CN108048335B - Noval strain grassland white mushroom No. 2 of Mongolian tricholoma mongolicum and breeding method thereof - Google Patents
Noval strain grassland white mushroom No. 2 of Mongolian tricholoma mongolicum and breeding method thereof Download PDFInfo
- Publication number
- CN108048335B CN108048335B CN201711421978.5A CN201711421978A CN108048335B CN 108048335 B CN108048335 B CN 108048335B CN 201711421978 A CN201711421978 A CN 201711421978A CN 108048335 B CN108048335 B CN 108048335B
- Authority
- CN
- China
- Prior art keywords
- strain
- days
- white mushroom
- mushroom
- new
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
- C12N1/145—Fungal isolates
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/645—Fungi ; Processes using fungi
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H15/00—Fungi; Lichens
-
- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F17/00—Preparation of fertilisers characterised by biological or biochemical treatment steps, e.g. composting or fermentation
-
- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F3/00—Fertilisers from human or animal excrements, e.g. manure
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
- Y02W30/00—Technologies for solid waste management
- Y02W30/40—Bio-organic fraction processing; Production of fertilisers from the organic fraction of waste or refuse
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Biochemistry (AREA)
- Mycology (AREA)
- General Health & Medical Sciences (AREA)
- General Engineering & Computer Science (AREA)
- Botany (AREA)
- Virology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Medicinal Chemistry (AREA)
- Biomedical Technology (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Developmental Biology & Embryology (AREA)
- Environmental Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Molecular Biology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention belongs to the field of edible fungus new strain domestication methods, and particularly discloses a new Mongolian tricholoma mongolicum strain prairie white mushroom No. 2 and a breeding method thereof. The preservation number of the Mongolian tricholoma mongolicum new strain prairie white mushroom No. 2 strain is CGMCC No.15084, the Mongolian tricholoma mongolicum new strain has the characteristics of strong stress resistance, stable genetic property, high quality and delicious taste, and a new high-quality edible fungus product is provided for people. The domestication method provided by the invention not only protects the endangered and extincted Lepista sordida, which is a rare species, but also realizes the resource utilization of agricultural and animal husbandry waste by cultivating the new strain of the prairie white mushroom No. 2, promotes the sustainable development of agricultural economy, expands new economic growth points of agricultural and animal husbandry, is an important ring for developing the garden economy and the grass and livestock circular economy of the pastoral area, and carries out the entrepreneurial and employment on the prairie white mushroom No. 2 cultivated by using the waste of the agricultural and animal husbandry area, the idle cattle and sheep pen and other facilities, and the farmer becomes rich.
Description
Technical Field
The invention belongs to the field of domestication methods of new strains of edible fungi, and particularly relates to a new strain of tricholoma mongolicum chen, prairie white mushroom No. 2 and a domestication method thereof.
Background
Lepista sordida (Lepista sordida) belongs to Agaricales (Agaricales), Tricholomataceae (Tricholomataceae), and Lepista (Lepista). Lepista sordida, called Cleohedra petthey, are mainly distributed on the Silibinoguo and Colqin grasslands of inner Mongolia. The Lepista sordida has high protein content and complete amino acid types, particularly contains rich calcium, iron, carotene, nicotinic acid, trace elements such as copper, zinc, fluorine and iodine, so that the Lepista sordida has the effects of nourishing blood, benefiting spirit, tonifying liver and five internal organs, is favorable for treating diseases such as anemia, metrorrhagia and metrostaxis, prolonged illness and weak constitution, mental fatigue and amnesia after being eaten for a long time, and is a precious edible fungus resource. At present, the degradation of grassland is increasingly serious, and in addition, the natural living conditions are increasingly worsened due to excessive ingestion of people.
Therefore, the new strain of the Mengolia tricholoma mongolicum Murr No. 2 suitable for artificial cultivation is urgently needed to be provided, the rare species of the endangered and extincted grassland white mushroom No. 2 is protected while the original flavor of the wild Lepista sordida is kept, and the farming and animal husbandry waste recycling can be realized by cultivating the new strain of the grassland white mushroom No. 2,
disclosure of Invention
In order to solve the problems in the prior art, the invention aims to provide a new strain of the tricholoma mongolicum 2 and a breeding method thereof.
In order to realize the purpose of the invention, the technical scheme of the invention is as follows:
in the first aspect, the invention provides a grassland white mushroom No. 2, which is identified as Lepista sordida, and is preserved in the China general microbiological culture Collection center (CGMCC for short, the address: Beijing university Hokko No.1 of North Chen West Lu No. 3 of Indonesia area, China academy of sciences, microbiological research institute, postal code: 100101), the preservation date is 2017, 12 months and 7 days, and the preservation number is CGMCC No. 15084.
In the grassland white mushroom No. 2, the pileus of the fruiting body is white, the edge of the flat hemispherical pileus at the initial stage is curled inwards, the pileus is in a bamboo hat-shaped edge water stain shape and black white at the mature stage, the diameter of the pileus of the mature fruiting body is 3.5-8.7 cm, and the pileus is white and thin; the fungus folds are white, straight or bent, slightly thin and unequal in length; the stipe is thick and white, the length is 3.5-7.6 cm, the thickness is 0.2-2.0 cm, the internal is solid, and the stipe is bent and grows normally close to the basal part (see figure 2).
The coding gene sequence of the 5.8S ribosome of the strain is shown in SEQ ID NO.1, and the agarose gel electrophoresis picture after the sequence is amplified is shown in figure 3.
The prairie white mushroom No. 2 provided by the invention is obtained by domesticating wild lepista sordida, retains the original flavor of the wild lepista sordida, and can realize artificial cultivation.
In a second aspect, the present invention provides a method for domesticating a grassland white mushroom No. 2 strain, comprising the steps of:
1) preparing a mother seed: culturing mother seeds by taking wild Lepista sordida fruiting bodies as materials at 21-24 ℃ until hyphae grow on a culture medium;
2) propagation: transferring the mother seeds obtained in the step 1) to a culture medium, and culturing at 21-24 ℃ until hyphae grow over;
3) preparing an original seed: transferring the mother seeds expanded and propagated in the step 2) into a vessel filled with a stock culture material, and culturing for 33-38 days at a constant temperature of 20-23 ℃ until hyphae overgrow the vessel to finish the production of the stock seeds;
4) preparing cultivars: and (3) implanting the stock seeds prepared in the step 3) into a vessel filled with culture material of the cultivated species, and culturing for 28-33 days at a constant temperature of 20-23 ℃ until hyphae overgrow the vessel to finish the preparation of the cultivated species.
Wherein, the step 1) uses a tissue separation method or a basidiospore collection method to prepare strains:
the tissue separation method comprises the following steps: cutting off fresh and wild Lepista sordida fruiting bodies under aseptic conditions, selecting mushroom flesh tissues at the junction of pileus and stipe, transferring the mushroom flesh tissues into a vessel filled with a culture medium, and culturing for 24-27 days in a constant-temperature incubator at 21-24 ℃;
the basidiospore collection method comprises the following steps: under aseptic condition, passing fresh and wild Lepista sordida fruiting bodies through iron wires, hanging the plectrum of the Lepista sordida fruiting bodies downwards in a container to collect basidiospores, picking a little basidiospores into a test tube, adding sterile water to dilute to 100 mu L of basidiospores containing 40-50 basidiospores, and obtaining basidiospore suspension; sucking basidiospore suspension, dripping the basidiospore suspension on a culture medium, uniformly coating, and culturing in a constant-temperature incubator at 21-24 ℃ for 24-27 days.
Wherein, the culture medium in the step 1) is a PDA improved culture medium;
the culture medium in the step 2) is a PDA improved culture medium or a PDA improved liquid culture medium; when the PDA improved culture medium is adopted, culturing the seeds on the PDA improved culture medium for 24-27 days; when the PDA improved liquid culture medium is adopted, culturing for 13-16 days in the PDA improved liquid culture medium at a rotating speed of 120-140 r/min;
the formula of the PDA improved culture medium is as follows: 15 g/L-200 g/L of carbon source, 10g/L of sucrose, 10g/L of glucose, 1.2g/L of yeast powder, 1.2g/L of peptone, 0.5g/L of magnesium sulfate, 1g/L of dipotassium hydrogen phosphate and 10g/L of agar;
the formula of the PDA improved liquid culture medium is as follows: 15 g/L-200 g/L of carbon source, 10g/L of sucrose, 10g/L of glucose, 1.2g/L of yeast powder, 1.2g/L of peptone, 0.5g/L of magnesium sulfate, 1g/L of dipotassium hydrogen phosphate and 0.6g/L of agar;
the carbon source is one or more of potato, sweet potato, bran, whole wheat flour and bean cake flour.
The formula of the stock culture material in the step 3) is as follows: whole wheat or grain, potassium dihydrogen phosphate and magnesium sulfate;
the mass of the potassium dihydrogen phosphate is 0.2 percent of the mass of whole wheat grains or grains;
the mass of the magnesium sulfate is 0.1% of the mass of whole wheat grains or grains;
the preparation method of the stock culture material comprises the following steps: soaking whole wheat grains or grains in a ratio of 1: 1.2-1: 1.5 of the mass of feed water for 8-12 hours, adding potassium dihydrogen phosphate and magnesium sulfate while soaking to fully dissolve the whole wheat grains or grains, boiling the whole wheat grains or grains after soaking until no hard core exists, controlling water, adjusting the pH value to 7.5-8 by using lime, and sterilizing.
Wherein the culture material of the cultivar in the step 4) comprises 20-30 parts of wheat grains and 70-80 parts of caragana microphylla powder by mass;
the preparation method of the culture material for the cultivated species comprises the following steps: soaking wheat grains for 8-12 hours according to the mass ratio of 1: 1.3-1: 2 of feed water, adding potassium dihydrogen phosphate and magnesium sulfate which are 0.2% and 0.1% of the mass of the wheat grains respectively while soaking the wheat grains to fully dissolve the wheat grains, boiling the wheat grains until no hard core exists but the wheat grains do not crack, and removing water; the caragana microphylla powder is soaked in water according to the mass ratio of 1:1.2, then is uniformly mixed with wheat grains, is adjusted to pH 7.5-8 by lime, and is sterilized.
The caragana microphylla powder is caragana microphylla branches of the overground part of caragana microphylla, and is crushed into particles of 0.5-1.0 cm. The caragana microphylla powder has high protein content, is rich in mineral elements, vitamins and the like, and is rich in nutrition.
In a third aspect, the present invention also provides a method for artificially cultivating grassland white mushroom No. 2, comprising the steps of:
(a) stacking and fermenting plant waste and livestock manure;
(b) uniformly spreading the grassland white mushroom No. 2 or other cultivated species obtained by domestication by the domestication method on the fermented material, spreading a layer of fermented material on the fermented material, then spreading a layer of cultivated species, spreading each layer of the fermented material with the thickness of 10-15 cm, mixing and flattening surface layer strains and cultivated species compost, finishing to form turtle back type fruiting rows with unlimited length, the bottom width of 1.3-1.5 m, the top width of 1.2-1.3 m and the height of 20-24 cm, and culturing mycelia for 50-60 days.
Preferably, the stockpile fermentation is as follows: uniformly mixing plant waste and livestock manure according to the mass ratio of 4: 6-5: 5 to form a fermentation pile, and fermenting in a clean and ventilated place;
the plant waste is selected from one or more of grass seeds, caragana microphylla branches and leaves, rice straws, corn stalks and wheat straws.
The composting fermentation is as follows: the method comprises the steps of building a fermentation pile with the bottom width of 1.2-1.4 m, the top width of 1.1-1.3 m and the height of 1.1-1.3 m, punching holes on the pile by using wood rods with the diameter of 6-8 cm until the bottom of the pile is reached, wherein the hole interval is 40-60 cm, keeping the pile temperature for 24 hours when the pile is heated to 65 ℃, turning the pile for the first time, continuing punching and fermenting, keeping the pile temperature for 24 hours when the pile is heated to 65 ℃, keeping the pile for the second time, keeping the pile for 12 hours when the temperature is raised to 65 ℃ again after 2 days, turning the pile for the third time, and adjusting the water content to be about 60-65% to be used for seeding.
Further, after the mycelium is full of materials, covering garden soil or turfy soil with the thickness of 4-6 cm on the surface, spraying water for moisturizing when the surface soil is white before fruiting, and controlling the temperature to be 21-23 ℃; and covering soil for 40-45 days, allowing grassland white mushroom No. 2 primordium to appear, keeping the soil moist, and harvesting after 9-12 days.
Further, after the first crop of mushrooms is harvested, the surface is leveled and lightly pressed for 2 days, fruiting management is continued after the growth of the mycelia is recovered, and second crop of mushroom primordia appear 15-18 days after the first crop of mushrooms are harvested, and 3 crops of mushrooms can be harvested in total.
The invention has the beneficial effects that:
the invention provides a new strain of wild lepista sordida, a domestication method and a cultivation method thereof. The domestication method not only protects Lepista sordida, a rare and cherished wild species, from extinction, maintains the ecosystem of the symbiosis of the pasture and the grassland mushrooms, but also provides edible fungus products with excellent taste for people.
The new strain of the grassland white mushroom No. 2 provided by the invention has strong stress resistance, stable genetic property, high quality and delicious taste, and provides a new high-quality edible mushroom product for people. The prairie white mushroom No. 2 obtained by the method retains the special flavor and nutritional ingredients of the wild Lepista sordida, and the product produced in cattle and sheep pens in pastoral areas in summer is fresher in taste and is deeply loved by people.
The invention not only protects the rare species of the prairie white mushroom No. 2 which is endangered to be extinct, but also realizes the resource utilization of the agricultural and animal husbandry waste by cultivating the new strain of the prairie white mushroom No. 2, changes waste into valuable, expands the new economic growth point of the agricultural and animal husbandry, is an important ring for developing the pastoral garden economy and the grass and livestock circular economy, and carries out the entrepreneurial and employment on the prairie white mushroom No. 2 cultivated by using the waste of the agricultural and animal husbandry, the idle cattle and sheep pen and other facilities, and the farmer becomes rich. Realizes the comprehensive utilization of resources and promotes the sustainable development of the new grass, livestock and bacteria industry ecological cycle economy.
Drawings
FIG. 1 shows Lepista sordida wild strain.
FIG. 2 shows the strain "Shiraia white mushroom No. 2" according to the present invention.
FIG. 3 is the amplified electrophoresis chart of the 5.8S ribosome coding gene sequence of the white grassland mushroom No. 2 according to the present invention.
Detailed Description
Preferred embodiments of the present invention will be described in detail with reference to the following examples. It is to be understood that the following examples are given for illustrative purposes only and are not intended to limit the scope of the present invention. Various modifications and alterations of this invention will become apparent to those skilled in the art without departing from the spirit and scope of this invention.
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
The PDA modified medium used in this example was: the temperature is between 121 and 125 ℃ (the pressure is 1.1 kg/cm)2～1.5kg/cm2) Sterilizing for 30 min, and containing sweet potato 200g/L, sucrose 10g/L, glucose 10g/L, yeast powder 1.2g/L, peptone 1.2g/L, magnesium sulfate 0.5g/L, dipotassium hydrogen phosphate 1g/LL, agar 10 g/L.
1) Preparing a mother seed: collecting robust and wild Lepista sordida to prepare mother seeds, cutting fresh and wild Lepista sordida fruiting bodies under aseptic condition, selecting mushroom flesh tissues at the junction of pileus stipe, transferring into a vessel filled with PDA improved culture medium, putting into a constant temperature incubator for culturing, and culturing at 21-24 ℃ for 24-27 days until hypha grows over the incubator.
2) Propagation: transferring the mother seeds to a PDA improved culture medium, and culturing for 24-27 days at 21-24 ℃ until hyphae grow over.
3) Preparing an original seed: transplanting the obtained expanded breeding mother seeds into a vessel filled with a culture material, culturing for 33-38 days under the constant temperature condition of 20-23 ℃, and growing hyphae over the vessel to finish the stock seed preparation.
4) Preparing cultivars: and (3) planting the obtained stock seeds into a vessel filled with culture materials, placing the vessel at the constant temperature of 20-23 ℃ for culturing for 28-33 days, and growing hypha in the vessel to finish the production of the cultivated seeds.
The preparation method of the stock culture material comprises the following steps: soaking whole wheat grain or corn in water at a ratio of 1:1.5 for 10 hr, adding 0.2% potassium dihydrogen phosphate and 0.1% magnesium sulfate to dissolve completely, boiling to no hard core but not cracking, removing water, adjusting pH to 7.5 with 2% lime, placing into a container, and standing at 121-125 deg.C (1.1 kg/cm pressure)2～1.5kg/cm2) Sterilizing for 2.5h, and cooling to obtain the final product.
The cultivation material for the cultivated species comprises 20 parts of wheat grains and 80 parts of caragana microphylla powder by mass; the preparation method of the culture material for the cultivated species comprises the following steps: soaking wheat grains for 8-12 hours according to the mass ratio of 1: 1.3-1: 2 of feed water, adding potassium dihydrogen phosphate and magnesium sulfate which are 0.2% and 0.1% of the mass of the wheat grains respectively while soaking the wheat grains to fully dissolve the wheat grains, boiling the wheat grains until no hard core exists but the wheat grains do not crack, and removing water; the caragana microphylla powder is soaked in water according to the mass ratio of 1:1.2, then is uniformly mixed with wheat grains, the pH value is adjusted to 7.5-8, and the sterilization is carried out.
Example 2
The PDA modified medium used in this example was: at a temperature of 121About 125 ℃ (pressure 1.1 kg/cm)2～1.5kg/cm2) Sterilizing for 30 minutes for 100/L of bran, 10g/L of sucrose, 10g/L of glucose, 1.2g/L of yeast powder, 1.2g/L of peptone, 0.5g/L of magnesium sulfate, 1g/L of dipotassium hydrogen phosphate and 0.6g/L of agar.
1) Preparing a mother seed: collecting robust and wild Lepista sordida to prepare mother seeds, using iron wires to penetrate through fresh and wild Lepista sordida fruiting bodies under aseptic conditions, enabling mushroom folds of the mushroom folds to be downwards hung in a wide-mouth bottle to collect basidiospores, picking a small amount of basidiospores to be placed in a test tube, adding sterile water to dilute the basidiospores to 100 mu L containing 40-50 basidiospores, and obtaining basidiospore suspension. And (3) sucking basidiospore suspension, dropwise adding the basidiospore suspension into a modified culture medium filled with PDA, uniformly coating, and culturing at the temperature of 21-24 ℃ for 24-27 days until hyphae overgrow the device.
2) Propagation: transferring the mother strain to an improved PDA liquid culture medium, and culturing the cultured bacterium balls at the rotating speed of 130r/min for 12-16 days at the temperature of 21-24 ℃ to uniformly grow the bacterium balls in a vessel, thereby completing the production of the liquid strain.
3) Preparing an original seed: transplanting the obtained expanded breeding mother seeds into a vessel filled with a culture material, culturing for 33-38 days under the constant temperature condition of 20-23 ℃, and growing hyphae over the vessel to finish the stock seed preparation.
4) Preparing cultivars: and (3) planting the obtained stock seeds into a vessel filled with culture materials, placing the vessel at the constant temperature of 20-23 ℃ for culturing for 28-33 days, and growing hypha in the vessel to finish the production of the cultivated seeds.
The preparation method of the stock culture material comprises the following steps: soaking whole wheat grain or corn in water at a ratio of 1:1.3 for 12 hr, adding 0.2% potassium dihydrogen phosphate and 0.1% magnesium sulfate to dissolve completely, boiling to no hard core but not cracking, removing water, adjusting pH to 7.5 with 2% lime, placing into a container, and standing at 121-125 deg.C (1.1 kg/cm pressure)2～1.5kg/cm2) Sterilizing for 2.5h, and cooling to obtain the final product.
The culture material for the cultivated species comprises 30 parts of wheat grains and 70 parts of caragana microphylla powder by mass; the preparation method of the culture material for the cultivated species comprises the following steps: soaking wheat grains for 8-12 h according to the mass ratio of 1:1.3 of the feed water, adding potassium dihydrogen phosphate and magnesium sulfate which are 0.2% and 0.1% of the mass of the wheat grains respectively while soaking the wheat grains to fully dissolve the wheat grains, boiling the wheat grains until no hard core exists but the wheat grains do not crack, and removing water; soaking caragana microphylla powder in water at a ratio of 1:1.2, mixing with wheat grains, adjusting pH to 7.5, and sterilizing.
Example 3
The PDA modified medium used in this example was: the PDA improved culture medium comprises: the temperature is between 121 and 125 ℃ (the pressure is 1.1 kg/cm)2～1.5kg/cm2) The potato powder is sterilized for 30 minutes and contains 200g/L of potato, 10g/L of cane sugar, 10g/L of glucose, 1.2g/L of yeast powder, 1.2g/L of peptone, 0.5g/L of magnesium sulfate, 1g/L of dipotassium hydrogen phosphate and 10g/L of agar.
1) Preparing a mother seed: collecting robust and wild Lepista sordida to prepare mother seeds, cutting fresh and wild Lepista sordida fruiting bodies under aseptic condition, selecting mushroom flesh tissues at the junction of pileus stipe, transferring into a vessel filled with PDA improved culture medium, putting into a constant temperature incubator for culturing, and culturing at 21-24 ℃ for 24-27 days until hypha grows over the incubator.
2) Propagation: transferring the mother seeds to a PDA improved culture medium, and culturing for 24-27 days at 21-24 ℃ until hyphae grow over.
3) Preparing an original seed: transplanting the obtained expanded breeding mother seeds into a vessel filled with wheat grain compost, placing the vessel under the constant temperature condition of 20-23 ℃ for culturing for 33-38 days, and growing hypha over the vessel to finish the stock seed preparation.
4) Preparing cultivars: and (3) planting the obtained stock seeds into a vessel filled with culture materials, placing the vessel at the constant temperature of 20-23 ℃ for culturing for 28-33 days, and growing hypha in the vessel to finish the production of the cultivated seeds.
The preparation method of the stock culture material comprises the following steps: soaking whole wheat grain or corn in water at a ratio of 1:1.3 for 11 hr, adding 0.2% potassium dihydrogen phosphate and 0.1% magnesium sulfate to dissolve completely, boiling to no hard core but not cracking, removing water, adjusting pH to 7.5 with 2% lime, placing into a container, and standing at 121-125 deg.C (1.1 kg/cm pressure)2～1.5kg/cm2) Sterilizing for 2.5h, and cooling to obtain the final product.
The culture material for the cultivated species comprises 25 parts of wheat grains and 75 parts of caragana microphylla powder by mass; the preparation method of the culture material for the cultivated species comprises the following steps: soaking wheat grains for 11h according to the mass ratio of 1:1.3 of the feed water, adding potassium dihydrogen phosphate accounting for 0.2% of the mass of the wheat grains and magnesium sulfate accounting for 0.1% of the mass of the wheat grains while soaking to fully dissolve the wheat grains, boiling the wheat grains until no hard core exists but the wheat grains are not cracked after soaking, and removing water; soaking caragana microphylla powder in water at a ratio of 1:1.2, mixing with wheat grains, adjusting pH to 8, and sterilizing.
Example 4
The PDA modified medium used in this example was: the temperature is between 121 and 125 ℃ (the pressure is 1.1 kg/cm)2～1.5kg/cm2) The mixture is sterilized for 30 minutes and contains 20g/L of bean cake powder, 10g/L of cane sugar, 10g/L of glucose, 1.2g/L of yeast powder, 1.2g/L of peptone, 0.5g/L of magnesium sulfate, 1g/L of dipotassium hydrogen phosphate and 0.6g/L of agar.
1) Preparing a mother seed: collecting robust and wild Lepista sordida to prepare mother seeds, using iron wires to penetrate through fresh and wild Lepista sordida fruiting bodies under aseptic conditions, enabling mushroom folds of the mushroom folds to be downwards hung in a wide-mouth bottle to collect basidiospores, picking a small amount of basidiospores to be placed in a test tube, adding sterile water to dilute the basidiospores to 100 mu L containing 40-50 basidiospores, and obtaining basidiospore suspension. And (3) sucking basidiospore suspension, dropwise adding the basidiospore suspension into a modified culture medium filled with PDA, uniformly coating, and culturing at the temperature of 21-24 ℃ for 24-27 days until hyphae overgrow the device.
2) Propagation: transferring the mother strain to an improved PDA liquid culture medium, and culturing the cultured bacterium balls at the rotating speed of 130r/min for 13-16 days at the temperature of 21-24 ℃ to uniformly grow over a vessel, thereby completing the production of the liquid strain;
3) preparing an original seed: transplanting the obtained expanded breeding mother seeds into a vessel filled with wheat grain compost, and culturing for 33-38 days at a constant temperature of 20-23 ℃ until hyphae grow over the vessel.
4) Preparing cultivars: and (3) planting the obtained stock seeds into a vessel filled with the culture material, and culturing for 28-33 days at a constant temperature of 20-23 ℃ until hyphae grow over the vessel.
The culture medium of steps 3) and 4) was the same as in example 1.
5) Fermentation: adding water into pre-ground and sieved sheep manure and fresh and mildew-free caragana microphylla powder branches and leaves according to the proportion of 4:6, fully pre-wetting, alternately and uniformly paving sheep manure and grass to form a pile with the bottom width of 1.3m, the upper width of 1.2m, the height of 1.2m and the length of no limit, punching holes to the bottom of the material by using a wood stick with the diameter of 6-8 cm at the hole distance of 50cm, keeping the pile for 24 hours when the temperature of the pile rises to 65 ℃, performing first pile turning, continuously punching and fermenting, keeping the pile temperature to 65 ℃ for 24 hours to perform second pile turning, raising the temperature again after 2 days to 65 ℃, and performing third pile turning after 12 hours, wherein the water content is adjusted to be about 60-65%, and then the pile can be used for seeding.
6) Domestication and cultivation: spreading the fermented culture material on the ground of a cleaned sheepfold in a pasturing area, uniformly spreading the cultivated species on the culture material, spreading a layer of fermented culture material on the ground, spreading a layer of cultivated species, mixing and flattening the surface layer strain and the culture material, arranging the mixture into a turtle back type fruiting row with the bottom width of 1.4m, the top width of 1.3m and the height of about 23cm, and culturing the mycelia for 50-60 days. After the mycelium grows over the culture material, covering 5cm of moist soil on the surface, spraying water to preserve moisture when the surface soil is white before fruiting, and controlling the temperature at 21-23 ℃. And (3) covering soil for 40-45 days, allowing the grassland white mushroom No. 2 primordium to appear, harvesting after 9-12 days, and scientifically managing fruiting to harvest 3 batches of mushrooms.
Example 5
The PDA modified medium used in this example was: sterilizing the culture medium containing 15g/L of whole wheat flour, 10g/L of sucrose, 10g/L of glucose, 1.2g/L of yeast powder, 1.2g/L of peptone, 0.5g/L of magnesium sulfate, 1g/L of dipotassium hydrogen phosphate and 10g/L of agar for 30 minutes at the temperature of 121-125 ℃ (the pressure is 1.1kg/cm 2-1.5 kg/cm 2).
1) Preparing a mother seed: collecting robust and wild Lepista sordida to prepare mother seeds, cutting fresh and wild Lepista sordida fruiting bodies under aseptic condition, selecting mushroom flesh tissues at the junction of pileus stipe, transferring into a vessel filled with PDA improved culture medium, putting into a constant temperature incubator for culturing, and culturing at 21-24 ℃ for 24-27 days until hypha grows over the incubator.
2) Propagation: transferring the mother seeds to a PDA improved culture medium, and culturing for 24-27 days at 21-24 ℃ until hyphae grow over.
3) Preparing an original seed: transplanting the obtained expanded breeding mother seeds into a vessel filled with wheat grain compost, placing the vessel under the constant temperature condition of 20-23 ℃ for culturing for 33-38 days, and growing hypha over the vessel to finish the stock seed preparation.
4) Preparing cultivars: and (3) planting the obtained stock seeds into a vessel filled with culture materials, placing the vessel at the constant temperature of 20-23 ℃ for culturing for 28-33 days, and growing hypha in the vessel to finish the production of the cultivated seeds.
The preparation method of the stock culture material comprises the following steps: soaking whole wheat grain or corn in water at a ratio of 1:1.3 for 11 hr, adding 0.2% potassium dihydrogen phosphate and 0.1% magnesium sulfate to dissolve completely, boiling to no hard core but not cracking, removing water, adjusting pH to 7.5 with 2% lime, placing into a container, and standing at 121-125 deg.C (1.1 kg/cm pressure)2～1.5kg/cm2) Sterilizing for 2.5h, and cooling to obtain the final product.
The culture material for the cultivated species comprises 25 parts of wheat grains and 75 parts of caragana microphylla powder by mass; the preparation method of the culture material for the cultivated species comprises the following steps: soaking wheat grains for 11h according to the mass ratio of 1:1.3 of the feed water, adding potassium dihydrogen phosphate accounting for 0.2% of the mass of the wheat grains and magnesium sulfate accounting for 0.1% of the mass of the wheat grains while soaking to fully dissolve the wheat grains, boiling the wheat grains until no hard core exists but the wheat grains are not cracked after soaking, and removing water; soaking caragana microphylla powder in water at a ratio of 1:1.2, mixing with wheat grains, adjusting pH to 8, and sterilizing.
5) Fermentation: adding water into pre-ground sheep manure and fresh caragana microphylla powder branches and leaves in a ratio of 1:1, fully pre-wetting, alternately and uniformly paving sheep manure and grass to form a pile with the bottom width of 1.2m, the top width of 1.1m, the height of 1.1m and the length of no limit, punching holes to the bottom of the pile by using a wood stick with the diameter of 6-8 cm at a distance of 40cm, keeping the pile for 24 hours when the temperature of the pile rises to 65 ℃, performing first pile turning, continuously punching and fermenting, keeping the pile temperature to 65 ℃ for 24 hours to perform second pile turning, raising the temperature again after 2 days, carrying out third pile turning after 12 hours, and adjusting the water content to be about 60-65% to be used for seeding.
6) Domestication and cultivation: spreading the fermented culture material on the ground of a cleaned sheepfold in a pasturing area, uniformly spreading the cultivated species on the culture material, spreading a layer of fermented culture material on the ground, spreading a layer of cultivated species, mixing and flattening the surface layer strain and the culture material, arranging the mixture into a turtle back type fruiting row with the bottom width of 1.3m, the top width of 1.2m and the height of about 23cm, and culturing the mycelia for 50-60 days. After the mycelium grows over the culture material, covering 5cm of moist soil on the surface, spraying water to preserve moisture when the surface soil is white before fruiting, and controlling the temperature at 21-23 ℃. And (3) covering soil for 40-45 days, allowing the grassland white mushroom No. 2 primordium to appear, harvesting after 9-12 days, and scientifically managing fruiting to collect 3 batches of mushrooms.
The invention successfully realizes the artificial domestication and cultivation of the grassland white mushroom No. 2, not only protects the endangered Lepista sordida which is a rare species, but also makes full use of the rich wastes such as cattle and sheep manure, grass dregs and the like in the pastoral area to cultivate the grassland white mushroom No. 2, realizes the comprehensive utilization of resources, promotes the cyclic utilization and sustainable development of the emerging industry of grass, livestock and fungi, and can increase the income of local herdsmen.
It should be understood that the technical solutions of the above embodiments, in which the amounts of reagents or raw materials used are proportionally increased or decreased, are substantially the same as those of the above embodiments.
Although the invention has been described in detail hereinabove with respect to a general description and specific embodiments thereof, it will be apparent to those skilled in the art that modifications or improvements may be made thereto based on the invention. Accordingly, such modifications and improvements are intended to be within the scope of the invention as claimed.
Sequence listing
<110> inner Mongolia university of academy of sciences of agriculture and animal husbandry in inner Mongolia autonomous region
<120> New strain grassland white mushroom No. 2 of Mongolian tricholoma mongolicum and breeding method thereof
<141> 2017-12-19
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 652
<212> DNA
<213> Lepista sordida (Lepista sordida)
<400> 1
acttggttgg gttgtgctgg cttttcggag catgtgcatg cctagcgcca tttttaccac 60
ctgtgcacat tttgtagatt tgaaacaatt ctcgaggaaa ctcggtttga ggaatgctgt 120
gcgaaagctt agcttttctt gtgtttcaag tctatgtttt tatatatacc ccataagaat 180
gtaatagaat gtcattaatg ggctttgttg cctttaaatt aatacaactt tcaacaacgg 240
atctcttggt tctcgcatcg atgaagaacg cagcgaaatg cgataagtaa tgtgaattgc 300
agaattcagt gaatcatcga atctttgaac gcaccttgcg ctccttggta ttccgaggag 360
catgcctgtt tgagtgtcat taaattctca acctttccag cttttgcaag ttggattggc 420
ttggatgtgg agggttttgc gggcttctca gaagtcggct cctcttaaat gcattagcag 480
aacctttgtg gaccagcttt ggtgtgataa ttatctatgc cattgttgta aagcagcttt 540
tatatggggt tcagcttcta atagtccatt gacttggaca atttctgaca ttttgacctc 600
aaatcaggta ggactacccg ctgaacttaa gcatatcaaa agccggaggg aa 652
Claims (3)
1. The new strain of the tricholoma mongolicum heim, the prairie white mushroom No. 2 strain, is characterized in that the tricholoma mongolicum heim is classified and named as Lepista sordida, and the preservation number of the tricholoma mongolicum heim is CGMCC No. 15084.
2. The method for cultivating a prairie white mushroom No. 2 strain according to claim 1, comprising the steps of:
(a) stacking and fermenting plant waste and livestock manure;
the composting fermentation is as follows: uniformly mixing plant waste and livestock manure according to the mass ratio of 4: 6-5: 5 to form a fermentation pile, and fermenting in a clean and ventilated place;
the plant waste is selected from one or more of grass seeds, caragana microphylla branches and leaves, rice straws, corn stalks and wheat straws;
(b) uniformly spreading the prairie white mushroom No. 2 strain of claim 1 on the fermented material, spreading a layer of the fermented material on the fermented material, then spreading a layer of cultivated species, spreading each layer of the fermented material with the thickness of 10-15 cm, mixing and flattening the surface layer strain and the culture material, finishing into turtle back type fruiting rows with the width of 1.3-1.5 m at the bottom, the width of 1.2-1.3 m at the top and the height of 20-24 cm, and culturing mycelia for 50-60 days;
after the mycelium is full of the materials, covering garden soil or turfy soil with the thickness of 4-6 cm on the surface, spraying water to preserve moisture when the surface soil is white before fruiting, and controlling the temperature to be 21-23 ℃; and covering soil for 40-45 days, allowing grassland white mushroom No. 2 primordium to appear, keeping the soil moist, and harvesting after 9-12 days.
3. The cultivation method according to claim 2, wherein the first crop of mushrooms is harvested, the surface of the first crop of mushrooms is leveled and lightly pressed for 2 days, fruiting management is continued after the growth of mycelia is recovered, and second crop of mushroom primordia appear 15-18 days after the first crop of mushrooms are harvested, and 3 crops of mushrooms can be harvested.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711421978.5A CN108048335B (en) | 2017-12-25 | 2017-12-25 | Noval strain grassland white mushroom No. 2 of Mongolian tricholoma mongolicum and breeding method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711421978.5A CN108048335B (en) | 2017-12-25 | 2017-12-25 | Noval strain grassland white mushroom No. 2 of Mongolian tricholoma mongolicum and breeding method thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN108048335A CN108048335A (en) | 2018-05-18 |
| CN108048335B true CN108048335B (en) | 2021-02-05 |
Family
ID=62131711
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201711421978.5A Active CN108048335B (en) | 2017-12-25 | 2017-12-25 | Noval strain grassland white mushroom No. 2 of Mongolian tricholoma mongolicum and breeding method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108048335B (en) |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108823107A (en) * | 2018-07-13 | 2018-11-16 | 迁西县林中宝生物科技有限公司 | A kind of white chestnut mushroom kind and selection using ARTP induced-mutation technique breeding |
| CN109706085A (en) * | 2018-12-25 | 2019-05-03 | 内蒙古自治区农牧业科学院 | No. 7 bacterial strains of one plant of Tricholoma mongolicum prairie Tricholomataceae and its acclimation method |
| CN111742778B (en) * | 2019-03-29 | 2022-04-12 | 中国科学院微生物研究所 | Lepista sordida strain and method for cultivating fruiting bodies by using liquid strain of Lepista sordida strain |
| CN110771426B (en) * | 2019-10-21 | 2021-08-27 | 郭红伟 | Stable culture method of phellinus igniarius strains |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1545841A (en) * | 2003-12-10 | 2004-11-17 | 方在吉 | Method for domesticating prairie Tricholomataceae and method for cultivating the same |
| CN102860218A (en) * | 2012-10-03 | 2013-01-09 | 徐国元 | Field cultivation method for grassland white mushrooms |
| CN103242091A (en) * | 2013-05-11 | 2013-08-14 | 徐国元 | Formula of prairie white mushroom nutrition fertilizer and preparation method thereof |
| CN103708968A (en) * | 2013-12-31 | 2014-04-09 | 贵州省生物研究所 | Lepista sordida first-class strain solid culture medium and strain rapid cultivating method |
| CN103947454A (en) * | 2014-05-08 | 2014-07-30 | 鲁东大学 | Artificial culturing method of lepista sordida mycelium and culturing medium thereof |
| CN107396751A (en) * | 2017-08-01 | 2017-11-28 | 内蒙古自治区农牧业科学院 | The black mushroom artificial cultivation method in grassland |
-
2017
- 2017-12-25 CN CN201711421978.5A patent/CN108048335B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1545841A (en) * | 2003-12-10 | 2004-11-17 | 方在吉 | Method for domesticating prairie Tricholomataceae and method for cultivating the same |
| CN102860218A (en) * | 2012-10-03 | 2013-01-09 | 徐国元 | Field cultivation method for grassland white mushrooms |
| CN103242091A (en) * | 2013-05-11 | 2013-08-14 | 徐国元 | Formula of prairie white mushroom nutrition fertilizer and preparation method thereof |
| CN103708968A (en) * | 2013-12-31 | 2014-04-09 | 贵州省生物研究所 | Lepista sordida first-class strain solid culture medium and strain rapid cultivating method |
| CN103947454A (en) * | 2014-05-08 | 2014-07-30 | 鲁东大学 | Artificial culturing method of lepista sordida mycelium and culturing medium thereof |
| CN107396751A (en) * | 2017-08-01 | 2017-11-28 | 内蒙古自治区农牧业科学院 | The black mushroom artificial cultivation method in grassland |
Non-Patent Citations (2)
| Title |
|---|
| Successful cultivation of a valuable wild strain of Lepista sordida from Thailand;Benjarong Thongbai et al.;《Mycological Progress》;20170106;第16卷(第04期);全文 * |
| 一株野生花脸香蘑的生物学特性及其栽培;周会明等;《食用菌学报》;20170131;第24卷(第01期);全文 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN108048335A (en) | 2018-05-18 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106350470B (en) | The production method and its application of bacillus amyloliquefaciens and microbial inoculum and the microbial inoculum | |
| CN108048335B (en) | Noval strain grassland white mushroom No. 2 of Mongolian tricholoma mongolicum and breeding method thereof | |
| CN107396751B (en) | Artificial cultivation method for grassland black mushroom | |
| CN106358751A (en) | Method for cultivating morchella | |
| CN106187515B (en) | Utilize the hickory chick nutrient bag and preparation method thereof of edible fungi residue production | |
| CN109456105A (en) | A kind of microbial bacterial agent method of preparation and use for alleviating solanaceous crops continuous cropping obstacle | |
| CN103382139A (en) | Agaricus bisporus (Lange) Sing culture medium | |
| CN104920068A (en) | Dictyophora rubrovalvata cultivation method | |
| CN106747820A (en) | A kind of method of culture medium of edible fungus, the preparation method of compost and culture edible mushroom | |
| CN108174744A (en) | Wild Lepista mucla (Bull.:Fr.) Cooke domesticating cultivation method | |
| CN103910581B (en) | A kind of multidimensional bisporous mushroom cultivation base stock and preparation method thereof | |
| CN107258328A (en) | A kind of cultural method of hickory chick | |
| CN107646616A (en) | A kind of preparation method of floriculture substrate | |
| CN106034742A (en) | Method for producing big Clitocybe by using mulberry stems, sugarcane bagasse and silkworm excrement | |
| CN107652093A (en) | A kind of preparation method of masson pine seedling medium | |
| CN105493894A (en) | Cultivation method for implanting mulberry trees with straw mushroom | |
| CN112715272A (en) | Under-forest out-of-season cultivation method for stropharia rugoso-annulata | |
| CN1490280A (en) | Microorganic fertilizer against various bacteria and its preparation and use | |
| CN107950288A (en) | A kind of planting technique of straw mushroom | |
| CN108293586A (en) | A kind of cultural method of agaricus bisporus | |
| CN107964513B (en) | Noval strain grassland white mushroom No.1 of Mongolian tricholoma mongolicum and breeding method thereof | |
| CN107950315A (en) | A kind of set cultivating method of capsicum and tomato | |
| CN1545845A (en) | Intercropping cultivation method for maize and pleurotu ostreatus | |
| CN103598000A (en) | Ecological and efficient bamboo fungus cultivation method | |
| CN106688622A (en) | Culture medium formula of artificially cultivated armillaria mellea and cultivation method |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |