CN114181837A - New selenium-rich kuchen mushroom strain and artificial cultivation method thereof - Google Patents

Abstract

The invention discloses a novel selenium-rich kuchen mushroom strain and an artificial cultivation method thereof. Kuhn-h-mai HMGIM-W140033 deposited at 17 th 9 th 2021 at the guangdong province collection of microorganisms and cell cultures (GDMCC) address: building 5 of first furnance, large yard, 100, building 59, Guangdong province, Guangzhou, China, zip code: 510070, accession number: GDMCC No: 61940. according to the invention, by collecting new wild Coprinus pedunculatus strains and confirming the classification of the new wild Coprinus pedunculatus strains, and through the research of artificial domestication, the new Coprinus pedunculatus strains which have high temperature resistance, are suitable for cultivation in south, have strong activity and high contents of polysaccharide, selenium and potassium, wherein the selenium content is higher than that of most medicinal and edible fungi are developed.

Description

New selenium-rich kuchen mushroom strain and artificial cultivation method thereof

Technical Field

The invention belongs to the field of rare edible fungus cultivation. In particular to a novel selenium-rich kui-en mushroom strain and an artificial cultivation method thereof.

Background

Edible fungi are internationally praised as 21 st century healthy food, are nutritional, health-care and safe, are not only combined components of a meat-vegetable-mushroom dietary mode recommended by the world health organization, but also important raw materials of functional food, dietary nutritional supplement, biological medicine and cosmetics, and internationally take edible and medicinal fungi as materials of clothing cases and green buildings. China is the country with the most abundant edible and medicinal fungus resources and is the country which knows, picks, eats and cultivates edible fungi at the earliest. Domestic edible fungi rapidly develop, and become the fifth main agricultural product which is only inferior to grains, oil, fruits and vegetables in the agricultural field from 2012. In 2019, the total yield of domestic edible fungi reaches 3933.87 ten thousand tons, the yield value is 3126.67 hundred million yuan, and the edible fungi accounts for more than 75 percent of the whole world. The edible fungi is also a high-efficiency and high-value agricultural industry in the planting industry, has the characteristics of no competition with people for grains, no competition with grains for land, no competition with fertilizer and no competition with agriculture, and has quick investment return and high return rate. The edible fungi is balanced in nutrition and rich in functional components, plays an important role in accurate nutrition and chronic disease prevention and control, and is a new choice for the development of a large health industry. The edible fungus cultivation takes agricultural wastes such as straws and the like as substrates, realizes the resource utilization of the wastes, solves the problem of rural environmental pollution, and plays an important role in environmental protection and circular economy.

Kuehneromyces pilosus (Schaeff.) Singer & A.H.Sm is belonging to Basidiomycota of Basidiomycota, Hymenomycetes, Agaricales, Strophariaceae, Kuehneromyces. Kuen mushroom (Fr.) kurz is also called Fugu mushroom (Fr.) kurz or Kuen mushroom (Fr.) kurz. The strain is nutritious, has high contents of protein and crude polysaccharide, and contains various minerals. The Mustelus tussah is advocated as one of the edible fungi with rich nutrition and delicious taste by relevant German experts. In addition, it has high medicinal value, such as anti-influenza virus, tumor cell proliferation inhibition, etc. Although the kuran mushroom has high nutritional value and medicinal value, due to the fact that regional distribution of kuran mushroom is narrow, the kuran mushroom is recorded and distributed in northeast, north China, northwest China, China and other regions, wild resources are scarce, and reports on research on the kuran mushroom are few. At the end of the 19 th century, workers at the institute of edible fungi in Hebei province briefly introduced the cultivation method of Kuen mushroom (the introduction of Kuen mushroom cut-log cultivation in Germany, Bo Dong, 1996). In addition, Ganyuppeng studied the ontogeny of 5 species of the Coprinaceae family including Coprinus comatus. Foreign research has focused more on the functional studies of the metabolite content and species of different tissues of Coprinus comatus and its extracts. At present, no Coprinus pedunculatus is sold in the domestic market, and no more researches on the breeding and cultivation of the strain and the active ingredients and functions of the strain are carried out by an edible fungus related research team.

Disclosure of Invention

The first purpose of the invention is to provide kuh-kuim-W140033 which has higher fruiting temperature and can be used for fruiting in southern areas, such as Guangzhou and other areas, and is preserved in Guangdong province microbial culture Collection (GDMCC) at 2021, 9 and 17 days, address: building 5 of first furnance, large yard, 100, building 59, Guangdong province, Guangzhou, China, zip code: 510070, accession number: GDMCC No: 61940.

the second purpose of the invention is to provide an artificial cultivation method of the kuhn-mushroom HMGIM-W140033, which comprises the following steps:

inoculating a growing mother strain of Coprinus comatus HMGIM-W140033 into a stock culture medium in a culture bag for stock culture, and obtaining a stock strain after hypha is full of materials; inoculating the stock seed into a production seed culture medium in a culture bag for production seed culture, and obtaining the production seed after the hypha is full of the material; inoculating the production seeds into a culture medium of a culture material in a culture bag for culture, and carrying out culture management to obtain sporocarps;

the stock culture medium comprises: comprises 38-42% of cottonseed hulls, 35-40% of sawdust, 18-20% of bran and 1-2% of calcium carbonate according to mass fraction;

the production seed culture medium comprises: comprises 38-42% of cottonseed hulls, 35-40% of sawdust, 18-20% of bran and 1-2% of calcium carbonate according to mass fraction;

the culture medium of the cultivation material is as follows: comprises, by mass, 31-35% of cottonseed hulls, 52-56% of wood chips, 8-10% of bran and 1-2% of calcium carbonate.

Preferably, the stock culture is performed in dark at a constant temperature of 25 ℃.

Preferably, the production seed culture is performed in dark at a constant temperature of 25 ℃.

Preferably, the cultivation management is that after production seed inoculation, the production seed is cultivated in a light-proof culture room with the temperature of 25 +/-1 ℃ and the relative air humidity of 60-70%; after hypha in the cultivation bag grows to fill the cultivation material in the bag and is subjected to after-ripening treatment, opening a cover of the cultivation bag to remove fungus rings, vertically arranging the cultivation bag, placing the cultivation bag at a shading position at 25 ℃, keeping the relative humidity of 95%, replacing fresh air for 2 times every day, maintaining for 1 hour every time for 3 to 4 days, controlling the temperature to be 20 to 22 ℃, keeping the relative humidity of air between 80 and 90 percent, illuminating for 13 hours every day, increasing ventilation to enable the concentration of carbon dioxide in the air to be less than 5 percent, and growing primordium to obtain the sporocarp after continuous cultivation.

Preferably, the preparation method of the production mother strain comprises the following steps: inoculating Coprinus comatus HMGIM-W140033 to a comprehensive PDA culture medium, culturing at a constant temperature of 25 ℃ in dark, and obtaining a production mother strain after hyphae grow over a slope.

The invention also provides application of Coprinus comatus HMGIM-W140033 in preparation of selenium-rich products.

Preferably, the selenium-enriched product is a selenium-enriched food.

The technical parameters of the cultivation process of kuhn mushroom HMGIM-W140033 provided by the invention are as follows:

1. hypha culture

Culturing at 25 deg.C under constant temperature and shading, and culturing for 30 days.

2. After ripening

After the mycelium matured, the mixture was placed in a shade at 25 ℃ for 15-30 days.

3. Fruiting body

The temperature is controlled to be 20-22 ℃, the relative humidity of air is 80-90%, and the illumination is carried out for 13 h/day. The oxygen demand is small (carbon dioxide concentration in air is less than 5%).

Fruiting status

1. And (3) fruiting period: the variety has regular fruiting period of about 63 days.

2. Domesticated fruiting body trait (fig. 2): the diameter of the pileus is 2.5-6cm, the pileus is flat and hemispherical, convex, and the pileus is gradually flat and smooth, the middle part is light yellow, and the edge is yellow brown. The mushroom flesh is white or brownish. The fungiform fold is slightly downward, slightly dense, thin, wide, nearly white in the early stage and rusty brown in the later stage. The length of the stipe is 3-10cm, the thickness is 0.5-0.8cm, the upper part and the lower part are as thick as each other, the upper part is light yellow, the lower part is yellow brown, the interior is soft and becomes hollow, the fungus ring grows at the front end of the stipe, and the film quality is high, so the fungus ring is easy to fall off. The part below the fungus ring is provided with scales. The fruit body after artificial domestication has good commodity characters and regular development.

The kuhn mushroom has the characteristics of beautiful appearance, delicious taste, rich endogenous organic selenium content and crude polysaccharide content and the like, and researches prove that the extract of the kuhn mushroom has the effects of resisting influenza viruses, inhibiting the proliferation of tumor cells and the like. The former research uses stumps and cut-logs for cultivation, the optimal temperature for forming the fruit bodies is 14-18 ℃, and the fruiting temperature is low. The new strain used in the invention is collected from Shaosuan city, Guangdong province, and can be used for normal fruiting in Guangzhou area in a mixed material bag cultivation mode, and compared with the edible and medicinal nutritional ingredients in China edible and medicinal mycology, the selenium content in the kuen mushroom is higher than 99 percent. Therefore, the kuhn mushroom can be used as a high-quality food source for supplementing organic selenium for human bodies, and a new high-valued edible mushroom variety is provided for southern areas. The invention obtains the pure strain of the excellent Coprinus pedunculatus fruiting body collected from the wild environment through separation and purification, and obtains the artificially cultivated fruiting body through the research of the artificial domestication cultivation process, thereby providing an important way for the utilization of the new strain of Coprinus pedunculatus.

According to the invention, a new strain of the wild Coprinus pedunculatus, which is high in temperature resistance, suitable for cultivation in the south, strong in activity and high in polysaccharide, selenium and potassium content, is developed through collecting the new strain of the wild Coprinus pedunculatus and confirming the classification of the new strain and through the research of artificial domestication.

Kuehneromyes mutabilis HMGIM-W140033 deposited at 17 months 9/2021 at the guangdong province collection of microorganisms (GDMCC) address: building 5 of first furnance, large yard, 100, building 59, Guangdong province, Guangzhou, China, zip code: 510070, accession number: GDMCC No: 61940.

description of the drawings:

FIG. 1 is a field collection of Kuhnella pedunculata (Kuehneromyces mutubiis) HMGIM-W140033;

FIG. 2 is an artificially domesticated fruit body of wild Kuhne pedunculata HMGIM-W140033.

The specific implementation mode is as follows:

the following examples are further illustrative of the present invention and are not intended to be limiting thereof.

Example 1:

in the investigation of large-scale wild edible and medicinal fungi resources developed nationwide, the group collects kuhn mushroom in Guangdong and Hubei. When large-scale fungus resources are collected and investigated in eight-ridge national natural reserve of Binggang, Guangdong province, Shaoguan city, 3-21 Japan research team members Huhui-Pink and Cao Kerun in 2014, a Coprinus peduncularis specimen (figure 1) is obtained, and the flora grows on sunward rotten wood.

Separating a culture medium: adding rich comprehensive PDA (calculated by mass percent, comprising 20 percent of potato, 2 percent of glucose, 2 percent of agar, 0.5 percent of peptone, 0.3 percent of monopotassium phosphate, 0.15 percent of magnesium sulfate, 10.001 percent of vitamin B and the balance of water), firstly cleaning and peeling the potato, weighing 200g of the potato, cutting the potato into small blocks, adding water, boiling the small blocks thoroughly (boiling for 20-30 minutes and being punctured by a glass rod), filtering the small blocks by eight layers of gauze, heating the filtrate, adding other components, continuously heating and stirring the mixture uniformly, slightly cooling the mixture, supplementing the water to 1000 ml, subpackaging the test tubes, carrying out moist heat sterilization at the high temperature and the high pressure of 121 ℃ under the atmospheric pressure of 0.11MPa for 20min, taking out, cooling and placing the test tubes into an inclined plane.

The collected wild fruiting body is wiped with 75% alcohol under aseptic condition, torn, and inoculated with 0.2-0.5mm × 0.2-0.5mm mushroom flesh tissue in aseptic manner. Culturing in 25 deg.C incubator at constant temperature in dark, and transferring after mycelia grow over the inclined plane. The original strain HMGIM-W140033 was thus obtained by tissue isolation.

The morphological characteristics of the strain HMGIM-W140033 are as follows: the diameter of the pileus is 2-6 cm, the pileus is hemispherical or convex lens-shaped, the pileus is gradually flattened, the middle part of the pileus is protruded, and the edge of the pileus is inwards curled; slightly sticky when wet, watery, smooth or with unnoticeable white fibrils, yellow brown to dark brown, often reddish brown in the middle, with translucent streaks when wet at the edges. The mushroom flesh is white to light yellow brown. The fungus fold grows directly or slightly and is light rusty brown. The stipe is 4-10 cm long, 0.2-1 cm in diameter, medium, cylindrical and equally thick, or gradually thins towards the base; the part above the fungus ring is nearly white to yellow brown and is powdery; dark brown below the fungus ring, with reverse-rolled off-white to brown flakes; the base of the stipe has no attachment or white flocculent hypha, and the interior of the stipe becomes hollow after being soft. Superior bacteria ring and membranous. Basidiospore 5.5-7.5 multiplied by 3.5-4.5 mu m, oval or oval, smooth and light rust color. The plant is grown on the fallen wood or stump of the broad-leaved tree.

Molecular biological identification of the strain HMGIM-W140033:

fresh hyphae are ground by liquid nitrogen, and DNA genome is extracted by an Ezup column type fungal genome DNA extraction kit (product number SK8259, manufactured by Biotechnology engineering (Shanghai) Co., Ltd.). The ITS-PCR experiment of the material was performed by using fungal ribosomal intergenic region universal primer ITS1/ITS4(ITS1: TCCGTAGGTGAACCTGCGG, ITS4: TCCTCCGCTTATTGATATGC, synthesized by BioBiotechnology, Inc., Shanghai), amplification was performed on a BioRad PCR instrument, and the composition of the PCR reaction solution (50. mu.l in total) was:

the relevant reagent (product number P111-01) was purchased from Biotech GmbH, Nanjing Novowed. The reaction conditions are that the reaction is carried out for 5min at 94 ℃; reacting at 94 ℃ for 30sec, at 55 ℃ for 30sec, at 72 ℃ for 1min, and performing 32 cycles; the reaction was carried out at 72 ℃ for 10 min. The PCR product is directly checked for bidirectional sequencing and completed by Huada gene.

The sequencing results were as follows:

CGGAAGGATCATTATTGAATGAACTTGGTAGTGGTTGTAGCTGGCTCTCTCGAGA GCATGTGCACGCCCTCCCATCTTTATCTTTCCACCTGTGCACTTCTTGTAGACTTTGGGA TTTGATTATCCGAGGCAACTCGGTCGTGAGGAGCTGCTGTAGCAATACGGCTCTCCTTG AACAAGTCTTTCAGAGTCTACGTTTATCATATACCCCATAGTATGTAACAGAATGTATAC GATGAGCCTTAGTGCTTATAAAACTTATACAACTTTCAACAACGGATCTCTTGGCTCTC GCATCGATGAAGAACGCAGCGAAATGCGATAAGTAATGTGAATTGCAGAATTCAGTGA ATCATCGAATCTTTGAACGCACCTTGCGCTCCTTGGTATTCCGAGGAGCATGCCTGTTT GAGTGTCATTAAATTCTCAACCTTTATTAGCTTTTGCTGATAATGGCTTGGACTTGGGG GTCTTTTGCCGGCTTTTACTAGTCAGCTCCCCTTAAACGCATTAGCCGGTCACCTCTGG TGGAACCGTCTATTGGTGTGATAATTATCTACGCCGTGGACATCTGCCCTTGGAGTAGG TGTGCTGCTTCTAACCGTCTGTTCATTCGGACAATACATATGACATTTTGACCTCAAATC AGGTAGGACTACCCGCTGAACTTAAGCATATC (shown in SEQ ID NO. 1).

Sequencing results sequence alignment was performed at NCBI and found to be up to 99.7% similar to Kuehneromyces mutabilis.

Through morphological and molecular biological identification, the macroscopic characteristics and the microscopic characteristics of the fungus specimen are consistent with those of Kuehn kuhn fungi (Kuehn eromyces mutabilis), the identification result is the Kuehn fungi (Kuehn eromyces mutabilis), the fungus specimen is named as the Kuehn fungi (Kuehn eromyces mutabilis) HMGIM-W140033, and the fungus specimen is deposited in Guangdong province microorganism culture collection center (GDMCC) at 2021, 9 and 17 days, and the address: building 5 of first furnance, large yard, 100, building 59, Guangdong province, Guangzhou, China, zip code: 510070, accession number: GDMCC No: 61940.

example 2

First, artificial cultivation

1 production of mother seed

The production mother culture medium formula comprises: comprehensive PDA (potato 20% + glucose 2% + agar 2% + potassium dihydrogen phosphate 0.3% + magnesium sulfate 0.15% + vitamin B10.001%, the rest is water).

The method comprises the steps of firstly cleaning and peeling the potatoes, weighing 200g of the potatoes, cutting the potatoes into small pieces, adding water, boiling the small pieces thoroughly (boiling for 20-30 minutes and being punctured by a glass rod), filtering the small pieces with eight layers of gauze, heating, adding other components, continuously heating, stirring, uniformly mixing, slightly cooling, supplementing water to 1000 ml, subpackaging test tubes, carrying out moist heat sterilization at the atmospheric pressure of 0.11MPa and the high temperature and the high pressure of 121 ℃ for 20min, taking out, cooling and placing the test tubes into an inclined plane. After cooling, the successfully separated strain Coprinus pedunculatus (Kuehneromyces mutabilis) HMGIM-W140033 is inoculated in an aseptic operation. Culturing in 25 deg.C incubator at constant temperature in dark, and transferring after mycelia grow over the inclined plane. The time for the mother seeds to grow is between 10 and 15 days.

2. Preparation of stock seed

Stock culture medium: the formula is as follows: 40% of cottonseed hulls, 40% of sawdust, 19% of bran and 1% of calcium carbonate.

Weighing cottonseed hulls in a required proportion, soaking the cottonseed hulls in water overnight, mixing sawdust, bran and calcium carbonate in proportion, filling the mixture into transparent polypropylene strain bags with the high temperature resistance of 13cm multiplied by 25 cm, and folding each bag to contain dry materials of 250-fold and 300 g. After the materials are filled, a small wood bar is used for punching a hole in the bag material, the hole is deep to the bottom of the bag, then a plastic ring is sleeved on the bag opening, and a matched cover is buckled, so that the prepared original seed bag material is obtained. Performing moist heat sterilization at the atmospheric pressure of 0.147MPa and the high temperature and the high pressure of 128 ℃ for 90min, taking out, cooling, performing aseptic operation, and inoculating the strain to a production mother strain. When inoculating, the mother seed block is embedded into the raw seed block. Placing the inoculated stock in an incubator at 25 deg.C for dark culture at constant temperature, and inoculating into a culture bag after the hypha is full of the material (about 30 days).

3. Production of production seeds

The preparation process is the same as that of the original strain, but the used bag is a high-temperature resistant transparent polypropylene strain bag of 15cm multiplied by 30 cm. The amount of the dry materials in each bag is 350-400 g. The full production time is about 30 d.

4. Cultivation bag manufacturing

Culture medium of cultivation material: comprises 34 percent of cotton seed hulls, 56 percent of wood chips, 9 percent of bran and 1 percent of calcium carbonate according to mass fraction.

Weighing cottonseed hulls in a required proportion, soaking the cottonseed hulls in water overnight, mixing sawdust, bran and calcium carbonate in proportion, filling the mixture into transparent polypropylene strain bags with the high temperature resistance of 15cm multiplied by 30cm, and folding each bag to contain 300-350g of dry materials. After the materials are filled, a small wood bar is used for punching a hole in the bag material, the hole is deep to the bottom of the bag, then a plastic ring is sleeved on the opening of the bag, and a matched cover is buckled, so that the finished cultivation bag material is obtained. Performing moist heat sterilization at the atmospheric pressure of 0.147MPa and the high temperature and the high pressure of 128 ℃ for 90min, taking out, cooling, performing sterile operation, and inoculating the seeds. The full time is about 30 d.

5. Cultivation management

Inoculating the culture bag, and culturing at 25 + -1 deg.C in a dark place in a culture room with air relative humidity of 60-70%. After hypha in the cultivation bag grows over the cultivation material in the bag and is subjected to after-ripening treatment (the cultivation material is placed in a shading position at 25 ℃ for 15-30 days), a cover of the cultivation bag is opened to remove fungus rings, the cultivation bags are vertically arranged (gaps are reserved between the bags), the cultivation bags are placed in the shading position at 25 ℃ for keeping 95% of relative humidity, fresh air is replaced 2 times every day, 1 hour is carried out each time, after 3-4 days, the temperature is controlled to be 20-22 ℃, the relative humidity of air is 80-90%, illumination is carried out for 13 hours every day, ventilation is increased, the concentration of carbon dioxide in the air is smaller than 5%, and primordium can grow out after 1-2 days. The temperature and humidity are kept, illumination is provided, and when the pileus begins to be flat, the sporocarp is mature.

Secondly, the summary of Kuhnella pedunculata (Kuehneromyces mutubiis) HMGIM-W140033 is as follows:

1. character of fruiting body of Kuen mushroom HMGIM-W140033 after artificial domestication

(1) The strain can produce mushroom normally at 20-22 deg.C without low temperature stimulation, and has advantages of uniform fruiting body development, strong stress resistance, cultivation yield of 100%, and good activity. The temperature for forming the fruiting body of the kuchenkia pedunculata reported at present is 14-18 ℃.

(2) The mushroom meat is thick, the texture is crisp, the taste is delicious, and the mushroom meat has good commodity characteristics.

2. Determination of nutrient content of strain

The method comprises the steps of carrying out multiple nutrient component measurement on the cultured fruiting body, and taking two bulk edible mushrooms of needle mushroom and shiitake mushroom as a reference. As shown in Table 1, the polysaccharide, potassium and selenium contents of Kun's mushroom were higher than those of Flammulina velutipes. The polysaccharide of edible and medicinal fungi has the functions of regulating immunity, resisting tumors, reducing blood sugar and the like, and the polysaccharide of kunjin mushroom has proved to have the activity of inhibiting the proliferation of various tumor cells, so the kunjin mushroom rich in the polysaccharide can be used as a high-quality food source and has higher medicine development value.

TABLE 1 comparison of nutrient composition of Kun Ensula Aurantha

Note: the data of the flammulina velutipes and the lentinus edodes are respectively derived from 'analysis and evaluation of nutrient components of flammulina velutipes of different varieties' and 'comparison of nutrient components of lentinus edodes in four large producing areas in China'. The detection method of the lentinan is a potassium permanganate titration method. The rest are the same as in the table.

The macromineral potassium is one of essential elements of human body, participates in the metabolic process of glycogen and regulates cell metabolism; regulating osmotic pressure, and keeping acid-base balance in vivo; maintaining an agonist function of the neuromuscular system; repair and protect the skin. The potassium content in the kuen mushroom is 2 times and 1.6 times of that in the flammulina velutipes and the shiitake mushrooms respectively. Therefore, kuhn's mushroom can be a good food source for supplementing potassium.

Selenium is one of essential trace elements of human body, widely exists in all organs except fat of human body, forms active centers of a plurality of oxidases, and enhances the oxidation resistance of organism; selenium also can act synergistically with vitamin E to enhance the antioxidant effect of vitamin E. Selenium can enhance the anticancer effect of lymphocytes and the resistance of the body to diseases by activating certain enzymes of lymphocytes and the production of immunoglobulin and antibody. In addition, selenium can obviously inhibit the growth and the reproduction of tumor cells. The selenium content in the kuen mushroom is 0.0601mg/kg, which is 3.3 times of that in the flammulina velutipes, and compared with the edible and medicinal nutritional ingredients in the Chinese edible and medicinal mycology, the selenium content in the kuen mushroom is higher than 99 percent. Therefore, the kuhn mushroom can be used as a high-quality food source for supplementing organic selenium for human bodies.

The contents of other nutrients such as protein, magnesium, calcium, iron, zinc, etc. are in reasonable ranges, although they have no obvious advantages, compared with those of enoki mushroom and shiitake mushroom. In conclusion, the kuhn mushroom is a high-quality edible mushroom rich in selenium and potassium and has good edible and potential medicinal development values.

Sequence listing

<110> institute of microbiology, academy of sciences of Guangdong province (center for microbiological analysis and detection of Guangdong province)

<120> novel selenium-rich kuchen mushroom strain and artificial cultivation method thereof

<160> 1

<170> SIPOSequenceListing 1.0

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<212> DNA

<213> Kuen mushroom Cauca petioli (Kuehneromyes mutubilis)

<400> 1

cggaaggatc attattgaat gaacttggta gtggttgtag ctggctctct cgagagcatg 60

tgcacgccct cccatcttta tctttccacc tgtgcacttc ttgtagactt tgggatttga 120

ttatccgagg caactcggtc gtgaggagct gctgtagcaa tacggctctc cttgaacaag 180

tctttcagag tctacgttta tcatataccc catagtatgt aacagaatgt atacgatgag 240

ccttagtgct tataaaactt atacaacttt caacaacgga tctcttggct ctcgcatcga 300

tgaagaacgc agcgaaatgc gataagtaat gtgaattgca gaattcagtg aatcatcgaa 360

tctttgaacg caccttgcgc tccttggtat tccgaggagc atgcctgttt gagtgtcatt 420

aaattctcaa cctttattag cttttgctga taatggcttg gacttggggg tcttttgccg 480

gcttttacta gtcagctccc cttaaacgca ttagccggtc acctctggtg gaaccgtcta 540

ttggtgtgat aattatctac gccgtggaca tctgcccttg gagtaggtgt gctgcttcta 600

accgtctgtt cattcggaca atacatatga cattttgacc tcaaatcagg taggactacc 660

cgctgaactt aagcatatc 679

Claims (8)

1. Coprinus pedunculatus (Kuehneromyces mutubilis) HMGIM-W140033 with accession number: GDMCC No: 61940.

2. the artificial cultivation method of Coprinus pedunculatus HMGIM-W140033 of claim 1, comprising the steps of:

inoculating a growing mother strain of Coprinus comatus HMGIM-W140033 into a stock culture medium in a culture bag for stock culture, and obtaining a stock strain after hypha is full of materials; inoculating the stock seed into a production seed culture medium in a culture bag for production seed culture, and obtaining the production seed after the hypha is full of the material; inoculating the production seeds into a culture medium of a culture material in a culture bag for culture, and carrying out culture management to obtain sporocarps;

the stock culture medium comprises: comprises 38-42% of cottonseed hulls, 35-40% of sawdust, 18-20% of bran and 1-2% of calcium carbonate according to mass fraction;

the production seed culture medium comprises: comprises 38-42% of cottonseed hulls, 35-40% of sawdust, 18-20% of bran and 1-2% of calcium carbonate according to mass fraction;

the culture medium of the cultivation material is as follows: comprises, by mass, 31-35% of cottonseed hulls, 52-56% of wood chips, 8-10% of bran and 1-2% of calcium carbonate.

3. The artificial cultivation method according to claim 2, wherein the stock culture is performed at a constant temperature of 25 ℃ in the dark.

4. The artificial cultivation method according to claim 2, wherein the production seed culture is a dark culture at a constant temperature of 25 ℃.

5. The artificial cultivation method according to claim 2, wherein the cultivation management is that the production seeds are inoculated and cultivated in a cultivation room with the temperature of 25 ℃ ± 1 ℃ and the relative air humidity of 60-70% in a dark place; after hypha in the cultivation bag grows to fill the cultivation material in the bag and is subjected to after-ripening treatment, opening a cover of the cultivation bag to remove fungus rings, vertically arranging the cultivation bag, placing the cultivation bag at a shading position at 25 ℃, keeping the relative humidity of 95%, replacing fresh air for 2 times every day, maintaining for 1 hour every time for 3 to 4 days, controlling the temperature to be 20 to 22 ℃, keeping the relative humidity of air between 80 and 90 percent, illuminating for 13 hours every day, increasing ventilation to enable the concentration of carbon dioxide in the air to be less than 5 percent, and growing primordium to obtain the sporocarp after continuous cultivation.

6. The artificial cultivation method as claimed in claim 2, wherein the production mother culture is prepared by: inoculating Coprinus comatus HMGIM-W140033 to a comprehensive PDA culture medium, culturing at a constant temperature of 25 ℃ in dark, and obtaining a production mother strain after hyphae grow over a slope.

7. The use of Coprinus pedunculata HMGIM-W140033 of claim 1 for preparing a selenium enriched product.

8. The use of claim 7, wherein the selenium enriched product is a selenium enriched food.

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