CN1580234A - Culture medium for cutivating cordyceps militaris and its method - Google Patents

Abstract

The invention relates to a kind of culture medium for cultivating insect grass and the preparation method. The culture medium mainly contains slabby admixture which is composed by aging corn and chrysalis. The weight ratio in the admixture follows this. Carbon is 0.4%-1.0%, while nitrogen is 0.02%-0.05%. The surroundings will change when bacterium grown up in alimentation period of growth and anaphase by this method, so the technique for preventing and diffusing light is needed. Thus, the content of sterine, nucleoside and organic acid in the insect grass is the same to or more than content in the wild. At the same time, the output of sporocarp can increase by 10%-15%.

Description

Cultivate substratum and the method thereof of Cordyceps militaris (L.) Link.

Technical field:

The present invention relates to a kind of method that is used to cultivate the substratum of Cordyceps militaris (L.) Link. and utilizes this substratum cultivation Cordyceps militaris (L.) Link..

Background technology:

Cordyceps militaris (L.) Link. has another name called Cordyceps militaris, is the type species of Cordyceps.Cordyceps militaris is very similar to Cordyceps sinensis, all is associations of animal and plant, so both have identical or close medicine components.Because kind of nucleoside compound, 6 kinds of dipeptide compounds, Cordyceps polysaccharide, multivitamin and inorganic salt etc. surplus kind of the sterols and 10 surplus containing 10 in the Chinese caterpillar fungus, and contain 20 multiple amino acids and nucleic acid, these enrich the isostatic nutrition person that is specially adapted to the immunologic hypofunction, hyperlipidaemia and after being ill the empty patient of body take, its uniqueness antibiotic, antiviral, anticancer, and be that any food is all incomparable improving aspect the body resistance against diseases.

Yet wild Chinese caterpillar fungus far can not satisfy human needs, manually excavates in a large number and has now caused its serious scarcity, and it is human problems of always exploring that old friend worker cultivates Chinese caterpillar fungus class species.Up to the present, Cordyceps militaris is the Cordyceps sinensis of unique tamed and dociled training, is possessing under the certain condition, and the efficacy component of the Cordyceps militaris of cultivation can be similar to or be higher than wild cordyceps.

Recently, the Cordyceps militaris aspect that people's exploitation possesses the industrialization working condition has accumulated considerable experience, for example, in CN1274006A disclosed " method of artificially cultivating cordyceps ", just adopt and insert the fungi kind with rice 20-25%, corn 10-15%, paddy sugar 4-8%, silkworm chrysalis 1-2% and be made in the substratum of the 50-55% nutritive medium that accounts for weight quota by water, glucose, peptone, potassium primary phosphate, sal epsom, triammonium citrate, vitamins B, and then under conditions such as nearly 10 ℃ of lucifuge and illumination and day and night temperature stimulating growth.

In addition, CN1421524A also discloses the patented technology that a kind of name is called " prescription of Cordyceps militaris (L.) Link. growth matrix and compound method ".Its growth matrix is that substratum also is made up of glucose, peptone, citric acid, sal epsom, vitamins B, water, pupa worm powder and brown rice.

In addition, CN1148623A also discloses the method for cultivating Cordyceps militaris (L.) Link. mycelia powder that is specifically designed to of a kind of " northern Cordyceps mycelium fermentation technology ".In this method, with the pupa toothed burclover spore bacterial classification that obtains in the Cordyceps militaris fruiting body, 7 days bevel bacterial classifications of constant temperature culture on the PDA slant medium, again this bacterial classification is put into and shaken bottle vibration 3-4 days, skillful good bacterial classification is put into temperature 25-27 ℃, and air quantity 1: 0.4 is in 38-45 hour the jar of pressure 0.05Mpa cultivation, inoculation is under these conditions made dry strain through dehydration, drying more at last.

In fact, in the technology of cultivation Cordyceps militaris, expanding species is the technical scheme that people generally adopt.Usually the process of expanding species can be as big as and is divided into three phases, and the fs is to plant the mother of Cordyceps militaris to dig a fritter mycelia and put on the medium slant and cultivate, i.e. the mycelium culture stage; Mycelia is put into the substratum of liquid after mycelia such as subordinate phase are covered with again, impel fungal growth by shaking of shaking table, this stage is the strain cultivation stage; Phase III puts into liquid spawn substratum again and cultivates the Chinese caterpillar fungus stage.In above-mentioned three phases, all need have substratum to cultivate, and the nutrition component of substratum and cultivation environment are the technical problems of most critical.

Practice shows, in the Chinese caterpillar fungus that nurtures active drug content whether balanced with substratum in the ratio of contained carbon nitrogen element direct relation is arranged, because organic carbon is to generate dipeptides, polysaccharide, comprise that the 3-Desoxyadenosine is a cordycepin, nitrogen is again the important element of Chinese caterpillar fungus growth, and is nucleic acid, amino acid whose integral part.Nourish and grow poorly as too high easily the causing of carbon content in the substratum, mycelial growth is unable, and output is reduced; And nitrogen content is too high in the substratum, can cause poor growth, hinder fruit body development and growth, and some sterol and ucleosides nutrition are significantly reduced, and nitrogen content is low excessively, is unfavorable for the growth of mycelia, the sporophore slight of stature; Secondly, bacterial classification is incomplete same at its required carbon nitrogen content of different growth phases, and as relatively low in the required carbon content of vegetative growth phase, and in the sporophore growth stage, carbon content requires higher relatively; In addition, selenium content can play very important effect to the pharmaceutical use that improves Chinese caterpillar fungus.

Aspect the cultivation environment, temperature and day and night temperature are the very important extraneous factors of stimulating growth, and at different growth phases, envrionment temperature, illumination, humidity and day and night temperature also are variable quantities, and these also are the problems that prior art is ignored.

As seen, in existing disclosed Cordyceps militaris culture technology, do not obtain enough attention for the particularly dynamic carbon-nitrogen ratio of carbon-nitrogen ratio, just embodied the compatibility of aspects such as inorganic salt such as phosphorus, calcium, potassium and nutrition in its described substratum, and ignored Chinese caterpillar fungus grow necessary internal factor and external conditions, like this, promptly influenced the quantum of output of Cordyceps militaris, simultaneously, also influenced the quality of Cordyceps militaris to a great extent.

Summary of the invention:

The purpose of this invention is to provide and a kind ofly adjust in the substratum carbon nitrogen content growing environment when, solve the Cordyceps militaris (L.) Link. contained drug component of cultivation and improve substratum and the method thereof of the cultivation Cordyceps militaris (L.) Link. of output according to different growth phases.

The substratum that is used to cultivate Cordyceps militaris (L.) Link. proposed by the invention, mainly comprise the mixture of viscous form of making by ripe cereal and silkworm chrysalis in this substratum, and in this mixture, be added with the inorganic salt of phosphorous, calcium, potassium, magnesium ion, the weight quota of carbon accounts for the 0.4%-1.0% of gross weight in the described mixture, and the weight quota of nitrogen element accounts for the 0.02%-0.05% of gross weight.Wherein, the phosphorus in the mixture, potassium ion can form nucleic acid, promote cell fission, and calcium ion can promote sporophore growth, reduce disease; Carbon-nitrogen ratio should be about 20: 1 in the mixture, and the low important drugs compositions such as dipeptides, polysaccharide, cordycepin that are unfavorable for of carbon element content generate, and the weight quota of carbon is too high, can hinder the cell fission of mycelia, promptly is unfavorable for the formation of sporophore; Carbon-nitrogen ratio is unreasonable in the mixture, is that the Cordyceps militaris pharmaceutical compositions of being cultivated is compared serious unbalance important factor with wild Chinese caterpillar fungus.

Per kilogram also should contain the selenium element of 0.04-0.06mg in the described mixture.Described cereal preferably rice.

Utilize above-mentioned substratum to cultivate the method for Cordyceps militaris (L.) Link. specially, comprise the liquid Cordyceps militaris spawn is injected the container that above-mentioned substratum is housed under aseptic condition, bacterial classification vegetative growth phase envrionment temperature remain on 12 ℃-20 ℃, the scheduled time gives lucifuge and gives the scattered light irradiation at interval simultaneously, should remain on 16 ℃-25 ℃ in late growing stage stage environment temperature, relative humidity is 85%-95%.Bacterial classification vegetative growth phase envrionment temperature be lower than 12 ℃, the poor growth of mycelia, and be higher than 25 ℃, then mycelia is prone to the early ageing phenomenon; And be lower than 12 ℃ in sporophore late growing stage stage environment temperature, and cause sporophore growth slower equally, be higher than 25 ℃, sporophore growth easily takes place withered.

Also comprise the step that contains organic carbon element material to the media surface injection in the bacterial classification late growing stage stage.

Describedly contain organic carbon material and can constitute by Liquid Glucose.

Described vegetative growth phase environment optimum temps is at 14 ℃-18 ℃, and in late growing stage stage environment optimum temps at 18 ℃-22 ℃.

Remain on growth under the lucifuge environment during the described growth about its first five day, remained on the environment growth down of scattered light irradiation in back about five days.

In the substratum proposed by the invention, correctly adjust carbon-nitrogen ratio, causing has 11 kinds of sterols, nucleosides and organic acid content to reach or be higher than the content of wild Chinese caterpillar fungus in the Chinese caterpillar fungus, and has kept the high-content of 3-Desoxyadenosine, can make fruiting body yield increase 10%-15% simultaneously.

Embodiment

Embodiment 1: in the culture medium preparation, its weight quota of at different levels parts is: select rice 41% for use, and dry product silkworm chrysalis 2.5%, peptone 0.25%, inorganic salt 0.25%, glucose 0.6%, surplus is a water.As calculated, the mutual content of carbon unit is 0.74%, and nitrogen content is 0.034%.The liquid Cordyceps militaris spawn is injected the container that above-mentioned substratum is housed under aseptic condition, bacterial classification vegetative growth phase envrionment temperature remain on about 16 ℃, and in the first five day lucifuge, gave scattered light in back five days and shine; Should remain on about 20 ℃ in late growing stage stage environment temperature, relative humidity is 85%, with 0.5% glucose solution media surface is sprayed simultaneously, with supplementary carbon source.

Claims (8)

1, is used to cultivate the substratum of Cordyceps militaris (L.) Link., mainly comprise the mixture of viscous form of making by ripe cereal and silkworm chrysalis in this substratum, and in this mixture, be added with the inorganic salt of phosphorous, calcium, potassium, magnesium ion, it is characterized in that: the weight quota of carbon accounts for the 0.4%-1.0% of gross weight in the described mixture, and the weight quota of nitrogen element accounts for the 0.02%-0.05% of gross weight.

2, substratum according to claim 1 is characterized in that: per kilogram also should contain the selenium element of 0.04-0.06mg in the described mixture.

3, substratum according to claim 1 and 2 is characterized in that: described cereal preferably rice.

4, utilize above-mentioned substratum to cultivate the method for Cordyceps militaris (L.) Link. specially, it is characterized in that: the liquid Cordyceps militaris spawn is injected the container that above-mentioned substratum is housed under aseptic condition, bacterial classification vegetative growth phase envrionment temperature remain on 12 ℃-20 ℃, the scheduled time gives lucifuge and gives the scattered light irradiation at interval simultaneously, should remain on 16 ℃-25 ℃ in late growing stage stage environment temperature, relative humidity is 85%-95%.

5, method according to claim 4 is characterized in that: also comprise to media surface in the bacterial classification late growing stage stage and spray the step that contains organic carbon element material.

6, method according to claim 5 is characterized in that: describedly contain organic carbon material and can be made of Liquid Glucose.

7, method according to claim 4 is characterized in that: described vegetative growth phase environment optimum temps is at 14 ℃-18 ℃, and in late growing stage stage environment optimum temps at 18 ℃-22 ℃.

8, method according to claim 4 is characterized in that: during the described growth, remained on growth under the lucifuge environment in its about preceding ten days, remain on the environment growth down of scattered light irradiation about back ten days.