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

Tricholoma mongolicum new strains prairie Tricholomataceae 2 and its selection Download PDF

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CN108048335A
CN108048335A CN201711421978.5A CN201711421978A CN108048335A CN 108048335 A CN108048335 A CN 108048335A CN 201711421978 A CN201711421978 A CN 201711421978A CN 108048335 A CN108048335 A CN 108048335A
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wheat
pda
cultigen
quality
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CN108048335B (en
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郭九峰
孙国琴
王润元
王勇
李亚娇
王海燕
庞杰
边淑萍
于传宗
金焕林
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Inner Mongolia University
Inner Mongolia Academy of Agricultural and Animal Husbandry Sciences
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Inner Mongolia Academy of Agricultural and Animal Husbandry Sciences
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    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00Microorganisms, 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
    • C12N1/14Fungi; Culture media therefor
    • C12N1/145Fungal isolates
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12RINDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00Microorganisms ; Processes using microorganisms
    • C12R2001/645Fungi ; Processes using fungi
    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01HNEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H15/00Fungi; Lichens
    • CCHEMISTRY; METALLURGY
    • C05FERTILISERS; MANUFACTURE THEREOF
    • C05FORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
    • C05F17/00Preparation of fertilisers characterised by biological or biochemical treatment steps, e.g. composting or fermentation
    • CCHEMISTRY; METALLURGY
    • C05FERTILISERS; MANUFACTURE THEREOF
    • C05FORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
    • C05F3/00Fertilisers from human or animal excrements, e.g. manure
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00Microorganisms, 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
    • C12N1/14Fungi; Culture media therefor
    • YGENERAL 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
    • Y02TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02WCLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
    • Y02W30/00Technologies for solid waste management
    • Y02W30/40Bio-organic fraction processing; Production of fertilisers from the organic fraction of waste or refuse

Abstract

The invention belongs to edible mushroom new strains acclimation method field, Tricholoma mongolicum new strains prairie Tricholomataceae 2 and its selection are specifically disclosed.The deposit number of No. 2 strains of Tricholoma mongolicum new strains prairie Tricholomataceae of the present invention is CGMCC No.15084, has the characteristics that resistance is strong, heritability is stable, high-quality, delicious, and a kind of new high-quality edible mushroom production is provided for people.Acclimation method provided by the present invention not only protects this rareness species of endangered Lepista sordida; it can also realize that farming and animal husbandry changing waste into resources utilizes by cultivating No. 2 new strains of prairie Tricholomataceae; promote the sustainable development of agricultural economy; 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; to being started an undertaking and being obtained employment using facility cultivations prairie Tricholomataceaes 2 such as farming and pastoral area discarded object, idle cattle and sheep circles, build up the family fortunes.

Description

Tricholoma mongolicum new strains prairie Tricholomataceae 2 and its selection
Technical field
The invention belongs to edible mushroom new strains acclimation method fields, specifically, are related to a Tricholoma mongolicum new strains grass Former mushroom No. 2 and its acclimation method in vain.
Background technology
Lepista sordida (Lepista sordida) belongs to Agaricales (Agaricales), Tricholomataceae (tricholomataceae), Lepista lentinus (Lepista).Lepista sordida is called the cylinder Guo that white flower face is mainly distributed on the Inner Mongol It strangles, on Horqin Caoyuan.Lepista sordida protein content is high, and amino acid classes are more complete, particularly containing abundant calcium, iron, Carrotene and niacin and trace copper, zinc, fluorine, iodine etc. make Lepista sordida have the work(of blood-nourishing, benefit god, tonifying liver and the five internal organs Effect, often feeding are conducive to treat the illnesss such as anaemia uterine bleeding, weakness due to chronic disease, god is tired, forgetful, are a kind of domestic fungus resources of preciousness.Mesh Presteppe degeneration getting worse, in addition people excessively search for food, Natural Survival condition is caused to deteriorate further.
Therefore, it is badly in need of providing a Tricholoma mongolicum new strains prairie Tricholomataceae 2 suitable for artificial cultivation, is retaining wild flowers While face perfume (or spice) mushroom original flavor, No. 2 this rareness species of endangered prairie Tricholomataceae are protected, moreover it is possible to by cultivating grassland White No. 2 new strains of mushroom realize that farming and animal husbandry changing waste into resources utilizes,
The content of the invention
In order to solve the problems in the existing technology, the object of the present invention is to provide a Tricholoma mongolicum new strains grassland Mushroom No. 2 and its selection in vain.
In order to realize the object of the invention, technical scheme is as follows:
In a first aspect, the present invention provides one plant of prairie Tricholomataceaes 2, Lepista sordida (Lepista is identified as Sordida), it is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center (abbreviation CGMCC, address:Beijing The institute 3 of city Chaoyang District North Star West Road 1, Institute of Microorganism, Academia Sinica, postcode:100101), preservation date is On December 7th, 2017, deposit number are CGMCC No.15084.
The prairie Tricholomataceae 2, fructification cap is white, and initial stage, flat hemispherical cap edge was involute, until maturity period bacterium Lid is white in the water stain shape crow in bamboo hat shape edge, mature sporophore 3.5~8.7cm of bacteria cover diameter, and bacterial context white is thin;Lamella white, Growing straight or curved life, slightly dilute, Length discrepancy;It is thick in stem, white, long 3.5~7.6cm, thick 0.2~2.0cm, interior reality, close to base portion Normal curved life (see Fig. 2).
The ribosomal coding gene sequences of 5.8S of the bacterial strain are as shown in SEQ ID NO.1, after the sequence amplification Agarose gel electrophoresis figure is as shown in Figure 3.
Prairie Tricholomataceae 2 provided by the present invention is by obtaining, remaining with wild to the domestication of wild Lepista sordida The original flavor of Lepista sordida, and can realize artificial cultivation.
Second aspect, the present invention provides a kind of methods for taming No. 2 bacterial strains of prairie Tricholomataceae, include the following steps:
1) parent species make:At 21 DEG C~24 DEG C parent species are cultivated using wild Lepista sordida fructification as material, until culture medium On cover with mycelia;
2) expand numerous:Parent species obtained by step 1) are transferred on culture medium, are cultivated at 21 DEG C~24 DEG C, until covering with mycelia;
3) original seed makes:Parent species of the step 2) expansion after numerous are moved into the vessel equipped with Primary spawn material, 20 DEG C~23 It is cultivated 33~38 days under DEG C constant temperature, until mycelia covers with vessel and completes original seed and makes;
4) cultigen makes:By step 3) make original seed implantation equipped with cultigen compost vessel in, 20 DEG C~ It is cultivated 28~33 days under 23 DEG C of constant temperature, until mycelia, which covers with vessel, completes cultigen making.
Wherein, the step 1) makes strain with tissue isolation or basidiospore collecting method:
The tissue isolation is:The Lepista sordida fructification of fresh wild is aseptically cut, chooses cap bacterium The meat bacteria organization of handle intersection, move into equipped with culture medium vessel in, in 21 DEG C~24 DEG C of constant incubator cultivate 24~ 27 days;
The basidiospore collecting method is:The Lepista sordida fructification of fresh wild is aseptically passed through with iron wire, is made Its lamella, which is hung on downwards, collects basidiospore in container, a little basidiospore of picking adds sterile water to be diluted to 100 μ L and contain into test tube 40~50 basidiospore, obtain basidiospore suspension;Basidiospore suspension is drawn to be added dropwise on culture medium, and even spread, 21 DEG C~ Culture 24~27 days in 24 DEG C of constant incubator.
Wherein, the culture medium in step 1) is PDA improved culture mediums;
Culture medium described in step 2) improves fluid nutrient medium for PDA improved culture mediums or PDA;It is improved when using PDA During culture medium, cultivated 24~27 days in PDA improved culture mediums;When using PDA improvement fluid nutrient mediums, liquid is improved in PDA With 120~140r/min rotating speeds culture 13~16 days in body culture medium;
The formula of the PDA improved culture mediums is:Carbon source 15g/L~200g/L, sucrose 10g/L, glucose 10g/L, ferment Female powder 1.2g/L, peptone 1.2g/L, magnesium sulfate 0.5g/L, dipotassium hydrogen phosphate 1g/L, agar 10g/L;
The formula of PDA improvement fluid nutrient medium is:Carbon source 15g/L~200g/L, sucrose 10g/L, glucose 10g/ L, dusty yeast 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 the one or more in potato, sweet potato, wheat bran, the full powder of wheat, beancake powder.
Wherein, the formula of the Primary spawn material in step 3) is:Whole wheat granular or grain, potassium dihydrogen phosphate and sulfuric acid Magnesium;
The quality of the potassium dihydrogen phosphate is the 0.2% of whole wheat granular or grain quality;
The quality of the magnesium sulfate is the 0.1% of whole wheat granular or grain quality;
The production method of the Primary spawn material is:By whole wheat granular or grain to expect water quality as 1:1.2~1:1.5 ratio Example impregnates 8~12h, and potassium dihydrogen phosphate and magnesium sulfate are added in while immersion, it is made fully to dissolve, and is boiled after immersion to without hard Water is removed in core, control, and pH to 7.5~8, sterilizing are adjusted with lime.
Wherein, the cultigen compost in step 4) includes wheat that mass parts are 20~30 parts and 70~80 parts Caragana microphylla powder;
The production method of the cultigen compost is:By wheat to expect water quality as 1:1.3~1:2 ratio impregnates 8 ~12h adds in the potassium dihydrogen phosphate and 0.1% magnesium sulfate of opposite wheat quality 0.2% while immersion, makes it fully molten Solution, boils to no hard core after immersion but does not split, and water is removed in control;To expect water quality as 1:1.2 ratio is soaked in water caragana microphylla powder, then It is uniformly mixed with wheat, pH to 7.5~8, sterilizing is adjusted with lime.
The caragana microphylla powder is the caragana microphylla branch of caragana microphylla aerial part, is ground into 0.5-1.0 centimetres of particle.Caragana microphylla amyloid proteins matter Content is high, full of nutrition rich in mineral matter element, vitamin etc..
The third aspect, the present invention also provides a kind of methods of artificial cultivation prairie Tricholomataceae 2, include the following steps:
(a) plant waste and excrement of animals windrow are fermented;
(b) by prairie Tricholomataceae provided by the present invention 2 or other tamed via acclimation method of the present invention Cultigen uniformly on the material of paving after fermentation, repaves the material after one layer of fermentation above, then spreads last layer cultigen, after fermentation Every layer of material paving 10cm~15cm it is thick, surface layer strain and cultigen compost puddle pressing, be organized into bottom width 1.3m~1.5m, The unlimited curvature of the spinal column type fruiting row of upper width 1.2m~1.3m, the length of high 20cm~24cm, carries out Mycelium culture 50~60 days.
Preferably, the windrow fermentation is:Plant waste and excrement of animals are pressed 4:6~5:5 mass ratio mixing is equal It is even to build up fermentation heap, it ferments at clean ventilation;
One or more of the plant waste in careless knot, caragana microphylla powder branches and leaves, straw, maize straw, wheat straw.
The windrow ferments:Build up the fermentation of bottom width 1.2m~1.4m, upper width 1.1m~1.3m, high 1.1m~1.3m Heap punches on material heap hole with the wooden stick of diameter 6cm~8cm to expecting bottom, hole spacing 40cm~60cm, when heap temperature is raised to 65 DEG C Keep 24 hours, carry out first time turning, continue punching fermentation, heap temperature be raised to 65 DEG C keep again 24 it is small when turned over for the second time Heap, heat up again after 2 days 65 DEG C when keep 12 it is small when after third time turning, conditioning can be used to 60%~65% or so Sowing.
Further, after the long full with substance of mycelium, the rural area soil or turfy soil of 4cm~6cm thickness are covered on surface, before fruiting Surface soil bleaches, moisturizing of spraying water, and temperature is controlled at 21 DEG C~23 DEG C;40~45 days after earthing, there are No. 2 former bases of prairie Tricholomataceae, Ground moistening is kept, can be harvested after 9~12 days.
Further, surface evening is gently pressed dry 2 days after harvesting first batch of mushroom, continued after making mycelium restoration ecosystem Management of producing mushroom harvests after first batch of mushroom and regrowth hair mushroom former base occurs on the 15th~18 day, can adopt 3 batches of mushrooms altogether.
The beneficial effects of the present invention are:
The present invention provides a kind of wild Lepista sordida new strains and its acclimation method and cultural method.The acclimation method is not This rare wildlife species cherished of Lepista sordida is only protected to exempt from extinction, maintain herbage and the ecological body of grassland mushroom symbiosis System, moreover it is possible to provide taste splendid edible fungus for people.
New strains resistance is strong, heritability is stable, high-quality, delicious for prairie Tricholomataceae 2 provided by the present invention, is people A kind of new high-quality edible mushroom product is provided.The prairie Tricholomataceae 2 obtained by the present invention, it is peculiar to maintain wild Lepista sordida Flavor and nutritional ingredient, the product taste that cattle and sheep circle of the summer in pastoral area is produced is fresher, very popular.
The present invention not only protects No. 2 this rareness species of endangered prairie Tricholomataceae, moreover it is possible to white by cultivating grassland No. 2 new strains of mushroom realize that farming and animal husbandry changing waste into resources utilizes, and turn waste into wealth, have expanded peasants and herdsmen's new growth engines, are hairs Pastoral area courtyard economy and an important ring for grass poultry circular economy are opened up, to being planted using facilities such as farming and pastoral area discarded object, idle cattle and sheep circles Training prairie Tricholomataceae 2 is started an undertaking and is obtained employment, and is built up the family fortunes.It realizes the comprehensive utilization of resource, promotes emerging《Grass, poultry, bacterium》Production The sustainable development of industry ecological circulation economy.
Description of the drawings
Fig. 1 is Lepista sordida wild strain.
Fig. 2 is No. 2 bacterial strains of prairie Tricholomataceae of the present invention.
Fig. 3 is the amplification electrophoretogram of the 5.8S ribosomes coding gene sequences of prairie Tricholomataceae 2 of the present invention.
Specific embodiment
The preferred embodiment of the present invention is described in detail below in conjunction with embodiment.It is it will be appreciated that following real Providing merely to play the purpose of explanation for example is applied, is not used to limit the scope of the present invention.The skill of this field Art personnel can carry out various modifications and replace to the present invention in the case of without departing substantially from spirit of the invention and spirit.
Experimental method used in following embodiments is conventional method unless otherwise specified.
The materials, reagents and the like used in the following examples is commercially available unless otherwise specified.
Embodiment 1
PDA improved culture mediums are used in the present embodiment:Through 121 DEG C~125 DEG C (pressure 1.1kg/cm of temperature2~ 1.5kg/cm2) sterilize 30 minutes 200g/L containing sweet potato, sucrose 10g/L, glucose 10g/L, dusty yeast 1.2g/L, peptone 1.2g/L, magnesium sulfate 0.5g/L, dipotassium hydrogen phosphate 1g/L, the culture medium of agar 10g/L.
1) parent species make:The healthy and strong wild Lepista sordida of acquisition makes parent species, aseptically cuts fresh wild Lepista sordida fructification chooses the meat bacteria organization of cap stem intersection, moves into the vessel equipped with PDA improved culture mediums, puts Enter culture in constant incubator, cultivated 24~27 days at a temperature of 21 DEG C~24 DEG C, until mycelia covers with device.
2) expand numerous:Parent species are transferred in PDA improved culture mediums, and 24~27 days are cultivated at 21 DEG C~24 DEG C to covering with bacterium Silk.
3) original seed makes:The obtained numerous parent species of expansion are implanted into the vessel equipped with compost, are placed on 20 DEG C~23 DEG C Constant temperature under cultivate 33~38 days, mycelia cover with vessel complete original seed making.
4) cultigen makes:By in vessel of the obtained original seed implantation equipped with compost, 20 DEG C~23 DEG C of perseverance is placed on It is cultivated 28~33 days under the conditions of temperature, mycelia covers with vessel and completes cultigen making.
The production method of the Primary spawn material is:By whole wheat granular or grain to expect water quality as 1:1.5 ratio is impregnated 10h impregnates and adds in 0.2% potassium dihydrogen phosphate, 0.1% magnesium sulfate simultaneously it is made fully to dissolve, boiled after immersion to no hard core but It 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 1.1kg/ of temperature cm2~1.5kg/cm2) sterilizing 2.5h, it can be aseptically inoculated with after cooling.
The cultigen compost includes the wheat and 80 parts of caragana microphylla powder that mass parts are 20 parts;The cultigen compost Production method be:By wheat to expect water quality as 1:1.3~1:2 ratio impregnates 8~12h, and immersion while adds in opposite The potassium dihydrogen phosphate of wheat quality 0.2% and 0.1% magnesium sulfate, make it fully dissolve, and are boiled after immersion to no hard core but not It splits, water is removed in control;To expect water quality as 1:1.2 ratio is soaked in water caragana microphylla powder, then is uniformly mixed with wheat, adjust pH to 7.5~8, sterilizing.
Embodiment 2
PDA improved culture mediums are used in the present embodiment:Through 121 DEG C~125 DEG C (pressure 1.1kg/cm of temperature2~ 1.5kg/cm2) sterilize 30 minutes 100/L containing wheat bran, sucrose 10g/L, glucose 10g/L, dusty yeast 1.2g/L, peptone 1.2g/L, magnesium sulfate 0.5g/L, dipotassium hydrogen phosphate 1g/L, the culture medium of agar 0.6g/L.
1) parent species make:The healthy and strong wild Lepista sordida of acquisition makes parent species, is aseptically passed through with iron wire fresh Wild Lepista sordida fructification makes its lamella hang on downwards and basidiospore is collected in wide-mouth bottle, a little basidiospore of picking as In test tube, sterile water is added to be diluted to 100 μ L containing 40~50 basidiospore, obtains basidiospore suspension.Basidiospore suspension is drawn to be added dropwise In moving into equipped with PDA improved culture mediums, and even spread, cultivated 24~27 days at a temperature of 21 DEG C~24 DEG C, until mycelia is long Full device.
2) expand numerous:Parent species are transferred in improvement PDA liquid medium, at 21 DEG C~24 DEG C, are trained with 130r/min rotating speeds Foster 12~16 days culture bacterium balls uniformly cover with vessel and complete liquid spawn making.
3) original seed makes:The obtained numerous parent species of expansion are implanted into the vessel equipped with compost, are placed on 20 DEG C~23 DEG C Constant temperature under cultivate 33~38 days, mycelia cover with vessel complete original seed making.
4) cultigen makes:By in vessel of the obtained original seed implantation equipped with compost, 20 DEG C~23 DEG C of perseverance is placed on It is cultivated 28~33 days under the conditions of temperature, mycelia covers with vessel and completes cultigen making.
The production method of the Primary spawn material is:By whole wheat granular or grain to expect water quality as 1:1.3 ratio is impregnated 12h impregnates and adds in 0.2% potassium dihydrogen phosphate, 0.1% magnesium sulfate simultaneously it is made fully to dissolve, boiled after immersion to no hard core but It 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 1.1kg/ of temperature cm2~1.5kg/cm2) sterilizing 2.5h, it can be aseptically inoculated with after cooling.
The cultigen compost includes the wheat and 70 parts of caragana microphylla powder that mass parts are 30 parts;The cultigen compost Production method be:By wheat to expect water quality as 1:1.3 ratio impregnates 8~12h, and immersion while adds in opposite wheat matter The potassium dihydrogen phosphate and 0.1% magnesium sulfate of amount 0.2%, make it fully dissolve, are boiled after immersion to no hard core but do not split, and control Remove water;To expect water quality as 1:1.2 ratio is soaked in water caragana microphylla powder, then is uniformly mixed with wheat, adjusts pH to 7.5, sterilizing.
Embodiment 3
PDA improved culture mediums are used in the present embodiment:PDA improved culture mediums are:Through 121 DEG C~125 DEG C (pressures of temperature Power 1.1kg/cm2~1.5kg/cm2) sterilize 30 minutes 200g/L containing potato, sucrose 10g/L, glucose 10g/L, dusty yeast 1.2g/L, peptone 1.2g/L, magnesium sulfate 0.5g/L, dipotassium hydrogen phosphate 1g/L, agar 10g/L.
1) parent species make:The healthy and strong wild Lepista sordida of acquisition makes parent species, aseptically cuts fresh wild Lepista sordida fructification chooses the meat bacteria organization of cap stem intersection, moves into the vessel equipped with PDA improved culture mediums, puts Enter culture in constant incubator, cultivated 24~27 days at a temperature of 21 DEG C~24 DEG C, until mycelia covers with device.
2) expand numerous:Parent species are transferred in PDA improved culture mediums, and 24~27 days are cultivated at 21 DEG C~24 DEG C to covering with bacterium Silk.
3) original seed makes:The obtained numerous parent species of expansion are implanted into the vessel equipped with wheat compost, be placed on 20 DEG C~ It is cultivated 33~38 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, 20 DEG C~23 DEG C of perseverance is placed on It is cultivated 28~33 days under the conditions of temperature, mycelia covers with vessel and completes cultigen making.
The production method of the Primary spawn material is:By whole wheat granular or grain to expect water quality as 1:1.3 ratio is impregnated 11h impregnates and adds in 0.2% potassium dihydrogen phosphate, 0.1% magnesium sulfate simultaneously it is made fully to dissolve, boiled after immersion to no hard core but It 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 1.1kg/ of temperature cm2~1.5kg/cm2) sterilizing 2.5h, it can be aseptically inoculated with after cooling.
The cultigen compost includes the wheat and 75 parts of caragana microphylla powder that mass parts are 25 parts;The cultigen compost Production method be:By wheat to expect water quality as 1:1.3 ratio impregnates 11h, and immersion while adds in opposite wheat quality 0.2% potassium dihydrogen phosphate and 0.1% magnesium sulfate, make it fully dissolve, and are boiled after immersion to no hard core but not split, control is gone Water;To expect water quality as 1:1.2 ratio is soaked in water caragana microphylla powder, then is uniformly mixed with wheat, adjusts pH to 8, sterilizing.
Embodiment 4
PDA improved culture mediums are used in the present embodiment:Through 121 DEG C~125 DEG C (pressure 1.1kg/cm of temperature2~ 1.5kg/cm2) sterilize 30 minutes 20g/L containing beancake powder, sucrose 10g/L, glucose 10g/L, dusty yeast 1.2g/L, peptone 1.2g/L, magnesium sulfate 0.5g/L, dipotassium hydrogen phosphate 1g/L, agar 0.6g/L.
1) parent species make:The healthy and strong wild Lepista sordida of acquisition makes parent species, is aseptically passed through with iron wire fresh Wild Lepista sordida fructification makes its lamella hang on downwards and basidiospore is collected in wide-mouth bottle, a little basidiospore of picking as In test tube, sterile water is added to be diluted to 100 μ L containing 40~50 basidiospore, obtains basidiospore suspension.Basidiospore suspension is drawn to be added dropwise In moving into equipped with PDA improved culture mediums, and even spread, cultivated 24~27 days at a temperature of 21 DEG C~24 DEG C, until mycelia is long Full device.
2) expand numerous:Parent species are transferred in improvement PDA liquid medium, at 21~24 DEG C, with 130r/min rotating speed cultures 13~16 days culture bacterium balls uniformly cover with vessel and complete liquid spawn making;
3) original seed makes:The obtained numerous parent species of expansion are implanted into the vessel equipped with wheat compost, be placed on 20 DEG C~ It is cultivated 33~38 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, 20 DEG C~23 DEG C of perseverance is placed on It is cultivated 28~33 days under the conditions of temperature, until mycelia covers with vessel.
Step 3) and compost 4) are the same as embodiment 1.
5) ferment:Caragana microphylla powder branches and leaves that the sheep manure of sieving and fresh nothing are gone mouldy are pulverized in advance with 4:6 ratio adds water abundant It prewets, 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 The wooden stick of~8cm burrows to material bottom, and hole is away from 50cm, when heap temperature is raised to 65 DEG C, is kept for 24 hours, carries out first time turning, after Continuous punching fermentation, heap temperature be raised to 65 DEG C keep again 24 it is small when carry out second of turning, heat up again after 2 days 65 DEG C, 12 it is small when after Third time turning, conditioning can be used to sow 60%~65% or so.
6) domesticating and cultivating:The compost fermented is layered on to the western sheep cleaned up to enclose the land face, cultigen is uniformly spread On compost, one layer of compost fermented is repaved above, then spreads last layer cultigen, and surface layer strain and compost puddle pressure It is flat, it is organized into the long unlimited curvature of the spinal column type fruiting row of bottom width 1.4m, upper width 1.3m, high 23cm or so, progress Mycelium culture 50~ 60 days.After mycelium covers with compost, in the soil of surface covering 5cm moistenings, fruiting front surface soil is sprayed water moisturizing when bleaching, temperature Degree control is at 21 DEG C~23 DEG C.40~45 days after earthing, there are No. 2 former bases of prairie Tricholomataceae, can be harvested after 9~12 days, science 3 batches of mushrooms can be harvested by carrying out management of producing mushroom.
Embodiment 5
PDA improved culture mediums are used in the present embodiment:Through 121 DEG C~125 DEG C of temperature (pressure 1.1kg/cm2~ 30 minutes 1.5kg/cm2) are sterilized containing the full powder 15g/L of wheat, sucrose 10g/L, glucose 10g/L, dusty yeast 1.2g/L, peptone 1.2g/L, magnesium sulfate 0.5g/L, dipotassium hydrogen phosphate 1g/L, the culture medium of agar 10g/L.
1) parent species make:The healthy and strong wild Lepista sordida of acquisition makes parent species, aseptically cuts fresh wild Lepista sordida fructification chooses the meat bacteria organization of cap stem intersection, moves into the vessel equipped with PDA improved culture mediums, puts Enter culture in constant incubator, cultivated 24~27 days at a temperature of 21 DEG C~24 DEG C, until mycelia covers with device.
2) expand numerous:Parent species are transferred in PDA improved culture mediums, and 24~27 days are cultivated at 21 DEG C~24 DEG C to covering with bacterium Silk.
3) original seed makes:The obtained numerous parent species of expansion are implanted into the vessel equipped with wheat compost, be placed on 20 DEG C~ It is cultivated 33~38 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, 20 DEG C~23 DEG C of perseverance is placed on It is cultivated 28~33 days under the conditions of temperature, mycelia covers with vessel and completes cultigen making.
The production method of the Primary spawn material is:By whole wheat granular or grain to expect water quality as 1:1.3 ratio is impregnated 11h impregnates and adds in 0.2% potassium dihydrogen phosphate, 0.1% magnesium sulfate simultaneously it is made fully to dissolve, boiled after immersion to no hard core but It 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 1.1kg/ of temperature cm2~1.5kg/cm2) sterilizing 2.5h, it can be aseptically inoculated with after cooling.
The cultigen compost includes the wheat and 75 parts of caragana microphylla powder that mass parts are 25 parts;The cultigen compost Production method be:By wheat to expect water quality as 1:1.3 ratio impregnates 11h, and immersion while adds in opposite wheat quality 0.2% potassium dihydrogen phosphate and 0.1% magnesium sulfate, make it fully dissolve, and are boiled after immersion to no hard core but not split, control is gone Water;To expect water quality as 1:1.2 ratio is soaked in water caragana microphylla powder, then is uniformly mixed with wheat, adjusts pH to 8, sterilizing.
5) ferment:The caragana microphylla powder branches and leaves that the sheep manure and fresh nothing pulverized in advance are gone mouldy are with 1:1 ratio adds water fully to prewet, 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~8cm Wooden stick burrow to material bottom, hole is away from 40cm, when heap temperature is raised to 65 DEG C, is kept for 24 hours, carries out first time turning, continue to punch Fermentation, heap temperature be raised to 65 DEG C keep again 24 it is small when carry out second of turning, heat up again after 2 days 65 DEG C, 12 it is small when after third time Turning, conditioning can be used to sow 60%~65% or so.
6) domesticating and cultivating:The compost fermented is layered on to the western sheep cleaned up to enclose the land face, cultigen is uniformly spread On compost, one layer of compost fermented is repaved above, then spreads last layer cultigen, and surface layer strain and compost puddle pressure It is flat, it is organized into the long unlimited curvature of the spinal column type fruiting row of bottom width 1.3m, upper width 1.2m, high 23cm or so, progress Mycelium culture 50~ 60 days.After mycelium covers with compost, in the soil of surface covering 5cm moistenings, fruiting front surface soil is sprayed water moisturizing when bleaching, temperature Degree control is at 21 DEG C~23 DEG C.40~45 days after earthing, there are No. 2 former bases of prairie Tricholomataceae, can be harvested after 9~12 days, science 3 batches of mushrooms can be adopted by carrying out management of producing mushroom.
The present invention successfully realizes the artificial domesticating cultivation of prairie Tricholomataceae 2, not only protects endangered paint face This rareness species of fragrant mushroom, and cattle and sheep excrement, the careless bits the like waste that pastoral area is made full use of to enrich, carry out cultivation prairie Tricholomataceae It No. 2, realizes the comprehensive utilization of resource, promotes emerging《Grass, poultry, bacterium》Industry recycle and sustainable development, moreover it is possible to increase The income of local herdsman.
It should be appreciated that after the dosage of above-described embodiment agents useful for same or raw material is carried out equal proportion expansion or is reduced Technical solution, it is substantially identical with above-described embodiment.
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 made some modifications or improvements, 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>University of the Inner Mongol of Inner Mongolia Autonomous Region Academy of Agricultural and Livestock Husbandry
<120>Tricholoma mongolicum new strains prairie Tricholomataceae 2 and its selection
<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 (10)

1. No. 2 strains of Tricholoma mongolicum new strains prairie Tricholomataceae, which is characterized in that its deposit number is CGMCC No.15084.
2. a kind of selection of No. 2 strains of prairie Tricholomataceae, which is characterized in that include the following steps:
1) parent species make:At 21 DEG C~24 DEG C parent species are cultivated using wild Lepista sordida fructification as material, until long on culture medium Full mycelia;
2) expand numerous:Parent species obtained by step 1) are transferred on culture medium, are cultivated at 21 DEG C~24 DEG C, until covering with mycelia;
3) original seed makes:Parent species of the step 2) expansion after numerous are moved into the vessel equipped with Primary spawn material, in 20 DEG C~23 DEG C perseverances It is cultivated 33~38 days under the conditions of temperature, until mycelia, which covers with vessel, completes original seed making;
4) cultigen makes:In vessel of the original seed implantation equipped with cultigen compost that step 3) is made, at 20 DEG C~23 DEG C Constant temperature under cultivate 28~33 days, until mycelia cover with vessel complete cultigen make.
3. according to the method described in claim 2, it is characterized in that, the step 1) tissue isolation or basidiospore collecting method Make strain:
The tissue isolation is:The Lepista sordida fructification of fresh wild is aseptically cut, cap stem is chosen and hands over Meat bacteria organization at boundary moves into the vessel equipped with culture medium, is cultivated 24~27 days in 21 DEG C~24 DEG C of constant incubator;
The basidiospore collecting method is:The Lepista sordida fructification of fresh wild is aseptically passed through with iron wire, makes its bacterium Pleat, which is hung on downwards, collects basidiospore in container, a little basidiospore of picking into test tube, add sterile water be diluted to 100 μ L containing 40~ 50 basidiospore, obtain basidiospore suspension;Basidiospore suspension is drawn to be added dropwise on culture medium, and even spread, 21 DEG C~24 DEG C Constant incubator in culture 24~27 days.
4. according to the method in claim 2 or 3, which is characterized in that the culture medium in step 1) is PDA improvement cultures Base;
Culture medium described in step 2) improves fluid nutrient medium for PDA improved culture mediums or PDA;When using PDA improvement cultures During base, cultivated 24~27 days in PDA improved culture mediums;When using PDA improvement fluid nutrient mediums, in PDA improvement liquid trainings It supports in base with 120~140r/min rotating speeds culture 13~16 days;
The formula of the PDA improved culture mediums is:Carbon source 15g/L~200g/L, sucrose 10g/L, glucose 10g/L, dusty yeast 1.2g/L, peptone 1.2g/L, magnesium sulfate 0.5g/L, dipotassium hydrogen phosphate 1g/L, agar 10g/L;
The formula of PDA improvement fluid nutrient medium is:Carbon source 15g/L~200g/L, sucrose 10g/L, glucose 10g/L, ferment Female 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 the one or more in 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 formula of the Primary spawn material in step 3) is:Entirely Wheat or grain, potassium dihydrogen phosphate and magnesium sulfate;
The quality of the potassium dihydrogen phosphate is the 0.2% of whole wheat granular or grain quality;
The quality of the magnesium sulfate is the 0.1% of whole wheat granular or grain quality;
The production method of the Primary spawn material is:By whole wheat granular or grain to expect water quality as 1:1.2~1:1.5 ratio leaching 8~12h is steeped, potassium dihydrogen phosphate and magnesium sulfate are added in while immersion, it is made fully to dissolve, is boiled after immersion to no hard core, control Water is removed, adjusts pH to 7.5~8, sterilizing.
6. according to the method described in claim 2, it is characterized in that, the cultigen compost in step 4) includes mass parts For 20~30 parts of wheats and 70~80 parts of caragana microphylla powder;
The production method of the cultigen compost is:By wheat to expect water quality as 1:1.3~1:2 ratio immersion 8~ 12h adds in the potassium dihydrogen phosphate and 0.1% magnesium sulfate of opposite wheat quality 0.2% while immersion, it is made fully to dissolve, It is boiled after immersion to no hard core but not split, water is removed in control;To expect water quality as 1:1.2 ratio is soaked in water caragana microphylla powder, then with Wheat is uniformly mixed, and adjusts pH to 7.5~8, sterilizing.
7. a kind of cultural method of prairie Tricholomataceae 2, which is characterized in that include the following steps:
(a) plant waste and excrement of animals windrow are fermented;
(b) by No. 2 cultigens of the prairie Tricholomataceae described in claim 1 uniformly material of paving after fermentation, one layer is repaved above Material after fermentation, then last layer cultigen is spread, every layer of paving 10cm~15cm of material after fermentation is thick, surface layer strain and compost Pressing is puddled, is organized into the unlimited curvature of the spinal column type fruiting of bottom width 1.3m~1.5m, upper width 1.2m~1.3m, the length of high 20cm~24cm Row, carries out Mycelium culture 50~60 days.
8. the method according to the description of claim 7 is characterized in that windrow fermentation is:By plant waste and excrement of animals By 4:6~5:5 mass ratio, which is uniformly mixed, builds up fermentation heap, ferments at clean ventilation;
One or more of the plant waste in careless knot, caragana microphylla powder branches and leaves, straw, maize straw, wheat straw.
9. the method according to the description of claim 7 is characterized in that after the long full with substance of mycelium, 4cm~6cm is covered on surface Thick rural area soil or turfy soil, fruiting front surface soil bleach, moisturizing of spraying water, and temperature is controlled at 21 DEG C~23 DEG C;40 after earthing~ 45 days, there are No. 2 former bases of prairie Tricholomataceae, keep ground moistening, can be harvested after 9~12 days.
10. according to the method described in claim 9, it is characterized in that, harvesting first batch of mushroom after surface evening is gently pressed dry 2 days, Continue management of producing mushroom after making mycelium restoration ecosystem, harvest after first batch of mushroom and regrowth hair mushroom former base occur on the 15th~18 day, 3 batches of mushrooms can be adopted altogether.
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