CN104012298B - The complete production of hybrid seeds technique of edible fungus liquid kind submerged fermentation and culture medium prescription thereof - Google Patents
The complete production of hybrid seeds technique of edible fungus liquid kind submerged fermentation and culture medium prescription thereof Download PDFInfo
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Abstract
The invention provides the culture medium prescription of the complete production of hybrid seeds of a kind of edible fungus liquid kind submerged fermentation, it comprises shaking flask kind culture medium prescription and fermentation tank kind culture medium prescription. This shaking flask kind culture medium prescription mainly adds farina, in order to replace the potato slices in traditional shaking flask kind culture medium prescription. This fermentation tank kind culture medium prescription mainly adds farina, in order to replace the materials such as corn flour in traditional zymotic tank kind culture medium prescription, dregs of beans, wheat bran. The present invention also provides a kind of complete production of hybrid seeds technique of edible fungus liquid kind submerged fermentation that uses above-mentioned formula. Technique scheme is mainly to introduce farina, and shaking flask kind is similar with the culture medium prescription of fermentation tank kind, and in production of hybrid seeds process, is convenient to that observations, strain quality equilibrium, raw material are easy to processing, utilization rate is higher.
Description
Technical field
The present invention relates to a kind of edible mashroom cultivating technique and culture medium prescription thereof, relate in particular to the complete production of hybrid seeds technique of a kind of edible fungus liquid kind and culture medium prescription thereof.
Background technology
In edible mushroom, contain bioactivator as: macromolecule polysaccharide, β-glucose and RNA complex, natural organic germanium, nucleic acid degradation product, Cyclic Nucleotides cAMP and triterpene compound etc. are to safeguarding that health has important value. Health food, health drink, wine and medicine taking edible mushroom as raw material production and processing be and input health-product market clinical for medical treatment in a large number. Therefore, edible mushroom is subject to the people of various countries' attention day by day as a kind of pollution-free food. So the demand of edible mushroom is also increasing, the production of hybrid seeds cultivation technique of edible mushroom is also subject to great attention.
The making of edible fungus species is one of industrialized important technology of Edible Fungi. Existing edible fungi liquid strain manufacturing technology mainly comprises test tube kind, shaking flask kind, fermentation tank kind three phases; There is the problems such as formula variation is large, inconvenience is observed, strain quality is unbalanced, raw material intractability is large, utilization rate is low in the making aspect of shaking flask kind and fermentation tank kind wherein.
Summary of the invention
By Given this, necessary culture medium prescription and the production of hybrid seeds technique that a kind of complete production of hybrid seeds of edible fungus liquid kind submerged fermentation that can overcome the problems referred to above is provided.
Technical scheme of the present invention is mainly that farina is introduced in the culture medium prescription of the complete production of hybrid seeds of submerged fermentation, make shaking flask kind similar with the culture medium prescription of fermentation tank kind, make the problems such as the formula variation that aspect exists is large, inconvenience is observed, strain quality is unbalanced, raw material intractability is large, utilization rate is low to solve edible mushroom shaking flask kind and fermentation tank kind.
In order to achieve the above object, the invention provides the culture medium prescription of the complete production of hybrid seeds of a kind of edible fungus liquid kind submerged fermentation, comprising: edible mushroom test tube kind culture medium prescription, edible mushroom shaking flask kind culture medium prescription and edible fungus fermented tank kind culture medium prescription, wherein,
Described shaking flask kind culture medium prescription comprises: in every 1200 ml distilled waters, contain 15 grams of (g) ~ 20g of brown granulated sugar, glucose 15g ~ 20g, farina 5g ~ 7g, peptone 5g ~ 7g, potassium dihydrogen phosphate 0.75g ~ 1.25g, magnesium sulfate 0.45g ~ 0.55g, yeast extract 0.45g ~ 0.55g, calcium chloride 0.25g ~ 0.35g;
Described fermentation tank kind culture medium prescription is: in every 800 premium on currency, contain 3 kilograms of (kg) ~ 5kg of cane sugar powder, brown granulated sugar 2kg ~ 4kg, glucose 0.5kg ~ 1kg, farina 2kg ~ 3kg, corn flour 0.5kg ~ 1.5kg, peptone 0.5kg ~ 1.5kg, potassium dihydrogen phosphate 200g ~ 300g, magnesium sulfate 100g ~ 150g, defoamer 80g ~ 120g, rapeseed oil 500ml, 2 bag wheat bran material bag 1kg/ bags.
Based on the culture medium prescription of the complete production of hybrid seeds of above-mentioned edible fungus liquid kind submerged fermentation, described test tube kind culture medium prescription is: potato 280g ~ 320g is liquor in 1500 ml distilled waters, glucose 27.5g ~ 32.5g, peptone 4.5g ~ 5.5g, potassium dihydrogen phosphate 1.5g ~ 2.5g, magnesium sulfate 0.75g ~ 1.25g, calcium chloride 0.45g ~ 0.55g, Cobastab1Powder 90mg ~ 110mg, agar powder 17.5g ~ 22.5g.
Based on above-mentioned, described edible mushroom comprises pleurotus eryngii, asparagus, mushroom, Ji mushroom, flat mushroom, elegant precious mushroom etc. The bacterial classification of edible mushroom, in manufacturing process, need to be controlled pH value. The kind difference of edible mushroom, needs the scope of the pH value of controlling also different. As, in the time that edible mushroom is mushroom, need in the formula of shaking flask kind, add a certain amount of NaOH, solution initial pH value is reached at 8.5-9.5, as, adding about 0.5g NaOH to make solution initial pH value is 9.0; In the time that edible mushroom is pleurotus eryngii, do not need to regulate in addition pH value, use the pH value of the culture medium of above-mentioned each formula to be enough to be used in cultivating pleurotus eryngii quel strains.
The present invention also provides a kind of production of hybrid seeds technique of the culture medium prescription that uses the complete production of hybrid seeds of above-mentioned edible fungus liquid kind submerged fermentation, comprising:
Test tube kind manufacture craft;
Shaking flask kind manufacture craft, comprising:
(21) component in above-mentioned shaking flask kind culture medium prescription is dissolved in and in 1200 ml waters, forms a culture medium solution, and this culture medium solution is packed in shaking flask and sealing;
(22) the described shaking flask that culture medium is housed is carried out to sterilizing, cooling processing; And
(23) edible mushroom test tube kind is accessed in cooled shaking flask kind culture medium and cultivated, and cultivate into edible mushroom shaking flask kind;
Fermentation tank kind manufacture craft, use volume is the fermentation tank of 1000 liters, comprising:
(31) by the each components dissolved in the formula of above-mentioned fermentation tank kind culture medium in fermentation tank to prepare fermentation tank kind culture medium;
(32) fermentation tank that contains fermentation tank kind culture medium is carried out to sterilizing, cooling processing; And
(33) edible mushroom shaking flask kind is accessed and in above-mentioned fermentation tank, cultivate into fermentation tank kind, and inoculate into bag.
Based on above-mentioned, described step (22) comprises step by step following:
(221) the described shaking flask that culture medium is housed is placed at 121 DEG C ~ 123 DEG C to autoclaving 35 ~ 40 minutes, wherein, can in high-pressure sterilizing pot, carries out sterilization treatment; And (222) carry out high wind pouring processing to the shaking flask kind culture medium after sterilizing; Particularly, the shaking flask kind culture medium juxtaposition of taking out after sterilizing enters in superclean bench, cooling as early as possible with high wind pouring promotion, shortens cool time, can when minimizing nutritional labeling is destroyed, reduce pollution risk.
Based on above-mentioned, described step (23) comprises step by step following:
(231), in the time that shaking flask temperature is down to 20 DEG C, adopt ultraviolet irradiation transfer room and superclean bench 45 ~ 55 minutes to reduce space miscellaneous bacteria radix, to reduce pollution risk;
(232) test tube kind is cut into the bacterial classification piece of mung bean size, in every bottle of shaking flask culture medium, access 5 ~ 7 test tube kind bacterial classifications;
(233) postvaccinal shaking flask is left standstill to 48 ~ 72 hours in super-clean bench and still keep clear state to culture medium, and occur without abnormal conditions, as bubble, become muddy; And
(234) in test tube kind described in constant temperature 20-22 DEG C stir culture, obtain above-mentioned shaking flask kind. Wherein, it should be noted that, within the cultivation shaking flask kind phase, in shaking flask, culture medium still keeps clear state, do not occur bubble, become the abnormal conditions such as muddiness, and mycelial growth rate is normal, mycelia amount increases gradually, bacterium fluid viscosity increases gradually, can be used as shaking flask bacterial classification and use. In addition, according to the difference of edible mushroom kind, the incubation time of this step is also different, as pleurotus eryngii and mushroom etc. can be cultivated 5 ~ 8 days; Flat mushroom, Ji mushroom etc. can be cultivated 4 ~ 5 days.
Based on above-mentioned, described step (31) comprises step by step following:
(311) cane sugar powder, brown granulated sugar and glucose are dissolved in the water of 85 DEG C ~ 95 DEG C and form the first material liquid.
Particularly, 3kg ~ 5kg cane sugar powder, 2kg ~ 4kg brown granulated sugar and 0.5kg ~ 1kg glucose being dissolved in to the temperature of about 10 liters is in the water of 85 DEG C ~ 95 DEG C.
(312) farina, corn flour, peptone, potassium dihydrogen phosphate, magnesium sulfate, defoamer are dissolved in the warm water of 35 DEG C ~ 55 DEG C and form the second material liquid.
Particularly, it is in the warm water of 35 DEG C ~ 55 DEG C that 2kg ~ 3kg farina, 0.5kg ~ 1.5kg corn flour, 0.5kg ~ 1.5kg peptone, 200g ~ 300g potassium dihydrogen phosphate, 100g ~ 150g magnesium sulfate and 80g ~ 120g defoamer are dissolved in to about 35 ~ 45 intensification degree, and repeatedly stir to accelerate raw material and fully dissolve, occur until no longer include bulk particle in the second material liquid.
(313) above-mentioned 2 bag wheat bran material bag 1kg/ bags are placed in the fermentation tank of half tank water and take out this 2 bag wheat bran material bag 1kg/ bag after liquor, and add first and second material liquid to prepare fermentation tank kind culture medium.
Particularly, above-mentioned 2 bag wheat bran material bags are added in fermentation tank from fermentation tank charging aperture, and in fermentation tank, have half tank temperature to be approximately the water of 100 DEG C; Be 123 DEG C ~ 126 DEG C these fermentation tanks of Steam Heating to passing into temperature in fermentation tank, when fermentation pot liquid is during in fluidized state, when there is the large vapour that continues to fling upwards at an i.e. inoculation mouthful valve place, stop passing into steam to fermentation tank, pass into clean cold air to fermentation tank body; Then from fermentation tank, take out wheat bran material bag, described first and second material liquid is delivered in fermentation tank, obtain described fermentation tank kind culture medium; When this first and second material liquid is to after carrying in fermentation tank, seal this fermentation tank; Wherein, necessary fastening in this fermentation tank, otherwise in follow-up cooling procedure, can cause that suck-back pollutes.
Based on above-mentioned, described step (32) comprises step by step following:
(321) pass into steam to increase the pressure of fermentation tank to fermentation tank, in the time that the pressure of fermentation tank rises to 0.11Mpa, discharge cold air residual in fermentation tank; Wherein, the cold air in fermentation tank must thoroughly be discharged, otherwise easily causes fermentation tank sterilizing not thorough, thereby causes pollution;
(322), in the time that the pressure of fermentation tank rises to 0.13Mpa, the condensed water in discharge filter is filter sterilised;
(323) filter sterilised, after approximately 20 ~ 25 minutes, passes into clean cold air to dry up filter to fermentation tank; Discharge the steam of fermentation tank with step-down simultaneously; And
(325), when fermentation tank Pressure Drop is during to 0.06-0.1Mpa, slightly open the lower valve discharge 30 ~ 60 seconds of fermentation tank; Discharge completes, and passes into water quench.
Based on above-mentioned, described step (33) comprises step by step following:
(331) in the time that culture medium in fermentation tank is cooled to 20 DEG C ~ 25 DEG C, stop passing into cooling water; Formaldehyde cotton balls plug, in fermentation tank inoculation mouth, is fixed to the alcohol sliver of 99% in its outside with copper wire; Light alcohol sliver, under flame protection, by the culture medium in shaking flask kind access fermentation tank;
(332), in the time that the temperature of fermentation tank maintains 20 DEG C ~ 22 DEG C, cultivate shaking flask kind and within 5 ~ 7 days, grow up to fermentation tank kind; Wherein, at fermentation tank kind growing period, do not occur that peculiar smell, variable color, mycelia the situation such as melt, through sterile sampling, that sediments microscope inspection is confirmed is qualified, can be used as and produce by kind, inoculate into bag; And
(333) employing temperature is that the steam of 110 DEG C ~ 115 DEG C is inoculation pipeline and inoculating gun sterilizing 8-12 minute, then inoculates into bag.
In addition, when inoculate bag complete after, also further comprise to fermentation tank and continue supercharging to remove remaining bacterial classification, in water pipe can,douche, especially on visor mouth, coil pipe and the equipotential of charging aperture top.
Based on above-mentioned, described test tube kind manufacture craft comprises the following steps:
(11) make test tube kind culture medium: by 280g ~ 320g potato slices, and in 1500ml distilled water liquor, filter and retain potato filtrate; In above-mentioned potato filtrate, add glucose 27.50g ~ 32.5g, peptone 4.5g ~ 5.5g, potassium dihydrogen phosphate 1.5g ~ 2.5g, magnesium sulfate 0.75g ~ 1.25g, calcium chloride 0.45g ~ 0.55g, Cobastab1Powder 90mg ~ 110mg is also stirred to abundant dissolving and obtains test tube kind culture medium premixed liquid; Under 300 DEG C of constant temperature, agar powder 17.5g ~ 22.5g is added in described test tube kind culture medium premixed liquid and be stirred to agar powder and fully dissolve and prepare test tube kind culture medium;
(12) by described test tube kind culture medium difference injecting tube, stoppered test tube mouth; And at 121 DEG C ~ 123 DEG C, this test tube is carried out to autoclaving and process 25 ~ 30 minutes; Wherein, can adopt syringe by test tube kind culture medium injecting tube;
(13) pendulum inclined-plane, cooling;
(14) access respectively edible mushroom tissue to every test tube kind culture medium and separate 1 of kind of piece, and at 22 DEG C ~ 24 DEG C, cultivate this tissue separation kind of piece to being about to cover with test tube, as the first test tube kind; Wherein, according to the difference of edible mushroom kind, the time that cultured tissue separates kind is also different, need cultivate 12 ~ 15 days as pleurotus eryngii and asparagus, and mushroom need be cultivated 15 ~ 20 days, and flat mushroom kind need be cultivated 10 ~ 13 days;
(15) above-mentioned the first test tube kind is carried out to test tube kind tube and expand numerous processing, and at 22 DEG C ~ 24 DEG C, cultivate again this first test tube kind, as the second test tube kind, as the kind of setting out of shaking flask kind; Equally, according to the difference of edible mushroom kind, it is also different that test tube kind tube expands numerous incubation time again, that is, the time of cultivating the first test tube kind is also different, as pleurotus eryngii and asparagus need be cultivated 12 ~ 15 days again, mushroom need be cultivated 15 ~ 20 days, and flat mushroom kind need be cultivated 10 ~ 13 days. Preferably, in the time that mycelia is about to full test tube, therefrom choose without assortedly dying, pure white dense, the moderate test tube kind of most advanced and sophisticated neat, long speed, can be used as kind of the use of setting out of shaking flask kind.
Be appreciated that the difference according to the concrete kind of edible mushroom, concrete production of hybrid seeds technique is also variant, and as in making mushroom shaking flask kind process, need in step (21), add a certain amount of NaOH to regulate ph is 8.5-9.5; As, adding about 0.5g NaOH to make solution initial pH value is 9.0.
The culture medium prescription of the complete production of hybrid seeds of edible fungus liquid kind submerged fermentation provided by the invention and the relative prior art of production of hybrid seeds technique have the following advantages:
First, in the formula of edible mushroom shaking flask kind and fermentation tank kind culture medium, all introduce farina, farina is soluble in water, and the links such as traditional potato is peeled, section, liquor have been removed in the introducing of farina, simplify traditional edible fungus liquid culture growth medium manufacturing process.
Secondly, in the formula of edible mushroom shaking flask kind and fermentation tank kind culture medium, all introduce farina, and starch materials using farina as production of hybrid seeds edible fungus species, compared with the traditional bulky grain such as corn flour, bean cake powder starch materials, the introducing of farina has improved transparency and the observability of culture medium raw material, is conducive to very first time discovery pollution condition in the complete production of hybrid seeds technique of liquid strain submerged fermentation.
Again, raw material compared to corn flour, bean cake powder etc. taking amylose as main component, farina is mainly amylopectin laxer on molecular structure, this makes in Starch Hydrolysis process, hydrogen proton is more prone to enter starch molecule inside and reacts, thereby thereby greatly having shortened Starch Hydrolysis is the time that glucose is absorbed by hyphal cell; And under identical production of hybrid seeds condition, make shaking flask kind and fermentation tank kind reach 100% density time shorten 1 ~ 3 day, improve the uniformity of shaking flask kind and fermentation tank bacterial classification cell age, inoculate after bacterium rod, made to have sent out bacterium purseful time shorten 7 ~ 13 days.
Further, in the formula of the culture medium of edible mushroom test tube kind and shaking flask kind, all introduce calcium chloride, especially in test tube kind culture medium prescription, introduce calcium chloride and can supplement the calcium of edible mushroom, make edible mushroom outward appearance good.
Detailed description of the invention
Below by detailed description of the invention, technical scheme of the present invention is described in further detail.
The production of hybrid seeds technique of the culture medium prescription of the complete production of hybrid seeds of embodiment 1-pleurotus eryngii liquid strain
A, pleurotus eryngii test tube kind culture medium prescription and production of hybrid seeds technique thereof
Formula
Potato 300g is liquor, glucose 30g, peptone 5g, potassium dihydrogen phosphate 2g, magnesium sulfate 1g, calcium chloride 0.5g, vitamin B1 powder 100mg, agar powder 20g in 1500 ml distilled waters
Test tube kind production of hybrid seeds technique
A1, potato 300g section, pours liquor in distilled water 1500ml into, gets potato filtrate stand-by; In potato filtrate, dissolve in glucose 30g, peptone 5g, potassium dihydrogen phosphate 2g, magnesium sulfate 1g, calcium chloride 0.5g, vitamin B1 powder 100mg, is stirred to abundant dissolving, obtains test tube kind culture medium premixed liquid;
A2, test tube kind culture medium premixed liquid is refunded in stainless-steel pan, under 300 DEG C of constant temperature, dissolve in agar powder 20g, and constantly at the uniform velocity stir and promote that agar powder fully dissolves, obtain test tube kind culture medium; In vitro inject 10ml test tube kind culture medium, the use silica gel plug jam-pack mouth of pipe with syringe to every while hot;
A3, at 121 DEG C ~ 123 DEG C, by the test tube sterilizing that contains test tube kind culture medium 28 minutes; Then take out test tube and set inclined-plane; Be cooled to 20 DEG C stand-by;
A4, choose the fresh pleurotus eryngii kind that kind of property is good, the pleurotus eryngii tissue that accesses respectively mung bean size for every invisible spectro culture medium separates 1 of kind of piece;
A5, postvaccinal test tube is lain in constant incubator, at 23 DEG C, cultivate 12 days;
A6, select growing way best test tube kind, the bacterial classification piece of switching mung bean size, in new test-tube culture medium, and is cultivated the test tube kind of 12 days at 23 DEG C; Therefrom choose without assorted dying, pure white dense, the moderate kind of setting out as shaking flask kind of most advanced and sophisticated neat, long speed.
B, pleurotus eryngii shaking flask kind culture medium prescription and production of hybrid seeds technique thereof
Formula
Embodiment test group: brown granulated sugar 15g, glucose 15g, farina 5g, peptone 5g, potassium dihydrogen phosphate 1g, magnesium sulfate 0.5g, yeast extract 0.5g, calcium chloride 0.3g.
Control group one: glucose 30g, corn flour 5g, peptone 5g, potassium dihydrogen phosphate 1g, magnesium sulfate 0.5g, yeast extract 0.5g.
Control group two: glucose 30g, dregs of beans 5g, peptone 5g, potassium dihydrogen phosphate 1g, magnesium sulfate 0.5g, yeast extract 0.5g.
Control group three: glucose 30g, wheat bran 5g, peptone 5g, potassium dihydrogen phosphate 1g, magnesium sulfate 0.5g, yeast extract 0.5g.
Shaking flask kind production of hybrid seeds technique
B1, will be above each group of formula raw material dissolve in respectively in 1200ml warm water, uniform stirring promotes fully to dissolve and obtains shaking flask kind culture medium; This shaking flask kind culture medium is distributed in 2 1000ml triangular flasks, adds 600ml shaking flask kind culture medium to each triangular flask, put into respectively 1 of 2.5cm stirrer; Cover tampon, build with eight layers of gauze, tighten bottleneck with cotton thread; Sterilizing 35 minutes at 123 DEG C;
B2, shaking flask kind culture medium after sterilizing is completed are inserted in superclean bench, with high wind drench promote cooling as early as possible; In the time that the temperature of shaking flask kind culture medium is down to 20 DEG C, get the second test tube kind expanding after numerous tube, for every bottle of shaking flask culture medium accesses respectively the pleurotus eryngii quel strains piece of 5-7 piece mung bean size;
B3, postvaccinal shaking flask kind leave standstill after 72h in super-clean bench, postvaccinal shaking flask kind be placed on magnetic stirring apparatus, and kick-down, 20 DEG C of cultivations of constant temperature obtain expanding the shaking flask kind after numerous, as the kind of setting out of fermentation tank kind.
C, pleurotus eryngii fermentation tank kind culture medium prescription and production of hybrid seeds technique thereof
Formula
Embodiment test group: cane sugar powder 5kg, brown granulated sugar 4kg, glucose 1kg, farina 2kg, corn flour 1kg, peptone 1kg, potassium dihydrogen phosphate 200g, magnesium sulfate 100g, defoamer 100g, rapeseed oil 500ml, install 1kg wheat bran/bag * 2 bundles, tighten sack and fasten stand-by with nylon rope;
Control group one: cane sugar powder 5kg, brown granulated sugar 4kg, glucose 1kg, corn flour 3kg, peptone 1kg, potassium dihydrogen phosphate 200g, magnesium sulfate 100g, defoamer 100g, rapeseed oil 500ml, wheat bran 2kg;
Control group two: cane sugar powder 5kg, brown granulated sugar 4kg, glucose 1kg, dregs of beans 3kg, peptone 1kg, potassium dihydrogen phosphate 200g, magnesium sulfate 100g, defoamer 100g, rapeseed oil 500ml, wheat bran 2kg;
Production of hybrid seeds technique
C1, the liquid spawn fermenting tank that is 1000L to volume add half tank water, and are heated to 100 DEG C;
C2, general above each group of formula raw material are fully dissolved and are added in fermentation tank, pass into 126 DEG C of steam and heat up;
C3, in the time that fermentation tank pressure rises to 0.11Mpa, half-open drain tap is to discharge residual cold air in fermentation tank;
C4, in the time that fermentation tank pressure rises to 0.13Mpa, the condensed water generating in discharge filter be filter sterilised;
After C5, filter sterilised approximately 20 minutes, in fermentation tank, pass into clean cold air to dry up filter;
C6, when the Pressure Drop of fermentation tank is during to 0.1Mpa, slightly open the about 60s of tank body lower valve discharge;
C7, discharge complete, and pass into water quench;
C7, in the time that culture medium in fermentation tank is cooled to 20 DEG C, stop passing into cooling water; Formaldehyde cotton balls plug, in fermentation tank inoculation mouth, is fixed to the alcohol sliver of 99% in its outside with copper wire; Light alcohol sliver, under flame protection, access each 1 bottle of the pleurotus eryngii shaking flask bacterial classification that 6 days cell age growing ways are good; At 22 DEG C of indoor temperatures, cultivate, through sterile sampling, that sediments microscope inspection is confirmed is qualified, three group of formula are respectively inoculated into 600 bags, bag, and at 22 DEG C of room temperatures, constant temperature culture is sent out bacterium.
Result of the test
Pleurotus eryngii shaking flask kind formula contrast test
| Classification | Transparency | Visible time/the h of mycelia | Density reaches 100% time/h |
| Embodiment test group | +++ | 48 | 120h |
| Control group 1 | ++ | 72 | 144h |
| Control group 2 | ++ | 72 | 168h |
| Control group 3 | + | 72 | 144h |
Pleurotus eryngii fermentation tank kind formula contrast test
| Classification | Viscosity | Mycelial density | Mobility | The mycelia time of revealing the exact details/sky | Send out bacterium purseful time/sky |
| Embodiment test group | +++ | +++ | +++ | 7~9 | 15~18 |
| Control group 1 | ++ | ++ | ++ | 12~15 | 22~25 |
| Control group 2 | + | ++ | + | 13~17 | 23~28 |
As can be seen here:
(1) shaking flask kind and fermentation tank kind production of hybrid seeds link, removed the links such as traditional potato is peeled, section, liquor, simplifies traditional edible fungus liquid culture growth medium manufacturing process;
(2) in shaking flask kind and fermentation tank kind contrast test, the farina of embodiment test group has substituted the bulky grain starch materials such as the corn flour, bean cake powder, wheat bran of control group, thereby improve transparency and the observability of culture medium raw material, be conducive to find pollution condition in the very first time;
(3) shaking flask production of hybrid seeds control group kind is mainly with corn flour, bean cake powder, the amyloses such as wheat bran are the raw material of main component, farina in embodiment test group is mainly amylopectin laxer on molecular structure, this makes in Starch Hydrolysis process, hydrogen proton is more prone to enter starch molecule inside and reacts, thereby it is the time that glucose is absorbed by hyphal cell thereby greatly shortened Starch Hydrolysis, the time that embodiment test group shaking flask kind and fermentation tank bacterial classification reach 100% density shortened to about 120 hours by about 168 hours, 48 hours are shortened,
(4) improve the uniformity of shaking flask kind and fermentation tank bacterial classification cell age, inoculate after bacterium rod, send out the bacterium purseful time by within 25-28 days, reducing to 15-18 days.
The production of hybrid seeds technique of the culture medium prescription of the complete production of hybrid seeds of embodiment 2-mushroom liquid strain
A, mushroom test tube kind culture medium prescription and production of hybrid seeds technique thereof
In the present embodiment, mushroom test tube kind culture medium prescription and production of hybrid seeds technique thereof are identical with embodiment 1 pleurotus eryngii test tube kind culture medium prescription and production of hybrid seeds technique thereof.
B, mushroom shaking flask kind culture medium prescription and production of hybrid seeds technique thereof
Formula
Embodiment test group: brown granulated sugar 15g, glucose 15g, farina 5g, peptone 5g, potassium dihydrogen phosphate 1g, magnesium sulfate 0.5g, yeast extract 0.5g, calcium chloride 0.3g, NaOH 0.5g.
Control group one: glucose 30g, wheat bran 10g, peptone 5g, potassium dihydrogen phosphate 1g, magnesium sulfate 0.5g, yeast extract 0.5g, calcium chloride 0.3g, NaOH 0.5g.
Control group two: glucose 30g, corn flour 10g, peptone 5g, potassium dihydrogen phosphate 1g, magnesium sulfate 0.5g, yeast extract 0.5g, calcium chloride 0.3g, NaOH 0.5g.
In the present embodiment, mushroom shaking flask kind production of hybrid seeds technique is identical with pleurotus eryngii shaking flask kind production of hybrid seeds technique in embodiment 1.
C, mushroom ferment tank kind culture medium prescription and production of hybrid seeds technique thereof
Formula
In the present embodiment, the formula of mushroom ferment tank kind embodiment test group is identical with the formula of pleurotus eryngii fermentation tank kind embodiment test group in embodiment 1.
Control group one: glucose 6.4kg, dregs of beans 2.4kg, corn flour 1kg, peptone 1kg, potassium dihydrogen phosphate 200g, magnesium sulfate 100g, defoamer 100g, rapeseed oil 500ml, install 1kg wheat bran/bag * 2 bundles, tighten sack and fasten stand-by with nylon rope.
Control group two: wheat bran 3.6kg, corn flour 2kg, peptone 1kg, potassium dihydrogen phosphate 200g, magnesium sulfate 100g, defoamer 100g, rapeseed oil 500ml, wheat bran 2kg directly put into tank and use, and wrap up without gauze.
In the present embodiment, in mushroom ferment tank kind production of hybrid seeds technique and embodiment 1, pleurotus eryngii fermentation tank kind production of hybrid seeds technique is basic identical, and difference is, the inoculation of mushroom enters different that bag and the inoculation of pleurotus eryngii enter bag. In the present embodiment, the inoculation of mushroom enters bag and is: will respectively organize liquid spawn above and inoculate respectively into each 500 bags of 18 × 33cm polypropylene strain bag lentinus edodes strain stick, at 24 DEG C of room temperatures, constant temperature culture is sent out bacterium.
Result of the test
Mushroom shaking flask kind formula contrast test
| Classification | Transparency | Visible time/the h of mycelia | Density reaches 100% time/h |
| Embodiment test group | +++ | 60 | 144 |
| Control group 1 | + | 96 | 216 |
| Control group 2 | ++ | 72 | 168 |
Mushroom ferment tank kind formula contrast test
| Classification | Viscosity | Mycelial density | Mobility | Send out bacterium purseful time/sky |
| Embodiment test group | +++ | +++ | +++ | 15~23 |
| Control group 1 | + | ++ | ++ | 19~28 |
| Control group 2 | ++ | ++ | + | 23~30 |
As can be seen here:
Compared with shaking flask production of hybrid seeds control group kind, embodiment test group shaking flask kind reach 100% density time shorten 1 ~ 3 day; The uniformity of shaking flask kind and fermentation tank bacterial classification cell age is improved, and inoculates after bacterium rod, and a bacterium purseful time can shorten 7-10 days.
In addition, the present embodiment further proves that the introducing of farina not only can simplify traditional edible fungus liquid culture growth medium manufacturing process; But also improved transparency and the observability of culture medium raw material, be conducive to find pollution condition in the very first time of edible fungus liquid culture growth medium manufacturing process.
Finally should be noted that: above embodiment is only in order to illustrate that technical scheme of the present invention is not intended to limit; Although the present invention is had been described in detail with reference to preferred embodiment, those of ordinary skill in the field are to be understood that: still can modify or part technical characterictic is equal to replacement the specific embodiment of the present invention; And not departing from the spirit of technical solution of the present invention, it all should be encompassed in the middle of the technical scheme scope of request protection of the present invention.
Claims (10)
1. a culture medium prescription for the complete production of hybrid seeds of edible fungus liquid kind submerged fermentation, comprising: edible mushroom test tube kind culture medium prescription, edible mushroom shaking flask kind culture medium prescription and edible fungus fermented tank kind culture medium prescription, is characterized in that:
Described edible mushroom shaking flask kind culture medium prescription comprises: in every 1200 ml distilled waters, contain 15 ~ 20 grams of brown granulated sugars, 15 ~ 20 grams of glucose, 5 ~ 7 grams of farinas, 5 ~ 7 grams of peptones, 0.75 ~ 1.25 gram of potassium dihydrogen phosphate, 0.45 ~ 0.55 gram, magnesium sulfate, 0.45 ~ 0.55 gram of yeast extract, 0.25 ~ 0.35 gram, calcium chloride;
Described edible fungus fermented tank kind culture medium prescription is: in every 800 premium on currency, contain 3 ~ 5 kilograms of cane sugar powders, 2 ~ 4 kilograms of brown granulated sugars, 0.5 ~ 1 kilogram of glucose, 2 ~ 3 kilograms of farinas, 0.5 ~ 1.5 kilogram of corn flour, 0.5 ~ 1.5 kilogram of peptone, 200 ~ 300 grams of potassium dihydrogen phosphates, 100 ~ 150 grams, magnesium sulfate, 80 ~ 120 grams of defoamers, 500 milliliters of rapeseed oils, 1 kilogram/bag of 2 bag wheat bran material bags.
2. the culture medium prescription of the complete production of hybrid seeds of edible fungus liquid kind submerged fermentation according to claim 1, it is characterized in that, described edible mushroom test tube kind culture medium prescription is: 280 ~ 320 grams of liquors in 1500 ml distilled waters of potato, 27.5 ~ 32.5 grams of glucose, 4.5 ~ 5.5 grams of peptones, 1.5 ~ 2.5 grams of potassium dihydrogen phosphates, 0.75 ~ 1.25 gram, magnesium sulfate, 0.45 ~ 0.55 gram, calcium chloride, Cobastab190 ~ 110 milligrams, powder, 17.5 ~ 22.5 grams of agar powders.
3. the culture medium prescription of the complete production of hybrid seeds of edible fungus liquid kind submerged fermentation according to claim 1, is characterized in that, described edible mushroom is pleurotus eryngii, asparagus, mushroom, Ji mushroom, flat mushroom or elegant precious mushroom.
4. a production of hybrid seeds technique for the culture medium prescription of the complete production of hybrid seeds of edible fungus liquid kind submerged fermentation described in right to use requirement 1 to 3 any one, comprising:
Test tube kind manufacture craft;
Shaking flask kind manufacture craft, comprising:
(21) the each component in described shaking flask kind culture medium prescription is dissolved in and in 1200 ml waters, forms a shaking flask kind culture medium, and this shaking flask kind culture medium is packed in shaking flask and sealing;
(22) the described shaking flask that culture medium is housed is carried out to sterilizing, cooling processing; And
(23) edible mushroom test tube kind is accessed in cooled shaking flask kind culture medium and cultivated, and cultivate into edible mushroom shaking flask kind;
Fermentation tank kind manufacture craft, use volume is the fermentation tank of 1000 liters, comprising:
(31) by the each components dissolved in the formula of described fermentation tank kind culture medium in fermentation tank to prepare fermentation tank kind culture medium;
(32) fermentation tank that contains fermentation tank kind culture medium is carried out to sterilizing, cooling processing; And
(33) edible mushroom shaking flask kind is accessed and in above-mentioned fermentation tank, cultivate into fermentation tank kind, and inoculate into bag.
5. production of hybrid seeds technique according to claim 4, is characterized in that, described step (22) comprising:
To the described shaking flask that culture medium is housed autoclaving 35 ~ 40 minutes at 121 DEG C ~ 123 DEG C; And
Shaking flask kind culture medium after sterilizing is carried out to high wind and drench processing.
6. production of hybrid seeds technique according to claim 5, is characterized in that, described step (23) comprising:
In the time that shaking flask temperature is down to 20 DEG C, adopt ultraviolet irradiation transfer room and superclean bench 45 ~ 55 minutes;
Test tube kind is cut into the bacterial classification piece of mung bean size, in every bottle of shaking flask culture medium, access 5-7 piece test tube kind bacterial classification, and standing 48-72 hour still keeps clear state to culture medium; And
In test tube kind described in 20 DEG C ~ 22 DEG C stir culture of constant temperature, obtain described shaking flask kind.
7. production of hybrid seeds technique according to claim 6, is characterized in that, described step (31) comprising:
Cane sugar powder, brown granulated sugar and glucose are dissolved in the water of 85 DEG C ~ 95 DEG C and form the first material liquid;
Farina, corn flour, peptone, potassium dihydrogen phosphate, magnesium sulfate, defoamer are dissolved in the warm water of 35 DEG C ~ 55 DEG C and form the second material liquid; And
Described 2 bag wheat bran material bags are placed in the fermentation tank of half tank water and take out this 2 bag wheat bran material bag after liquor, and add first and second material liquid to prepare fermentation tank kind culture medium.
8. production of hybrid seeds technique according to claim 7, is characterized in that, described step (32) comprising:
Pass into steam to increase the pressure of fermentation tank to fermentation tank, in the time that the pressure of fermentation tank rises to 0.11Mpa, discharge cold air residual in fermentation tank;
In the time that the pressure of fermentation tank rises to 0.13Mpa, the condensed water in discharge filter is filter sterilised;
After filter sterilised 20 ~ 25 minutes, pass into clean cold air to dry up filter to fermentation tank, discharge the steam of fermentation tank with step-down simultaneously; And
In the time of fermentation tank Pressure Drop to 0.06 ~ 0.1Mpa, open fermentation tank lower valve discharge 30 ~ 60 seconds, discharge completes, and passes into water quench.
9. production of hybrid seeds technique according to claim 8, is characterized in that, described step (33) comprising:
In the time that culture medium in fermentation tank is cooled to 20 DEG C ~ 25 DEG C, stop passing into cooling water; Formaldehyde cotton balls plug, in fermentation tank inoculation mouth, is fixed to the alcohol sliver of 99% in its outside with copper wire; Light alcohol sliver, under flame protection, by the culture medium in shaking flask kind access fermentation tank;
In the time that the temperature of fermentation tank maintains 20 DEG C ~ 22 DEG C, cultivate shaking flask kind and within 5 ~ 7 days, grow up to fermentation tank kind; And
Employing temperature is that the steam of 110 DEG C ~ 115 DEG C is inoculation pipeline and inoculating gun sterilizing 8-12 minute, then inoculates into bag.
10. production of hybrid seeds technique according to claim 9, is characterized in that, described test tube kind manufacture craft comprises the following steps:
By 280 ~ 320 grams of potato slices, and in 1500ml distilled water liquor, filter and retain potato filtrate; In this potato filtrate, add 27.5 ~ 32.5 grams of glucose, 4.5 ~ 5.5 grams of peptones, 1.5 ~ 2.5 grams of potassium dihydrogen phosphates, 0.75 ~ 1.25 gram, magnesium sulfate, 0.45 ~ 0.55 gram, calcium chloride, Cobastab190 ~ 110 milligrams, powder is also stirred to abundant dissolving and obtains test tube kind culture medium premixed liquid; Under 300 DEG C of constant temperature, 17.5 ~ 22.5 grams of agar powders are added in described test tube kind culture medium premixed liquid and be stirred to agar powder and fully dissolve and prepare test tube kind culture medium;
In described test tube kind culture medium difference injecting tube, stoppered test tube mouth; And at 121 DEG C ~ 123 DEG C, this test tube is carried out to autoclaving and process 25 ~ 30 minutes;
Pendulum inclined-plane, cooling;
In every test tube kind culture medium, access 1 of edible mushroom tissue separation kind of piece, and at 22 DEG C ~ 24 DEG C, cultivate this tissue separation kind of piece to being about to cover with this test tube, as the first test tube kind; And
Described the first test tube kind is carried out to test tube kind tube and expand numerous processing, and at 22 DEG C ~ 24 DEG C, cultivate again this first test tube kind, as the kind of setting out of shaking flask kind.
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| CN104498370A (en) * | 2014-12-11 | 2015-04-08 | 常盛杰 | Preparation method of culture medium for mother seeds of lentinus edodes |
| CN104996166A (en) * | 2015-07-01 | 2015-10-28 | 天津齐心菌类种植有限公司 | Cultivation method for liquid strains |
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| CN105385607A (en) * | 2015-12-18 | 2016-03-09 | 湖北五林中地农业科技有限公司 | Lentinus edodes liquid submerged fermentation culture medium formula and fermentation technology |
| CN105693330A (en) * | 2016-02-04 | 2016-06-22 | 湖南省宇秀生物科技有限公司 | High-transparency PDA (potato dextrose agar) culture medium preparing method |
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| CN112616560A (en) * | 2019-09-24 | 2021-04-09 | 吉林益隆长白山实业有限公司 | Mushroom liquid strain culture material and preparation method thereof |
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| CN112005813A (en) * | 2020-09-09 | 2020-12-01 | 济南蓬生农业科技有限公司 | Liquid strain production process and purity detection method |
| CN112088720A (en) * | 2020-09-19 | 2020-12-18 | 灌南云农食用菌研究所(有限合伙) | Pleurotus eryngii liquid culture medium and pleurotus eryngii liquid culture production process |
| CN114009272B (en) * | 2021-10-11 | 2023-07-21 | 中国农业科学院农业资源与农业区划研究所 | Liquid edible fungus culture solution and preparation method thereof |
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