CN106431572A - Lucid-ganoderma cultivation method for increasing ganoderan and selenium content - Google Patents

Lucid-ganoderma cultivation method for increasing ganoderan and selenium content Download PDF

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Publication number
CN106431572A
CN106431572A CN201610870053.8A CN201610870053A CN106431572A CN 106431572 A CN106431572 A CN 106431572A CN 201610870053 A CN201610870053 A CN 201610870053A CN 106431572 A CN106431572 A CN 106431572A
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ganoderma
temperature
day
parts
greenhouse
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苏华
陆兵
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Huai-An Natural Health-Care Food Development Co Ltd Guilin
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Huai-An Natural Health-Care Food Development Co Ltd Guilin
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    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01GHORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
    • A01G18/00Cultivation of mushrooms
    • CCHEMISTRY; METALLURGY
    • C05FERTILISERS; MANUFACTURE THEREOF
    • C05DINORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C; FERTILISERS PRODUCING CARBON DIOXIDE
    • C05D3/00Calcareous fertilisers
    • CCHEMISTRY; METALLURGY
    • C05FERTILISERS; MANUFACTURE THEREOF
    • C05DINORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C; FERTILISERS PRODUCING CARBON DIOXIDE
    • C05D9/00Other inorganic fertilisers
    • C05D9/02Other inorganic fertilisers containing trace elements

Abstract

The invention discloses a lucid-ganoderma cultivation method for increasing the ganoderan and selenium content. The lucid-ganoderma cultivation method includes the steps of 1) medium preparing, wherein a medium is prepared from, by weight, 75 parts of wood flour, 20 parts of wheat bran, 2 parts of corn flour, 1 part of lime, 1 part of gypsum and 1 part of sugar, the ratio of a material to water is adjusted by selenium-rich nutrient liquid to be 1:(1.3-1.5); 2) material stirring, bagging, sterilizing and inoculating; 3) spawn running, wherein after inoculating is carried out, lucid ganoderma is put into a greenhouse and subjected to spawn running, at first, the temperature of the greenhouse is controlled to be 26 DEG C to 28 DEG C, and on the 20th day after inoculating is carried out, the temperature of the greenhouse is increased to be 34 DEG C, the temperature is kept for 20 h to 24 h, and then the temperature of the greenhouse is recovered to be 26 DEG C to 28 DEG C; 4) lucid ganoderma discharging, wherein after hyphas overgrows a bag, the lucid ganoderma enters the lucid ganoderma discharging stage, the temperature of the greenhouse is increased to be 34 DEG C on the 5th day, the 10th day, the 15th day, the 20th day and the 25th day of the lucid ganoderma discharging stage of the lucid ganoderma respectively, the temperature is kept for 20 h to 24 h, and then the temperature of the greenhouse is recovered to be 26 DEG C to 28 DEG C.

Description

Improve the ganoderma lucidum cultivation method of ganoderan and Se content
Technical field
The present invention relates to fungus growing technique field, more particularly, to improve the cultivation of glossy ganoderma side of ganoderan and Se content Method.
Background technology
Ganoderma (scientific name:Ganoderma Lucidum Karst), also known as Ganoderma lucidum seu Japonicum, Herba mesonae chinensis, the Ganoderma lucidum (Leyss. Ex Fr.) Karst. that has another name called, red sesame, wood spirit Sesame, Ganoderma, Ganoderma lucisum, Ganoderma lucidum seu Japonicum, are the Herbs of Polyporaceae plant Ganoderma lucidum (Leyss. Ex Fr.) Karst. or Ganoderma, are the son of On Polyporaceae Ganoderma Entity.Profile is in umbrella, cap kidney shape, semicircle or subcircular.Ganoderma is as the Chinese tradition having thousands of years medicinal history Valuable ingredient of Chinese medicine, has invigorating QI and tranquilization, effect of relieving cough and asthma, for dizziness sleeplessness, shortness of breath and palpitation, asthenia cough with asthma.Ganoderma possesses Very high medical value, through scientific research institution's many decades modern pharmacology research confirm, Ganoderma for strengthen body immunity, Adjust blood glucose, control blood pressure, adjuvant therapy chemicotherapy, hepatoprotective, the aspect such as hypnotic is respectively provided with significant curative effect.Ganoderma is many Sugar is one of principle active component of glossy ganoderma medicinal function, is primarily present in glossy ganoderma cell wall inwall, and most of is beta glucan, Its monosaccharide forms in addition to containing a large amount of glucoses, also contains a small amount of arabinose, xylose, fucose, rhamnose, galactose and sweet Dew sugar etc., they are mostly bonded with glucosides such as (1-3), (1-4) and (1-6), have extensive biological activity, such as strengthen immunity The effects such as power, removing human free radical, protection cardiovascular, radioprotective, the liver protecting, antitumor.Triterpeness are also having of Ganoderma Effect one of composition, contained by Ganoderma, triterpeness are no less than 100 kinds of, wherein based on tetracyclic triterpenes, the bitterness of Ganoderma and contained triterpene Class is relevant, has cytotoxicity to human hepatoma cell, also can suppress the release of histamine, has liver protection and antiallergic etc. and makees With.
At present, the technology such as artificial generation (bottle) the material cultivation of Ganoderma, fermenting and producing is in succession successful, wherein, fermenting and producing High to equipment requirements, investment is big, is unfavorable for wide popularization and application;Ganoderma Lucidum is long-standing in China, from cultivation basswood To the development of substituting stuff cultivation technology, promote Ganoderma artificial culture on a large scale.As Chinese patent CN103724115A, disclose one Plant Lucid ganoderma pocket planting fungus bag, be mainly formulated using following raw materials by weight:Mulberry wood chips 75-80%, cotton seed hullss 10- 16%th, Testa Tritici 3-5%, Gypsum Fibrosum powder 0.8-1.2%, water content 55-65%, pH value 6.5-7.5, pack.Required for Ganoderma growth Nutrient substance and other edible fungi on the whole for unanimous on the whole, such as carbon nutrition, nitrogen nutrition, vitamin and mineral Deng in the proportioning of Ganoderma substituting stuff cultivation culture medium, traditional major ingredient has cotton seed hullss, weed tree sawdust, agricultural crop straw etc., adjuvant There are Testa Tritici, wooden bran, Semen Maydis powder etc., separately have the additives such as Gypsum Fibrosum, calcium superphosphate, white sugar.Artificial substituting stuff cultivation is mainly with wood flour, cotton The agricultural leftover bits and pieces such as seed shell, bagasse, crop material and waste residue waste material make culturing raw material, easy and simple to handle, and small investment is it is easy to generally push away Wide application, but waste residue waste material complicated component, have may contain harmful components, unfavorable to ganoderma lucidum product quality control.
Selenium is trace element needed by human.Selenium participates in multiple selenoenzymes and Selenonic protein in synthesized human.Wherein paddy Guang Sweet peptide peroxidase, is catalyzed hydroperoxides in vivo or lipid peroxide is changed into water or various alcohols, eliminates Free radical, to biomembranous attack, protects biomembrane from oxidative damage;Selenium participates in constituting the de- iodine enzyme of iodate thyroid amino acid.? Internal selenium and Vitamin E are worked in coordination with, and can protect cell membrane, prevent the oxidation of unsaturated fatty acid.Trace selenium has protective effect on cancer risk And liver-protective effect.Selenium can improve human immunity, promotes the propagation of lymphocyte and the synthesis of antibody and immunoglobulin. Selenium has the obvious effect suppressing and protecting to kinds cancers such as colon cancer, skin carcinoma, hepatocarcinoma, breast carcinoma, and it is in body Mesostate methyl enol there is stronger active anticancer.Selenium is sought with Vitamin E, garlicin, linoleic acid, germanium, zinc etc. Foster element has collaborative antioxidative effect, increases antioxidant activity.Meanwhile, selenium has the work mitigating and alleviating heavy metal toxicity With.
But, the Ganoderma quantity of growth is few naturally, and containing substantial amounts of crude fibre and other impurity, and ganoderan contains Measure not high (usually 2-5%), be not easy to directly as health food or pharmaceutical raw material, therefore, development can improve ganoderan Studying with the artificial culture of the Ganoderma of Se content and cultivation technique is the key of cultivation of glossy ganoderma industrialization.
Content of the invention
It is an object of the invention to provide improving the ganoderma lucidum cultivation method of ganoderan and Se content, Ganoderma can be effectively improved In main active ganoderan and selenium content.
The ganoderma lucidum cultivation method improving ganoderan and Se content of the present invention, is Lai real by following technical step Existing:
1) preparation of culture medium:It is made up of the raw material of following weight parts:75 parts of wood flour, 20 parts of Testa Tritici, 2 parts of Semen Maydis powder, stone 1 part of ash, 1 part of Gypsum Fibrosum, 1 part of sugar, adjusting material-water ratio using Selenium-rich nutrient solution is 1:1.3-1.5;
2) spice, pack, sterilizing, inoculation;
3) send out bacterium:After inoculation, put into booth and send out bacterium, just start, control greenhouse temperature at 26-28 DEG C, after inoculation the 20th My god, greenhouse temperature is brought up to 34 DEG C, after keeping 20-24h, recovers greenhouse temperature to 26-28 DEG C;
4) go out sesame:After the long purseful of mycelia, Ganoderma enters out the sesame phase, respectively Ganoderma go out the 5th day of the sesame phase, the 10th day, Respectively greenhouse temperature is brought up to 34 DEG C within 15th day, the 20th day, the 25th day, recover greenhouse temperature after keeping 20-24h to 26-28 ℃.
Step 1 of the present invention) described in Selenium-rich nutrient solution, be prepared by the following preparation method:
1. raw material prepares:Prepare 100 parts of pure water, 1.0 parts of L- Selenomethionine, 5 parts of Semen Maydis powder;
2. after raw material 1. mixes by step, put into steam explosion tank, pressure 4MPa, be heated to 150 DEG C, keep 2min, Then in 1-3s, quick release pressure obtains serosity, at 25 DEG C by the hay bud that percent by volume is that maturation is cultivated in 10% access Spore bacillus liquid spawn, puts in fermentation tank, is passed through filtrated air in fermentation tank, is stirred fermenting, speed of agitator 250r/ Min, fermentation time is 4h, and temperature keeps 35-37 DEG C, after fermentation ends, rapidly temperature is risen to 70 DEG C and keeps 1.0h inactivation, obtain Arrive.
The cultural method of described bacillus subtilises liquid spawn is:Using triangular flask strain cultivation, below attach most importance to Amount percent, by the albumen processing slurry 30.0-35.0%, KH2P041.0%, MgS042.0%, NaCl0.5% put into In triangular flask, stir, adjust pH to 5.6,125 DEG C of sterilizing 10min, be cooled to 25 DEG C, access withered through slant culture Careless Bacillus, 28 DEG C of shaking table cultures 60-100h, for fermentation tank inoculation.
Step 2) described in spice, pack, sterilizing, inoculation, with reference to prior art method.
Compared with prior art, the invention has the advantages that:
1st, the high temperatures mushroom of Ganoderma, all can grow between 15-35 DEG C, and thermophilic is 25-30 DEG C.Sporophore is in 10-32 All can grow in the range of DEG C, but the optimum temperature of former base differentiation and fruit body development is 25-28 DEG C.Less than 25 DEG C, sporophore Poor growth, cot color and luster is also poor, and higher than 35 DEG C, sporophore can be dead.Prior art, during Ganoderma sends out bacterium and goes out sesame, one As be to control greenhouse temperature in room temperature or 26-28 DEG C.The innovative point of the present invention is, sends out bacterium period and goes out sesame in certain In period, the temperature improving culture room, to 34 DEG C, keeps a period of time, gives Ganoderma high temperature stimulation, after intensification stimulation terminates and extensive Again to normal growth temperature.
2nd, ganoderan is very sensitive to temperature, is heat-sensitive substance, and temperature raises, and can cause the degraded of ganoderan, many Glycosidic bond easy fracture in sugared body and form multiple sugar monomers, and sugar monomer be do not have activated.The present invention passes through many experiments, grinds Study carefully the impact to ganoderma polyoses content for the high-temperature time, strict control gives the time of Ganoderma high temperature stimulation, the time is short, Ganoderma The content of polysaccharide does not significantly improve, time length, can cause the decomposition of ganoderan, and the present invention passes through interim high temperature Stimulate, ganoderma polyoses content can be improved with effectively utilizes high temperature, turn avoid the decomposition to ganoderan for the long-time high temperature.
3rd, by the Ganoderma of non-high-temperature process and the Ganoderma through high-temperature process, the Ganoderma of 100 bacterium bags is sampled detecting, Detection project is ganoderan.From the point of view of the testing result of polysaccharide, through high-temperature process ganoderma polyoses content than unprocessed Ganoderma polyoses content significantly improve.
4th, Selenium-rich nutrient solution of the prior art, usually using inorganic selenium, particularly sodium selenite and sodium selenate, sub- selenium Sour potassium and potassium selenate etc., obtain from the side-product of metal mineral reserves, and inorganic selenium have larger toxicity, and are difficult to be absorbed, as Nutritional solution, assimilation effect is bad.The Selenomethionine that the present invention uses, is a kind of organic selenium source, compared with inorganic selenium, Selenomethionine There is safe and efficient advantage, it can substitute some methionine and be attached to one reversible selenium depots of formation on body protein, And there is not toxic hazard.
5th, Selenomethionine is that in nature, selenium is present in one of plant and feed grain seleno amino with organic form The plant growings such as acid, frumentum and forage crop are not necessarily required to Selenomethionine, but with Methionine synthesis they can utilize ring Selenite particularly in soil and selenate synthesis Selenomethionine in border.Available selenium in soil is more, in plant Selenomethionine content is higher.At present, Selenomethionine is mainly derived from natural selenium yeast, is being primarily present of organic selenium in selenium yeast Form, but Selenomethionine has dense yeast taste, directly water-soluble rear sprinkling, and absorption efficiency is low and influences whether finally to produce The taste and flavor of product.Bacillus subtillis, are one kind of Bacilluss, and bacillus subtilises thalline itself synthesizes alphalise starch The enzymes such as enzyme, protease, Digestive Enzyme, cellulase, are equivalent to compound enzyme, and the present invention is by Selenomethionine mixing Ko subtilis bar Bacterium after fermentation, is fermented using the zymasises that Selenomethionine product carries, and is on the one hand beneficial to selenium element and enters Ganoderma, on the other hand With Methionine synthesis they can be using the selenite particularly in soil and selenate synthesis Selenomethionine in environment, also Can be effectively improved, dissolve yeast flavor.
6th, steam explosion be using high-temperature high-pressure steam process biomass material, and realized by moment pressure leak process right The surface texture change of raw material, Component seperation and hemicellulose degradation, are generally used for corn straw, straw and Wooden Biomass etc. The pretreatment of raw material.The present invention passes through " to put into steam explosion tank, pressure 4MPa, be heated to 150 DEG C, keep 2min, then in 1- Pressure is reduced to 0 by moment by quick release pressure in 3s ", on the one hand can change to sterilize by the moment of pressure, another Aspect can effectively refine, disperse Selenomethionine, advantageously in the absorption of Ganoderma.
Specific embodiment
With embodiment, the invention will be further described below, but the invention is not limited in these embodiments.
Embodiment 1-5:
In order to verify the technique effect of the present invention, 2016, produce the product of 5 batches using the technology of the present invention, each The method of batch is as follows:
1) preparation of culture medium:It is made up of the raw material of following weight parts:Wood flour 75kg, Testa Tritici 20kg, Semen Maydis powder 2kg, stone Grey 1kg, Gypsum Fibrosum 1kg, sugared 1kg, adjusting material-water ratio using Selenium-rich nutrient solution is 1:1.3-1.5;
2) spice, pack, sterilizing, inoculation;
3) send out bacterium:After inoculation, put into booth and send out bacterium, just start, control greenhouse temperature at 26-28 DEG C, after inoculation the 20th My god, greenhouse temperature is brought up to 34 DEG C, after keeping 20-24h, recovers greenhouse temperature to 26-28 DEG C;
4) go out sesame:After the long purseful of mycelia, Ganoderma enters out the sesame phase, respectively Ganoderma go out the 5th day of the sesame phase, the 10th day, Respectively greenhouse temperature is brought up to 34 DEG C within 15th day, the 20th day, the 25th day, recover greenhouse temperature after keeping 20-24h to 26-28 ℃;
Step 1) described in Selenium-rich nutrient solution, be prepared by the following preparation method:
1. raw material prepares:Prepare 100 parts of pure water, 1.0 parts of L- Selenomethionine, 5 parts of Semen Maydis powder;
2. after raw material 1. mixes by step, put into steam explosion tank, pressure 4MPa, be heated to 150 DEG C, keep 2min, Then in 1-3s, quick release pressure obtains serosity, at 25 DEG C by the hay bud that percent by volume is that maturation is cultivated in 10% access Spore bacillus liquid spawn, puts in fermentation tank, is passed through filtrated air in fermentation tank, is stirred fermenting, speed of agitator 250r/ Min, fermentation time is 4h, and temperature keeps 35-37 DEG C, after fermentation ends, rapidly temperature is risen to 70 DEG C and keeps 1.0h inactivation, obtain Arrive.
Comparative example 1:
1) preparation of culture medium:It is made up of the raw material of following weight parts:Wood flour 75kg, Testa Tritici 20kg, Semen Maydis powder 2kg, stone Grey 1kg, Gypsum Fibrosum 1kg, sugared 1kg, material-water ratio is 1:1.3-1.5;
2) spice, pack, sterilizing, inoculation;
3) send out bacterium:After inoculation, put into booth and send out bacterium, just start, control greenhouse temperature at 26-28 DEG C;
4) go out sesame:After the long purseful of mycelia, Ganoderma enters out the sesame phase, controls greenhouse temperature at 26-28 DEG C.
Comparative example 2:
1) preparation of culture medium:It is made up of the raw material of following weight parts:Wood flour 75kg, Testa Tritici 20kg, Semen Maydis powder 2kg, stone Grey 1kg, Gypsum Fibrosum 1kg, sugared 1kg, material-water ratio is 1:1.3-1.5;
2) spice, pack, sterilizing, inoculation;
3) send out bacterium:After inoculation, put into booth and send out bacterium, just start, control greenhouse temperature at 26-28 DEG C;
4) go out sesame:After the long purseful of mycelia, Ganoderma enters out the sesame phase, controls greenhouse temperature at 34 DEG C.
Comparative example 3:
1) preparation of culture medium:It is made up of the raw material of following weight parts:Wood flour 75kg, Testa Tritici 20kg, Semen Maydis powder 2kg, stone Grey 1kg, Gypsum Fibrosum 1kg, 1 part of sugar, material-water ratio is 1:1.3-1.5;
2) spice, pack, sterilizing, inoculation;
3) send out bacterium:After inoculation, put into booth and send out bacterium, just start, control greenhouse temperature at 26-28 DEG C, after inoculation the 10th My god, greenhouse temperature is brought up to 34 DEG C, after keeping 20-24h, recovers greenhouse temperature to 26-28 DEG C;
4) go out sesame:After the long purseful of mycelia, Ganoderma enters out the sesame phase, goes out carrying greenhouse temperature on the 20th day of sesame phase in Ganoderma Height, to 34 DEG C, recovers greenhouse temperature to 26-28 DEG C after keeping 20-24h.
Result:
Each batch stochastic sampling, after testing, result see table:

Claims (2)

1. the ganoderma lucidum cultivation method of raising ganoderan and Se content is it is characterised in that be to be realized by following technical step 's:
1) preparation of culture medium:It is made up of the raw material of following weight parts:75 parts of wood flour, 20 parts of Testa Tritici, 2 parts of Semen Maydis powder, Calx 1 Part, 1 part of Gypsum Fibrosum, 1 part of sugar, adjusting material-water ratio using Selenium-rich nutrient solution is 1:1.3-1.5;
2) spice, pack, sterilizing, inoculation;
3) send out bacterium:After inoculation, put into booth and send out bacterium, just start, control greenhouse temperature at 26-28 DEG C, the 20th day after inoculation, will Greenhouse temperature brings up to 34 DEG C, recovers greenhouse temperature to 26-28 DEG C after keeping 20-24h;
4) go out sesame:After the long purseful of mycelia, Ganoderma enters out the sesame phase, respectively Ganoderma go out the 5th day of the sesame phase, the 10th day, the 15th My god, respectively greenhouse temperature is brought up to 34 DEG C within the 20th day, the 25th day, keep 20-24h after recover greenhouse temperature to 26-28 DEG C;
Step 1) described in Selenium-rich nutrient solution, be prepared by the following preparation method:
1. raw material prepares:Prepare 100 parts of pure water, 1.0 parts of L- Selenomethionine, 5 parts of Semen Maydis powder;
2. after raw material 1. mixes by step, put into steam explosion tank, pressure 4MPa, be heated to 150 DEG C, keep 2min, then In 1-3s, quick release pressure obtains serosity, at 25 DEG C by the bacillus subtilis that percent by volume is that maturation is cultivated in 10% access Bacteria liquid strain, puts in fermentation tank, is passed through filtrated air in fermentation tank, is stirred fermenting, and speed of agitator 250r/min sends out The ferment time is 4h, and temperature keeps 35-37 DEG C, after fermentation ends, rapidly temperature is risen to 70 DEG C and keeps 1.0h inactivation, obtain.
2. the ganoderma lucidum cultivation method improving ganoderan and Se content according to claim 1 it is characterised in that:Step 1) The cultural method of described bacillus subtilises liquid spawn is:Using triangular flask strain cultivation, below for weight percent Number, by the albumen processing slurry 30.0-35.0%, KH2PO41.0%, MgSO42.0%, NaCl0.5% put into triangular flask In, stir, adjust pH to 5.6,125 DEG C of sterilizing 10min, be cooled to 25 DEG C, access the hay spore through slant culture Bacillus species, 28 DEG C of shaking table cultures 60-100h, for fermentation tank inoculation.
CN201610870053.8A 2016-09-30 2016-09-30 Lucid-ganoderma cultivation method for increasing ganoderan and selenium content Pending CN106431572A (en)

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CN107646531A (en) * 2017-09-30 2018-02-02 安徽利民生物科技股份有限公司 A kind of preparation method of Se-rich lucid ganoderma culture medium
CN108739058A (en) * 2018-05-12 2018-11-06 安徽过湾农业科技有限公司 A kind of preparation method of Se-rich lucid ganoderma culture medium
CN112592414A (en) * 2021-03-03 2021-04-02 江西绿博士生物科技有限公司 Extraction of rangooncreeper fruit polysaccharide and application of rangooncreeper fruit polysaccharide in preparation of organic selenium fertilizer by edible fungus planting

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CN102628023A (en) * 2012-03-12 2012-08-08 鹤山市新的生物制品有限公司 Bacillus subtilis.XD08B having high ganoderma lucidum polysaccharide yield, feeding microbial preparation prepared from bacillus subtilis.XD08B and preparation method of the feeding microbial preparation
CN104892097A (en) * 2015-05-11 2015-09-09 苏州葛家坞生物科技有限公司 Liquid culture medium of selenium-rich edible fungus and preparation method for liquid culture medium
CN105315081A (en) * 2015-11-29 2016-02-10 湖南省致远农业科技发展有限公司 Nutrient medium for industrially cultivating lucid ganoderma and preparation method

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CN102628023A (en) * 2012-03-12 2012-08-08 鹤山市新的生物制品有限公司 Bacillus subtilis.XD08B having high ganoderma lucidum polysaccharide yield, feeding microbial preparation prepared from bacillus subtilis.XD08B and preparation method of the feeding microbial preparation
CN104892097A (en) * 2015-05-11 2015-09-09 苏州葛家坞生物科技有限公司 Liquid culture medium of selenium-rich edible fungus and preparation method for liquid culture medium
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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107646531A (en) * 2017-09-30 2018-02-02 安徽利民生物科技股份有限公司 A kind of preparation method of Se-rich lucid ganoderma culture medium
CN108739058A (en) * 2018-05-12 2018-11-06 安徽过湾农业科技有限公司 A kind of preparation method of Se-rich lucid ganoderma culture medium
CN112592414A (en) * 2021-03-03 2021-04-02 江西绿博士生物科技有限公司 Extraction of rangooncreeper fruit polysaccharide and application of rangooncreeper fruit polysaccharide in preparation of organic selenium fertilizer by edible fungus planting

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Application publication date: 20170222