is divided into pre-fermentation and post-fermentation, wherein the pre-fermentation comprises the following steps: pre-wetting crop straws and cow dung 3-4 days before stacking, wherein the water content is 65% -70%, stacking and fermenting in the north-south direction, monitoring the temperature of a stack body, turning the stack body once when the internal temperature of the stack body begins to fall after reaching more than 70 ℃, and completing the pre-fermentation after 3-4 times of stack turning.

The post-fermentation comprises the following steps: fermenting again the cultivation material after the previous fermentation, raising the temperature inside the stack to 58-62 ℃ and keeping for 12-24h, then ventilating to lower the temperature of the stack to 50-52 ℃ and keeping for 4-6d, and then lowering the temperature of the stack to normal temperature (25-28 ℃). And (5) finishing the secondary fermentation after the completion of the fermentation.

The cultivation material after secondary fermentation has no ammonia smell and feces smell, and can be seen as white actinomycete layer with obvious material surface, the water content of the cultivation material after secondary fermentation is adjusted to 65% (mass fraction), and the pH value is adjusted to 7.0-7.5, thus obtaining the cultivation material.

The cultivation mode in the step (4) can be specifically layer frame cultivation.

The layer shelf cultivation comprises the following steps: the specification of the shelf is 3-5 layers, the height of the shelf is 0.6m, the width of the shelf is 1.5m, and the distance between the bottom layer and the ground is 0.3m (the length is determined according to the requirement); the cultivation material is spread on each layer of bed frame for cultivation and fruiting.

Second, condition optimization

The following factors in the cultivation process are optimized respectively: and (4) groping the culture temperature of the stock and preparing the formula of the stock culture medium.

1. Determination of stock culture temperature

Inoculating mycelia of Lepista nuda DCH560(CGMCC No.15376) obtained by tissue separation on PDA culture medium by punching according to the amount of inoculation blocks with the diameter of 1.0cm, culturing at different temperatures (15, 20, 22, 25 and 30 ℃) in dark, measuring the growth speed of the mycelia during the culture, and observing the growth condition of the mycelia.

The experiment was repeated three times and the quantitative data results averaged.

As shown in Table 1, it can be seen that the growth rate of mycelia is higher under the condition of 22 ℃ than under other temperature conditions by using Lepista nuda DCH560(CGMCC No. 15376).

Table 1 shows the measurement results of the growth rate of hyphae at different temperatures (unit: mm/d)

Note: the same letter in the same column indicates no significant difference in the multiplex assay (P ═ 0.05).

2. Determination of stock culture Medium

Inoculating the same batch of mother seeds of the lepista nuda DCH560(CGMCC No.15376) cultured under the same condition into 750mL culture bottles of different stock culture media (the formula is shown below), continuously culturing in a constant-temperature incubator at 22 ℃ in a dark place with the air relative humidity of 60-70%, and observing and counting the germination time of hyphae and the time that the hyphae overgrows the bottles.

The stock culture media tested were three as follows (fig. 4, a wheat grain cow dung medium, B wheat grain medium, C wood chip medium):

formula 1: cottonseed hull wood chip culture medium

40% of cottonseed hulls, 50% of sawdust, 8% of wheat bran, 1% of gypsum and 1% of cane sugar;

and (2) formula: wheat grain culture medium

98% of wheat grains, 1% of gypsum and 1% of quicklime;

and (3) formula: wheat grain and cow dung culture medium

93% of wheat grains, 5% of cow dung, 1% of gypsum and 1% of quicklime;

the parts of each substance in the above formula are parts by weight (dry weight). On the basis of the formulas, water is added for humidifying until the moisture content is 65 mass percent, and three stock culture mediums to be tested are obtained.

The quicklime and the gypsum powder are produced by chemical reagents Limited, which are far from Tianjin.

The results show (fig. 4) that the lepista nuda DCH560(CGMCC No.15376) has a relatively short germination time, a relatively high hypha growth rate, a relatively strong hypha, and a relatively good growth status in the stock culture media (named wheat grain culture medium and wheat grain fecal culture medium) corresponding to the formulas 2 and 3. And hyphae grow slowly and stop growing gradually on the wood chip culture medium of the formula 1.

Thirdly, the fruiting body characteristics obtained by cultivating the lepista nuda DCH560(CGMCC No.15376)

According to the method one, fruiting bodies are obtained.

Lepista nuda DCH560(CGMCC No.15376) not only successfully cultivated fruiting bodies, but also basidiomycetes and basidiospores (FIG. 8, A B: basidiospores, C-basidiomycetes on basidiomycetes) were observed. The cultured fruiting body pileus has diameter of 3-12cm, purple pileus, inward rolling edge, thick and light purple mushroom flesh, light color after maturation, purple mushroom fold, and strong fragrance. The hyphae are thin to slightly thick, moderately branched and bent, some hyphae are expanded and arranged in a interweaving way, the diameter of the hyphae is usually 4-10 mu m, and the diameter of the expanded hyphae reaches 15 mu m. The mycelial hyphae in the plementum is colorless, thin-walled, less branched, frequently separated, slightly bent and regularly arranged, and has the diameter of 2.5-6 μm; the sub-parenchyma layer has no capsule; the Basidion is in the shape of a stick, and has 2-4 small stems with a size of (24.0-26.0) × (5.0-7.0) μm. The basidiospores are oblong, colorless, thin to slightly thick, and have a size of (6.7-9.0) × (4.0-5.5) μm.

Sequence listing

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Claims (9)

1. Lepista nuda (Lepista nuda) DCH560 with the preservation number of CGMCC No. 15376.

2. Use of the lepista nuda of claim 1 as a parent for cross breeding.

3. A fruit body obtained by cultivating the Lepista nuda of claim 1.

4. A mycelium obtained by culturing the Lepista nuda of claim 1.

5. The method for cultivating fruit body according to claim 3, comprising the steps of: sequentially carrying out stock preparation, cultivated species preparation, inoculation and spawn running, soil covering, fruiting management and fruiting on the mycelia of the lepista nuda as claimed in claim 1 to obtain fruiting bodies.

6. The method of claim 5, wherein:

the preparation method of the stock comprises the following steps: inoculating the mycelium of the lepista nuda of claim 1 into a stock culture medium, and culturing at 15-25 ℃ in the dark, wherein the relative air humidity is 60% -70% until the mycelium overgrows the culture medium to obtain a stock;

or, the method for preparing the cultivar comprises the following steps: inoculating the stock seeds into a culture medium, and culturing at 15-25 ℃ in a dark place until hyphae overgrow the culture medium to obtain culture seeds, wherein the relative air humidity is 60% -70%;

or, the inoculation and spawn running method comprises the following steps: inoculating the cultivated species into a cultivation material, and culturing at 15-25 ℃ in the dark, wherein the relative humidity of air is 65% -75% until hypha overgrows the cultivation material;

or, the soil covering method comprises the following steps: in the inoculation and spawn running steps, after hyphae grow over the surface of the cultivation material, covering soil on the surface of the cultivation material, and culturing in a dark place at 15-25 ℃, wherein the relative humidity of air is 70-75%, and the relative concentration of carbon dioxide in the air is 0.1-0.5% until the hyphae grow to the surface of a soil layer;

or, the fruiting management method comprises the following steps: when the hypha obtained in the soil covering step grows to the surface of the soil layer, supplementing a layer of fine soil, maintaining the temperature at 15-23 ℃, the illumination intensity at 500-;

or, the fruiting method comprises the following steps: and transferring the mushroom primordium obtained in the fruiting management step to the condition that the ambient temperature is 18-20 ℃, the relative air humidity is 75-90% and the relative carbon dioxide concentration is 0.05-0.15% for culturing to obtain the fruiting body.

7. The method of claim 6, wherein:

the stock culture medium and the cultivated species culture medium are both wheat grain culture media or wheat grain cow dung culture media;

the wheat grain culture medium is prepared by mixing raw materials consisting of 98% of wheat grains, 1% of gypsum and 1% of lime by mass with water, wherein the water content of the raw materials is 65% by mass;

the wheat grain and cow dung culture medium is prepared by mixing raw materials consisting of 93% of wheat grains, 5% of cow dung, 1% of gypsum and 1% of lime with water, wherein the water content is 65% by weight.

8. The method according to claim 6 or 7, characterized in that:

in the preparation method of the stock, the light-tight culture at 15-25 ℃ is the light-tight culture at 22 ℃.

9. Use of the fruit body of claim 3 in food processing.

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