CN104557315B - Elegant precious mushroom liquefaction special bacteria culture medium and corresponding cultivation - Google Patents
Elegant precious mushroom liquefaction special bacteria culture medium and corresponding cultivation Download PDFInfo
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- LDXJRKWFNNFDSA-UHFFFAOYSA-N 2-(2,4,6,7-tetrahydrotriazolo[4,5-c]pyridin-5-yl)-1-[4-[2-[[3-(trifluoromethoxy)phenyl]methylamino]pyrimidin-5-yl]piperazin-1-yl]ethanone Chemical compound C1CN(CC2=NNN=C21)CC(=O)N3CCN(CC3)C4=CN=C(N=C4)NCC5=CC(=CC=C5)OC(F)(F)F LDXJRKWFNNFDSA-UHFFFAOYSA-N 0.000 description 2
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- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
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Abstract
The invention discloses a kind of elegant precious mushroom liquefaction special bacteria culture medium, it is composed of the following components in parts by weight：45 50 parts of cornstarch, 12 parts of glucose, 15 20 parts of Semen Tritici aestivi fiber element, 20 25 parts of fine bran, 1 1.5 parts of yeast extract, 1 1.5 parts of beef peptone, 23 parts of potassium dihydrogen phosphate, 1 1.5 parts of magnesium sulfate, 100 parts of water.The present invention also discloses the elegant precious mushroom liquefaction Spawn incubation method carried out using above-mentioned culture medium simultaneously, and elegant precious mushroom parent species are followed the steps below successively：Elegant precious mushroom is liquefied special bacteria culture medium autoclave sterilization, in accessing elegant precious mushroom parent species under aseptic condition in culture medium after sterilization, 23~25 DEG C of dark culturings 19~21 days；The liquefaction special solid strain of gained is diluted processing, elegant precious mushroom liquefaction strain is obtained.
Description
Technical field
The present invention relates to a kind of elegant precious mushroom liquefaction special bacteria formula and cultivation.
Background technology
Edible mushroom is China's most one of strong industry of modern agricultural development feature.Be not only because China be the world most
Big Edible Fungi and country of consumption, Edible Fungi have a large capacity and a wide range, and more importantly mushroom industry in biotechnology
Good carrier and prominent position in industrialization and agricultural sustainable development.The mushroom industry system of current China, there is a lot
The key element of world market economy is not suitable with, wherein most importantly intensive, the low degree of specialized division of labor of Industry Management, production factors
Fall behind, production technology is perfect not to the utmost, lack the support of key technology, wherein most representational is the intensive efficiently numerous of strain
Technology is educated, the main bottleneck faced into industry.The traditional three-level solid spawn generally used at present breeds technique, production efficiency
Low, cultivation cycle is long, and strain contamination rate is high, it is impossible to breaks through craftization, workshop-based poorly efficient production model, makes Large-scale enterprises and casual household
The production of hybrid seeds is in same competition platform, and this is the main contributor for causing current Edible Fungi safety, quality accident to take place frequently, and is also food
With the not good enough main cause of the bacterium performance of enterprises.It is external generally to use liquid as current in Japan, South Korea's edible fungus industrial production
Body strain technology, and China lacks the successful experience of large-scale production and technology in this field, so strengthening strain in scale
The research and development applied in production, using modern biotechnology science and technology and biotechnology, realize that efficiently breeding for strain (bacterium bag) has been compeled
In the eyebrows and eyelashes.
Traditional three-level solid spawn, which breeds technique, 3 steps (test tube stock-bottled original seed-bottled cultigen)：1st grade is examination
Pipe parent species, are formulated as (200 grams of fresh potato of peeling, 20 grams of glucose, agar 20, water 1000ml), incubation time 7- based on PDA
15 days (different according to mushroom kind, elegant precious mushroom is generally 7-10), 1 parent species 5 bottles of original seed of switching.2nd grade is bottled original seed
(750ml), formula based on wood chip, bran mass (such as mushroom original seed be formulated thin wood chip 78%, wheat bran 20%, white sugar 1%,
Gypsum 1%, water content 60%), incubation time 45-60 days (different according to mushroom kind), the switchable 50 bottles of cultigens of 1 bottle of original seed.3rd
Level is bottled cultigen (750ml), and formula is based on wood chip, bran mass (with original seed formula, 35-45 days (roots of incubation time
It is different according to mushroom kind), 1 bottle of switchable 20 fruiting bag of cultigen (600 grams of composts of weight in wet base), every bag of fruiting bag needs solid spawn
30 grams.Above-mentioned 3 grades of kind technique whole cultivation cycle 87-120 days.Strain obtained by this method is according to conventional inoculation fruiting bag method
Contamination rate is generally 5-10% when cultivating fresh mushroom.Remarks explanation：Above-mentioned solid spawn is in original seed (the 2nd grade), cultigen (3rd level)
Stage is long due to incubation time, and culture environment is poor, adds and seals lack of standardization, can be contaminated in the finished product strain surface stealth of the full mycelia of hair
Bacterium, after the strain of this subclinical infection is vaccinated, can become dominant pollution (now miscellaneous bacteria is faster than hypha of edible fungus growth).Institute
It is very risky with existing solid spawn, and be difficult to avoid.
Existing edible fungus liquid fermented bacterium (also referred to as submerged fermentation) technological process is 3 steps (test tube stock-triangle at present
Bottle shaker fermentation liquid original seed-fermentation tank submerged fermentation liquid cultivation seed)：1st grade is parent species, (facture is the same) incubation time
7-15 days (different according to mushroom kind, elegant precious mushroom is generally 7-10 days), 1 parent species 2-3 bottles of triangular flask shaker fermentation liquid of switching are former
Plant (200ml).2nd grade be triangular flask shaker fermentation liquid original seed (200ml nutrient solutions are put into 500ml triangular flasks), formula with
Remove the peel based on fresh potato, glucose, yeast extract, peptone etc., 7-10 days shaking table culture time (different according to mushroom kind), 1 bottle
The switchable 2000ml of liquid original seed 3 grades of fermentation tank culture liquid (10% inoculum concentration).3rd level is planted for fermentation tank submerged fermentation liquid
Cultivate, formula needs special submerged fermentation tank or fermentation system, incubation time 3- with triangular flask shaking table liquid pedigree seed culture medium, fermentation
5 days (different according to mushroom kind), switchable 100 fruiting bags of 1 liter of liquid spawn.Aforesaid liquid strain breeds technique whole process culture week
17-25 days phase.Contamination rate is generally 2-5% when strain obtained by this method cultivates fresh mushroom according to conventional inoculation fruiting bag method.
Remarks explanation：Strain purity is high on liquid fermentation strain technology theory, and the contamination rate of large scale fermentation production is between 1%-5%
(in 3 grades of fermentation tanks), add microbiological contamination (based on bacillary) often phase after fermentation, conventional is difficult to find in X -ray inspection X, causes to use
The inoculation of liquid fermentation strain produces the production accident of high-volume fruiting bag pollution, and lesson is painful.In addition, liquid spawn is due to culture
Need to use the nutritional ingredients such as the organic nitrogen of high concentration and sugar source, these component residues in bacterium solution by access fruiting bag, due to operation
When unavoidably there is miscellaneous bacteria to bring into, so residual nutrition just become the hotbed of miscellaneous bacteria, add fruiting bag inoculation after dirt
Contaminate risk.This is that liquid fermentation strain is unable to large-scale application in the subject matter of Edible Fungi.
The content of the invention
The technical problem to be solved in the present invention is to provide a kind of elegant precious mushroom that cultivation cycle is short, strain quality is high liquefaction is special
Bacterium culture medium and corresponding cultivation.
In order to solve the above-mentioned technical problem, the present invention provides a kind of elegant precious mushroom liquefaction special bacteria culture medium, and it is by following
The composition composition of parts by weight：
As the improvement of the elegant precious mushroom liquefaction special bacteria culture medium of the present invention, it is composed of the following components in parts by weight：
The preparation method of the present invention also simultaneously there is provided above-mentioned culture medium, comprises the following steps：
Cornstarch, fine bran (wheat bran), Semen Tritici aestivi fiber element is well mixed, obtain dispensing I；
Glucose, yeast extract, beef peptone, potassium dihydrogen phosphate, magnesium sulfate are added to the water, are sufficiently stirred for (making Portugal
Grape sugar, yeast extract, beef peptone, potassium dihydrogen phosphate, magnesium sulfate are dissolved in the water), obtain dispensing II；
After dispensing I and dispensing II are sufficiently mixed, elegant precious mushroom liquefaction special bacteria culture medium is obtained.
The elegant precious mushroom liquefaction Spawn incubation method of the invention also provided simultaneously using the progress of above-mentioned culture medium, elegant precious mushroom is female
Plant and follow the steps below successively：
1), strain makes：
Elegant precious mushroom liquefaction special bacteria culture medium is fitted into blake bottle, microporous barrier ventilating cover, autoclave sterilization is covered
(autoclave sterilization being carried out i.e. under 121 DEG C, 0.11Mpa 90 minutes), obtains sterilizing wild Oryza species；
Remarks explanation：Shanghai Song Tuo Industrial Co., Ltd.s ZP14-200 gas-permeable flasks are can select, it is saturating that it carries microporous barrier
Gas lid.
According to every 200g elegant precious mushroom liquefaction special bacteria culture medium correspondence 4.8~5.2g (preferably 5g) elegant precious mushroom parent species
Inoculum concentration, in accessing elegant precious mushroom parent species under aseptic condition in culture medium after sterilization, 23~25 DEG C (preferably 24 DEG C) are dark
19~21 days (preferable 20 days) are cultivated, must liquefy special solid strain；Now mycelia sends out full full bottle；
Remarks explanation：
1st, elegant precious mushroom parent species can be obtained according to routine techniques；
2nd, can be to step 1 in order to prove the purity and kind property of liquefaction special bacteria obtained by the present invention) obtained by liquefaction it is special
Solid spawn carries out following examine：
1., purity check, including mould is examined, bacteriologic test, using microscopic examination and Conventional bacteria test stone；
The detection species of mould is：Trichoderma, mould；
The detection species of bacterium is：Bacillus subtilis.
2., vitality test, using ttc methods；
3., organoleptic examination：Including being formed and (that is, being formed without immature mushroom flower bud) without former base, cultivate bottle cap complete seal, mark
Sign correct etc..
2), prepared by liquefaction strain：
Special solid strain will be liquefied aseptically first through high speed homogenization (under 8000~10000 revs/min of rotating speed
Homogeneous 1~1.5 minute), then according to 1:100 dilution factor (that is, adds 99 in the liquefaction special solid strain of 1 parts by weight
The sterilized water of parts by weight；Remarks explanation：Belong to two grades of dilutions) dilute, the gains (pH value is 6.5-7.0 naturally) after dilution exist
In thinning tank in temperature be 20-23 DEG C, ventilation ratio be 1：Liquefied under conditions of 0.4 (v/v/min) 4~6 minutes and (be preferably 5 points
Clock)；Obtain elegant precious mushroom liquefaction strain.
Elegant precious mushroom liquefaction strain obtained by the present invention, liquefaction strain mycelia fragment is more, and good dispersion degree is (through 400 power microscopes
Detection, there is hyphal cell 100 in each visual field), it is energetic that (TTC- dehydrogenases reducing process is detected：0.2g testing samples+2ml
2h is dyed in 0.5%TTC-PBS (PH=8.0), 40 DEG C of waters bath with thermostatic control, adds 5ml absolute ethyl alcohols room temperature extraction 1h, and extract is inhaled
Light value OD485 values, remarks：0.45-0.55 is qualified), in 30ml/ bottles of inoculum concentration, satisfied bacterium germination effect can be obtained.Remarks
Explanation：30ml/ bottles of inoculum concentration is directed to access 30ml liquefaction bacterium solution (that is, this hair in each elegant precious mushroom fruiting bag to be seeded
The elegant precious mushroom liquefaction strain of bright gained), gained is elegant precious mushroom fresh mushroom product after cultivation.
The elegant precious mushroom of the present invention liquefies Spawn incubation method compared with traditional three-level solid spawn breeds technique, with following skill
Art advantage：
The technique of the present invention is 2 big steps (test tube stock-bottled liquefaction Special seed), and the 1st step is that (facture is with above-mentioned for parent species
Prior art)；2nd step is liquefaction special bacteria (200ml), and then incubation time 20 days or so need to only be diluted 5 points of liquefaction
Clock or so.Therefore, technique whole cultivation cycle 27-30 days.Every bottle liquefaction special solid strain (200 grams) through liquefaction, into
20 liters of bacterium solutions, switchable more than 600 fruiting bags (every bag connects 30ml bacterium solutions), every bag of fruiting bag needs 0.3 gram of solid spawn.Inoculation
Efficiency is 100 times of conventional solid strain.
The elegant precious mushroom liquefaction Spawn incubation method of the present invention is distinguished with the maximum with existing liquid fermentation：Using 2 step kinds
Source cultivation, the cycle is short, technique is simple；Simultaneously because sowing quantity is the 1/100 of solid spawn, so can be to every before strain use
Bottle introduces a collection carries out the inspection of purity, vigor and kind property, so that (liquid spawn is online using not accomplishing preceding for the quality of standard bacteria introduces a collection
Examine, dangerous)；3rd be the present invention liquefaction strain fruiting bag when without culture (liquid spawn need 3-5 days fermentation trains
Support), using simple, low equipment investment, more crucially bacterium solution is all without culture medium (only sterilized water) containing only pure mycelia
Prevent bacterium solution after inoculation and, with secondary contact scar caused by abundant nutrition, improve purity (this of inoculation yield rate and fruiting bag
Do not accomplish in solid three-class strain and liquid fermentation strain).
Liquefaction special solid strain obtained by the present invention can preserve 30 under 5 DEG C of environment (the dry refrigerator of cleaning)
My god, and aforesaid liquid fermented bacterium can not be preserved, and be used immediately after the completion of fermentation.
Beneficial effects of the present invention are as follows：
The elegant precious mushroom liquefaction special bacteria and liquefaction inoculation technique of the present invention, due to good dispersion, mycelia is energetic, can be more
The fast full cultivating bag of hair, the dark culturing time of liquefaction strain purseful time (that is, step 1)) shorten 1/3 or so than solid spawn；
And pollution rate is low, bacterium bag weight-loss ratio is low after purseful, and mycelia is energetic.Yield and quality is brighter than the advantage being inoculated with using solid spawn
It is aobvious.Both the cultivation of introduces a collection had been solved, technique is simplified, inoculation consumption has been reduced, it is even more important that it efficiently solves bacterium
Silk liquefy culture medium in size it is uneven the problem of, improve the quality and vigor of bacterium solution, and met quick inoculation will
Ask, it is overall to reduce inoculation link cost more than 50%.
In summary, elegant precious mushroom strain quality responses key technology, bacterium bottle (bag) scale are efficiently bred through inventor
New industrial research has been carried out, a kind of elegant precious mushroom liquefaction special bacteria and corresponding culture technique has been invented, has been produced with the invention
Liquefying, the special solid strain cycle is short, and mycelial growth is vigorous, energetic, and production cost is low, and culture bottle or cultivation are used for after liquefaction
Bag is trained, multiple spot bacterium germination after inoculation, mycelial growth is rapid, and high yield rate, inoculation efficiency is 50 times -100 times of conventional solid strain,
Be it is a kind of efficiently, stably, reliable strain pattern, particularly suitable elegant precious mushroom batch production intensive production mode pattern.
Embodiment
Embodiment 1, a kind of elegant precious mushroom liquefaction special bacteria culture medium, it is composed of the following components in parts by weight：
The preparation method of above-mentioned culture medium is to carry out following steps successively：
In rustless steel container, cornstarch, fine bran (wheat bran), Semen Tritici aestivi fiber element are mixed in dry conditions
Uniformly, dispensing I is obtained；
Glucose, yeast extract, beef peptone, potassium dihydrogen phosphate, magnesium sulfate are added to the water, are sufficiently stirred for (making Portugal
Grape sugar, yeast extract, beef peptone, potassium dihydrogen phosphate, magnesium sulfate are dissolved in the water), obtain dispensing II；
After dispensing I and dispensing II are sufficiently mixed, elegant precious mushroom liquefaction special bacteria culture medium is obtained.
Remarks explanation：Elegant precious mushroom liquefaction special bacteria culture medium is now with the current.
Embodiment 2, the elegant precious mushroom liquefaction Spawn incubation method carried out using the culture medium of the gained of embodiment 1, elegant precious mushroom is female
Plant and follow the steps below successively：
1), strain makes：
The elegant precious mushroom now prepared liquefaction special bacteria culture medium 200g is dispensed into 200ml special culture bottle immediately,
Microporous barrier ventilating cover (from Shanghai Song Tuo Industrial Co., Ltd.s ZP14-200 gas-permeable flasks) is covered, in 121 DEG C, 0.11Mpa
Lower progress autoclave sterilization 90 minutes；Must be sterilized wild Oryza species；
In elegant precious mushroom parent species 5 grams of (solid parent species), 24 DEG C of dark are accessed under aseptic condition in above-mentioned sterilizing wild Oryza species
Culture 20 days, must liquefy special solid strain；Now mycelia sends out full full bottle.
Through examining, purity is 100%；
After testing, the mould such as trichoderma, mould is not measured；
After testing, the bacterium such as bacillus subtilis is not measured；
The vigor data obtained by the detection of ttc methods are used for OD485 values 0.49；
Organoleptic examination result is：Formed without former base, cultivate bottle cap complete seal, label is correct.
2), prepared by liquefaction strain：
Liquefaction special solid strain aseptically first (is liquefied through high speed homogenization in homogeneous under 10,000 revs/min of rotating speed
1 minute), then according to 1:100 dilution factor (belonging to two grades of dilutions) dilution, the gains (pH6.5, mycelium) after dilution exist
In thinning tank in temperature be 20 DEG C, ventilation ratio be 1：Liquefied 5 minutes under conditions of 0.4 (v/v/min)；Obtain elegant precious mushroom liquefaction bacterium
Kind.
Experiment 1, the elegant precious mushroom of the present invention liquefied strain and existing solid spawn, liquid fermentation strain are according to conventional
It is inoculated with fruiting bag method and cultivates elegant precious mushroom fresh mushroom, acquired results contrast such as table 1 below：
Table 1, elegant precious mushroom liquefaction strain are compared with solid spawn, liquid fermentation strain inoculation fruiting bag effect
From the data comparison of above-mentioned table 1, it is known that, elegant precious mushroom liquefaction strain of the present invention is far superior to consolidating obtained by prior art
Body strain and liquid fermentation strain.
Comparative example 1-1,
The formula of elegant precious mushroom liquefaction special bacteria culture medium in embodiment 1 is made into following change：
Cancel the use of 20 parts of Semen Tritici aestivi fiber element, and cornstarch is increased to 66 parts by 46 parts accordingly；Remaining is equal
In embodiment 1.
Comparative example 1-2,
The formula of elegant precious mushroom liquefaction special bacteria culture medium in embodiment 1 is made into following change：
" 20 parts of Semen Tritici aestivi fiber element " is made into " 20 parts of lignin ", remaining is equal to embodiment 1.
Comparative example 1-3,
The formula of elegant precious mushroom liquefaction special bacteria culture medium in embodiment 1 is made into following change：
" 20 parts of Semen Tritici aestivi fiber element " is made into " 20 parts of wood chip ", remaining is equal to embodiment 1.
It is used for elegant precious mushroom liquefaction Spawn incubation described in embodiment 2 using above-mentioned comparative example 1-1~comparative example 1-3 culture medium
Method, in order that step 1) realize that time of the mycelia hair completely needed for full bottle, and the elegant precious mushroom of gained liquefy strain according to above-mentioned reality
The conventional inoculation fruiting bag method for testing 1 cultivates elegant precious mushroom fresh mushroom, and acquired results are contrasted as described in Table 2：
Table 2
| Project | Embodiment | Comparative example 1-1 | Comparative example 1-2 | Comparative example 1-3 |
| Strain liquefied fraction % | 100 | 100 | 90 | 66 |
| Pollution rate % | 0 | 12 | 10 | 15 |
| Mycelia purseful time d | 20 | 35 | 31 | 29 |
| Weight-loss ratio % | 1 | 2 | 2 | 2 |
| Mycelia bulk properties | It is dense | Typically | Typically | It is denseer |
| The head damp mushroom formation time (my god) | 25 | 39 | 33 | 38 |
| G/ bags of per unit area yield | 259 | 211 | 225 | 203 |
| Biological efficiency % | 86 | 70 | 75 | 68 |
Comparative example 2-1,
By " the strain preparation of 2), liquefying in embodiment 2：" make following change：
By dilution factor by " 1:100 " make " 1 into:1”；Remaining is equal to embodiment 2.
As a result it is：Strain can not carry out follow-up liquefaction, into viscose shape.The strain that liquefies fails！
Comparative example 2-2,
By " the strain preparation of 2) liquefying in embodiment 2：" make following change：
By dilution factor by " 1:100 " make " 1 into:200”；Remaining is equal to embodiment 2.
Comparative example 2-3, by embodiment 2 " 2) liquefy strain prepare：" make following change：
By ventilation ratio by " 1：0.4 " makes " 1 into：0.2”；Remaining is equal to embodiment 2.
Comparative example 2-4, by embodiment 2 " 2) liquefy strain prepare：" make following change：
By ventilation ratio by " 1：0.4 " makes " 1 into：0.6”；Remaining is equal to embodiment 2.
Comparative example 3-1,
" 1), strain makes " in embodiment 2 is made into following change：
Cultivation temperature is made into " 26 DEG C " by " 24 DEG C "；Remaining is equal to embodiment 2.
Comparative example 3-2,
" 1), strain makes " in embodiment 2 is made into following change：
Cultivation temperature is made into " 22 DEG C " by " 24 DEG C "；Remaining is equal to embodiment 2.
Elegant precious mushroom liquefaction strain obtained by above-mentioned comparative example 2-2~comparative example 3-2 is connect according to the conventional of above-mentioned experiment 1
Plant fruiting bag method and cultivate elegant precious mushroom fresh mushroom, acquired results are contrasted as described in Table 3：
Table 3
Remarks explanation：
By the step 1 of embodiment 2) obtained by liquefaction special solid strain in being preserved under 5 DEG C of environment (cleaning dry refrigerator)
30 days, then proceed by follow-up step 2).
It is fresh that the elegant precious mushroom liquefaction strain of gained cultivates elegant precious mushroom according to the conventional inoculation fruiting bag method described in above-mentioned experiment 1
Mushroom, acquired results are substantially with the result obtained by " liquefaction strain (present invention) " in table 1 (gap is no more than 5%).
Finally, in addition it is also necessary to it is noted that listed above is only several specific embodiments of the invention.Obviously, this hair
It is bright to be not limited to above example, there can also be many deformations.One of ordinary skill in the art can be from present disclosure
All deformations for directly exporting or associating, are considered as protection scope of the present invention.
Claims (2)
- The special bacteria culture medium 1. elegant precious mushroom liquefies, it is characterized in that being composed of the following components in parts by weight：
- The Spawn incubation method 2. the elegant precious mushroom carried out using the culture medium described in claim 1 is liquefied, it is characterized in that：Elegant precious mushroom is female Plant and follow the steps below successively：1), strain makes：Elegant precious mushroom liquefaction special bacteria culture medium is fitted into blake bottle, microporous barrier ventilating cover is covered, autoclave sterilization must go out Bacterium wild Oryza species；According to the inoculum concentration of every 200g elegant precious mushroom liquefaction special bacteria culture medium correspondence 4.8~5.2g elegant precious mushroom parent species, Yu Wu Access elegant precious mushroom parent species under the conditions of bacterium in culture medium after sterilization, 23~25 DEG C of dark culturings 19~21 days must liquefy special solid Body strain；2), prepared by liquefaction strain：Special solid strain will be liquefied aseptically first through high speed homogenization, then according to 1:100 dilution, dilution Gains afterwards in thinning tank in temperature be 20-23 DEG C, ventilation ratio be 1：Liquefied 4~6 points under conditions of 0.4 (v/v/min) Clock；Obtain elegant precious mushroom liquefaction strain.
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