CN108048335A - Tricholoma mongolicum new strains prairie Tricholomataceae 2 and its selection - Google Patents

Abstract

The invention belongs to the field of domestication methods of new strains of edible fungi, and specifically discloses a new strain of Tricholoma mongolicum, Grassland White Mushroom No. 2 and a breeding method thereof. The preservation number of the No. 2 strain of Tricholoma mongolicum strain of the present invention is CGMCC No. 15084, which has the characteristics of strong stress resistance, stable genetic performance, high quality and delicious taste, and provides people with a new high-quality edible Bacteria. The domestication method provided by the present invention not only protects the rare species of the endangered Pleurotus chinensis, but also realizes the resource utilization of agricultural and animal husbandry waste through the cultivation of the new strain No. It has created a new economic growth point for farmers and herdsmen, and is an important part of the development of garden economy in pastoral areas and the circular economy of grass and livestock. For the use of waste in agricultural and pastoral areas, idle cattle and sheep pens and other facilities to cultivate Grassland White Mushroom No. 2 for entrepreneurship and employment, Get rich.

Description

A New Strain of Tricholoma mongolica and Its Breeding Method

technical field

The invention belongs to the field of domestication methods of new strains of edible fungi, and in particular relates to a new strain of Tricholoma mongolicum, Grassland White Mushroom No. 2 and its domestication method.

Background technique

Lepista sordida belongs to the order Agaricales, the family Tricholomataceae, and the genus Lepista. The flower face mushroom is also called the white flower face, and it is mainly distributed on the grasslands of Xilin Gol and Horqin in Inner Mongolia. The protein content of the flower face mushroom is high, and the amino acid types are relatively complete, especially rich in calcium, iron, carotene, niacin and trace elements such as copper, zinc, fluorine, iodine, etc. The effect of the liver and the five internal organs, regular consumption is beneficial to the treatment of anemia, metrorrhagia, protracted illness, physical weakness, fatigue, forgetfulness and other diseases. It is a valuable edible fungus resource. At present, grassland degradation is becoming more and more serious, coupled with people's over-eating of food, resulting in worsening natural living conditions.

Therefore, there is an urgent need to provide a new strain of Tricholoma mongolicum that is suitable for artificial cultivation, Grassland White Mushroom No. 2. While retaining the original flavor of the wild Amanita chinensis, protect the endangered Grassland White Mushroom No. 2, a rare species that can be cultivated. The new strain of Grassland White Mushroom No. 2 realizes the resource utilization of agricultural and animal husbandry waste,

Contents of the invention

In order to solve the problems existing in the prior art, the object of the present invention is to provide a new strain of Tricholoma mongolica No. 2 and its breeding method.

In order to realize the object of the invention, the technical scheme of the present invention is as follows:

In the first aspect, the present invention provides No. 2 Grassland White Mushroom, identified as Lepistasordida, which is preserved in the General Microbiology Center of China Microbiological Culture Collection Management Committee (abbreviated as CGMCC, address: West Beichen, Chaoyang District, Beijing) No. 3, No. 1 Road, Institute of Microbiology, Chinese Academy of Sciences, postal code: 100101), the preservation date is December 7, 2017, and the preservation number is CGMCC No.15084.

The prairie white mushroom No. 2 has a white fruiting body cap, and the edge of the flat hemispherical cap is rolled inward at the early stage, and the water-stained edge of the cap is in the shape of a bamboo hat at the mature stage. The diameter of the mature fruiting body cap is 3.5 to 8.7 cm , the flesh is white and thin; the gills are white, straight or curved, slightly thinner, and unequal in length; the stipe is medium-thick, white, 3.5-7.6cm long, 0.2-2.0cm thick, solid inside, often curved near the base (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 image of the amplified sequence is shown in FIG. 3 .

The Prairie White Mushroom No. 2 provided by the present invention is obtained through domestication of the wild variegated mushroom, which retains the original flavor of the wild variegated mushroom, and can be cultivated artificially.

In a second aspect, the present invention provides a method for acclimating the No. 2 strain of White Mushroom Prairie, comprising the steps of:

1) Production of the mother seed: use the fruiting body of wild Amanita chinensis as the material to cultivate the mother seed at 21°C to 24°C until the culture medium is covered with mycelium;

2) Propagation: transfer the mother species obtained in step 1) to the culture medium, and cultivate it at 21°C to 24°C until it is covered with hyphae;

3) Production of original seed: transfer the mother seed after step 2) into a vessel containing the original seed culture material, and cultivate it at a constant temperature of 20°C to 23°C for 33 to 38 days until the mycelia cover the vessel to complete the original seed. kind of production;

4) Production of cultivars: put the original plants produced in step 3) into a vessel containing cultivar compost, and cultivate them at a constant temperature of 20°C to 23°C for 28 to 33 days until the mycelium is overgrown with the vessel to complete the cultivar make.

Wherein, described step 1) makes bacterial classification with tissue separation method or basidiospore collection method:

The tissue separation method is as follows: under aseptic conditions, cut the fresh wild fruiting body of Amanita variegata, select the bacterial flesh tissue at the junction of the cap and stipe, transfer it into a container with a culture medium, and heat it at 21°C to 24°C. Cultivate in a constant temperature incubator for 24 to 27 days;

The method for collecting basidiospores is as follows: under aseptic conditions, use an iron wire to pass through the fruiting bodies of fresh wild Amanita chinensis, make its gills hang downwards in the container to collect basidiospores, pick a little basidiospores into the test tube, add Dilute with sterile water to 100 μL containing 40-50 basidiospores to obtain a basidiospore suspension; absorb the basidiospore suspension and add it dropwise on the medium, and evenly spread it, and cultivate it in a constant temperature incubator at 21°C-24°C for 24-24°C. 27 days.

Wherein, the medium described in step 1) is PDA improved medium;

The medium described in step 2) is PDA improved medium or PDA improved liquid medium; when adopting PDA improved medium, cultivate 24～27 days on PDA improved medium; when adopting PDA improved liquid medium, Cultivate in PDA modified liquid medium at 120-140r/min for 13-16 days;

The formula of the PDA improved medium is: carbon source 15g/L～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/L, agar 10g/L;

The formula of the PDA improved liquid medium is: carbon source 15g/L～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/L, agar 0.6g/L;

The carbon source is one or more of potatoes, sweet potatoes, bran, whole wheat flour, and bean cake flour.

Wherein, the formula of the original seed compost in step 3) is: whole wheat grains or grains, potassium dihydrogen phosphate and magnesium sulfate;

The quality of the potassium dihydrogen phosphate is 0.2% of the whole grain or grain quality;

The quality of the magnesium sulfate is 0.1% of whole grain or grain quality;

The preparation method of the original seed compost is as follows: soak whole wheat grains or grains with the ratio of 1:1.2 to 1:1.5 of water quality for 8 to 12 hours, and add potassium dihydrogen phosphate and magnesium sulfate while soaking, so that It is fully dissolved, soaked and boiled until there is no hard core, controlled to remove water, adjusted pH to 7.5-8 with lime, and sterilized.

Wherein, the cultivar compost in step 4) includes 20-30 parts of wheat grains and 70-80 parts of caragana powder in parts by mass;

The preparation method of the cultivation material for cultivars is as follows: soak the wheat grains at a ratio of 1:1.3 to 1:2 for 8 to 12 hours, and add 0.2% potassium dihydrogen phosphate and 0.1 % magnesium sulfate, to make it fully dissolve, after soaking, boil until there is no hard core but not cracked, control the water; soak the caragana powder with water at a ratio of 1:1.2, and then mix it with wheat grains evenly, Adjust the pH to 7.5-8 with lime and sterilize.

The Caragana powder is Caragana branch of the aerial part of Caragana, which is crushed into 0.5-1.0 cm particles. Caragana powder is high in protein content, rich in mineral elements, vitamins, etc., and rich in nutrition.

In a third aspect, the present invention also provides a method for artificially cultivating Grassland White Mushroom No. 2, comprising the following steps:

(a) Fermentation of plant waste and livestock manure;

(b) Evenly spread the prairie white mushroom No. 2 provided by the present invention or other cultivars domesticated through the domestication method described in the present invention on the fermented material, spread a layer of fermented material on it, and then sprinkle One layer of cultivated species, the fermented material is spread on each layer with a thickness of 10cm~15cm, the surface layer of bacteria and cultivated species culture materials are mixed and flattened, and the bottom width is 1.3m~1.5m, the upper width is 1.2m~1.3m, and the height is 20cm. ～24cm long turtle-back-shaped fruiting rows with unlimited length, the mycelium is cultured for 50～60 days.

Preferably, the compost fermentation is as follows: plant waste and livestock manure are uniformly mixed in a mass ratio of 4:6 to 5:5 to form a fermentation pile, and fermented in a clean and ventilated place;

The plant waste is selected from one or more of grass knots, caragana powder branches and leaves, rice straw, corn straw, and wheat straw.

The fermentation of the pile is as follows: a fermentation pile with a bottom width of 1.2m to 1.4m, an upper width of 1.1m to 1.3m, and a height of 1.1m to 1.3m is built, and a wooden stick with a diameter of 6cm to 8cm is used to punch holes in the pile until At the bottom of the material, the distance between the holes is 40cm to 60cm. When the stack temperature rises to 65°C, keep it for 24 hours, perform the first turn over, continue to perforate and ferment, and keep the stack temperature at 65°C and keep it for 24 hours for the second turn. After 2 days, raise the temperature again at 65°C and keep it for 12 hours, then turn the pile for the third time, and adjust the water content at about 60% to 65% to be used for sowing.

Further, after the mycelium is covered with the material, cover the surface with 4cm-6cm thick pastoral soil or peat soil, spray water to keep the surface soil white before fruiting, and control the temperature at 21°C-23°C; After ~45 days, the No. 2 primordium of Grassland White Mushroom appears, keep the soil moist, and it can be harvested after 9 to 12 days.

Further, after harvesting the first crop of mushrooms, the surface was flattened and lightly pressed and dried for 2 days, so that the mycelium resumed growth and then the fruiting management continued. The second crop of mushrooms appeared on the 15th to 18th day after the first crop of mushrooms was harvested. The original base, a total of 3 crops of mushrooms can be harvested.

The beneficial effects of the present invention are:

The present invention provides a new strain of wild fenugreek and its domestication method and cultivation method. The domestication method not only protects the rare and rare wild species of Pleurotus chinensis from extinction, maintains the symbiotic ecosystem of grass and grassland mushrooms, but also provides people with edible mushroom products with excellent taste.

The new grassland white mushroom No. 2 strain provided by the invention has strong stress resistance, stable genetic performance, high quality and delicious taste, and provides people with a new high-quality edible fungus product. The Grassland White Mushroom No. 2 obtained by the present invention retains the unique flavor and nutritional components of the wild Pleurotus chinensis, and the product produced in cattle and sheep pens in pastoral areas in summer has a fresher taste and is deeply loved by people.

The present invention not only protects the rare species of Prairie White Mushroom No. 2, which is on the verge of extinction, but also realizes the resource utilization of agricultural and animal husbandry waste by cultivating the new strain of Prairie White Mushroom No. 2, turning waste into treasure, and expanding new opportunities for farmers and herdsmen. The point of economic growth is an important part of the development of courtyard economy and circular economy of grassland and livestock in pastoral areas. The use of waste from agricultural and pastoral areas, idle cattle and sheep pens and other facilities to cultivate Grassland White Mushroom No. 2 is used to start a business and get a job, and make a fortune. Realize the comprehensive utilization of resources, and promote the sustainable development of the emerging "grass, livestock, fungus" industry ecological circular economy.

Description of drawings

Fig. 1 is the wild strain of Amanita chinensis.

Fig. 2 is the No. 2 bacterial strain of Prairie White Mushroom according to the present invention.

Fig. 3 is an amplified electrophoresis image of the 5.8S ribosome-encoded gene sequence of No. 2 grassland mushroom of the present invention.

Detailed ways

Preferred embodiments of the present invention will be described in detail below in conjunction with examples. It should be understood that the following examples are given for the purpose of illustration only, and are not intended to limit the scope of the present invention. Those skilled in the art can make various modifications and substitutions to the present invention without departing from the purpose and spirit of the present invention.

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

The materials and reagents used in the following examples can be obtained from commercial sources unless otherwise specified.

Example 1

The PDA modified medium used in this example is: sterilized at a temperature of 121°C to 125°C (pressure of 1.1kg/cm ² to 1.5kg/cm ² ) for 30 minutes, containing 200g/L of sweet potato, 10g/L of sucrose, and 10g of glucose /L, yeast powder 1.2g/L, peptone 1.2g/L, magnesium sulfate 0.5g/L, dipotassium hydrogen phosphate 1g/L, agar 10g/L.

1) Production of mother species: Collect strong and wild Amanita variegata to make mother species, cut open the fruiting body of fresh wild Amanita variegata under aseptic conditions, select the fungus meat tissue at the junction of the cap and stipe, and transfer it into a PDA-improved Put it into a constant temperature incubator in a medium container and cultivate it at a temperature of 21°C to 24°C for 24 to 27 days until the mycelium is overgrown.

2) Propagation: the mother species is transferred to the improved PDA medium, and cultivated at 21°C-24°C for 24-27 days until the hyphae are covered.

3) Production of original seeds: Transplant the obtained multiplied mother species into a container containing culture material, place it at a constant temperature of 20°C to 23°C and cultivate it for 33 to 38 days. .

4) Production of cultivars: Put the obtained original plants into a container filled with compost, place them under a constant temperature condition of 20°C to 23°C and cultivate them for 28 to 33 days, and the production of cultivars is completed when the mycelia cover the container.

The preparation method of the original seed compost is as follows: soak whole wheat grains or grains with the ratio of 1:1.5 of water quality for 10 hours, add 0.2% potassium dihydrogen phosphate and 0.1% magnesium sulfate to fully dissolve them while soaking, After soaking, boil until there is no hard core but no cracking, remove the water, adjust the pH to 7.5 with 2% lime, put it into a container, and pass through the temperature of 121°C to 125°C (pressure of 1.1kg/cm ² to 1.5kg/cm2 ² ) Sterilize for 2.5 hours, and inoculate under aseptic conditions after cooling.

The cultivar compost includes 20 parts by mass of wheat grains and 80 parts of caragana powder; the preparation method of the cultivar compost is: the wheat grains are mixed with a feed water quality of 1:1.3 to 1:2 Proportional soaking for 8 to 12 hours, while soaking, add 0.2% potassium dihydrogen phosphate and 0.1% magnesium sulfate relative to the mass of the wheat kernels to make them fully dissolve, boil until there is no hard core but not cracked after soaking, and then remove the water; Soak caragana powder with water at a ratio of 1:1.2, mix it with wheat grains evenly, adjust the pH to 7.5-8, and sterilize.

Example 2

The PDA modified medium used in this example is: sterilized at a temperature of 121°C to 125°C (pressure of 1.1kg/cm ² to 1.5kg/cm ² ) for 30 minutes, containing bran 100/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/L, agar 0.6g/L.

1) Preparation of mother species: collect robust and wild Amanita chinensis to make mother species, and under aseptic conditions, use iron wire to pass through the fruiting bodies of fresh wild Amanita variegata, so that the gills hang downwards in jars to collect basidiospores , pick a few basidiospores into a test tube, add sterile water to dilute to 100 μL containing 40 to 50 basidiospores, and obtain a basidiospore suspension. Take the basidiospore suspension and add it dropwise to the modified medium with PDA, spread it evenly, and cultivate it at a temperature of 21°C to 24°C for 24 to 27 days until the mycelium is overgrown.

2) Propagation: transfer the mother species to the improved PDA liquid medium, and culture at 21°C to 24°C at a speed of 130r/min for 12 to 16 days. The cultured balls evenly cover the vessel to complete the production of liquid strains.

3) Production of original seeds: Transplant the obtained multiplied mother species into a container containing culture material, place it at a constant temperature of 20°C to 23°C and cultivate it for 33 to 38 days. .

4) Production of cultivars: Put the obtained original plants into a container filled with compost, place them under a constant temperature condition of 20°C to 23°C and cultivate them for 28 to 33 days, and the production of cultivars is completed when the mycelia cover the container.

The preparation method of the original seed compost is as follows: soak the whole wheat grains or grains at a ratio of 1:1.3 to water quality for 12 hours, add 0.2% potassium dihydrogen phosphate and 0.1% magnesium sulfate to fully dissolve them while soaking, After soaking, boil until there is no hard core but no cracking, remove the water, adjust the pH to 7.5 with 2% lime, put it into a container, and pass through the temperature of 121°C to 125°C (pressure of 1.1kg/cm ² to 1.5kg/cm2 ² ) Sterilize for 2.5 hours, and inoculate under aseptic conditions after cooling.

The cultivar compost includes 30 parts by mass of wheat grains and 70 parts of caragana powder; the preparation method of the cultivar compost is: soak the wheat grains at a ratio of 1:1.3 to the quality of water for 8- 12 hours, while soaking, add 0.2% potassium dihydrogen phosphate and 0.1% magnesium sulfate relative to the mass of wheat grains to make them fully dissolve, boil until there is no hard core but not cracked after soaking, and control the water; Soak caragana powder in water at a ratio of 1:1.2, mix it with wheat grains evenly, adjust the pH to 7.5, and sterilize.

Example 3

The PDA modified medium used in this example is: the PDA modified medium is: sterilized at a temperature of 121°C to 125°C (pressure of 1.1kg/cm ² to 1.5kg/cm ² ) for 30 minutes, containing 200g/L of potatoes and sucrose 10g/L, glucose 10g/L, yeast powder 1.2g/L, peptone 1.2g/L, magnesium sulfate 0.5g/L, dipotassium hydrogen phosphate 1g/L, agar 10g/L.

1) Production of mother species: Collect strong and wild Amanita variegata to make mother species, cut open the fruiting body of fresh wild Amanita variegata under aseptic conditions, select the fungus meat tissue at the junction of the cap and stipe, and transfer it into a PDA-improved Put it into a constant temperature incubator in a medium container and cultivate it at a temperature of 21°C to 24°C for 24 to 27 days until the mycelium is overgrown.

2) Propagation: the mother species is transferred to the improved PDA medium, and cultivated at 21°C-24°C for 24-27 days until the hyphae are covered.

3) Production of original seed: Transplant the obtained multiplied mother seed into a container containing wheat grain compost, place it at a constant temperature of 20°C to 23°C and cultivate it for 33 to 38 days. kind of production.

4) Production of cultivars: Put the obtained original plants into a container filled with compost, place them under a constant temperature condition of 20°C to 23°C and cultivate them for 28 to 33 days, and the production of cultivars is completed when the mycelia cover the container.

The preparation method of the original seed compost is as follows: soak whole wheat grains or grains with the ratio of 1:1.3 to water quality for 11 hours, add 0.2% potassium dihydrogen phosphate and 0.1% magnesium sulfate to fully dissolve them while soaking, After soaking, boil until there is no hard core but no cracking, remove the water, adjust the pH to 7.5 with 2% lime, put it into a container, and pass through the temperature of 121°C to 125°C (pressure of 1.1kg/cm ² to 1.5kg/cm2 ² ) Sterilize for 2.5 hours, and inoculate under aseptic conditions after cooling.

The cultivar compost comprises 25 parts by mass of wheat grains and 75 parts of caragana powder; the preparation method of the cultivar compost is: soaking the wheat grains for 11 hours at a ratio of 1:1.3 to the quality of feed water, While soaking, add 0.2% potassium dihydrogen phosphate and 0.1% magnesium sulfate relative to the mass of the wheat kernels to make them fully dissolve, boil until there is no hard core but not cracked after soaking, and control the water; the quality of the material water is 1 : Soak caragana powder with water at a ratio of 1.2, mix it with wheat grains evenly, adjust the pH to 8, and sterilize.

Example 4

The PDA modified medium used in this example is: sterilized at a temperature of 121°C to 125°C (pressure of 1.1kg/cm ² to 1.5kg/cm ² ) for 30 minutes, containing 20g/L of bean cake powder, 10g/L of sucrose, and glucose 10g/L, yeast powder 1.2g/L, peptone 1.2g/L, magnesium sulfate 0.5g/L, dipotassium hydrogen phosphate 1g/L, agar 0.6g/L.

1) Preparation of mother species: collect robust and wild Amanita chinensis to make mother species, and under aseptic conditions, use iron wire to pass through the fruiting bodies of fresh wild Amanita variegata, so that the gills hang downwards in jars to collect basidiospores , pick a few basidiospores into a test tube, add sterile water to dilute to 100 μL containing 40 to 50 basidiospores, and obtain a basidiospore suspension. Take the basidiospore suspension and add it dropwise to the modified medium with PDA, spread it evenly, and cultivate it at a temperature of 21°C to 24°C for 24 to 27 days until the mycelium is overgrown.

2) Propagation: transfer the mother species to the improved PDA liquid medium, and culture at 21-24°C at a speed of 130r/min for 13-16 days to culture the balls to evenly cover the vessel to complete the production of liquid strains;

3) Production of original seeds: Transplant the obtained multiplied female species into a vessel containing wheat grain compost, place it at a constant temperature of 20° C. to 23° C., and cultivate it for 33 to 38 days until the mycelia cover the vessel.

4) Production of cultivars: Put the obtained original plants into a container filled with compost, place them under a constant temperature condition of 20° C. to 23° C., and cultivate them for 28 to 33 days until the mycelia cover the container.

Step 3) and 4) culture material is the same as embodiment 1.

5) Fermentation: pre-crushed and screened sheep manure and fresh non-mildew caragana powder branches and leaves are fully pre-wetted with water at a ratio of 4:6, and the sheep manure and grass are alternately and evenly spread, and the bottom width is 1.3m, and the top For a pile with a width of 1.2m, a height of 1.2m, and an unlimited length, use a wooden stick with a diameter of 6cm to 8cm to make holes to the bottom of the material, with a distance of 50cm. When the temperature of the pile rises to 65°C, keep it for 24 hours and perform the first turning Pile, continue to perforate and ferment, raise the temperature of the pile to 65°C and keep it for 24 hours to turn over the pile for the second time, raise the temperature again to 65°C after 2 days, turn over the pile for the third time after 12 hours, adjust the water content at about 60% to 65% Ready to use for sowing.

6) Domestication and cultivation: spread the fermented compost on the cleaned ground of sheep pens in pastoral areas, spread the cultivars evenly on the compost, spread a layer of fermented compost on top, and then sprinkle a layer of cultivars, Mix and flatten the surface strains and compost, organize them into turtle-shaped fruiting rows with a bottom width of 1.4m, an upper width of 1.3m, and a height of about 23cm. After the mycelium is covered with compost, cover the surface with 5cm of moist soil, spray water to keep the surface soil white before fruiting, and keep the temperature at 21°C to 23°C. 40 to 45 days after covering with soil, No. 2 primordium of Prairie White Mushroom will appear, and it can be harvested after 9 to 12 days, and 3 crops of mushrooms can be harvested after scientific fruiting management.

Example 5

The PDA modified medium used in this example is: sterilized at a temperature of 121°C to 125°C (pressure of 1.1kg/cm2 to 1.5kg/cm2) for 30 minutes, containing 15g/L of whole grain powder, 10g/L of sucrose, and glucose 10g/L, yeast powder 1.2g/L, peptone 1.2g/L, magnesium sulfate 0.5g/L, dipotassium hydrogen phosphate 1g/L, agar 10g/L.

1) Production of mother species: Collect strong and wild Amanita variegata to make mother species, cut open the fruiting body of fresh wild Amanita variegata under aseptic conditions, select the fungus meat tissue at the junction of the cap and stipe, and transfer it into a PDA-improved Put it into a constant temperature incubator in a medium container and cultivate it at a temperature of 21°C to 24°C for 24 to 27 days until the mycelium is overgrown.

2) Propagation: the mother species is transferred to the improved PDA medium, and cultivated at 21°C-24°C for 24-27 days until the hyphae are covered.

3) Production of original seed: Transplant the obtained multiplied mother seed into a container containing wheat grain compost, place it at a constant temperature of 20°C to 23°C and cultivate it for 33 to 38 days. kind of production.

4) Production of cultivars: Put the obtained original plants into a container filled with compost, place them under a constant temperature condition of 20°C to 23°C and cultivate them for 28 to 33 days, and the production of cultivars is completed when the mycelia cover the container.

The preparation method of the original seed compost is as follows: soak whole wheat grains or grains with the ratio of 1:1.3 to water quality for 11 hours, add 0.2% potassium dihydrogen phosphate and 0.1% magnesium sulfate to fully dissolve them while soaking, After soaking, boil until there is no hard core but no cracking, remove the water, adjust the pH to 7.5 with 2% lime, put it into a container, and pass through the temperature of 121°C to 125°C (pressure of 1.1kg/cm ² to 1.5kg/cm2 ² ) Sterilize for 2.5 hours, and inoculate under aseptic conditions after cooling.

The cultivar compost comprises 25 parts by mass of wheat grains and 75 parts of caragana powder; the preparation method of the cultivar compost is: soaking the wheat grains for 11 hours at a ratio of 1:1.3 to the quality of feed water, While soaking, add 0.2% potassium dihydrogen phosphate and 0.1% magnesium sulfate relative to the mass of the wheat kernels to make them fully dissolve, boil until there is no hard core but not cracked after soaking, and control the water; the quality of the material water is 1 : Soak caragana powder with water at a ratio of 1.2, mix it with wheat grains evenly, adjust the pH to 8, and sterilize.

5) Fermentation: The pre-crushed sheep manure and fresh non-mildew caragana powder branches and leaves are fully pre-wetted with water in a ratio of 1:1, and the sheep manure and grass are alternately and evenly spread, and the bottom width is 1.2m, and the top width is 1.1m. m, 1.1m in height, and unlimited in length, use a wooden stick with a diameter of 6cm to 8cm to make holes to the bottom of the material, the distance between the holes is 40cm, when the temperature of the pile rises to 65°C, keep it for 24 hours, and perform the first turning. Continue to perforate and ferment, raise the pile temperature to 65°C and keep it for 24 hours for the second turning, then raise the temperature again to 65°C after 2 days, turn the pile for the third time after 12 hours, and adjust the water content to about 60% to 65%. for sowing.

6) Domestication and cultivation: spread the fermented compost on the cleaned ground of sheep pens in pastoral areas, spread the cultivars evenly on the compost, spread a layer of fermented compost on top, and then sprinkle a layer of cultivars, Mix and flatten the surface strains and compost, arrange them into turtle-shaped fruiting rows with a bottom width of 1.3m, an upper width of 1.2m, and a height of about 23cm. After the mycelium is covered with compost, cover the surface with 5cm of moist soil, spray water to keep the surface soil white before fruiting, and keep the temperature at 21°C to 23°C. 40 to 45 days after covering with soil, No. 2 primordium of Prairie White Mushroom appears, which can be harvested after 9 to 12 days, and 3 crops of mushrooms can be harvested after scientific fruiting management.

The present invention successfully realizes the artificial domestication and cultivation of Prairie White Mushroom No. 2, which not only protects the rare species of the endangered Pleurotus chinensis, but also makes full use of wastes such as cow and sheep dung and grass residues in pastoral areas to cultivate Prairie White Mushroom. Mushroom No. 2 realizes the comprehensive utilization of resources, promotes the recycling and sustainable development of the emerging "grass, livestock, and fungus" industries, and can also increase the income of local herdsmen.

It should be understood that the technical solutions obtained by proportionally expanding or reducing the amount of reagents or raw materials used in the above examples are substantially the same as those of the above examples.

Although the present invention has been described in detail with general descriptions and specific embodiments above, it is obvious to those skilled in the art that some modifications or improvements can be made on the basis of the present invention. Therefore, the modifications or improvements made on the basis of not departing from the spirit of the present invention all belong to the protection scope of the present invention.

sequence listing

<110> Inner Mongolia Academy of Agriculture and Animal Husbandry Sciences Inner Mongolia University

<120> A New Strain of Tricholoma mongolica and Its Breeding Method

<141> 2017-12-19

<160> 1

<170> SIPOSequenceListing 1.0

<210> 1

<211> 652

<212>DNA

<213> Lepista sordida

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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 (10)

1. A new strain of Tricholoma mongolica strain No. 2 of Tricholoma mongolica, characterized in that its preservation number is CGMCC No.15084. 2. a method for selecting and breeding No. 2 bacterial classification of prairie white mushroom, is characterized in that, comprises the steps: 1) Production of the mother seed: use the fruiting body of wild Amanita chinensis as the material to cultivate the mother seed at 21°C to 24°C until the culture medium is covered with mycelium; 2) Propagation: transfer the mother species obtained in step 1) to the culture medium, and cultivate it at 21°C to 24°C until it is covered with hyphae; 3) Production of original seed: transfer the mother seed after step 2) into a vessel containing the original seed culture material, and cultivate it at a constant temperature of 20°C to 23°C for 33 to 38 days until the mycelia cover the vessel to complete the original seed. kind of production; 4) Production of cultivars: put the original plants produced in step 3) into a vessel containing cultivar compost, and cultivate them at a constant temperature of 20°C to 23°C for 28 to 33 days until the mycelium is overgrown with the vessel to complete the cultivar make. 3. method according to claim 2, is characterized in that, described step 1) makes bacterial classification with tissue separation method or basidiospore collection method: The tissue separation method is as follows: under aseptic conditions, cut the fresh wild fruiting body of Amanita variegata, select the bacterial flesh tissue at the junction of the cap and stipe, transfer it into a container with a culture medium, and heat it at 21°C to 24°C. Cultivate in a constant temperature incubator for 24 to 27 days; The method for collecting basidiospores is as follows: under aseptic conditions, use an iron wire to pass through the fruiting bodies of fresh wild Amanita chinensis, make its gills hang downwards in the container to collect basidiospores, pick a little basidiospores into the test tube, add Dilute with sterile water to 100 μL containing 40-50 basidiospores to obtain a basidiospore suspension; absorb the basidiospore suspension and add it dropwise on the medium, and evenly spread it, and cultivate it in a constant temperature incubator at 21°C-24°C for 24-24°C. 27 days. 4. according to the described method of claim 2 or 3, it is characterized in that, the described substratum in step 1) is PDA improved substratum; The medium described in step 2) is PDA improved medium or PDA improved liquid medium; when adopting PDA improved medium, cultivate 24～27 days on PDA improved medium; when adopting PDA improved liquid medium, Cultivate in PDA modified liquid medium at 120-140r/min for 13-16 days; The formula of the PDA improved medium is: carbon source 15g/L～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/L, agar 10g/L; The formula of the PDA improved liquid medium is: carbon source 15g/L～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/L, agar 0.6g/L; The carbon source is one or more of potatoes, sweet potatoes, bran, whole wheat flour, and bean cake flour. 5. The method according to claim 2, characterized in that, the formula of the original seed culture material in step 3) is: whole grains or grains, potassium dihydrogen phosphate and magnesium sulfate; The quality of the potassium dihydrogen phosphate is 0.2% of the whole grain or grain quality; The quality of the magnesium sulfate is 0.1% of whole grain or grain quality; The preparation method of the original seed compost is as follows: soak whole wheat grains or grains with the ratio of 1:1.2 to 1:1.5 of water quality for 8 to 12 hours, and add potassium dihydrogen phosphate and magnesium sulfate while soaking, so that It is fully dissolved, soaked and boiled until there is no hard core, drained of water, adjusted to pH 7.5-8, and sterilized. 6. The method according to claim 2, characterized in that, the cultivar compost in step 4) comprises 20 to 30 parts of wheat grains and 70 to 80 parts of caragana powder in parts by mass; The preparation method of the cultivation material for cultivars is as follows: soak the wheat grains at a ratio of 1:1.3 to 1:2 for 8 to 12 hours, and add 0.2% potassium dihydrogen phosphate and 0.1 % magnesium sulfate, to make it fully dissolve, after soaking, boil until there is no hard core but not cracked, control the water; soak the caragana powder with water at a ratio of 1:1.2, and then mix it with wheat grains evenly, Adjust the pH to 7.5-8 and sterilize. 7. A cultivation method of Prairie White Mushroom No. 2, comprising the steps of: (a) Fermentation of plant waste and livestock manure; (b) spreading No. 2 cultivars of the prairie white mushroom described in claim 1 evenly on the fermented material, laying a layer of fermented material on top, and then spreading a layer of cultivars, each layer of the fermented material Spread 10cm to 15cm thick, mix and flatten the surface layer of bacteria and culture materials, arrange it into a turtle-back-shaped fruiting row with a bottom width of 1.3m to 1.5m, an upper width of 1.2m to 1.3m, and a height of 20cm to 24cm. , Carry out mycelium culture for 50-60 days. 8. The method according to claim 7, characterized in that, the fermentation of the stockpile is as follows: plant waste and livestock manure are uniformly mixed in a mass ratio of 4:6 to 5:5 to form a fermentation pile, and fermented in a clean and ventilated place ; The plant waste is selected from one or more of grass knots, caragana powder branches and leaves, rice straw, corn straw, and wheat straw. 9. The method according to claim 7, characterized in that, after the mycelium is covered with materials, the surface is covered with pastoral soil or peat soil with a thickness of 4cm to 6cm, and the surface soil turns white before mushrooming, then spray water to keep it moist. The temperature is controlled at 21°C to 23°C; 40 to 45 days after covering with soil, No. 2 primordium of Prairie White Mushroom will appear, keep the soil moist, and it can be harvested after 9 to 12 days. 10. The method according to claim 9, characterized in that after the first crop of mushrooms is harvested, the surface is leveled and lightly pressed and dried for 2 days, so that the mycelium resumes growth and then the mushroom fruiting management is continued, and the first crop of mushrooms is harvested On the 15th to 18th day after mushrooming, two crops of mushroom primordia appeared, and a total of three crops of mushrooms could be harvested.