CN108235900A - A kind of Tricholoma mongolicum new strains prairie Tricholomataceae 3 and its acclimation method - Google Patents
A kind of Tricholoma mongolicum new strains prairie Tricholomataceae 3 and its acclimation method Download PDFInfo
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Abstract
The present invention provides a kind of Tricholoma mongolicum new strains prairie Tricholomataceae 3, it is tamed by wild green hired thug, the present invention also provides wild green hired thug strain making and acclimation methods, this method makes including strain, expand numerous, original seed, cultigen makes and domestication, not only protect endangered this rareness species of green hired thug, it can also realize that farming and animal husbandry changing waste into resources utilizes by cultivating No. 3 new strains of prairie Tricholomataceae, turn waste into wealth, peasants and herdsmen's new growth engines is expanded, it is the important ring for developing pastoral area courtyard economy and grass poultry circular economy, using farming and pastoral area waste, the facility cultivations prairie Tricholomataceaes 3 such as idle cattle and sheep circle are started an undertaking and are obtained employment, build up the family fortunes.It realizes the comprehensive utilization of resource and conversion increment, promotes emerging《Grass, poultry, bacterium》The sustainable development of industry ecological circular economy.
Description
Technical field
The invention belongs to strain domestication method field, specifically, being related to a kind of Tricholoma mongolicum new strains prairie Tricholomataceae 3
And its acclimation method.
Background technology
Green hired thug category Agaricales (Agaricales), Tricholomataceae (tricholomataceae), Lepista lentinus (Lepista).
It is mainly distributed on Siklingelei in the Inner Mongol, on Horqin Caoyuan.Green hired thug contain abundant protein, vitamin and potassium,
The several mineral materials such as phosphorus, selenium, iron and polysaccharide, triterpene isoreactivity ingredient.Green hired thug meat fertilizer matter is tender, Yu Xiang is pure, excellent taste, is
Treasure in bacterium, while there is medical care effect, antitumor, hypoglycemic, blood fat etc., be China's preciousness domestic fungus resource.Mesh
Presteppe degeneration getting worse, in addition people excessively search for food, Natural Survival condition is caused to deteriorate further.The present invention is directed to wild blueness
Hired thug establish strain isolation and its acclimation method, and No. 3 new strains realities of prairie Tricholomataceae are obtained by this method habituation of wild blueness hired thug
Existing artificial cultivation not only protects this rare wildlife species of green hired thug from extinction, maintains herbage and grassland mushroom symbiosis
Ecosystem, moreover it is possible to provide taste splendid edible fungus for people.
At present, there is not yet the report in relation to wild green hired thug acclimation method.
Invention content
The object of the present invention is to provide a kind of Tricholoma mongolicum new strains prairie Tricholomataceae 3 and its acclimation methods.
In order to realize the object of the invention, Tricholoma mongolicum new strains prairie Tricholomataceae 3 provided by the invention, by wild green hired thug
(Lepista sp.) is tamed, and has been preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, ground
Location Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, Institute of Microorganism, Academia Sinica, postcode 100101, deposit number
CGMCC No.15083, preservation date on December 7th, 2017.
Wild blueness hired thug category Agaricales (Agaricales), Tricholomataceae (tricholomataceae), Lepista lentinus
(Lepista).The form of wild blueness hired thug is shown in Fig. 2.
The form that prairie Tricholomataceae 3 is following (Fig. 1)：Fructification cap pure white, initial stage hemispherical to maturity period cap are put down
Side edge upwarps, 10~22cm of mature sporophore bacteria cover diameter.The thicker white of bacterial context, initial stage edge are involute；Lamella white, it is thick
It is close, curved life, Length discrepancy；Thick in stem, white, long 3.5~7cm, thick 0.8~3.0cm, interior reality, base portion is slightly larger, and handle is from bottom to top
Cyan gloss gradually deepens.
The 5.8S rRNA coding gene sequences of prairie Tricholomataceae 3 are shown in SEQ ID NO:1, it is carried out using it as target sequence
PCR amplification, electrophoresis result are shown in Fig. 3,
The present invention also provides a kind of method for preparing the prairie Tricholomataceae 3 by wild green hired thug domestication, including parent species system
Make, expand the step of numerous, original seed makes, cultigen makes and tames, it is specific as follows：
1) parent species make：At 22 DEG C~24 DEG C parent species are cultivated using wild green hired thug fructification as material, until on culture medium
Cover with mycelia；
2) expand numerous：Step 1) parent species are transferred on culture medium, are cultivated at 22 DEG C~24 DEG C, until covering with mycelia；
3) original seed makes：Expand numerous parent species to move into the vessel equipped with compost, be cultivated under 21 DEG C~23 DEG C constant temperatures
32~37 days, until mycelia, which covers with vessel, completes original seed making；
4) cultigen makes：Original seed is moved into the vessel equipped with compost, is cultivated under 21 DEG C~23 DEG C of constant temperature
28~32 days, until mycelia, which covers with vessel, completes cultigen making；
5) it tames：Agriculture and forestry organic waste material and excrement of animals are built into heap fermentation, sowing is completed after fermentation, carries out management of producing mushroom.
Preferably, step 1) makes parent species using tissue isolation or basidiospore collecting method.
The tissue isolation is：Fresh wild blueness hired thug strain fructification is aseptically cut, chooses cap bacterium
The meat bacteria organization of handle intersection, move into equipped with culture medium vessel in, in 22 DEG C~24 DEG C of constant incubator cultivate 25~
28 days.
The basidiospore collecting method is：The strain fructification of fresh wild blueness hired thug is acquired, aseptically makes its bacterium
Pleat, which is hung on downwards, collects basidiospore in container, sterile water is added to be diluted to 100 μ L containing 50~60 basidiospore, basidiospore is obtained and hangs
Liquid；Basidiospore suspension is drawn to be added dropwise on culture medium, and even spread, cultivate 28 in 22 DEG C~24 DEG C of constant incubator~
30 days.
Preferably, the culture medium of step 1) is PDA improved culture mediums.
Preferably, step 2) is cultivated 25~28 days in PDA improved culture mediums or in PDA improves fluid nutrient medium
It is cultivated 12~15 days under 120~140r/min rotating speeds.
In the present invention, PDA improved culture medium formulas are：Carbon source 15g/L~200g/L, sucrose 10g/L, glucose 10g/L,
Dusty yeast 1.8g/L, peptone 1.8g/L, magnesium sulfate 0.5g/L, dipotassium hydrogen phosphate 1g/L and agar 10g/L.
PDA improves Liquid Culture based formulas：Carbon source 15g/L~200g/L, sucrose 10g/L, glucose 10g/L, yeast
Powder 1.8g/L, peptone 1.8g/L, magnesium sulfate 0.5g/L, dipotassium hydrogen phosphate 1g/L and agar 0.6g/L.
The carbon source is at least one of potato, sweet potato, wheat bran, the full powder of wheat, beancake powder etc..
Preferably, the step 3) compost is：Quality of material 0.2% as material and is accounted for using whole wheat granular and/or grain
Potassium dihydrogen phosphate and 0.1% magnesium sulfate；Its production method is：By material to expect water quality 1:1.3~1：1.5 ratios are impregnated
9~12h, immersion adds in potassium dihydrogen phosphate simultaneously, magnesium sulfate makes it fully dissolve, and is boiled after immersion to no hard core, and water is removed in control,
(using lime) adjusts pH to 7.5~8, then sterilizes.
Preferably, the step 4) compost is：Wheat and caragana microphylla powder and the biphosphate for accounting for quality of material 0.2%
Potassium and 0.1% magnesium sulfate, the wherein weight ratio of wheat and caragana microphylla powder are 3:7；Its production method is：By wheat to expect water quality
1：1.5~1:2 ratios impregnate 9~12h, and immersion adds in potassium dihydrogen phosphate simultaneously, magnesium sulfate makes it fully dissolve, and is boiled after immersion
It to no hard core but does not split, water is removed in control；By caragana microphylla powder to expect water quality 1:1.2 ratios impregnate, then with the wheat after above-mentioned control water
It is uniformly mixed, (using lime) adjusts pH to 7.5~8, then sterilizes.
In the present invention, caragana microphylla powder is the caragana microphylla branch of caragana microphylla aerial part, and crushed after being dried is into the caragana microphylla of 0.5-1.0cm
Grain.Caragana microphylla amyloid proteins matter content is high, full of nutrition rich in mineral matter element, vitamin etc..
In the present invention, the agriculture and forestry organic waste material in careless knot, caragana microphylla branches and leaves, straw, maize straw, wheat straw etc. extremely
Few one kind.The excrement of animals comes from sheep manure, cow dung, horsehit etc..
Preferably, heap fermentation is built described in step 5) is specially：After agriculture and forestry organic waste material is crushed, discarded according to sheep manure and agricultural
Amount of substance is than 4：6 be uniformly mixed build up fermentation heap, at clean ventilation ferment.
Preferably, step 5) builds up the fermentation heap of bottom width 1.2m~1.4m, upper width 1.1m~1.3m, high 1.1m~1.3m,
Hole is punched to bottom, hole spacing 40cm~50cm is expected in material heap with the wooden stick of diameter 6cm~8cm, and heap temperature is protected when being raised to 65 DEG C
Hold 24 hours, carry out first time turning, continue punching fermentation, heap temperature be raised to 65 DEG C keep again 24 hours carry out second of turning, 2
Heat up again after it 65 DEG C when kept for 12 hours after third time turning, conditioning is 60%~65%, the object after gained fermentation
Material can be used to sow.
Preferably, step 5) domestication has been divulged information surrounding enviroment cleaning in greenhouse, heliogreenhouse, storage cellar for storing things, cattle and sheep circle etc.
Place carries out.Material after fermentation is uniformly layered on to the cultivation room floor cleaned up, the cultigen that step 4) is obtained is uniform
On the material of paving after fermentation, the material after one layer of fermentation is repaved above, then spread last layer cultigen, every layer of material 10cm~
15cm is thick, and surface layer strain and material puddle pressing, be organized into bottom width 1.3m~1.5m, upper width 1.2m~1.3m, high 20cm~
24cm, long unlimited curvature of the spinal column type fruiting row, carry out Mycelium culture 50~55 days.
Preferably, step 5) is covered the rural area soil or turfy soil of 4cm~6cm thickness on surface, is gone out after the long full with substance of mycelium
Mushroom front surface soil bleaches, moisturizing of spraying water, and temperature is controlled at 21 DEG C~23 DEG C；40~45 days after earthing, it is former to there is the mushroom after domestication
Base keeps ground moistening, can be harvested after 10~12 days.
Surface evening is gently pressed dry 2 days after first batch of mushroom of harvesting, continues management of producing mushroom after making mycelium restoration ecosystem,
There is regrowth hair mushroom former base within the 15th~17 day after first batch of mushroom of harvesting, three batches of mushrooms can be adopted altogether.
The present invention also provides food, drug, health products, cosmetics and the feeds by No. 3 preparations of prairie Tricholomataceae.
The present invention also provides No. 3 applications in artificial cultivation of prairie Tricholomataceae.
The present invention further provides the grassland mushroom No. 3 in food, drug, health products, cosmetics and feed is prepared
Application.
No. 3 realization artificial cultivations of prairie Tricholomataceae are tamed by the method for the present invention, No. 3 new strains resistance of prairie Tricholomataceae are strong
It is (can be kept at subzero 30 DEG C activity, antifungi and bacterium performance are strong), stable, high-quality, delicious, for people provide it is a kind of newly
High-quality edible mushroom product.The prairie Tricholomataceae 3 maintains the distinctive flavor of wild Tricholoma mongolicum blueness hired thug and nutritional ingredient, summer
The product taste that cattle and sheep circle in pastoral area is produced is fresher, very popular.
Compared with prior art, the present invention has the following advantages：
Wild green hired thug acclimation method provided by the invention, the technology mainly include parent species make, expand numerous, original seed make,
Cultigen makes and domestication, and the artificial cultivation to wild green hired thug can be realized using the technology.Tame obtained prairie Tricholomataceae 3
Maintain the original flavor of wild green hired thug；The green hired thug new strains prairie Tricholomataceae 3 that the present invention successfully tames out simultaneously, no
But protect endangered wild this rareness species of green hired thug, moreover it is possible to realize agriculture by cultivating No. 3 new strains of prairie Tricholomataceae
Animal husbandry changing waste into resources utilizes, and turns waste into wealth, has expanded peasants and herdsmen's new growth engines, is development pastoral area courtyard economy and grass
An important ring for circular economy is raiseeed, is created using facility cultivations prairie Tricholomataceaes 3 such as farming and pastoral area waste, idle cattle and sheep circles
Industry and employment, build up the family fortunes.It realizes the comprehensive utilization conversion increment of resource, promotes emerging grass -- poultry -- bacterium industry ecological cycle warp
The sustainable development of Ji.
Description of the drawings
Fig. 1 is the aspect graph of prairie Tricholomataceae 3 of the present invention.
Fig. 2 is the aspect graph of wild green hired thug.
Fig. 3 is to carry out PCR expansions by target sequence of the 5.8S rRNAs coding gene sequence of prairie Tricholomataceae 3 of the present invention
The electrophoresis result of increasing.
Specific embodiment
The following examples are used to illustrate the present invention, but are not intended to limit the scope of the present invention..Unless otherwise specified, embodiment
Used in the conventional means that are well known to those skilled in the art of technological means, raw materials used is commercial goods.
1 cultivating bacterial spawn kind of embodiment makes
1) parent species make：The healthy and strong wild green hired thug of acquisition make parent species, aseptically cut the blueness of fresh wild
Hired thug fructification chooses the meat bacteria organization of cap stem intersection, moves into the vessel equipped with PDA improved culture mediums, is put into perseverance
It cultivates, is cultivated 25~28 days at a temperature of 22 DEG C~24 DEG C, until mycelia covers with device in warm incubator.
2) expand numerous：Parent species are transferred in the PDA culture medium of improvement, and 25~28 days are cultivated at 22 DEG C~24 DEG C to covering with bacterium
Silk.
PDA improved culture mediums are：Contain sweet potato 200g/L, sucrose 10g/L, glucose 10g/L, dusty yeast 1.8g/L, egg
The culture medium of peptone 1.8g/L, magnesium sulfate 0.5g/L, dipotassium hydrogen phosphate 1g/L, agar 10g/L in vain.Sterilising conditions are：121 DEG C~
125 DEG C of (pressure 1.1kg/cm2~1.5kg/cm2) sterilize 30 minutes.
3) original seed makes：The obtained numerous parent species of expansion are implanted into the vessel equipped with compost, are placed on 21 DEG C~23 DEG C
Constant temperature under cultivate 32~37 days, mycelia cover with vessel complete original seed making.
4) cultigen makes：By in vessel of the obtained original seed implantation equipped with compost, it is placed on 21 DEG C~23 DEG C of perseverance
It is cultivated 28~32 days under the conditions of temperature, mycelia covers with vessel and completes cultigen making.
Original seed wheat culture medium production method：By the wheat of wash clean and water with quality 1：1.4 ratios impregnate 12h, impregnate
Add in the potassium dihydrogen phosphate of wheat quality 0.2% simultaneously, 0.1% magnesium sulfate makes it fully dissolve, boiled after immersion to without hard
It core but does not split, water is removed in control, adjusts pH to 7.5 with 2% lime, vessel is put into, through 121 DEG C~125 DEG C (pressure of temperature
1.1kg/cm2~1.5kg/cm2) sterilizing 2.5h, it can be aseptically inoculated with after cooling.
Cultigen wheat caragana microphylla powder culture medium production method：Wheat and caragana microphylla powder, and account for the phosphoric acid of quality of material 0.2%
Potassium dihydrogen and 0.1% magnesium sulfate, the wherein weight ratio of wheat and caragana microphylla powder are 3:7；Its production method is：By wheat to expect water
Quality 1:2 ratios impregnate 10h, and immersion adds in potassium dihydrogen phosphate simultaneously, magnesium sulfate makes it fully dissolve, and are boiled after immersion to without hard
It core but does not split, water is removed in control；By caragana microphylla powder to expect water quality 1:1.2 ratios are impregnated, then are mixed with the wheat after above-mentioned control water
It is even, pH to 7.5~8 is adjusted, then sterilize 2.5h, can be aseptically inoculated with after cooling.
2 cultivating bacterial spawn kind of embodiment makes
1) parent species make：The healthy and strong wild green hired thug of acquisition make parent species, aseptically cut the blueness of fresh wild
Hired thug fructification chooses the meat bacteria organization of cap stem intersection, moves into equipped with PDA improved culture mediums (only by embodiment 1
Carbon source becomes potato 200g/L in PDA improved culture medium formulas) vessel in, be put into constant incubator and cultivate, 22 DEG C~
It is cultivated 25~28 days at a temperature of 24 DEG C, until mycelia covers with device.
2) expand numerous：Parent species are transferred in improvement PDA liquid medium, at 22 DEG C~24 DEG C, are trained with 140r/min rotating speeds
Foster 12~15 days culture bacterium balls uniformly cover with vessel and complete liquid spawn making.
PDA improves fluid nutrient medium：Contain potato 200g/L, sucrose 10g/L, glucose 10g/L, dusty yeast 1.8g/
L, peptone 1.8g/L, magnesium sulfate 0.5g/L, dipotassium hydrogen phosphate 1g/L.Sterilising conditions are：121 DEG C~125 DEG C (pressure 1.1kg/
cm2~1.5kg/cm2) sterilize 30 minutes.
3) original seed makes：The obtained numerous parent species of expansion are implanted into the vessel equipped with compost, are placed on 21 DEG C~23 DEG C
Constant temperature under cultivate 32~37 days, mycelia cover with vessel complete original seed making.
4) cultigen makes：By in vessel of the obtained original seed implantation equipped with compost, it is placed on 21 DEG C~23 DEG C of perseverance
It is cultivated 28~32 days under the conditions of temperature, mycelia covers with vessel and completes cultigen making.
Primary spawn material production method：By the wheat of wash clean and water with quality 1：The immersion 12h of 1.2 ratios impregnates same
When add in the potassium dihydrogen phosphate of wheat quality 0.2%, 0.1% magnesium sulfate makes it fully dissolve, boiled after immersion to no hard core
But it does not split, water is removed in control；PH to 7.5 is adjusted with 2% lime, vessel are put into, through 121 DEG C~125 DEG C (pressure 1.1kg/cm2
~1.5kg/cm2) sterilizing 2.5h, it can be aseptically inoculated with after cooling.
Cultigen wheat caragana microphylla powder culture medium production method：Wheat and caragana microphylla powder and the phosphoric acid for accounting for quality of material 0.2%
Potassium dihydrogen and 0.1% magnesium sulfate, the wherein weight ratio of wheat and caragana microphylla powder are 3:7；Its production method is：By wheat to expect water
Quality 1：1.5 ratios impregnate 12h, and immersion adds in potassium dihydrogen phosphate simultaneously, magnesium sulfate makes it fully dissolve, and is boiled after immersion to nothing
It hard core but does not split, water is removed in control；By caragana microphylla powder to expect water quality 1:1.2 ratios are impregnated, then are mixed with the wheat after above-mentioned control water
Uniformly, pH to 7.5~8 is adjusted, then sterilize 2.5h, can be aseptically inoculated with after cooling.
3 cultivating bacterial spawn kind of embodiment makes
1) parent species make：The healthy and strong wild green hired thug of acquisition make parent species, aseptically pass through fresh open country with iron wire
Raw green hired thug fructification makes its lamella hang on downwards and basidiospore is collected in wide-mouth bottle, and a little basidiospore of picking is as test tube
In, sterile water is added to be diluted to 100 μ L containing 50~60 basidiospore, obtains basidiospore suspension.Basidiospore suspension is drawn to be added dropwise in shifting
Enter equipped with PDA improved culture mediums (carbon source in the PDA improved culture medium formulas of embodiment 1 only is become wheat bran 100g/L), and
Even coating is cultivated 25~28 days at a temperature of 22 DEG C~24 DEG C, until mycelia covers with device.
2) expand numerous：Parent species are transferred in the good PDA culture medium changed, and 25~28 days are cultivated at 22 DEG C~24 DEG C to covering with bacterium
Silk.
PDA improved culture mediums are：Contain wheat bran 100/L, sucrose 10g/L, glucose 10g/L, dusty yeast 1.8g/L, albumen
Peptone 1.8g/L, magnesium sulfate 0.5g/L, dipotassium hydrogen phosphate 1g/L, agar 10g/L culture medium.Sterilising conditions are：121 DEG C~125
DEG C (pressure 1.1kg/cm2~1.5kg/cm2) sterilize 30 minutes.
3) original seed makes：The obtained numerous parent species of expansion are implanted into the vessel equipped with wheat compost, be placed on 21 DEG C~
It is cultivated 32~37 days under 23 DEG C of constant temperature, mycelia covers with vessel and completes original seed making.
4) cultigen makes：By in vessel of the obtained original seed implantation equipped with compost, it is placed on 21 DEG C~23 DEG C of perseverance
It is cultivated 28~32 days under the conditions of temperature, mycelia covers with vessel and completes cultigen making.
Original seed wheat culture medium production method：By the grain of wash clean and water with quality 1：1.3 ratios impregnate 12h, impregnate
Add in the potassium dihydrogen phosphate of wheat quality 0.2% simultaneously, 0.1% magnesium sulfate makes it fully dissolve, boiled after immersion to without hard
It core but does not split, water is removed in control, adjusts pH to 7.5 with 2% lime, vessel is put into, through 121 DEG C~125 DEG C (pressure of temperature
1.1kg/cm2~1.5kg/cm2) sterilizing 2.5h, it can be aseptically inoculated with after cooling.
Cultigen wheat caragana microphylla powder culture medium production method：Wheat and caragana microphylla powder and the phosphoric acid for accounting for quality of material 0.2%
Potassium dihydrogen and 0.1% magnesium sulfate, the wherein weight ratio of wheat and caragana microphylla powder are 3:7；Its production method is：By wheat to expect water
Quality 1:2 ratios impregnate 9h, and immersion adds in potassium dihydrogen phosphate simultaneously, magnesium sulfate makes it fully dissolve, and are boiled after immersion to without hard
It core but does not split, water is removed in control；By caragana microphylla powder to expect water quality 1:1.2 ratios impregnate, then with the well-done wheat after above-mentioned control water
It is uniformly mixed, adjusts pH to 7.5~8, vessel are put into, through 121 DEG C~125 DEG C (pressure 1.1kg/cm of temperature2~1.5kg/cm2)
Sterilize 2.5h, can be aseptically inoculated with after cooling.
The domestication of the wild green hired thug of embodiment 4
1) parent species make：The healthy and strong wild green hired thug of acquisition make parent species, aseptically pass through fresh open country with iron wire
Raw green hired thug fructification makes its lamella hang on downwards and basidiospore is collected in wide-mouth bottle, and a little basidiospore of picking is as test tube
In, sterile water is added to be diluted to 100 μ L containing 50~60 basidiospore, obtains basidiospore suspension.Basidiospore suspension is drawn to be added dropwise in shifting
Enter equipped with PDA improved culture mediums (carbon source in the PDA improved culture medium formulas of embodiment 1 only is become beancake powder 20g/L), and
Even coating is cultivated 25~28 days at a temperature of 22 DEG C~24 DEG C, until mycelia covers with device.
2) expand numerous：Parent species are transferred in improvement PDA liquid medium, at 22~24 DEG C, with 130r/min rotating speed cultures
12~15 days culture bacterium balls uniformly cover with vessel and complete liquid spawn making.
PDA improves fluid nutrient medium：Contain beancake powder 20g/L, sucrose 10g/L, glucose 10g/L, dusty yeast 1.8g/
L, the fluid nutrient medium of peptone 1.8g/L, magnesium sulfate 0.5g/L, dipotassium hydrogen phosphate 1g/L.Sterilising conditions are：121 DEG C~125
DEG C (pressure 1.1kg/cm2~1.5kg/cm2) sterilize 30 minutes.
3) original seed makes：The obtained numerous parent species of expansion are implanted into the vessel equipped with wheat compost, be placed on 21 DEG C~
It is cultivated 32~37 days under 23 DEG C of constant temperature, until mycelia covers with vessel.
4) cultigen makes：By in vessel of the obtained original seed implantation equipped with compost, it is placed on 21 DEG C~23 DEG C of perseverance
It is cultivated 28~32 days under the conditions of temperature, until mycelia covers with vessel.
Step 3) and compost 4) are the same as embodiment 1.
5) it ferments：The caragana microphylla branches and leaves that the sheep manure and fresh nothing pulverized in advance are gone mouldy are with 4:6 mass ratio adds water fully to prewet,
Alternating uniformly spreads sheep manure and grass, bottom width 1.3m, upper width 1.2m, high 1.2m, long unlimited heap is built up, with diameter 6cm~8cm
Wooden stick burrow to material bottom, hole is away from 50cm, when heap temperature is raised to 65 DEG C, is kept for 24 hours, carries out first time turning, continues punching hair
Ferment, heap temperature are raised to 65 DEG C and keep carrying out second of turning in 24 hours again, heat up after 2 days 65 DEG C, turned over for the third time after 12 hours again
Heap, conditioning can be used to sow 60%~65% or so.
6) it tames：The compost fermented is layered on to the western sheep cleaned up to enclose the land face, cultigen is uniformly sprinkling upon training
On nutriment, one layer of compost fermented is repaved above, then spread last layer cultigen, surface layer strain and compost puddle pressing,
The long unlimited curvature of the spinal column type fruiting row of bottom width 1.4m, upper width 1.3m, high 23cm or so is organized into, carries out Mycelium culture 50~55
My god.After mycelium covers with compost, in the soil of surface covering 5cm moistenings, fruiting front surface soil is sprayed water moisturizing when bleaching, temperature
Control is at 21 DEG C~23 DEG C.40~45 days after earthing, there are No. 3 former bases of prairie Tricholomataceae, can be harvested after 10~12 days, science into
Row management of producing mushroom can harvest 3 batches of mushrooms.
The domestication of the wild green hired thug of embodiment 5
1) parent species make：The healthy and strong wild green hired thug of acquisition make parent species, aseptically cut the blueness of fresh wild
Hired thug fructification chooses the meat bacteria organization of cap stem intersection, moves into the vessel equipped with PDA improved culture mediums, is put into perseverance
It cultivates, is cultivated 25~28 days at a temperature of 22 DEG C~24 DEG C, until mycelia covers with device in warm incubator.
2) expand numerous：Parent species are transferred in the PDA culture medium of improvement, and 25~28 days are cultivated at 22 DEG C~24 DEG C to covering with bacterium
Silk.
PDA improved culture mediums are：Containing the full powder 15g/L of wheat, sucrose 10g/L, glucose 10g/L, dusty yeast 1.8g/L,
Peptone 1.8g/L, magnesium sulfate 0.5g/L, dipotassium hydrogen phosphate 1g/L, agar 10g/L culture medium.Sterilising conditions are：121 DEG C~
125 DEG C of (pressure 1.1kg/cm2~1.5kg/cm2) sterilize 30 minutes.
3) original seed makes：The obtained numerous parent species of expansion are implanted into the vessel equipped with wheat compost, be placed on 21 DEG C~
It is cultivated 32~37 days under 23 DEG C of constant temperature, mycelia covers with vessel and completes original seed making.
4) cultigen makes：By in vessel of the obtained original seed implantation equipped with compost, it is placed on 21 DEG C~23 DEG C of perseverance
It is cultivated 28~32 days under the conditions of temperature, mycelia covers with vessel and completes cultigen making.
Original seed wheat culture medium production method：By the wheat of wash clean and water with quality 1：1.4 ratios impregnate 12h, impregnate
Add in the potassium dihydrogen phosphate of wheat quality 0.2% simultaneously, 0.1% magnesium sulfate makes it fully dissolve, boiled after immersion to without hard
It core but does not split, water is removed in control, adjusts pH to 7.5 with 2% lime, vessel is put into, through 121 DEG C~125 DEG C (pressure of temperature
1.1kg/cm2~1.5kg/cm2) sterilizing 2.5h, it can be aseptically inoculated with after cooling.
Cultigen wheat caragana microphylla powder culture medium production method：Wheat and caragana microphylla powder and the phosphoric acid for accounting for quality of material 0.2%
Potassium dihydrogen and 0.1% magnesium sulfate, the wherein weight ratio of wheat and caragana microphylla powder are 3:7；Its production method is：By the wheat of wash clean
Grain is with water with quality 1：The immersion 12h of 1.4 ratios impregnates the potassium dihydrogen phosphate for adding in wheat quality 0.2% simultaneously, 0.1%
Magnesium sulfate makes it fully dissolve, and is boiled after immersion to no hard core but not split, and water is removed in control；Simultaneously with caragana microphylla powder and water quality 1：
1.2 ratio is impregnated caragana microphylla powder and is impregnated with it completely to moisture, then caragana microphylla powder is uniformly mixed with well-done wheat, with 2%
Lime adjusts pH to 7.5, vessel is put into, through 121 DEG C~125 DEG C (pressure 1.1kg/cm of temperature2~1.5kg/cm2) sterilizing 2.5h,
It can be aseptically inoculated with after cooling.
5) it ferments：Caragana microphylla branches and leaves that the sheep manure of sieving and fresh nothing are gone mouldy are pulverized in advance with 4:6 mass ratio adds water abundant
It prewets, alternating uniformly spreads sheep manure and grass, bottom width 1.2m, upper width 1.1m, high 1.1m, long unlimited heap is built up, with diameter 6cm
The wooden stick of~8cm burrows to material bottom, and hole when heap temperature is raised to 65 DEG C, is kept for 24 hours away from 40cm, carries out first time turning, continued
Punching fermentation, heap temperature are raised to 65 DEG C and keep carrying out second of turning in 24 hours again, heat up again after 2 days 65 DEG C, the after 12 hours
Turning three times, conditioning can be used to sow 60%~65% or so.
6) it tames：The compost fermented is layered on to the western sheep cleaned up to enclose the land face, cultigen is uniformly sprinkling upon training
On nutriment, one layer of compost fermented is repaved above, then spread last layer cultigen, surface layer strain and compost puddle pressing,
The long unlimited curvature of the spinal column type fruiting row of bottom width 1.3m, upper width 1.2m, high 23cm or so is organized into, carries out Mycelium culture 50~55
My god.After mycelium covers with compost, in the soil of surface covering 5cm moistenings, fruiting front surface soil is sprayed water moisturizing when bleaching, temperature
Control is at 21 DEG C~23 DEG C.40~45 days after earthing, there are No. 3 former bases of prairie Tricholomataceae, can be harvested after 10~12 days, science into
Row management of producing mushroom can adopt 3 batches of mushrooms.
The present invention is successfully realized the artificial cultivation of wild green hired thug new strains prairie Tricholomataceae 3, not only protects and is on the verge of
This rareness species of the green hired thug of extinction, and cattle and sheep excrement, the careless bits the like waste for making full use of pastoral area abundant, are cultivated
Prairie Tricholomataceae 3, realizes the comprehensive utilization of resource, promote emerging grass poultry bacterium industry recycle and sustainable development,
The income of local herdsman can also be increased.
Although above the present invention is described in detail with a general description of the specific embodiments,
On the basis of the present invention, it can be modified or improved, this will be apparent to those skilled in the art.Cause
This, these modifications or improvements, belong to the scope of protection of present invention without departing from theon the basis of the spirit of the present invention.
Sequence table
<110>Inner Mongolia Autonomous Region Academy of Agricultural and Livestock Husbandry
<120>A kind of Tricholoma mongolicum new strains prairie Tricholomataceae 3 and its acclimation method
<130> KHP171118593.6
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 656
<212> DNA
<213>Tricholoma mongolicum (Lepista sp.)
<400> 1
tcatacttgg ttgggttgtg ctggcttttc ggagcatgtg catgcctagc gccattttta 60
ccacctgtgc acattttgta gatttgaaac aattctcgag gaaactcggt ttgaggaatg 120
ctgtgcgaaa gcttagcttt tcttgtgttt caagtctatg tttttatata taccccataa 180
gaatgtaata gaatgtcatt aatgggcttt gttgccttta aattaataca actttcaaca 240
acggatctct tggttctcgc atcgatgaag aacgcagcga aatgcgataa gtaatgtgaa 300
ttgcagaatt cagtgaatca tcgaatcttt gaacgcacct tgcgctcctt ggtattccga 360
ggagcatgcc tgtttgagtg tcattaaatt ctcaaccttt ccagcttttg caagttggat 420
tggcttggat gtggagggtt ttgcgggctt ctcagaagtc ggctcctctt aaatgcatta 480
gcagaacctt tgtggaccag ctttggtgtg ataattatct atgccattgt tgtaaagcag 540
cttttatatg gggttcagct tctaatagtc cattgacttg gacaatttct gacattttga 600
cctcaaatca ggtaggacta cccgctgaac ttaagcatat caaaagccgg agggaa 656
Claims (10)
- Tricholoma mongolicum 1. (Lepista sp.) new strains prairie Tricholomataceae 3, which is characterized in that deposit number CGMCC No.15083。
- 2. the method for prairie Tricholomataceae 3 as described in wild green hired thug domestication prepares claim 1, which is characterized in that including parent species It makes, expand the step of numerous, original seed makes, cultigen makes and tames, it is specific as follows：1) parent species make：At 22 DEG C~24 DEG C parent species are cultivated using wild green hired thug fructification as material, until being covered on culture medium Mycelia；2) expand numerous：Step 1) parent species are transferred on culture medium, are cultivated at 22 DEG C~24 DEG C, until covering with mycelia；3) original seed makes：Expand numerous parent species to move into the vessel equipped with compost, cultivate 32 under 21 DEG C~23 DEG C constant temperatures~ 37 days, until mycelia, which covers with vessel, completes original seed making；4) cultigen makes：Original seed move into equipped with compost vessel in, under 21 DEG C~23 DEG C of constant temperature cultivate 28~ 32 days, until mycelia, which covers with vessel, completes cultigen making；5) it tames：Agriculture and forestry organic waste material and excrement of animals are built into heap fermentation, sowing is completed after fermentation, carries out management of producing mushroom.
- 3. according to the method described in claim 2, it is characterized in that, step 1) collects legal system using tissue isolation or basidiospore Make parent species：The tissue isolation is：Fresh wild blueness hired thug fructification is aseptically cut, chooses cap stem intersection Meat bacteria organization, move into equipped with culture medium vessel in, cultivated 25~28 days in 22 DEG C~24 DEG C of constant incubator；The basidiospore collecting method is：Acquire fresh wild green hired thug strain fructification, aseptically make its lamella to Under hang on basidiospore collected in container, sterile water is added to be diluted to 100 μ L containing 50~60 basidiospore, obtains basidiospore suspension； It draws basidiospore suspension to be added dropwise on culture medium, and even spread, 28~30 is cultivated in 22 DEG C~24 DEG C of constant incubator My god.
- 4. according to the method described in claim 3, it is characterized in that, the culture medium of the step 1) is PDA improved culture mediums；Step It is rapid 2) be cultivated 25~28 days in PDA improved culture mediums or PDA improve fluid nutrient medium in 120~140r/min rotating speeds Lower culture 12~15 days；The PDA improved culture mediums formula is：Carbon source 15g/L~200g/L, sucrose 10g/L, glucose 10g/L, dusty yeast 1.8g/L, peptone 1.8g/L, magnesium sulfate 0.5g/L, dipotassium hydrogen phosphate 1g/L and agar 10g/L；The PDA improves Liquid Culture based formulas：Carbon source 15g/L~200g/L, sucrose 10g/L, glucose 10g/L, yeast Powder 1.8g/L, peptone 1.8g/L, magnesium sulfate 0.5g/L, dipotassium hydrogen phosphate 1g/L and agar 0.6g/L；The carbon source is at least one of potato, sweet potato, wheat bran, the full powder of wheat, beancake powder.
- 5. according to the method described in claim 2, it is characterized in that, the step 3) compost is：With whole wheat granular and/or grain For material and account for the potassium dihydrogen phosphate of quality of material 0.2% and 0.1% magnesium sulfate；Its production method is：By material to expect Water quality 1:1.3~1：1.5 ratios impregnate 9~12h, impregnate while add in potassium dihydrogen phosphate, magnesium sulfate makes it fully dissolve, leaching It is boiled after bubble to no hard core, water is removed in control, adjusts pH to 7.5~8, then sterilizes.
- 6. according to the method described in claim 2, it is characterized in that, the step 4) compost is：Wheat and caragana microphylla powder and The potassium dihydrogen phosphate of quality of material 0.2% and 0.1% magnesium sulfate are accounted for, the wherein weight ratio of wheat and caragana microphylla powder is 3:7；It is made It is as method：By wheat to expect water quality 1：1.5~1:2 ratios impregnate 9~12h, impregnate and add in potassium dihydrogen phosphate, sulfuric acid simultaneously Magnesium makes it fully dissolve, and is boiled after immersion to no hard core but not split, and water is removed in control；By caragana microphylla powder to expect water quality 1:1.2 ratio It impregnates, then is uniformly mixed with the wheat after above-mentioned control water, adjust pH to 7.5~8, then sterilize.
- 7. according to the method described in claim 2, it is characterized in that, step 5) is described builds heap fermentation and be specially：Agricultural are discarded After object crushes, according to sheep manure and agriculture and forestry organic waste material mass ratio 4：6 be uniformly mixed build up fermentation heap, at clean ventilation ferment；Its In, the agriculture and forestry organic waste material is selected from least one of careless knot, caragana microphylla branches and leaves, straw, maize straw, wheat straw.
- 8. the method according to the description of claim 7 is characterized in that step 5) build up bottom width 1.2m~1.4m, upper width 1.1m~ The fermentation heap of 1.3m, high 1.1m~1.3m punch in material heap hole with the wooden stick of diameter 6cm~8cm to expecting bottom, hole spacing 40cm~50cm, heap temperature are kept for 24 hours when being raised to 65 DEG C, carry out first time turning, continue punching fermentation, and heap temperature is raised to 65 DEG C Keep again 24 hours carry out second of turning, heat up again after 2 days 65 DEG C when keep 12 hours after third time turning, conditioning Material after 60%~65%, gained fermentation can be used to sow；The cultigen that step 4) is obtained uniformly on the material of paving after fermentation, repaves the material after one layer of fermentation, then spread above Last layer cultigen, every layer of material 10cm~15cm is thick, and surface layer strain and material puddle pressing, be organized into bottom width 1.3m~ 1.5m, upper width 1.2m~1.3m, high 20cm~24cm, long unlimited curvature of the spinal column type fruiting row, carry out Mycelium culture 50~55 days.
- 9. according to the method described in claim 8, it is characterized in that, step 5) covers 4cm after the long full with substance of mycelium on surface The rural area soil or turfy soil of~6cm thickness, fruiting front surface soil bleach, moisturizing of spraying water, and temperature is controlled at 21 DEG C~23 DEG C；Earthing 40~45 days afterwards, there is the mushroom former base after domestication, keep ground moistening, can be harvested after 10~12 days；Surface evening is gently pressed dry 2 days after first batch of mushroom of harvesting, continue management of producing mushroom after making mycelium restoration ecosystem, harvested There is regrowth hair mushroom former base within the 15th~17 day after first batch of mushroom, adopt three batches of mushrooms altogether.
- 10. No. 3 applications in food, drug, health products, cosmetics and feed is prepared of grassland mushroom described in claim 1.
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