CN104557211B - Mushroom liquefaction special bacteria culture medium and corresponding cultivation - Google Patents
Mushroom liquefaction special bacteria culture medium and corresponding cultivation Download PDFInfo
- Publication number
- CN104557211B CN104557211B CN201510028938.9A CN201510028938A CN104557211B CN 104557211 B CN104557211 B CN 104557211B CN 201510028938 A CN201510028938 A CN 201510028938A CN 104557211 B CN104557211 B CN 104557211B
- Authority
- CN
- China
- Prior art keywords
- mushroom
- culture medium
- parts
- strain
- liquefaction
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 235000001674 Agaricus brunnescens Nutrition 0.000 title claims abstract description 82
- 241000894006 Bacteria Species 0.000 title claims abstract description 54
- 239000001963 growth medium Substances 0.000 title claims abstract description 36
- 239000007787 solid Substances 0.000 claims abstract description 27
- 241000894007 species Species 0.000 claims abstract description 18
- 238000011534 incubation Methods 0.000 claims abstract description 13
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims abstract description 12
- 230000001954 sterilising effect Effects 0.000 claims abstract description 10
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 10
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims abstract description 8
- 238000004659 sterilization and disinfection Methods 0.000 claims abstract description 8
- 239000001888 Peptone Substances 0.000 claims abstract description 7
- 108010080698 Peptones Proteins 0.000 claims abstract description 7
- 229940041514 candida albicans extract Drugs 0.000 claims abstract description 7
- 239000000835 fiber Substances 0.000 claims abstract description 7
- 235000019319 peptone Nutrition 0.000 claims abstract description 7
- 210000000582 semen Anatomy 0.000 claims abstract description 7
- 239000012138 yeast extract Substances 0.000 claims abstract description 7
- 235000015278 beef Nutrition 0.000 claims abstract description 6
- 239000008103 glucose Substances 0.000 claims abstract description 6
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims abstract description 6
- 235000019341 magnesium sulphate Nutrition 0.000 claims abstract description 6
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims abstract description 6
- 235000019796 monopotassium phosphate Nutrition 0.000 claims abstract description 6
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 claims abstract description 6
- 229920002261 Corn starch Polymers 0.000 claims abstract description 5
- 239000008120 corn starch Substances 0.000 claims abstract description 5
- 229940099112 cornstarch Drugs 0.000 claims abstract description 5
- 239000010440 gypsum Substances 0.000 claims abstract description 5
- 229910052602 gypsum Inorganic materials 0.000 claims abstract description 5
- 238000012258 culturing Methods 0.000 claims abstract description 4
- 238000010790 dilution Methods 0.000 claims description 11
- 239000012895 dilution Substances 0.000 claims description 11
- 238000002360 preparation method Methods 0.000 claims description 5
- 238000009423 ventilation Methods 0.000 claims description 5
- 241000209094 Oryza Species 0.000 claims description 4
- 230000004888 barrier function Effects 0.000 claims description 4
- 239000002054 inoculum Substances 0.000 claims description 4
- 238000000265 homogenisation Methods 0.000 claims description 3
- 238000000034 method Methods 0.000 description 27
- 238000000855 fermentation Methods 0.000 description 24
- 230000004151 fermentation Effects 0.000 description 23
- 239000007788 liquid Substances 0.000 description 23
- 230000000052 comparative effect Effects 0.000 description 16
- 238000011081 inoculation Methods 0.000 description 16
- 238000012360 testing method Methods 0.000 description 11
- 238000005516 engineering process Methods 0.000 description 10
- 238000004519 manufacturing process Methods 0.000 description 10
- 230000008859 change Effects 0.000 description 9
- 239000000243 solution Substances 0.000 description 8
- 241000233866 Fungi Species 0.000 description 7
- 238000011109 contamination Methods 0.000 description 4
- 210000004209 hair Anatomy 0.000 description 4
- 239000002023 wood Substances 0.000 description 4
- VZSRBBMJRBPUNF-UHFFFAOYSA-N 2-(2,3-dihydro-1H-inden-2-ylamino)-N-[3-oxo-3-(2,4,6,7-tetrahydrotriazolo[4,5-c]pyridin-5-yl)propyl]pyrimidine-5-carboxamide Chemical compound C1C(CC2=CC=CC=C12)NC1=NC=C(C=N1)C(=O)NCCC(N1CC2=C(CC1)NN=N2)=O VZSRBBMJRBPUNF-UHFFFAOYSA-N 0.000 description 3
- 241000196324 Embryophyta Species 0.000 description 3
- NIPNSKYNPDTRPC-UHFFFAOYSA-N N-[2-oxo-2-(2,4,6,7-tetrahydrotriazolo[4,5-c]pyridin-5-yl)ethyl]-2-[[3-(trifluoromethoxy)phenyl]methylamino]pyrimidine-5-carboxamide Chemical compound O=C(CNC(=O)C=1C=NC(=NC=1)NCC1=CC(=CC=C1)OC(F)(F)F)N1CC2=C(CC1)NN=N2 NIPNSKYNPDTRPC-UHFFFAOYSA-N 0.000 description 3
- 238000001514 detection method Methods 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 238000007689 inspection Methods 0.000 description 3
- 235000016709 nutrition Nutrition 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 235000015099 wheat brans Nutrition 0.000 description 3
- 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
- YLZOPXRUQYQQID-UHFFFAOYSA-N 3-(2,4,6,7-tetrahydrotriazolo[4,5-c]pyridin-5-yl)-1-[4-[2-[[3-(trifluoromethoxy)phenyl]methylamino]pyrimidin-5-yl]piperazin-1-yl]propan-1-one Chemical compound N1N=NC=2CN(CCC=21)CCC(=O)N1CCN(CC1)C=1C=NC(=NC=1)NCC1=CC(=CC=C1)OC(F)(F)F YLZOPXRUQYQQID-UHFFFAOYSA-N 0.000 description 2
- 244000063299 Bacillus subtilis Species 0.000 description 2
- 235000014469 Bacillus subtilis Nutrition 0.000 description 2
- AFCARXCZXQIEQB-UHFFFAOYSA-N N-[3-oxo-3-(2,4,6,7-tetrahydrotriazolo[4,5-c]pyridin-5-yl)propyl]-2-[[3-(trifluoromethoxy)phenyl]methylamino]pyrimidine-5-carboxamide Chemical compound O=C(CCNC(=O)C=1C=NC(=NC=1)NCC1=CC(=CC=C1)OC(F)(F)F)N1CC2=C(CC1)NN=N2 AFCARXCZXQIEQB-UHFFFAOYSA-N 0.000 description 2
- 244000061456 Solanum tuberosum Species 0.000 description 2
- 235000002595 Solanum tuberosum Nutrition 0.000 description 2
- 241000223259 Trichoderma Species 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 238000004140 cleaning Methods 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000018109 developmental process Effects 0.000 description 2
- 239000006185 dispersion Substances 0.000 description 2
- 230000035784 germination Effects 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 230000035764 nutrition Effects 0.000 description 2
- 208000016261 weight loss Diseases 0.000 description 2
- 230000004580 weight loss Effects 0.000 description 2
- HMUNWXXNJPVALC-UHFFFAOYSA-N 1-[4-[2-(2,3-dihydro-1H-inden-2-ylamino)pyrimidin-5-yl]piperazin-1-yl]-2-(2,4,6,7-tetrahydrotriazolo[4,5-c]pyridin-5-yl)ethanone Chemical compound C1C(CC2=CC=CC=C12)NC1=NC=C(C=N1)N1CCN(CC1)C(CN1CC2=C(CC1)NN=N2)=O HMUNWXXNJPVALC-UHFFFAOYSA-N 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- 208000031504 Asymptomatic Infections Diseases 0.000 description 1
- 108020005199 Dehydrogenases Proteins 0.000 description 1
- 229920000297 Rayon Polymers 0.000 description 1
- 229910000831 Steel Inorganic materials 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- 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
- 239000008272 agar Substances 0.000 description 1
- 230000000721 bacterilogical effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 239000002361 compost Substances 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 235000019441 ethanol Nutrition 0.000 description 1
- 125000005909 ethyl alcohol group Chemical group 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 210000004709 eyebrow Anatomy 0.000 description 1
- 210000000720 eyelash Anatomy 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 210000003128 head Anatomy 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 238000011031 large-scale manufacturing process Methods 0.000 description 1
- 229920005610 lignin Polymers 0.000 description 1
- 238000009630 liquid culture Methods 0.000 description 1
- 238000011169 microbiological contamination Methods 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 125000001477 organic nitrogen group Chemical group 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 238000009738 saturating Methods 0.000 description 1
- 231100000241 scar Toxicity 0.000 description 1
- 238000011218 seed culture Methods 0.000 description 1
- 238000009331 sowing Methods 0.000 description 1
- 239000010959 steel Substances 0.000 description 1
- 239000004575 stone Substances 0.000 description 1
- 238000005728 strengthening Methods 0.000 description 1
- 238000012549 training Methods 0.000 description 1
- 230000000007 visual effect Effects 0.000 description 1
- 239000003643 water by type Substances 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05B—PHOSPHATIC FERTILISERS
- C05B7/00—Fertilisers based essentially on alkali or ammonium orthophosphates
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G18/00—Cultivation of mushrooms
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Mycology (AREA)
- Environmental Sciences (AREA)
- Mushroom Cultivation (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a kind of mushroom liquefaction special bacteria culture medium, it is composed of the following components in parts by weight：40 50 parts of cornstarch, 12 parts of glucose, 15 20 parts of Semen Tritici aestivi fiber element, 15 23 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, 2 parts of gypsum, 100 parts of water.The present invention further simultaneously discloses the mushroom liquefaction Spawn incubation method carried out using above-mentioned culture medium, follows the steps below successively：Mushroom is liquefied special bacteria culture medium autoclave sterilization, in accessing mushroom parent species under aseptic condition in culture medium after sterilization, 22~24 DEG C of dark culturings 25~26 days；The liquefaction special solid strain of gained is diluted processing, mushroom liquefaction strain is obtained.
Description
Technical field
The present invention relates to a kind of 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, mushroom is generally 12-15 days), 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, mushroom is generally 12-15 days), 1 parent species 2-3 bottles of triangular flask shaker fermentation liquid original seed of switching
(200ml).2nd grade is triangular flask shaker fermentation liquid original seed (200ml nutrient solutions are put into 500ml triangular flasks), is formulated to go
Based on skin fresh potato, glucose, yeast extract, peptone etc., 7-10 days shaking table culture time (different according to mushroom kind), 1 bottle of liquid
The switchable 2000ml of body original seed 3 grades of fermentation tank culture liquid (10% inoculum concentration).3rd level is fermentation tank submerged fermentation liquid culture
Kind, formula needs special submerged fermentation tank or fermentation system, incubation time 3-5 with triangular flask shaking table liquid pedigree seed culture medium, fermentation
My god (different according to mushroom kind), switchable 100 fruiting bags of 1 liter of liquid spawn.Aforesaid liquid strain breeds technique whole process culture week
22-30 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 mushroom that cultivation cycle is short, strain quality is high liquefaction special bacterium
Plant culture medium and corresponding cultivation.
In order to solve the above-mentioned technical problem, the present invention provides a kind of mushroom liquefaction special bacteria culture medium, and it is by following heavy
Measure the composition composition of part：
As the improvement of the 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, gypsum are well mixed, 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, mushroom liquefaction special bacteria culture medium is obtained.
The present invention also simultaneously there is provided using above-mentioned culture medium carry out mushroom liquefaction Spawn incubation method, by mushroom parent species according to
It is secondary to follow the steps below：
1), strain makes：
Mushroom liquefaction special bacteria culture medium is fitted into blake bottle, microporous barrier ventilating cover is covered, autoclave sterilization is (i.e.
Autoclave sterilization is carried out under 121 DEG C, 0.11Mpa 90 minutes), obtain 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.
Connecing for 4.8~5.2g (preferably 5g) mushroom parent species is corresponded to according to every 200g mushroom liquefaction special bacteria culture medium
Kind of amount, in accessing mushroom parent species, 22~24 DEG C of (preferably 23 DEG C) dark culturings 25 under aseptic condition in culture medium after sterilization
~26 days (preferably 25 days), must liquefy special solid strain；Now mycelia sends out full full bottle；
Remarks explanation：
1st, 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
Strain 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 (being formed without immature mushroom flower bud) without former base, cultivate bottle cap complete seal, label
It is 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 mushroom liquefaction strain.
Mushroom liquefaction strain obtained by the present invention, liquefaction strain mycelia fragment is more, and good dispersion degree (is examined through 400 power microscopes
Survey, 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：0.50-0.55 is qualified), in 30ml/ bottles of inoculum concentration, satisfied bacterium germination effect can be obtained.Remarks
Explanation：The liquefaction bacterium solution that 30ml/ bottles of inoculum concentration is directed to access 30ml in each mushroom fruiting bag to be seeded is (that is, of the invention
The mushroom liquefaction special bacteria of gained), gained is mushroom fresh mushroom product after cultivation.
The mushroom of the present invention liquefies Spawn incubation method compared with traditional three-level solid spawn breeds technique, with following technology
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 25 days or so need to only be diluted 5 points of liquefaction
Clock or so.Therefore, technique whole cultivation cycle 37-40 days.Every bottle of liquefaction special bacteria (200 grams) is through liquefaction, into 20 liters
Bacterium solution, 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
It is 100 times of conventional solid strain.
The mushroom liquefaction Spawn incubation method of the present invention and the maximum of existing liquid fermentation are distinguished：Trained using 2 step introduces a collections
The method of supporting, the cycle is short, technique is simple；Simultaneously because sowing quantity is the 1/100 of solid spawn, so can be to every bottle of kind before strain use
Source carries out the inspection of purity, vigor and kind property, so that (liquid spawn is not using accomplishing online inspection preceding for the quality of standard bacteria introduces a collection
Test, dangerous)；3rd is that without culture, (liquid spawn needs ferment for 3-5 days when liquefaction strain of the invention is inoculated with fruiting bag
Culture), using simple, low equipment investment, more crucially bacterium solution is containing only pure mycelia, without culture medium (only sterilized water), institute
There is bacterium solution after having prevented to be inoculated with secondary contact scar caused by abundant nutrition, to improve inoculation yield rate and the purity of fruiting bag
(this does 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 mushroom liquefaction special bacteria and liquefaction inoculation technique of the present invention, due to good dispersion, mycelia is energetic, can be faster
The 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, and 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, inventor efficiently breeds progress to mushroom strain quality responses key technology, bacterium bottle (bag) scale
New industrial research, has invented a kind of mushroom liquefaction special bacteria culture medium and corresponding culture technique, is produced with the invention
The special bacteria cycle is short, and mycelial growth is vigorous, energetic, and production cost is low, and culture bottle or cultivating bag, inoculation are used for after liquefaction
Multiple spot bacterium germination afterwards, mycelial growth is rapid, and high yield rate, inoculation efficiency is 50 times -100 times of conventional solid strain, is a kind of high
Effect, stably, reliable strain pattern, particularly suitable mushroom plant intensive production mode pattern.
Embodiment
Embodiment 1, a kind of 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, by cornstarch, fine bran (wheat bran), Semen Tritici aestivi fiber element, gypsum in drying condition
It is lower 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, mushroom liquefaction special bacteria culture medium is obtained.
Remarks explanation：Mushroom liquefaction special bacteria culture medium is now with the current.
Embodiment 2, the mushroom carried out using the culture medium of the gained of embodiment 1 are liquefied Spawn incubation method, by mushroom parent species according to
It is secondary to follow the steps below：
1), strain makes：
The mushroom now prepared liquefaction special bacteria culture medium 200g is dispensed into 200ml special culture bottle immediately, covered
Good microporous barrier ventilating cover (from Shanghai Song Tuo Industrial Co., Ltd.s ZP14-200 gas-permeable flasks), under 121 DEG C, 0.11Mpa
Carry out autoclave sterilization 90 minutes；Must be sterilized wild Oryza species；
In accessing 5 grams of parent species of mushroom (solid parent species) under aseptic condition in above-mentioned sterilizing wild Oryza species, 23 DEG C of dark are trained
Support 25 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 0.52；
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 mushroom liquefaction strain.
Experiment 1, strain and existing solid spawn, liquid fermentation strain connecing according to routine that the mushroom of the present invention liquefies
Plant fruiting bag method and cultivate mushroom fresh mushroom, acquired results contrast is as follows：
Table 1, 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, mushroom liquefaction strain of the present invention is far superior to the solid obtained by prior art
Strain and liquid fermentation strain.
Comparative example 1-1,
The formula of 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 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 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 mushroom liquefaction special bacteria training described in embodiment 2 using above-mentioned comparative example 1-1~comparative example 1-3 culture medium
The method of supporting, in order that step 1) realize that time of the mycelia hair completely needed for full bottle, and the mushroom of gained liquefy strain according to above-mentioned reality
The conventional inoculation fruiting bag method for testing 1 cultivates 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 | 65 |
| Pollution rate % | 0 | 10 | 13 | 15 |
| Mycelia purseful time d | 25 | 35 | 28 | 30 |
| 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) | 75 | 86 | 88 | 91 |
| G/ bags of per unit area yield | 201 | 148 | 159 | 131 |
| Biological efficiency % | 67 | 49 | 53 | 44 |
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 " 25 DEG C " by " 23 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 " 21 DEG C " by " 23 DEG C "；Remaining is equal to embodiment 2.
By conventional inoculation of the mushroom liquefaction strain obtained by above-mentioned comparative example 2-2~comparative example 3-2 according to above-mentioned experiment 1
Fruiting bag method cultivates mushroom fresh mushroom, and 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).
The mushroom liquefaction strain of gained cultivates mushroom fresh mushroom according to the conventional inoculation fruiting bag method described in above-mentioned experiment 1,
The basic result with obtained by " liquefaction strain (present invention) " in table 1 of acquired results (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 (3)
- The special bacteria culture medium 1. mushroom liquefies, it is characterized in that being composed of the following components in parts by weight：
- 2. the preparation method of culture medium as claimed in claim 1, it is characterized in that comprising the following steps：Cornstarch, fine bran, Semen Tritici aestivi fiber element, gypsum are well mixed, dispensing I is obtained；Glucose, yeast extract, beef peptone, potassium dihydrogen phosphate, magnesium sulfate are added to the water, are sufficiently stirred for, dispensing II is obtained；After dispensing I and dispensing II are sufficiently mixed, mushroom liquefaction special bacteria culture medium is obtained.
- The Spawn incubation method 3. the mushroom carried out using the culture medium described in claim 1 is liquefied, it is characterized in that：By mushroom parent species according to It is secondary to follow the steps below：1), strain makes：Mushroom liquefaction special bacteria culture medium is fitted into blake bottle, microporous barrier ventilating cover is covered, autoclave sterilization must sterilize Wild Oryza species；According to the inoculum concentration of every 200g mushroom liquefaction special bacteria culture medium correspondence 4.8~5.2g mushroom parent species, in sterile bar Access mushroom parent species under part in culture medium after sterilization, 22~24 DEG C of dark culturings 25~26 days, must liquefy special solid 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 mushroom liquefaction strain.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510028938.9A CN104557211B (en) | 2015-01-20 | 2015-01-20 | Mushroom liquefaction special bacteria culture medium and corresponding cultivation |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510028938.9A CN104557211B (en) | 2015-01-20 | 2015-01-20 | Mushroom liquefaction special bacteria culture medium and corresponding cultivation |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104557211A CN104557211A (en) | 2015-04-29 |
| CN104557211B true CN104557211B (en) | 2017-11-07 |
Family
ID=53074398
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510028938.9A Active CN104557211B (en) | 2015-01-20 | 2015-01-20 | Mushroom liquefaction special bacteria culture medium and corresponding cultivation |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104557211B (en) |
Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105399470A (en) * | 2015-11-30 | 2016-03-16 | 全椒县香妃农业专业合作社 | Lentinula edodes strain medium and preparing method thereof |
| CN107771614A (en) * | 2017-12-14 | 2018-03-09 | 全椒县香妃农业专业合作社 | A kind of mushroom strain nutrient medium |
| CN108076973B (en) * | 2018-01-15 | 2020-11-24 | 石家庄学院 | Production method of mushroom concentrated strain |
| CN108094051A (en) * | 2018-02-08 | 2018-06-01 | 贵州省生物研究所 | A kind of Coriaria sinica mushroom strain culture medium and preparation method thereof and the method for cultivating Coriaria sinica mushroom |
| CN112655462A (en) * | 2020-12-18 | 2021-04-16 | 廊坊师范学院 | Mushroom reduction strain and preparation and application thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102715014A (en) * | 2012-06-25 | 2012-10-10 | 浙江常山荣昇食用菌有限公司 | Preparation method for liquefied strain of edible mushroom |
| CN102972205A (en) * | 2012-12-10 | 2013-03-20 | 杨凌芝君生物科技有限责任公司 | Liquefying method and use method of solid strains of edible fungi and medicinal fungi |
-
2015
- 2015-01-20 CN CN201510028938.9A patent/CN104557211B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102715014A (en) * | 2012-06-25 | 2012-10-10 | 浙江常山荣昇食用菌有限公司 | Preparation method for liquefied strain of edible mushroom |
| CN102972205A (en) * | 2012-12-10 | 2013-03-20 | 杨凌芝君生物科技有限责任公司 | Liquefying method and use method of solid strains of edible fungi and medicinal fungi |
Also Published As
| Publication number | Publication date |
|---|---|
| CN104557211A (en) | 2015-04-29 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104557211B (en) | Mushroom liquefaction special bacteria culture medium and corresponding cultivation | |
| CN104541975B (en) | A kind of fast preparation method and application method of true pleurotus cornucopiae liquid spawn | |
| CN102154194B (en) | Preparation method for high-yield chlamydospore liquid fermentation from trichoderma on pilot plant test scale | |
| CN104928202A (en) | Fermentation culture method of bacillus | |
| CN110184201A (en) | A kind of true pleurotus cornucopiae bacterial strain and its selection | |
| CN104557315B (en) | Elegant precious mushroom liquefaction special bacteria culture medium and corresponding cultivation | |
| CN104541983B (en) | Seafood mushroom liquefaction special bacteria culture medium and corresponding cultivation | |
| Jin et al. | Development of Media and Automated Liquid Fermentation Methods to Produce Desiccation-Tolerant Propagules ofTrichoderma harzianum | |
| CN104620852B (en) | Mushroom class liquefaction Spawn incubation method | |
| CN103952332A (en) | Bacillus amyloliquefaciens ZH1 and its application | |
| CN109182151A (en) | The separating screening method of gingko endogenous fungus | |
| CN104557209B (en) | Pleurotus eryngii liquefaction special bacteria culture medium and corresponding cultivation | |
| CN101940131A (en) | Method for culturing cordyceps militaris by using drink bottle | |
| CN104557210B (en) | Asparagus liquefaction special bacteria culture medium and corresponding cultivation | |
| CN106416749A (en) | Method for reducing operational pollution during cultivation process of liquid spawn | |
| CN110036829A (en) | A kind of preparation method of oyster mushroom solid liquefaction strain | |
| CN106399123B (en) | Method for quickly producing seeds of edible fungi | |
| CN1952108A (en) | Long term storage method for large-size fungus species | |
| CN100532534C (en) | An epiphyte capable of digesting agar | |
| CN104737815A (en) | Cloth type strain and production method thereof | |
| CN108998404A (en) | A kind of conidial method of induction anthrax-bacilus generation | |
| CN105154340B (en) | A kind of selection of true pleurotus cornucopiae strain | |
| CN109761688A (en) | A kind of liquid fertilizer and preparation method thereof preventing tomato dehiscent fruit | |
| CN109956778A (en) | A kind of liquid fertilizer resistant to lodging and preparation method thereof | |
| CN109956801A (en) | A kind of liquid fertilizer and preparation method thereof for fruit corn flower bud development |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |