CN105009939A - Method for cultivating hedgehog hydnum by using wild jujube branch cuttings - Google Patents
Method for cultivating hedgehog hydnum by using wild jujube branch cuttings Download PDFInfo
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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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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05D—INORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C; FERTILISERS PRODUCING CARBON DIOXIDE
- C05D3/00—Calcareous fertilisers
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- C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
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- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
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Abstract
The present invention provides a method for cultivating hedgehog hydnum by using wild jujube branch cuttings. The method comprises the steps of (A) the preparation and cultivation of a liquid strain, (B) the preparation and cultivation of a grain strain, (C) the preparation of a culture medium for cultivation, and (D) the connection of the grain strain prepared in the step (B) into the culture medium prepared in the step (C) under a sterile condition to carry out cultivation, raising management and harvest, and the innovation points of the method are that the culture medium in the step (A), (B) or (C) comprises wild jujube branch cuttings. According to the method, forestry waste wild jujube branches are fully utilized, a circular economy mode is formed, the waste of natural resources and the pollution of an environment are avoided, and the cost of cultivation is reduced. By using the cultivation technology of cultivating the hedgehog hydnum by using the wild jujube branch cuttings, the economic and ecological benefits are significant, and the method has abroad application prospect.
Description
Technical field
The invention belongs to the technical field of edible fungus cultivation, being specifically related to a kind of the hedgehog hydnum composts or fertilisers of cultivating and the method for cultivating hedgehog hydnum thereof that comprise Ramulus Ziziphi Spinosae bits.
Hedgehog hydnum (Hericiumerinaceus) belongs to Basidiomycotina, Hymenomycetes, hedgehog hydnum Zoopagales, hedgehog hydnum Cordycepps, Hericium.Hydnum erinaceus sporophore meat, round and thick, obovate, shape as hedgehog hydnum, therefore is gained the name.Hericium erinaceus comprises rich in protein, fat, cellulose, polysaccharide, also containing 16 seed amino acids, wherein has eight kinds of essential amino acids, and various amino acid whose ratio and human body need close.Hericium erinaceus polysaccharide has anti-aging effects, strengthens immunologic function in addition, has protective effect to cellular damage, to cardiovascular and and the geriatric disease such as diabetes also have obvious preventive and therapeutic effect, there is medical science and pharmacological interest.Hericium erinaceus is very wide in China's distribution, and East, West, South, North all produces Hericium erinaceus, abound with in Heilungkiang, Jilin, Liaoning, Shanxi, Shaanxi, the provinces and regions such as Gansu.
Wild jujube is a kind of shrub of Rhamnaceae zizyphus.For a kind of wild resource that northern China is more common, be distributed in the Hills And Low Mountains of height above sea level 200-800 rice more, only for Hebei, 48.1% of Taihang Mountain, the Hebei gross area can be accounted for.The purposes of usual Ramulus Ziziphi Spinosae is: make bavin and burn, be used as hedge revetment, be used as the protecting wall in orchard, as the use of checking winds and fixing drifting sand.At present, not yet carry out the exploitation of wild jujube branch resource, its abundant wild resource can not fully be utilized, and causes great waste.With Ramulus Ziziphi Spinosae bits for culturing raw material, very large economy and ecological benefits will inevitably be produced, improve local economy structural adjustment, realizing will playing huge effect in recycling economy.
Chinese sorghum has long cultivation history in China, is the important cereal crops of China, and distribution is very wide, and all there is cultivation all parts of the country.Mainly be distributed in Heilungkiang, Hebei, Beijing, Shaanxi, Yunnan, Sichuan, Guizhou, Anhui, Fujian, Guangdong, Jiangxi, zhejiang and other places.Its cultivated area and output are only second to wheat, paddy rice, corn, barley, occupy the 5th.Its stalk is often by as fuel or on-site incineration, and not only multi-environment pollutes, and can not make full use of resource.Broomcorn straw crude fiber content is high, crude protein content is low, Hericium erinaceus as wood decay fungi secretes extracellular enzyme as laccase, cellulase, hemicellulase, lignin peroxidase etc. in straw substrate, can degraded cellulose, hemicellulose and lignin, raw fiber is converted into the high-quality protein that the mankind can eat again.After Ramulus Ziziphi Spinosae bits are pulverized, more common weed tree sawdust powder is thin, as mushroom cultivation matrix, after mixing water, affect respiration, if admixed as auxiliary material by broomcorn straw, not only can supplementary carbon source, because of its physical form, the gas permeability of Ramulus Ziziphi Spinosae bits major ingredient can also be improved, be used for cultivating Hericium erinaceus, can reduce costs, increase economic efficiency, can recycling economy be realized again.
Summary of the invention
The object of this invention is to provide and a kind ofly utilize Ramulus Ziziphi Spinosae to consider to be worth doing to cultivate the method for hedgehog hydnum, at environmentally safe and the resource recycling realizing agriculture and forestry organic waste material under the prerequisite of high yield utilize.
To achieve these goals, the invention provides a kind of Ramulus Ziziphi Spinosae that utilizes and consider the method for cultivating hedgehog hydnum to be worth doing, comprise the steps:
A. the preparation of liquid spawn and cultivation;
B. the preparation of grain spawn and cultivation;
C. the preparation of cultivation medium;
D., under aseptic condition, the medium that the grain spawn that prepared by step B access step C prepares, carries out cultivating, management of producing mushroom, to gather;
In steps A or/and step B or/and step C medium in containing Ramulus Ziziphi Spinosae bits.
The described Ramulus Ziziphi Spinosae that utilizes considers the method for cultivating hedgehog hydnum to be worth doing, in steps A hedgehog hydnum liquid spawn culture medium in as substrate for induction thing, add Ramulus Ziziphi Spinosae bits in advance; It optimizes liquid spawn culture medium formulation weight percentage: potato 20%, Ramulus Ziziphi Spinosae bits 5%, wheat bran 3%, glucose 2%, KH
2pO
40.3%, MgSO
40.15%, VB
110mg/L, carboxymethyl cellulose 0.3%, water 1000mL, pH nature.
The described Ramulus Ziziphi Spinosae that utilizes considers the method for cultivating hedgehog hydnum to be worth doing, and in described steps A, the preparation of liquid spawn is characterised in that:
A. the preparation of level liquid bacterial classification
Under aseptic condition, commercially available mother is planted in access level liquid medium, 25 DEG C, 160r/min constant-temperature shaking culture 7d, i.e. obtained level liquid bacterial classification;
Described level liquid bacterium culture medium comprises: by weight percentage, potato 20%, wheat bran 3%, glucose 2%, KH
2pO
40.3%, MgSO
40.15%, CMC 0.3%, VB
110mg/L;
B. the preparation of second-class liquid isolate
Described second-class liquid isolate culture medium prescription percentage by weight is: potato 20%, Ramulus Ziziphi Spinosae bits 5%, wheat bran 3%, glucose 2%, KH
2pO
40.3%, MgSO
40.15%, CMC0.3%, VB
110mg/L;
The preparation method of liquid spawn is that the 1000mL that potato, Ramulus Ziziphi Spinosae bits powder and wheat bran added water boils altogether, keeps micro-and boils 30 minutes, filter, get filtrate, add other component while hot, stir after melting, be settled to water 1000mL, packing, sterilizing after boiling, to be seeded;
Under aseptic condition, by level liquid bacterial classification with in the inoculum concentration access secondary liquid medium of 10%, 25 DEG C, 160r/min constant-temperature shaking culture 7d, i.e. obtained second-class liquid isolate.
The described Ramulus Ziziphi Spinosae that utilizes considers the method for cultivating hedgehog hydnum to be worth doing, in step B hedgehog hydnum grain spawn medium in also add Ramulus Ziziphi Spinosae bits in advance as substrate for induction thing; Its grain spawn culture medium prescription optimized: sorghum grain 84%, Ramulus Ziziphi Spinosae bits 15%, quicklime 1%.
The described Ramulus Ziziphi Spinosae that utilizes considers the method for cultivating hedgehog hydnum to be worth doing, and in described step B, the preparation of grain spawn is characterised in that:
Second-class liquid isolate access grain spawn medium steps A obtained, inoculum concentration is 10%, 22-28 DEG C and is cultured to full, obtains grain spawn.
The described Ramulus Ziziphi Spinosae that utilizes considers the method for cultivating hedgehog hydnum to be worth doing, and in step C, Ramulus Ziziphi Spinosae bits are as the major ingredient of cultivation hedgehog hydnum, and culture medium for cultivating weight composition comprises: Ramulus Ziziphi Spinosae bits 60%, cotton seed hull 19%, broomcorn straw 10%, wheat bran 10%, quicklime 1%.
The described Ramulus Ziziphi Spinosae that utilizes considers the method for cultivating hedgehog hydnum to be worth doing, in described step C, the preparation of Ramulus Ziziphi Spinosae bits medium is characterised in that: by Ramulus Ziziphi Spinosae, broomcorn straw is ground into wood chip shape, be equipped with cotton seed hull, wheat bran and quicklime, material-water ratio is 1: 1.4, piles vexed 30min, loads in container, sterilizing, for subsequent use after cooling.
The described Ramulus Ziziphi Spinosae that utilizes considers the method for cultivating hedgehog hydnum to be worth doing, and in step D, the inoculum concentration of grain spawn is 5-10%.
The described method utilizing Ramulus Ziziphi Spinosae to consider medium cultivation hedgehog hydnum to be worth doing, in liquid nutrient medium, Ramulus Ziziphi Spinosae bits used, pulverize and cross 80 mesh sieves in advance.
The described Ramulus Ziziphi Spinosae that utilizes considers the method for cultivating hedgehog hydnum to be worth doing, and in step C, the sterilising conditions of Ramulus Ziziphi Spinosae bits medium is 121 DEG C of autoclaving 150min or 100 DEG C normal-pressure sterilization 24h.
The described method utilizing Ramulus Ziziphi Spinosae to consider composts or fertilisers of cultivating cultivating black fungus to be worth doing, in grain spawn medium, Ramulus Ziziphi Spinosae bits used, pulverize in advance as wood chip shape.
Technological progress effect of the present invention shows the primary raw material using Ramulus Ziziphi Spinosae bits as cultivation hedgehog hydnum, and advantage mainly contains 3 points:
1, for edible mushroom, provide its growth needed for carbon source;
2, Ramulus Ziziphi Spinosae is not yet developed at present, aboundresources, reduces the cost of culturing edible fungus, improves its economic benefit.
3, from the angle of ecology, achieve " turning waste into wealth ", have found a kind of resource exploitation wild jujube forestry waste and the cheap culturing raw material of applicable Hericium erinaceus growth, not only take full advantage of abundant wild resource, and reduce cultivation cost, add economy and zoology benefit, have broad application prospects.
Embodiment
Below in conjunction with specific embodiment, content of the present invention is further described in detail.
Embodiment: utilize Ramulus Ziziphi Spinosae to consider the method for cultivating hedgehog hydnum to be worth doing, hedgehog hydnum bacterial classification is market purchasing;
Common process: mother plants-expand numerous-cotton seed hull original seed-Ramulus Ziziphi Spinosae bits or cotton seed hull cultivated species-Ramulus Ziziphi Spinosae bits fruiting bag-Fa tube reason-management of producing mushroom-harvesting.
Liquid spawn technique for applying: mother plants-expand the fermentation-grain spawn-Ramulus Ziziphi Spinosae bits fruiting bag-Fa tube reason-management of producing mushroom-harvesting of numerous-liquid spawn.
The liquid spawn technique that the present invention adds Ramulus Ziziphi Spinosae bits can shorten the whole production cycle more than 1/3rd, introduces liquid spawn technique below:
Steps A. the preparation of liquid spawn
First class inoculum culture medium prescription: potato 20%, wheat bran 2%, glucose 2%, KH
2pO
40.3%, MgSO
40.15%, VB
110mg/L, CMC 0.3%, pH nature; Preparation method is potato, wheat bran boils 30min altogether, and filter and remove residue obtains leaching liquor and adds other compositions.Use 500mL conical flask time prepared by level liquid bacterial classification, liquid amount is 200mL, after 121 DEG C of autoclaving 30min, accesses commercially available PDA slant strains under aseptic condition, and every bottle of inoculum concentration is 4 pieces of 0.5cm × 0.5cm bacterial classification blocks.After inoculation 25 DEG C, under the condition of 180r/min, constant-temperature shaking culture 7d.
Second-class liquid isolate culture medium prescription: potato 20%, Ramulus Ziziphi Spinosae bits 5%, wheat bran 3%, glucose 2%, KH
2pO
40.3%, MgSO
40.15%, CMC 0.3%, VB
110mg/L, pH nature; Preparation method is that Ramulus Ziziphi Spinosae bits were pulverized 80 mesh sieves and obtained powdery, and boil 30min altogether with potato, wheat bran, filter and remove residue obtains leaching liquor.
Second-class liquid isolate liquid amount specification, sterilising conditions and condition of culture are the same, and inoculum concentration is 10%.At second-class liquid isolate preparatory phase, in culture fluid, enzymatic determination result is as shown in table 1:
The enzyme live data of table 1 hedgehog hydnum
As shown in Table 1, laccase activity, the CMC enzyme of secondary liquid medium of the present invention are lived, all than the height of control group (not adding Ramulus Ziziphi Spinosae bits), add Ramulus Ziziphi Spinosae bits in visible liquid medium within and certain inducing action is played to the laccase of mycelia, the secretion of CMC enzyme, allow mycelia adapt to substrate (Ramulus Ziziphi Spinosae bits) in advance, thus accelerate mycelia growing speed and shortening its growth cycle in planting material.
Step B.The preparation of grain spawn
Grain spawn culture medium prescription: sorghum grain 84%, Ramulus Ziziphi Spinosae bits 15%, quicklime 1%.
Accurately take each constituent in medium, material-water ratio 1:1.3 carries out spice, and load 17cm × 32cm × 0.005cm polyethylene plastic bag, every packed siccative 300g, prepares 500 bags.To the second-class liquid isolate and control group liquid spawn (not adding Ramulus Ziziphi Spinosae bits) access grain spawn medium respectively that obtain be optimized, and inoculate 500 bags respectively, measure mycelium length, and time, purseful time completely be takeed in record.The advantage comparative test result of grain spawn medium is as shown in table 2:
The purseful time of table 2 hedgehog hydnum grain spawn
As shown in Table 2, mycelia is faster than the long speed of pure wheat medium in " wheat+Ramulus Ziziphi Spinosae bits " grain culture medium, this is because the interpolation of wheat gap Ramulus Ziziphi Spinosae bits, more be conducive to mycelial growth, and add Ramulus Ziziphi Spinosae bits in liquid medium within, enable the faster Adaptable growth substrate of mycelia, thus accelerate to send out bacterium speed, shorten the production cycle and enhance productivity.
C. the preparation of Ramulus Ziziphi Spinosae bits composts or fertilisers of cultivating
By weight, take Ramulus Ziziphi Spinosae bits 60%, broomcorn straw 10%, wheat bran 10%, cotton seed hull 19%, quicklime 1%, material-water ratio is that 1:1.3 carries out spice, after stirring, loads in container and carry out sterilizing and inoculation work after piling vexed 30min.
Loaded in Polythene Bag, Polypropylene Bag or vial by above-mentioned composts or fertilisers of cultivating, bore is not limit, conventional tying, autoclaving 121 DEG C, 150min; Normal-pressure sterilization 100 DEG C, 24h; Inoculation after cooling.
The consumption of Ramulus Ziziphi Spinosae bits has carried out analysis of experiments, specific as follows:
Accurately take each constituent by shown in table 3, mixed, add water, material-water ratio is 1:1.3, the composts or fertilisers of cultivating prepared is loaded in test tube, each formula repetition 5 times, 121 DEG C of autoclaving 150min.Grain spawn described in step B is accessed respectively in the test tube containing composts or fertilisers of cultivating, carry out test tube and to nourish and grow experiment.
Table 3 culture medium prescription (siccative, %)
Observe long speed and the growing way situation of mycelia in a test tube every 2 days, measure the long speed of mycelia with vernier calliper (accuracy 0.02mm).Growth rata and potential of hypha the results are shown in Table 4.
Growth rata and potential of hypha result is as shown in table 4
Table 4 hericium mycelium growing state
(note: list long speed and be mean+/-standard error; +++ represent that growth is fine and close, ++ represent that growth is finer and close ,+represent that growth is general)
As can be seen from Table 4, on pure Ramulus Ziziphi Spinosae bits medium, hericium mycelium growing way is general, and on pure broomcorn straw medium, the mycelium growth vigor of hedgehog hydnum is more weak; During increase along with Ramulus Ziziphi Spinosae bits addition, its growing way is better, and mycelia is pure white, fine and close, has comparativity with pure cotton seed hull medium, and mycelia grows the fastest on formula P6, all faster than pure Ramulus Ziziphi Spinosae bits P5 group, pure broomcorn straw group leader P0 speed.
Above-mentioned hedgehog hydnum biological transformation ratio result of the test is as shown in table 5.
The biological transformation ratio of table 5 hedgehog hydnum
As can be seen from Table 5, when P4, Ramulus Ziziphi Spinosae bits reach 60% as primary carbon source, the biological transformation ratio of Hericium erinaceus reaches 85.23%, higher than the biological transformation ratio of pure broomcorn straw; Although do not reach the biological transformation ratio 86.34% of the pure cotton seed hull of planting material major ingredient, there is comparativity.In sum, Ramulus Ziziphi Spinosae bits can partly replace cotton seed hull become hedgehog hydnum plantation in major ingredient, Ramulus Ziziphi Spinosae is considered this forestry waste to be worth doing and be have also been obtained recycling, significant to ecological agriculture construction.
D. inoculate, cultivate, gather
By cooled for sterilizing fruiting bag, under aseptic condition, press the inoculum concentration access step B gained grain spawn of 5-10%.Carry out lucifuge cultivation under the bacterium bag to connect kind can being placed in 22-25 DEG C of room temperature, bacterial classification sprouted as early as possible, field planting.After mycelia purseful, unlimited sack carries out reproductive growth management phase.When Hydnum erinaceus sporophore 7-8 is ripe, spheroid is substantially considerable, solid, and bacterium stings long 0.5-1cm, gathers before not launching spore.Gather and within first 6 hours, stop water spray and ventilate.Cut off from base portion when gathering, note clear residual mushroom handle at any time, what bag was planted tries not to cut plastic sack, prevents living contaminants.
In sum, Ramulus Ziziphi Spinosae bits of the present invention can be used for cultivating hedgehog hydnum, therefore, using the living resources of Ramulus Ziziphi Spinosae as reusable edible, for industrialization development, there is important innovative significance and realistic price, be suitable in conjunction with cell engineering directive breeding the strain excellent decomposing Ramulus Ziziphi Spinosae bits again, better effect will be produced.
Claims (10)
1. utilize Ramulus Ziziphi Spinosae to consider a method of cultivating hedgehog hydnum to be worth doing, comprise the steps:
A. the preparation of liquid spawn and cultivation;
B. the preparation of grain spawn and cultivation;
C. the preparation of cultivation medium;
D., under aseptic condition, the medium that the grain spawn that prepared by step B access step C prepares, carries out cultivating, management of producing mushroom, to gather;
It is characterized in that: in steps A or/and step B or/and step C medium in containing Ramulus Ziziphi Spinosae bits.
2. according to claim 1ly utilize Ramulus Ziziphi Spinosae to consider to be worth doing to cultivate the method for hedgehog hydnum, its feature comprises: in steps A hedgehog hydnum liquid spawn culture medium in add Ramulus Ziziphi Spinosae bits in advance as substrate for induction thing; It optimizes liquid spawn culture medium formulation weight percentage: potato 20%, Ramulus Ziziphi Spinosae bits 5%, wheat bran 3%, glucose 2%, KH
2pO
40.3%, MgSO
40.15%, VB
110mg/L, carboxymethyl cellulose 0.3%, water 1000mL, pH nature.
3. utilize Ramulus Ziziphi Spinosae to consider the method for cultivating hedgehog hydnum to be worth doing according to claim 2, in described steps A, the preparation of liquid spawn is characterised in that:
A. the preparation of level liquid bacterial classification
Under aseptic condition, commercially available mother is planted in access level liquid medium, 25 DEG C, 160r/min constant-temperature shaking culture 7d, i.e. obtained level liquid bacterial classification;
Described level liquid bacterium culture medium comprises: by weight percentage, potato 20%, wheat bran 3%, glucose 2%, KH
2pO
40.3%, MgSO
40.15%, CMC 0.3%, VB
110mg/L;
B. the preparation of second-class liquid isolate
Described second-class liquid isolate culture medium prescription percentage by weight is: potato 20%, Ramulus Ziziphi Spinosae bits 5%, wheat bran 3%, glucose 2%, KH
2pO
40.3%, MgSO
40.15%, CMC 0.3%, VB
110mg/L;
The preparation method of liquid spawn is that the 1000mL that potato, Ramulus Ziziphi Spinosae bits powder and wheat bran added water boils altogether, keeps micro-and boils 30 minutes, filter, get filtrate, add other component while hot, stir after melting, be settled to water 1000mL, packing, sterilizing after boiling, to be seeded;
Under aseptic condition, by level liquid bacterial classification with in the inoculum concentration access secondary liquid medium of 10%, 25 DEG C, 160r/min constant-temperature shaking culture 7d, i.e. obtained second-class liquid isolate.
4. according to claim 1ly utilize Ramulus Ziziphi Spinosae to consider to be worth doing to cultivate the method for hedgehog hydnum, its feature comprises: in step B hedgehog hydnum grain spawn medium in also as substrate for induction thing, add Ramulus Ziziphi Spinosae bits in advance; Its grain spawn culture medium prescription optimized: sorghum grain 84%, Ramulus Ziziphi Spinosae bits 15%, quicklime 1%.
5. utilize Ramulus Ziziphi Spinosae to consider the method for cultivating hedgehog hydnum to be worth doing according to claim 4, in described step B, the preparation of grain spawn is characterised in that:
Second-class liquid isolate access grain spawn medium steps A obtained, inoculum concentration is 10%, 22-28 DEG C and is cultured to full, obtains grain spawn.
6. the Ramulus Ziziphi Spinosae that utilizes according to claim 1 considers the method for cultivating hedgehog hydnum to be worth doing, its feature comprises: in step C, Ramulus Ziziphi Spinosae bits are as the major ingredient of cultivation hedgehog hydnum, and culture medium for cultivating weight composition comprises: Ramulus Ziziphi Spinosae bits 60%, cotton seed hull 19%, broomcorn straw 10%, wheat bran 10%, quicklime 1%.
7. utilize Ramulus Ziziphi Spinosae to consider the method for cultivating hedgehog hydnum to be worth doing according to claim 6, in described step C, the preparation of Ramulus Ziziphi Spinosae bits medium is characterised in that: by Ramulus Ziziphi Spinosae, broomcorn straw is ground into wood chip shape, be equipped with cotton seed hull, wheat bran and quicklime, material-water ratio is 1: 1.4, piles vexed 30min, loads in container, sterilizing, for subsequent use after cooling.
8. utilize Ramulus Ziziphi Spinosae to consider the method for cultivating hedgehog hydnum to be worth doing according to claim 1, it is characterized in that, in step D, the inoculum concentration of grain spawn is 5-10%.
9. utilize Ramulus Ziziphi Spinosae to consider the method for medium cultivation hedgehog hydnum to be worth doing according to claim 2, it is characterized in that, in liquid nutrient medium, Ramulus Ziziphi Spinosae bits used, pulverize and cross 80 mesh sieves in advance.
10. utilize Ramulus Ziziphi Spinosae to consider the method for cultivating hedgehog hydnum to be worth doing according to claim 1, it is characterized in that, in step C, the sterilising conditions of Ramulus Ziziphi Spinosae bits medium is 121 DEG C of autoclaving 150min or 100 DEG C normal-pressure sterilization 24h.
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| CN113383700A (en) * | 2021-06-30 | 2021-09-14 | 何妙仪 | Preparation method of organic selenium raw material |
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