CN109220530A - A kind of black fungus strain and its application - Google Patents
A kind of black fungus strain and its application Download PDFInfo
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- CN109220530A CN109220530A CN201811268406.2A CN201811268406A CN109220530A CN 109220530 A CN109220530 A CN 109220530A CN 201811268406 A CN201811268406 A CN 201811268406A CN 109220530 A CN109220530 A CN 109220530A
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- 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
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- 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
- A01G18/20—Culture media, e.g. compost
Abstract
The invention discloses a kind of black fungus strain, the strain is cloud 3, black fungus strain osmanthus, is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, and deposit number is CGMCC No.15367, and the deposit date is on March 22nd, 2018.The fructification of black fungus of the present invention is soaked rate up to 20.0 or more, much higher than the black fungus strain of existing Extend culture；Black fungus fructification iron-content of the invention is not less than 166mg/Kg, and polyoses content is not less than 4.15g/100g, and black fungus fructification iron-content of the invention and polyoses content are higher, is above the black fungus strain of existing Extend culture in Guangxi District.Black fungus single sheet of the present invention, no vein or few vein, auricle softness is flexible, and mouthfeel is lubricious to blurt out, and is loved by consumers.Fresh goods yellowish-brown, it is bright-coloured transparent；Dry product outside of belly dark brown, back side grey black, condition is fabulous, wide market.
Description
Technical field
The present invention relates to edible mushroom fields, and in particular to a kind of black fungus strain and its application.
Background technique
Black fungus is under the jurisdiction of Basidiomycotina, and heterobasidium Gammaproteobacteria, Auriculariale, Auriculariaceae, Auricularia is that China four is edible greatly
One of bacteria cultivation kind.Black fungus belongs to middle warm type edible fungi, has cold-resistant, fearness thermal property, main producing region is typically distributed across
Summer more nice and cool area.Guangxi is also one of the major production areas of China's black fungus, and the black fungus kind of cultivation is based on 916
It leads, Xin Ke and black third kind also have fragmentary distribution, and cultivar is very single, and essentially nonlocal kind, lack this earth culture
The black fungus kind educated.Guangxi black fungus is resourceful, and especially (local popular name is Baise cloud to the black fungus of Baise Area, Guangxi Zhuang Autonomous Region
Ear) it is distributed at most, it is best in quality.In recent years, Guangxi departments of government pays much attention to the protection and benefit of Baise cloud ear resource
With local scientific research department also increases using the wild cloud ear resource in Baise, tames and cultivate grinding for Guangxi native country black fungus kind
Dynamics is studied carefully, still, currently with the excellent black fungus kind pole screened, tame and cultivated from the cloud ear wild resource of Baise
It is few.
The rate that is soaked of the fructification of existing black fungus kind is generally 9~15, and sugar content is generally in 2.0~3.0g/
100g, iron-content are 30~80mg/Kg.The nutrition content and quality of existing black fungus do not protrude.
Summary of the invention
The technical issues of the present invention overcomes above-mentioned black fungus, providing can a kind of black fungus strain.
To solve the above problems, the present invention provides a kind of black fungus strain:
The specific name of black fungus strain provided by the invention are as follows: black fungus (Auricularia auricular-
Judae), belong to Basidiomycotina, heterobasidium Gammaproteobacteria, Auriculariale, Auriculariaceae, Auricularia, black fungus kind.The black fungus strain
Bacterial strain is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, address: Chaoyang District, Beijing City North Star west
The institute 3 of road 1, deposit number are CGMCC No.15367, and the deposit date is on March 12nd, 2018.
Black fungus strain hereditary difference of the present invention analysis: using osmanthus cloud 3,916, it is No. 10 new, No. 4 new, black three, hundred
Cloud 6, Au8129, Au149, YE2, black 29, HME2, Ty013, Ty041, Ty051, Ty063, YEIt 3Totally 16 bacterial strains, 13
The ISSR of ISSR primer pair black fungus strain hereditary difference of the present invention is analyzed, as a result as shown in Fig. 3-Figure 15,13 of the application
Primer can amplify stable polymorphic bands to 16 black fungus strains.DNA (ISSR) fingerprint polymorphism analysis of strain
The result shows that Gui Yun 3 and the banding pattern and quantity of other strains differ greatly.ISSR clustering (Fig. 2) shows: Gui Yun 3
Genetic similarity with other strains is 0.68 hereinafter, strain affiliation farther out, illustrates No. 3 bacterium of osmanthus cloud of the invention
Strain is larger with the hereditary difference of other black fungus strains, and No. 3 bacterial strains of black fungus osmanthus of the present invention cloud and new No. 4,916, hundred clouds
The antagonistic experiment result (such as Figure 16-Figure 18) of No. 6 bacterial strains shows No. 3 bacterial strains of Gui Yun and the main black fungus cultivation kind new 4 in Guangxi
Number, No. 6 bacterial strains of 916, hundred clouds have an apparent mycelia Antagonistic reaction, it is not affine between strain.By ISSR points of bacterial strain hereditary difference
The antagonistic experiment of analysis and bacterial strain is the result shows that black fungus strain of the invention is a new black fungus strain.
The morphological feature of black fungus of the invention is as follows: auricle size (long × wide) is about 4.21cm × 5.28cm；Ear
Base is smaller；Auricle thickness: 1.07mm；Fructification piece type are as follows: Dan Sheng, sheet；Ear arteries and veins is few, and fructification base portion has a small amount of big tendon and vessel.
Fruit-body color is yellowish-brown, and gum level is more, and elasticity is good for auricle, and auricle stockless, fructification edge is not split, and hymenophore is smooth
Or slightly gauffer；Infertile velvet hair is obvious；There is marrow layer in cross section；The produce agaric period -70 days 55 days, belong to mid-early maturity.
The colony morphology characteristic of black fungus of the invention on PDA solid medium is as follows: hypha form: villiform；Bacterium
Fall form: round；Mycelia density degree is medium；Aerial hyphae is more, flourishing；Colony edge is neat；Pigment is yellowish-brown；Mycelia is raw
Long speed: 9.84mm/d；Mycelia color is pure white.
Cultivation of auricularia auricula character of the invention is as follows: during anti-hybrid ability is；Heat-resisting ability is strong；Biological conversion rate is
120%~140%；Fructification is soaked rate not less than 20.0, and fructification is soft.
Black fungus strain of the present invention is by the wild black fungus screening in Baise, domestication and cultivates and obtains, screening, acclimation method
It is as follows:
(1) the wild black fungus in collected Baise is separated by tissue and purifying obtains parental strain Ty026；
(2) parental strain Ty026 carries out cultivation domestication, screening, obtains the first generation tissue separation strains 60 of Ty026 bacterial strain
It is a, it is denoted as F1Generation；The screening technique are as follows: filter out that auricle is thick and solid, be of moderate size, vein is less from parental strain Ty026
Fructification carry out tissue separation and purifying；
(3) to F1Cultivation domestication, screening are carried out for bacterial strain, second generation tissue separation strains 30 is obtained, is denoted as F2Generation；It is described
Screening technique are as follows: from F1For filtering out in bacterial strain, performance is stablized, yield is high, auricle is thick and solid, be of moderate size, son that vein is less is real
Body carries out tissue separation and purifying；
(4) to F2Cultivation domestication, screening are carried out for bacterial strain, third generation tissue separation strains 30 is obtained, is denoted as F3Generation；It is described
Screening technique are as follows: from F2For filtering out in bacterial strain, performance is stablized, yield is high, auricle is thick and solid, be of moderate size, son that vein is less is real
Body carries out tissue separation and purifying；
(5) to F3Cultivation domestication, screening are carried out for bacterial strain, obtains the black fungus strain；The screening technique are as follows: from F3
For filtering out in bacterial strain, auricle is thick and solid, is of moderate size, the bacterial strain that vein is less, yield is high and character is uniform and stable；
Wherein, for step (1) into rapid (5), the tissue Isolation and purification method of the fructification is equal are as follows: uses volume basis
Number carries out disinfection 1~3min to black fungus fructification surface for 70% ethanol solutions, cuts the tissue block of 2cm × 2cm, continues
1min is sterilized with the ethanol solution that percentage by volume is 70%, then using after aseptic water washing 2~3 times, is accessed after natural drying
In PAD culture medium, the constant temperature incubation at 25~28 DEG C is until mycelia covers with test tube；Picking test tube leading portion mycelium, is transferred to examination
Pipe slant medium culture selects transplanting culture 2~3 times repeatedly according to the method and obtains purifying bacterial strain；The test tube slant
Culture medium prescription are as follows: potato 200g, glucose 20g, agar 20g, water 1000ml；
Wherein, for step (1) into rapid (5), the cultivation of auricularia auricula acclimation method is all made of the cultivation of pocket type clinker, cultivation step
It is rapid as follows:
(1) prepared by bacterium bag: select the short bag of 17cm × 33cm × 0.004cm polyethylene dog-ear bag or 15cm × 55cm ×
The long bag of polyethylene dog-ear bag of 0.004cm, after culture material is packed, is placed in autoclave and sterilizes, into pot after temperature in 4 hours
100 DEG C are risen to, constant temperature sterilizes 12-16 hours, during which cannot cease fire；Kettle temperature is allowed to drop to 60 DEG C naturally while hot after sterilizing
Charge bar is taken out, and moves in cooling chamber or transfer room, is down to 28 DEG C hereinafter, being inoculated with by sterile working requirement to charge bar temperature；Inoculation
When, short bag is directly inoculated in sack, and long bag plays the inoculation of 3-4 hole in bag body, and bore dia is 1.5~2.0cm, and hole depth 2~
2.5cm；The culture material is by the following at being grouped as of parts by weight: 40~80 parts of weed tree sawdust, 10~15 parts of wheat bran, gypsum 0.5~1
Part, 0.5~1 part of lime, 0~1 part of sucrose, 0~25 part of cotton seed hulls, ramulus mori consider 0~42 part to be worth doing；
(2) cultural hypha: after inoculation, bacteria stick is placed in dark, dry and comfortable, draughty environment and carries out cultural hypha, training
Supporting space enrironment temperature is 25-28 DEG C, air humidity 70% hereinafter, 2 months cultural hypha time, during which according to environment temperature
Turning is carried out with hypha growth condition, dissipates heap and ventilation；
(3) acanthopore: when mycelia covers with bacterium bag, and having a small amount of black ear base to be formed, and when mycelia physiological maturity carries out acanthopore；It adopts
With de- bag acanthopore produce agaric is exempted from, every bag of 80-100, the short bag of acanthopore quantity, every bag of 150-200 of long bag is a, aperture 0.4cm, hole depth
0.5cm.Bacteria stick " well " font is stacked after acanthopore, opens simultaneously door and window, and bacteria 7-10 days；
(4) grand style: when bacteria stick majority acanthopore mouth have black ear base formation when, temperature stablize 22 DEG C it is below sunny or
In the case of cloudy weather, in greenhouse or outdoor carry out grand style；The east-west , Kui wide 1-1.2m of the Hao Kui , Kui of be before grand style, length is not
The face Xian , Kui furrow width 40cm , Kui limes；Short bag bacteria stick directly can uprightly be placed in the face Kui, and long bag bacteria stick need to set up frame in the face Kui
Son, allow bacteria stick lean to on shelf；When bacteria stick is put, two bacteria stick spacing are 10cm, in favor of ventilation and light；
(5) ear management: after grand style 2-3 days, start to carry out water management；Water spray sprays band using water mist, sprays 3~5 daily
It is secondary；After auricle length to 1cm or more, water spray number can be reduced, each amount of water sprayed is increased；When temperature is higher than 25 DEG C, spray water in the morning and evening,
It avoids spraying water at high temperature；It should stop within 1-2 days spraying water before harvesting, stop water spray 5~10 days after harvesting；When continuous more it rains,
It should be taken shelter from rain using film；
(6) it harvests: showing that agaric maturation can harvest when auricle is unfolded, basal part of the ear contraction attenuates；Cut off the water 1-2 days or rain after just
Fine, auricle harvests after receiving side；
(7) prevention and control of plant diseases, pest control: aiming at prevention, when preventing the living contaminants such as Neurospora, trichoderma from should produce preparation from bacteria stick
Start, carries out sterilizing pass, inoculation is closed, cultural hypha is closed；Anti- fluid stopping ear, prevents spray-water or soil from bringing line during produce agaric
Insect infection.
The fructification for the black fungus that black fungus breeding strain of the present invention obtains is soaked rate not less than 20.0, and iron-content is not less than
166 mg/Kg, polyoses content are not less than 4.15g/100g；The fructification of the black fungus be single sheet, no vein or few vein,
Auricle softness is flexible, is in yellowish-brown before unseasoned, bright-coloured transparent, outside of belly dark brown after drying, back side grey black.
Another object of the present invention also resides in the application of protection black fungus strain, application method are as follows: by the black fungus bacterial
Instant black fungus powder is made in the black fungus fructification that kind cultivation obtains, and the instant black fungus powder is prepared by the following method:
20~30min of high temperature and pressure boiling is carried out after black fungus is cleaned, the percent water of freeze-day with constant temperature to black fungus contains at 60 DEG C
When amount is 12~15%, carries out Chao Wei Fen crushed and obtain the instant black fungus powder.The instant black fungus powder that the present invention is prepared by
Solid-to-liquid ratio stands 1~3min volume expansion 200% or more after being mixed into paste for the ratio and water of 1g:5ml.
Instant black fungus powder of the invention can be made into it is instant brew food, such as: it is instant brew rice paste, it is instant brew batter,
It is instant to brew milk tea, instant soaking beverage, instant brew that instant noodles etc. are instant to brew food.Contain instant black fungus of the invention
Powder brews food iron-content height immediately, and nutritive value is high, brews instant black fungus powder mass percentage in food when instant
There is apparent swelling effect when higher than 5%, when brewing, the paste gumminess such as rice paste brewed is high, and stability is high, contains
Instant instant brew when instant noodles are brewed compared to common aspect of black fungus powder of the present invention obtains more instant noodles, has pressure
Contracting wheaten food, the effect being convenient for carrying.
The present invention has the advantages that compared with prior art
(1) cloud No. 3 black fungus in osmanthus of the present invention are a new black fungus strains, and black fungus fructification of the present invention is in monolithic
Shape, no vein or few vein, auricle softness is flexible, and mouthfeel is lubricious to blurt out, and is loved by consumers.Black fungus of the present invention is real
The unseasoned preceding yellowish-brown of body, bright-coloured transparent, outside of belly dark brown after drying, back side grey black, condition is fabulous, wide market.
(2) fructification of the black fungus provided by the invention rate that is soaked is high compared to the black fungus of other strains, is not less than
20.0；Black fungus fructification iron-content of the invention is not less than 166mg/Kg, much higher than the black wood of Extend culture existing in Guangxi District
Ear strain；The black fungus strain of black fungus fruitbody polysaccharide content of the invention also above Extend culture existing in Guangxi District, this hair
Bright black fungus fruitbody polysaccharide content is not less than 4.15g/100g.
Detailed description of the invention
Fig. 1 is the breeding family tree of black fungus of the present invention；
Fig. 2 is that the present invention is based on the clustering figures of the ISSR black fungus strain analyzed；
Fig. 3 is the PCR electrophoretogram that primer numbers are 1 in table 2；
Fig. 4 is the PCR electrophoretogram that primer numbers are 2 in table 2；
Fig. 5 is the PCR electrophoretogram that primer numbers are 3 in table 2；
Fig. 6 is the PCR electrophoretogram that primer numbers are 4 in table 2；
Fig. 7 is the PCR electrophoretogram that primer numbers are 5 in table 2；
Fig. 8 is the PCR electrophoretogram that primer numbers are 6 in table 2；
Fig. 9 is the PCR electrophoretogram that primer numbers are 7 in table 2；
Figure 10 is the PCR electrophoretogram that primer numbers are 8 in table 2；
Figure 11 is the PCR electrophoretogram that primer numbers are 9 in table 2；
Figure 12 is the PCR electrophoretogram that primer numbers are 10 in table 2；
Figure 13 is the PCR electrophoretogram that primer numbers are 11 in table 2；
Figure 14 is the PCR electrophoretogram that primer numbers are 12 in table 2；
Figure 15 is the PCR electrophoretogram that primer numbers are 13 in table 2；
In Fig. 3-Figure 15, swimming lane sequence from left to right successively are as follows: DNA Marker, Gui Yun No. 3,916, it is No. 10 new,
New No. 4, black three, hundred cloud 6, Au8129, Au149, YE2, black 29, HME2, TY013, TY041, TY051, TY063, YE days
3。
Figure 16 is the osmanthus cloud 3 antagonism figures with new No. 4 bacterial strains on culture dish, and lower section is No. 3 bacterial strains of osmanthus cloud in figure, under
No. 4 bacterial strains of Fang Weixin section；
Figure 17 is the osmanthus cloud 3 antagonism figures with 916 bacterial strains on culture dish, and lower section is No. 3 bacterial strains of osmanthus cloud in figure, and lower section is
916 bacterial strains；
Figure 18 is the antagonism figure of osmanthus cloud 3 with No. 6 bacterial strains of hundred clouds, and lower section is No. 3 bacterial strains of osmanthus cloud in figure, and lower section is hundred clouds 6
Number bacterial strain；
Figure 19 is osmanthus cloud 3 and new No. 4 subobject graphs, and the left side is 3 work song entity of osmanthus cloud in figure, and the right is new 4
Work song sterogram；
Figure 20 is the subobject graph of 916 with black 3, and the left side is 916 fructifications in figure, and the right is black 3 subobject graph；
Figure 21 is the bacteria stick figure of osmanthus cloud 3 with new No. 4, and the left side is No. 3 bacteria sticks of osmanthus cloud in figure, and the right is new No. 4 bacterium
Stick；
Specific embodiment
Below with reference to embodiment and test, the invention will be further described.
Embodiment 1
Black fungus (Auricularia auricular-judae) of the invention, belongs to Basidiomycotina, heterobasidium bacterium
Guiding principle, Auriculariale, Auriculariaceae, Auricularia, black fungus kind.The black fungus species strain is preserved in Chinese microorganism strain preservation pipe
Reason committee common micro-organisms center, address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, deposit number CGMCC
No.15367, the deposit date is on March 12nd, 2018.
The black fungus strain is by the wild black fungus screening in Baise, domestication and cultivates and obtains, and screening, acclimation method are as follows:
(1) the wild black fungus in collected Baise is separated by tissue and purifying obtains parental strain Ty026；
(2) parental strain Ty026 carries out cultivation domestication, screening, obtains the first generation tissue separation strains 60 of Ty026 bacterial strain
It is a, it is denoted as F1Generation；The screening technique are as follows: filter out that auricle is thick and solid, be of moderate size, vein is less from parental strain Ty026
Fructification carry out tissue separation and purifying；
(3) to F1Cultivation domestication, screening are carried out for bacterial strain, second generation tissue separation strains 30 is obtained, is denoted as F2Generation；It is described
Screening technique are as follows: from F1For filtering out in bacterial strain, performance is stablized, yield is high, auricle is thick and solid, be of moderate size, son that vein is less is real
Body carries out tissue separation and purifying；
(4) to F2Cultivation domestication, screening are carried out for bacterial strain, third generation tissue separation strains 30 is obtained, is denoted as F3Generation；It is described
Screening technique are as follows: from F2For filtering out in bacterial strain, performance is stablized, yield is high, auricle is thick and solid, be of moderate size, son that vein is less is real
Body carries out tissue separation and purifying；
(5) to F3Cultivation domestication, screening are carried out for bacterial strain, obtains the black fungus strain；The screening technique are as follows: from F3
For filtering out in bacterial strain, auricle is thick and solid, is of moderate size, the bacterial strain that vein is less, yield is high and character is uniform and stable；
Wherein, for step (1) into rapid (5), the tissue Isolation and purification method of the fructification is equal are as follows: uses volume basis
Number carries out disinfection 3min to black fungus fructification surface for 70% ethanol solutions, cuts the tissue block of 2cm × 2cm, continues to use
The ethanol solution that percentage by volume is 70% sterilizes 1min, then using after aseptic water washing 2 times, accesses PAD training after natural drying
It supports in base, the constant temperature incubation at 28 DEG C is until mycelia covers with test tube；Picking test tube leading portion mycelium, is transferred to test tube slant culture
Base culture selects transplanting culture 2 times repeatedly according to the method and obtains purifying bacterial strain；The test tube slant culture medium prescription are as follows:
Potato 200g, glucose 20g, agar 20g, water 1000ml；
Wherein, for step (1) into rapid (5), the cultivation of auricularia auricula acclimation method is all made of the cultivation of pocket type clinker, cultivation step
It is rapid as follows:
(1) prepared by bacterium bag: the short bag of 17cm × 33cm × 0.004cm polyethylene dog-ear bag is selected, after culture material is packed,
Be placed in autoclave and sterilize, into pot after in 4 hours temperature rise to 100 DEG C, constant temperature sterilizes 16 hours, during which cannot cease fire；Sterilizing
After allow kettle temperature to drop to 60 DEG C naturally to take out charge bar while hot, and move in cooling chamber or transfer room, be down to 28 to charge bar temperature
DEG C hereinafter, by sterile working requirement be inoculated with；When inoculation, short bag is directly inoculated in sack；The culture material is by the as follows of mass parts
At being grouped as: 80 parts of weed tree sawdust, 10 parts of wheat bran, 1 part of gypsum, 0.5 part of lime, 1 part of sucrose, ramulus mori consider 42 parts to be worth doing；
(2) cultural hypha: after inoculation, bacteria stick is placed in dark, dry and comfortable, draughty environment and carries out cultural hypha, training
Supporting space enrironment temperature is 28 DEG C, air humidity 70% hereinafter, 2 months cultural hypha time, during which according to environment temperature and
Hypha growth condition carries out turning, dissipates heap and ventilation；
(3) acanthopore: when mycelia covers with bacterium bag, and having a small amount of black ear base to be formed, and when mycelia physiological maturity carries out acanthopore；It adopts
With exempting from de- bag acanthopore produce agaric, 80, aperture 0.4cm, hole depth 0.5cm, short every bag of the bag of acanthopore quantity.Bacteria stick " well " font after acanthopore
It stacks, opens simultaneously door and window, bacteria 10 days；
(4) grand style: when bacteria stick majority acanthopore mouth have black ear base formation when, temperature stablize 22 DEG C it is below sunny or
In the case of cloudy weather, grand style is carried out in greenhouse；The east-west , Kui wide 1.2m of the Hao Kui , Kui of be before grand style, long unlimited , Kui ditch
The wide face 40cm , Kui limes；Short bag bacteria stick directly can uprightly be placed in the face Kui；When bacteria stick is put, two bacteria stick spacing are 10cm, with
Conducive to ventilation and light；
(5) ear management: after grand style 3 days, start to carry out water management；Water spray sprays band using water mist, daily spray 5 times；Ear
After length of a film to 1cm or more, water spray number can be reduced, increases each amount of water sprayed；When temperature is higher than 25 DEG C, sprays water, avoid in the morning and evening
It sprays water at high temperature；Harvesting should stop spraying water for first 2 days, stop water spray 10 days after harvesting；When continuous more it rains, film should be used
It takes shelter from rain；
(6) it harvests: showing that agaric maturation can harvest when auricle is unfolded, basal part of the ear contraction attenuates；Cut off the water 2 days or Harbeth,
Auricle harvests after receiving side；
(7) prevention and control of plant diseases, pest control: aiming at prevention, when preventing the living contaminants such as Neurospora, trichoderma from should produce preparation from bacteria stick
Start, carries out sterilizing pass, inoculation is closed, cultural hypha is closed；Anti- fluid stopping ear, prevents spray-water or soil from bringing line during produce agaric
Insect infection.
Embodiment 2
Black fungus (Auricularia auricular-judae) of the invention, belongs to Basidiomycotina, heterobasidium bacterium
Guiding principle, Auriculariale, Auriculariaceae, Auricularia, black fungus kind.The black fungus species strain is preserved in Chinese microorganism strain preservation pipe
Reason committee common micro-organisms center, address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, deposit number CGMCC
No.15367, the deposit date is on March 12nd, 2018.
The black fungus strain is screened by the wild black fungus in Baise, tamed and cultivated and obtained, and screening, acclimation method are the same as implementation
The difference of example 1 is:
For step (1) into rapid (5), the tissue Isolation and purification method of the fructification is equal are as follows: use percentage by volume for
70% ethanol solution carries out disinfection 1min to black fungus fructification surface, cuts the tissue block of 2cm × 2cm, continues to use volume
The ethanol solution that percentage is 70% sterilizes 1min, then using after aseptic water washing 3 times, accesses PAD culture medium after natural drying
In, the constant temperature incubation at 25 DEG C is until mycelia covers with test tube；Picking test tube leading portion mycelium, is transferred to test tube slant culture medium
Culture selects transplanting culture 3 times repeatedly according to the method and obtains purifying bacterial strain；The test tube slant culture medium prescription are as follows: horse
Bell potato 200g, glucose 20g, agar 20g, water 1000ml；
Step (1) is into rapid (5), and the cultivation of auricularia auricula acclimation method is all made of the cultivation of pocket type clinker, and cultivation step is such as
Under:
(1) prepared by bacterium bag: the long bag of 15cm × 55cm × 0.004cm polyethylene dog-ear bag is selected, after culture material is packed,
Be placed in autoclave and sterilize, into pot after in 4 hours temperature rise to 100 DEG C, constant temperature sterilizes 12 hours, during which cannot cease fire；Sterilizing
After allow kettle temperature to drop to 60 DEG C naturally to take out charge bar while hot, and move in cooling chamber or transfer room, be down to 28 to charge bar temperature
DEG C hereinafter, by sterile working requirement be inoculated with；When inoculation, 4 holes are made a call in bag body and are inoculated with, bore dia 2.0cm, hole depth 2.5cm；Institute
Culture material is stated by the following at being grouped as of mass parts: 40 parts of weed tree sawdust, 15 parts of wheat bran, 0.5 part of gypsum, 1 part of lime, cotton seed hulls 25
Part；
(2) cultural hypha: after inoculation, bacteria stick is placed in dark, dry and comfortable, draughty environment and carries out cultural hypha, training
Supporting space enrironment temperature is 25 DEG C, air humidity 70% hereinafter, 2 months cultural hypha time, during which according to environment temperature and
Hypha growth condition carries out turning, dissipates heap and ventilation；
(3) acanthopore: when mycelia covers with bacterium bag, and having a small amount of black ear base to be formed, and when mycelia physiological maturity carries out acanthopore；It adopts
With exempting from de- bag acanthopore produce agaric, acanthopore quantity every bag 200, aperture 0.4cm, hole depth 0.5cm.Bacteria stick " well " font heap after acanthopore
It puts, opens simultaneously door and window, bacteria 7 days；
(4) grand style: when bacteria stick majority acanthopore mouth have black ear base formation when, temperature stablize 22 DEG C it is below sunny or
In the case of cloudy weather, outdoor carry out grand style；The east-west , Kui wide 1m of the Hao Kui , Kui of be before grand style, long unlimited , Kui furrow width 40cm,
The face Kui limes；Bacteria stick Xu Kui sets up shelf in face, allow bacteria stick lean to on shelf；When bacteria stick is put, two bacteria stick spacing are
10cm, in favor of ventilation and light；
(5) ear management: after grand style 2 days, start to carry out water management；Water spray sprays band using water mist, daily spray 3 times；Ear
After length of a film to 1cm or more, water spray number can be reduced, increases each amount of water sprayed；When temperature is higher than 25 DEG C, sprays water, avoid in the morning and evening
It sprays water at high temperature；Harvesting should stop spraying water for first 1 day, stop water spray 5 days after harvesting；When continuous more it rains, film should be used
It takes shelter from rain；
(6) it harvests: showing that agaric maturation can harvest when auricle is unfolded, basal part of the ear contraction attenuates；Cut off the water 1-2 days or rain after just
Fine, auricle harvests after receiving side；
(7) prevention and control of plant diseases, pest control: aiming at prevention, when preventing the living contaminants such as Neurospora, trichoderma from should produce preparation from bacteria stick
Start, carries out sterilizing pass, inoculation is closed, cultural hypha is closed；Anti- fluid stopping ear, prevents spray-water or soil from bringing line during produce agaric
Insect infection.
Embodiment 3
Black fungus (Auricularia auricular-judae) of the invention, belongs to Basidiomycotina, heterobasidium bacterium
Guiding principle, Auriculariale, Auriculariaceae, Auricularia, black fungus kind.The black fungus species strain is preserved in Chinese microorganism strain preservation pipe
Reason committee common micro-organisms center, address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, deposit number CGMCC
No.15367, the deposit date is on March 12nd, 2018.
The black fungus strain is screened by the wild black fungus in Baise, tamed and cultivated and obtained, and screening, acclimation method are the same as implementation
The difference of example 1 is:
For step (1) into rapid (5), the tissue Isolation and purification method of the fructification is equal are as follows: use percentage by volume for
70% ethanol solution carries out disinfection 2min to black fungus fructification surface, cuts the tissue block of 2cm × 2cm, continues to use volume
The ethanol solution that percentage is 70% sterilizes 1min, then using after aseptic water washing 2 times, accesses PAD culture medium after natural drying
In, the constant temperature incubation at 27 DEG C is until mycelia covers with test tube；Picking test tube leading portion mycelium, is transferred to test tube slant culture medium
Culture selects transplanting culture 3 times repeatedly according to the method and obtains purifying bacterial strain；The test tube slant culture medium prescription are as follows: horse
Bell potato 200g, glucose 20g, agar 20g, water 1000ml；
Step (1) is into rapid (5), and the cultivation of auricularia auricula acclimation method is all made of the cultivation of pocket type clinker, and cultivation step is such as
Under:
(1) prepared by bacterium bag: the long bag of 15cm × 55cm × 0.004cm polyethylene dog-ear bag is selected, after culture material is packed,
Be placed in autoclave and sterilize, into pot after in 4 hours temperature rise to 100 DEG C, constant temperature sterilizes 15 hours, during which cannot cease fire；Sterilizing
After allow kettle temperature to drop to 60 DEG C naturally to take out charge bar while hot, and move in cooling chamber or transfer room, be down to 28 to charge bar temperature
DEG C hereinafter, by sterile working requirement be inoculated with；When inoculation, 3 holes are made a call in bag body and are inoculated with, bore dia 1.5cm, hole depth 2cm；It is described
Culture material is by the following at being grouped as of mass parts: 43 parts of weed tree sawdust, 12 parts of wheat bran, 0.8 part of gypsum, 0.8 part of lime, sucrose 0.5
Part, 13 parts of cotton seed hulls, ramulus mori consider 38 parts to be worth doing；
(2) cultural hypha: after inoculation, bacteria stick is placed in dark, dry and comfortable, draughty environment and carries out cultural hypha, training
Supporting space enrironment temperature is 26 DEG C, air humidity 70% hereinafter, 2 months cultural hypha time, during which according to environment temperature and
Hypha growth condition carries out turning, dissipates heap and ventilation；
(3) acanthopore: when mycelia covers with bacterium bag, and having a small amount of black ear base to be formed, and when mycelia physiological maturity carries out acanthopore；It adopts
With exempting from de- bag acanthopore produce agaric, acanthopore quantity every bag 150, aperture 0.4cm, hole depth 0.5cm.Bacteria stick " well " font heap after acanthopore
It puts, opens simultaneously door and window, bacteria 8 days；
(4) grand style: when bacteria stick majority acanthopore mouth have black ear base formation when, temperature stablize 22 DEG C it is below sunny or
In the case of cloudy weather, grand style is carried out in greenhouse；The east-west , Kui wide 1.1m of the Hao Kui , Kui of be before grand style, long unlimited , Kui ditch
The wide face 40cm , Kui limes；Short bag bacteria stick directly can uprightly be placed in the face Kui, and long bag bacteria stick need to set up shelf in the face Kui, allow bacterium
Stick lean to on shelf；When bacteria stick is put, two bacteria stick spacing are 10cm, in favor of ventilation and light；
(5) ear management: after grand style 3 days, start to carry out water management；Water spray sprays band using water mist, daily spray 4 times；Ear
After length of a film to 1cm or more, water spray number can be reduced, increases each amount of water sprayed；When temperature is higher than 25 DEG C, sprays water, avoid in the morning and evening
It sprays water at high temperature；Harvesting should stop spraying water for first 2 days, stop water spray 8 days after harvesting；When continuous more it rains, film should be used
It takes shelter from rain；
(6) it harvests: showing that agaric maturation can harvest when auricle is unfolded, basal part of the ear contraction attenuates；Cut off the water 2 days or Harbeth,
Auricle harvests after receiving side；
(7) prevention and control of plant diseases, pest control: aiming at prevention, when preventing the living contaminants such as Neurospora, trichoderma from should produce preparation from bacteria stick
Start, carries out sterilizing pass, inoculation is closed, cultural hypha is closed；Anti- fluid stopping ear, prevents spray-water or soil from bringing line during produce agaric
Insect infection.
The black fungus that above-mentioned plantation obtains is detected as follows:
(1) hereditary difference is analyzed
ISSR analysis to black fungus strain hereditary difference of the present invention, using 16 bacterial strains, 13 ISSR primers, 16 black
Fungus strain numbers 1,13 ISSR primer sequence such as the following table 2 of table specific as follows.
Table 1
Table 2
| Number | Primer | Primer sequence | Renaturation temperature |
| 1 | P3 | GA GA GA GA GA GA GA GA T | 53.1 |
| 2 | P4 | GA GA GA GA GA GA GA GAC | 56.0 |
| 3 | P5 | AG AG AG AG AG AG AG AGG | 56.0 |
| 4 | P6 | AG AG AG AG AG AG AG AGC | 56.0 |
| 5 | P9 | GSGTGTGTGTGTGTGTGT | 52.0 |
| 6 | P10 | AG AG AG AG AG AG AG AGT | 54.0 |
| 7 | P16 | TC TC TC TC TC TC TC TC C | 56.0 |
| 8 | P21 | AG AG AG AG AG AG AG AGYA | 54.0 |
| 9 | P22 | AG AG AG AG AG AG AG AGYC | 56.0 |
| 10 | P25 | GA GA GA GA GA GA GA GAYC | 56.0 |
| 11 | P26 | GA GA GA GA GA GA GA GAYT | 54.0 |
| 12 | P27 | GA GA GA GA GA GA GA GAYG | 56.0 |
| 13 | P32 | GA GA GA GA GA GA GA GAA | 54.0 |
By Fig. 3-Figure 15 it is found that 13 primers of the application can amplify stable band to 16 black fungus strains,
ISSR analysis to the hereditary difference of strain, the results showed that, Gui Yun 3 and the banding pattern and quantity of other strains differ greatly.
ISSR clustering, as a result as shown in the ISSR dendrogram of Fig. 2, the results showed that the hereditary phase of Gui Yun 3 and other strains
Like coefficient 0.68 hereinafter, strain affiliation farther out, illustrates No. 3 bacterial strains of osmanthus cloud of the invention and other black fungus strains
Hereditary difference is larger, is a new black fungus strain.
(2) antagonistic experiment
No. 3 bacterial strains of black fungus osmanthus of the present invention cloud with new No. 4, the antagonistic experiments of 916, hundred No. 6 bacterial strains of cloud respectively as Figure 16,
Figure 17, Figure 18.By Figure 16-Figure 18 it is found that No. 3 bacterial strains of osmanthus cloud of the present invention and the main black fungus cultivation kind in Guangxi it is No. 4 new, 916,
Hundred bacterial strains of cloud 6 have apparent mycelia Antagonistic reaction, and bacterial strain is not affine, illustrate the black fungus of osmanthus cloud 3 with existing Extend culture
Bacterial strain has significantly different, is a new black fungus strain.
(3) Morphometric analysis of black fungus strain of the present invention
By the left side in the new 4 work song entity, Figure 20 on the right in the 3 work song entity of osmanthus cloud and Figure 19 on the left side in Figure 19
916 fructifications, in Figure 20 black 3 fructification on the right compare and Figure 21 in the left side No. 3 bacteria sticks of osmanthus cloud and Figure 21 in the right
New No. 4 bacteria sticks compared to known to: osmanthus cloud 3 morphological features of the invention have with the morphological feature of other black fungus kinds
Different degrees of difference, cloud No. 3 black fungus single sheets in osmanthus of the present invention, no vein or few vein, auricle softness is flexible, and mouthfeel is sliding
Profit is blurted out, and is loved by consumers.Fresh goods yellowish-brown, it is bright-coloured transparent；Dry product outside of belly dark brown, back side grey black, condition is fabulous,
Wide market.
(4) rate that is soaked measurement
1, measuring method
Selection Ty071,916, the black fungus of tetra- strains of YE1, YE2 and black fungus osmanthus of the present invention cloud 3 carry out the rate that is soaked
Detection, the Ty071 of selection, 916, the cultural method of black fungus and the black fungus osmanthus of the present invention cloud No. 3 numbers of tetra- strains of YE1, YE2 it is equal
With embodiment 1.Measuring method are as follows: black fungus is weighed respectively and air-dries each 10g of product, is placed in clear water and impregnates 4-6 hours, is taken out, drip
Solid carbon dioxide point (blotting surface moisture with blotting paper) claims weight in wet base W1, then black fungus is spread out overnight (about 12h), is subsequently placed in 60 DEG C
Baking oven drying, claims dry weight W2, and calculating is soaked rate as W1:W2.Replication twice, first time measurement result such as the following table 3, and it is secondary
Measurement result such as the following table 4.
Table 3
Table 4
By table 3 and table 4 it is found that of the invention osmanthus cloud 3 rates that are soaked are up to 20.0 or more, significantly larger than other black fungus bacterials
Strain.
(5) iron and polyoses content of osmanthus cloud of the present invention 3 and other black fungus strains compare, as a result such as the following table 5.
Table 5
| Bacterial strain | Osmanthus cloud 3 | New No. 4 | Ty071 | 916 | YE1 | Black three |
| Polyoses content (g/100g) | 4.15 | 2.90 | 2.60 | 3.42 | 3.50 | 3.19 |
| Iron content (mg/Kg) | 166 | 85 | 53.3 | 36.3 | 71.1 | 37.8 |
As shown in Table 5, cloud 3 work song entity iron content 166mg/Kg in osmanthus of the present invention, much higher than the black fungus product of existing Extend culture
Kind, cloud 3 work song entity polyoses content in osmanthus of the invention is also above other comparison bacterial strains.
Prepared using the black fungus of the application and instant brew food: the food that specifically prepares and the preparation method is as follows:
(1) powder of edible fungus is prepared:
Embodiment 4
A kind of powder of edible fungus, is prepared by the following method: a kind of embodiment being planted after obtained black fungus is cleaned and is carried out
High temperature and pressure boiling 30min carries out Chao Wei Fen crushed when the percent water of freeze-day with constant temperature to black fungus is 12% at 60 DEG C
Obtain the powder of edible fungus.
Embodiment 5
A kind of powder of edible fungus, is prepared by the following method: a kind of embodiment being planted after obtained black fungus is cleaned and is carried out
High temperature and pressure boiling 20min carries out Chao Wei Fen crushed when the percent water of freeze-day with constant temperature to black fungus is 15% at 60 DEG C
Obtain the powder of edible fungus.
Embodiment 6
A kind of powder of edible fungus, is prepared by the following method: a kind of embodiment being planted after obtained black fungus is cleaned and is carried out
High temperature and pressure boiling 25min carries out Chao Wei Fen crushed when the percent water of freeze-day with constant temperature to black fungus is 13% at 60 DEG C
Obtain the powder of edible fungus.
(2) it prepares and instant brews food
Embodiment 7
One kind is instant to brew food, there is the following component of mass percentage meter: powder of edible fungus 5%, the core of embodiment 4
Peach powder 5%, corn flour 5%, rice meal 60%, black rice flour 20%, sugar 5%.
Embodiment 8
It is a kind of it is instant brews food, have a following component of mass percentage meter: the powder of edible fungus 80% of embodiment 4,
Rice meal 15%, sugar 5%.
(3) it prepares and instant brews instant noodles
Embodiment 9
One kind is instant to brew instant noodles, there is the following component of mass percentage meter: the powder of edible fungus of embodiment 4
20%, flour 80%.
Embodiment 10
One kind is instant to brew instant noodles, there is the following component of mass percentage meter: the powder of edible fungus of embodiment 4
50%, flour 50%.
The powder of edible fungus that embodiment 4, embodiment 4, embodiment 6 are prepared into is mixed in the ratio and water that solid-to-liquid ratio is 1g:5ml
Standing volume expands 200%, 250%, 227% respectively after conjunction stirs into paste.
The instant food that brews of embodiment 7 and embodiment 8 is brewed edible, is had when being brewed using 100 DEG C of boiled water obvious
Swelling effect, the paste foods gumminess such as rice paste brewed is high, and stability is high, iron-content is high, good health care effect.
The instant instant noodles that brew of embodiment 9, embodiment 10 are brewed edible, are obtained when being brewed compared to common aspect
The rate that is soaked of more instant noodles, embodiment 9 improves 14.5% compared to general aspect, embodiment 10 be soaked rate compared to
General aspect improves 10.2%, has the effect of compressing wheaten food, be convenient for carrying.
Above description is the detailed description for the present invention preferably possible embodiments, but embodiment is not limited to this hair
Bright patent claim, it is all the present invention suggested by technical spirit under completed same changes or modifications change, should all belong to
In the covered the scope of the patents of the present invention.
Claims (7)
1. a black fungus strain, which is characterized in that the strain is black fungus (Auricularia auricular-judae),
Its deposit number is CGMCC No.15367.
2. a kind of black fungus strain according to claim 1, which is characterized in that the black fungus strain is wild black by Baise
Agaric separation tames and cultivates and obtain.
3. a kind of black fungus strain according to claim 2, which is characterized in that the screening, acclimation method are as follows:
(1) the wild black fungus in collected Baise is separated by tissue and purifying obtains parental strain Ty026；
(2) parental strain Ty026 carries out cultivation domestication, screening, obtains the first generation tissue separation strains 60 of Ty026 bacterial strain, remembers
For F1Generation；The screening technique are as follows: filter out that auricle is thick and solid, be of moderate size, vein is less, and son is real from parental strain Ty026
Body carries out tissue separation and purifying；
(3) to F1Cultivation domestication, screening are carried out for bacterial strain, second generation tissue separation strains 30 is obtained, is denoted as F2Generation；The screening
Method are as follows: from F1For filtered out in bacterial strain performance is stablized, yield is high, auricle is thick and solid, be of moderate size, fructification that vein is less into
The separation of row tissue and purifying；
(4) to F2Cultivation domestication, screening are carried out for bacterial strain, third generation tissue separation strains 30 is obtained, is denoted as F3Generation；The screening
Method are as follows: from F2For filtered out in bacterial strain performance is stablized, yield is high, auricle is thick and solid, be of moderate size, fructification that vein is less into
The separation of row tissue and purifying；
(5) to F3Cultivation domestication, screening are carried out for bacterial strain, obtains the black fungus strain；The screening technique are as follows: from F3For bacterium
Filter out that auricle is thick and solid, is of moderate size, the bacterial strain that vein is less, yield is high and character is uniform and stable in strain.
4. a kind of black fungus strain according to claim 3, which is characterized in that for step (1) into rapid (5), the son is real
The tissue Isolation and purification method of body is equal are as follows: use percentage by volume for 70% ethanol solution to black fungus fructification surface into
Row 1~3min of disinfection, cuts the tissue block of 2cm × 2cm, continues to sterilize 1min with the ethanol solution that percentage by volume is 70%,
It again using after aseptic water washing 2~3 times, accesses in PAD culture medium after natural drying, the constant temperature incubation at 25~28 DEG C is until bacterium
Filament length expires test tube；Picking test tube leading portion mycelium, is transferred to test tube slant culture medium culture, selects transplanting repeatedly according to the method
It cultivates 2~3 times and obtains purifying bacterial strain；The test tube slant culture medium prescription are as follows: potato 200g, glucose 20g, agar
20g, water 1000ml.
5. a kind of black fungus strain according to claim 3, which is characterized in that step (1) is into rapid (5), the black wood
Ear cultivation domestication method is all made of the cultivation of pocket type clinker, and steps are as follows for cultivation:
(1) prepared by bacterium bag: select the short bag of 17cm × 33cm × 0.004cm polyethylene dog-ear bag or 15cm × 55cm ×
The long bag of polyethylene dog-ear bag of 0.004cm, after culture material is packed, is placed in autoclave and sterilizes, into pot after temperature liter in 4 hours
To 100 DEG C, constant temperature sterilizes 12-16 hours, during which cannot cease fire；It allows kettle temperature to drop to 60 DEG C naturally after sterilizing to take while hot
Charge bar out, and cooling chamber or transfer room are moved in, 28 DEG C are down to hereinafter, being inoculated with by sterile working requirement to charge bar temperature；When inoculation,
Short bag is directly inoculated in sack, and long bag is beaten 3-4 hole in bag body and is inoculated with, and bore dia is 1.5~2.0cm, 2~2.5cm of hole depth；Institute
Culture material is stated by the following at being grouped as of parts by weight: 40~80 parts of weed tree sawdust, 10~15 parts of wheat bran, 0.5~1 part of gypsum, lime
0.5~1 part, 0~1 part of sucrose, 0~25 part of cotton seed hulls, 0~42 part of ramulus mori bits；
(2) cultural hypha: after inoculation, bacteria stick is placed in dark, dry and comfortable, draughty environment and carries out cultural hypha, culture farm
Ground environment temperature is 25-28 DEG C, air humidity 70% hereinafter, 2 months cultural hypha time, during which according to environment temperature and bacterium
Silk growing state carries out turning, dissipates heap and ventilation；
(3) acanthopore: when mycelia covers with bacterium bag, and having a small amount of black ear base to be formed, and when mycelia physiological maturity carries out acanthopore；Using exempting from
De- bag acanthopore produce agaric, every bag of 80-100, the short bag of acanthopore quantity, every bag of 150-200, long bag, aperture 0.4cm, hole depth 0.5cm.
Bacteria stick " well " font is stacked after acanthopore, opens simultaneously door and window, and bacteria 7-10 days；
(4) grand style: below sunny or cloudy at 22 DEG C in temperature stabilization when bacteria stick majority acanthopore mouth has the formation of black ear base
Under weather condition, in greenhouse or outdoor carry out grand style；The east-west , Kui wide 1-1.2m of the Hao Kui , Kui of be before grand style, length is unlimited,
The face Kui furrow width 40cm , Kui limes；Short bag bacteria stick directly can uprightly be placed in the face Kui, and long bag bacteria stick need to set up shelf in the face Kui,
Allow bacteria stick lean to on shelf；When bacteria stick is put, two bacteria stick spacing are 10cm, in favor of ventilation and light；
(5) ear management: after grand style 2-3 days, start to carry out water management；Water spray sprays band using water mist, sprays 3~5 times daily；Ear
After length of a film to 1cm or more, water spray number can be reduced, increases each amount of water sprayed；When temperature is higher than 25 DEG C, sprays water, avoid in the morning and evening
It sprays water at high temperature；It should stop within 1-2 days spraying water before harvesting, stop water spray 5~10 days after harvesting；When continuous more it rains, Ying Cai
It is taken shelter from rain with film；
(6) it harvests: showing that agaric maturation can harvest when auricle is unfolded, basal part of the ear contraction attenuates；Cut off the water 1-2 days or Harbeth, ear
Piece harvests after receiving side；
(7) prevention and control of plant diseases, pest control: aiming at prevention, preventing the living contaminants such as Neurospora, trichoderma should be when bacteria stick production preparation,
Sterilizing is carried out to close, be inoculated with pass, cultural hypha pass；Anti- fluid stopping ear, prevents spray-water or soil from bringing nematode infections during produce agaric.
6. a kind of black fungus strain according to claim 1, which is characterized in that the black fungus breeding strain obtains black
The fructification of agaric is soaked rate not less than 20.0, and iron-content is not less than 166mg/Kg, and polyoses content is not less than 4.15g/100g；Institute
The fructification for stating black fungus is single sheet, no vein or few vein, and auricle softness is flexible, is in yellowish-brown before unseasoned, bright-coloured
It is transparent, outside of belly dark brown after drying, back side grey black.
7. a kind of application of any black fungus strain of claim 1-6, which is characterized in that by the black fungus bacterial planting
It trains obtained black fungus fructification and instant black fungus powder is made, the instant black fungus powder is prepared by the following method: will be black
Agaric carries out 20~30min of high temperature and pressure boiling after cleaning, and the percent water of freeze-day with constant temperature to black fungus is at 60 DEG C
When 12~15%, carries out Chao Wei Fen crushed and obtain the instant black fungus powder.
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