CN108770595A - A kind of Ecology soil covering culture method of cicada fungus - Google Patents

Abstract

The present invention discloses a kind of Ecology soil covering culture method of cicada fungus, it is intended to real Emulational culture is realized, to reduce the exploitation to wild cicada fungus, to slow down the pressure of cicada fungus wild resource shortage.Its step is inoculation cicada fungus bacterial strain（Senior general mountain cicada fungus）In triangular flask inclined-plane, constant temperature incubation 7 days, spore on sterile water elution inclined-plane, adjusts the concentration of spore suspension, and tussah chrysalis is injected with spore suspension, Nostoc commune Vanch is to white hypha of emerging at room temperature, then, by its earthing kind in flowerpot, tetra- processing of thickness of earth-fill cover 1cm, 2cm, 3cm and 4cm, each processing has 15 silkworm chrysalises, is cultivated at room temperature to fructification maturation.Record growing state simultaneously compares, and finally show that the effect of earthing 1cm and earthing 2cm are preferable.

Description

A kind of Ecology soil covering culture method of cicada fungus

Technical field

The present invention relates to the artificial cultivation technique fields of cordyceps sinensis, and in particular to a kind of Ecology earthing artificial cultivation of cicada fungus Method invention.

Background technology

Cicada fungus also known as cicada grass, cicada young pilose antler, Hu cicada etc. are the big cicada grass of Clavicipitaceae fungi（Cordyceps cicadae X. Q. Shing）Phorozoon Paecilomyces cicadae [Paecilomyces cicadae (Miquel) Samson], parasitic Yushan Hill cicada （Cicada flammata Dist）The entomogenous fungi complex of formation.Cicada fungus is the traditional Chinese medicine in China, stroma and host elder brother The corpse of worm（Sclerotium）All can be used as medicinal effects, there is the effect of dissipate wind-heat, relieving convulsion, improving eyesight, cure mainly children day hang, the morbid night crying of babies heart Throb with fear, expelling wind and relieving convulsion, measles, hot eyes, more tears.Cicada fungus has good nutritive value as edible and medical fungi, wherein containing glycogen, The chemical compositions such as mannitol, a variety of alkaloids, protein, a variety of amino acid and various trace elements, and carbohydrate and protein Content it is relatively high.By some pharmacological researches, show that cicada fungus has immunological regulation, antitumor, improvement renal function, adjusting fat The effects that class is metabolized.

And due to the cicada fungus shortage of resources of wild, people are more and more to the research of cicada fungus manually cultivated, but mesh Before realize only obtain fructification with solid medium culture or infect silkworm larva or silkworm chrysalis is obtained through Nostoc commune Vanch All and real Emulational culture is not implemented in the compound of the entomogenous fungi one obtained.And the present invention disseminates by way of injection Tussah chrysalis, later stage obtain the relatively high entomogenous fungi complex of yield cultivating by way of earthing again, realize really Emulational culture, and obtained best thickness of earth-fill cover, it lays the foundation to promote soil covering culture later.

Invention content

The technical problem to be solved by the present invention is to：The purpose of the present invention is to provide a kind of cultivations of the Ecology earthing of cicada fungus Culture method provides the best thickness of earth-fill cover of soil covering culture, realizes real Emulational culture, to reduce to wild cicada fungus Exploitation, to slow down the pressure of cicada fungus wild resource shortage.

The invention is realized in this way：

A kind of Ecology soil covering culture method of cicada fungus, is cultivated using following steps：

Step 1:, cicada fungus bacterial strain is connected in the triangular flask inclined-plane made of PDA solid mediums, cultivate 5- under 25 DEG C of constant temperature Extremely generate conidium within 10th；

Step 2: spore suspension is made with the spore in the sterile ultrapure water elution triangular flask of the Tween 80 added with 0.5%, use Blood counting chamber counts and adjusts spore suspension concentration to 0.8-1.2 × 107/mL；

Again fall to remain alcohol with aseptic water washing Step 3: cleaning silkworm chrysalis body surface with 75% alcohol；Then with matching in step 2 Set the spore suspension injection tussah chrysalis of concentration, every silkworm chrysalis injects 0.08-0.15mL, injection site be silkworm chrysalis wing just after Side and third link crosspoint；

Step 4: the silkworm chrysalis injected is positioned in the culture box to sterilize；Put one layer of cotton to sterilize in culture cassette bottom portion And the sterile water of appropriateness is added to soak cotton, then silkworm chrysalis is positioned on cotton, to keep humidity to be infected in favor of silkworm chrysalis；It puts It is placed in and cultivates at room temperature about 12-20 days, culture to silkworm chrysalis body surface grows white hypha；

Step 5: transplanting the silkworm chrysalis with white hypha in flowerpot, thickness of earth-fill cover is divided into the processing of at least two different depth, Each processing has identical quantity silkworm chrysalis, cultivates at room temperature, then records growing state, when culture to fructification grows simultaneously ripe, Cicada fungus can be harvested.

By adopting the above-described technical solution, compared with prior art, beneficial effects of the present invention：By testing final obtain Go out, the Ecology earthing artificial cultivation method for the cicada fungus which provides, the very high degree simulation wild situation of cicada fungus obtains The cicada fungus of the entomogenous fungi one similar with wild cicada fungus, and the cicada fungus of the cultivation method have been arrived, has gone out that careless high, fructification is long, Yield is high, and the length of fructification can reach 8.8cm, and the fructification fresh weight on every silkworm chrysalis can reach 2.73g, and dry weight can be with Reach 0.44g, industrialized development may be implemented, and the cultural method can mitigate foodstuff waste in the cultivation of ordinary solid culture medium The problem of, finally meet growing dietotherapy and demand clinically, exploitation of the people to wild cicada fungus is reduced, to protect The wild resource of cicada fungus.

Specific implementation mode

The following is a clear and complete description of the technical scheme in the embodiments of the invention, it is clear that described embodiment Only a part of the embodiment of the present invention, instead of all the embodiments；Based on the embodiments of the present invention, the common skill in this field The every other embodiment that art personnel are obtained without making creative work belongs to the model that the present invention protects It encloses.

The embodiment of the present invention：

A kind of Ecology soil covering culture method of cicada fungus, includes the following steps：

By cicada fungus bacterial strain（Senior general mountain cicada fungus）It is connected in the triangular flask inclined-plane made of PDA solid mediums, is trained under 25 DEG C of constant temperature About 7 are supported to generation conidium；

Spore suspension is made with spore of the sterile ultrapure water elution of the Tween 80 added with 0.5% in triangular flask, uses hemocytometer Number plate counting number simultaneously adjusts spore suspension concentration to 1 × 107/mL；

Silkworm chrysalis body surface is cleaned with 75% alcohol again to fall to remain alcohol with aseptic water washing.Then use the configuration in previous step good dense The spore suspension of degree injects tussah chrysalis, and every silkworm chrysalis injects 0.1mL, and injection site is silkworm chrysalis wing dead astern and third link Crosspoint；

The silkworm chrysalis injected is positioned in the culture box to sterilize.It puts the cotton that one layer sterilized and adds appropriateness in culture cassette bottom portion Sterile water cotton is soaked, then silkworm chrysalis is positioned on cotton, to keep certain humidity, is infected conducive to silkworm chrysalis.Then Placement is cultivated about 15 days at room temperature, and culture to silkworm chrysalis body surface grows white hypha；

Silkworm chrysalis with white hypha is transplanted in flowerpot, thickness of earth-fill cover makees tetra- processing of 1cm, 2cm, 3cm, 4cm respectively, often A processing has 15 silkworm chrysalises, cultivates at room temperature, growing state is then recorded, when culture to fructification grows simultaneously ripe, you can adopt Receive cicada fungus.

It is the record case of silkworm chrysalis growth course below：

After earthing culture 1 week, the fructification tip that had begun of earthing 1cm emits unearthed face, other thickness of earth-fill cover have no sub real Body is emerged；After earthing culture the 12nd day, earthing 1cm's has had 14 only fructifications to emit unearthed face about 1cm high；And 2cm's is rigid It begins with fructification tip and emits unearthed face；Earthing 3cm's and 4cm has no fructification tip；Earthing 1cm after earthing culture 20 days Silkworm chrysalis is emerged fructification, and most silkworm chrysalis length have reached 2cm or more；The fructification for having 5 silkworm chrysalises of earthing 2cm has been emerged There are about 1cm high, there is 7 fructification tips of having emerged；The fructification tip of only 7 silkworm chrysalises of earthing 3cm is emerged, and has one There are about 1cm high for fructification；It emerges at the fructification tip without a silkworm chrysalis of earthing 4cm；Show that the thicker fructification of earthing is emerged The time in native face is longer, and when thickness of earth covering reaches 4cm cordyceps sinensis fructification almost long face of not being unearthed, therefore earthing 4cm is not adopted The data of collected works physical length and yield directly can exclude the processing by observation；After earthing culture 30 days, earthing 1-3cm Deep fructification is mature on the whole, and the sporophore growth situation data record respectively handled is shown in Table 1.

As it can be seen from table 1 the growing state of different thickness of earth-fill cover fructifications is different, be embodied in earthing 1cm and The sub- true length gesture of 2cm is uniform and yield is higher；And to reach 3cm cordyceps sinensis " going out grass " irregular for thickness of earth-fill cover, and go out careless low, therefore it covers Native 3cm is not suitable for cultivation cicada fungus；Hardly " go out grass " when thickness of earth-fill cover reaches 4cm, therefore the more unsuitable cultivation cicada fungus of earthing 4cm； So after Integrated comparative, the thickness of earthing 1-2cm is all adapted for cultivating cicada fungus, but no more than 2cm, best earthing is thick Degree can be identified as 1.5cm.

The description of said program is the invention being intended to facilitate those of ordinary skill in the art to understand and use, and is familiar with The personnel of art technology obviously easily can make various modifications to embodiment, and therefore, the present invention is not limited to above-mentioned realities Scheme, those skilled in the art according to the method for the present invention, all should by improvement and modification made without departing from the scope of the present invention Within protection scope of the present invention.

Claims (1)

1. a kind of Ecology soil covering culture method of cicada fungus, it is characterised in that cultivated using following steps：

Step 1:, cicada fungus bacterial strain is connected in the triangular flask inclined-plane made of PDA solid mediums, cultivate 5- under 25 DEG C of constant temperature Extremely generate conidium within 10th；

Step 2: spore suspension is made with the spore in the sterile ultrapure water elution triangular flask of the Tween 80 added with 0.5%, use Blood counting chamber counts and adjusts spore suspension concentration to 0.8-1.2 × 107/mL；

Again fall to remain alcohol with aseptic water washing Step 3: cleaning silkworm chrysalis body surface with 75% alcohol；Then it uses in step 2 Configured the spore suspension injection tussah chrysalis of concentration, every silkworm chrysalis injects 0.08-0.15mL, injection site be silkworm chrysalis wing just Rear and third link crosspoint；

Step 4: the silkworm chrysalis injected is positioned in the culture box to sterilize；Put one layer of cotton to sterilize in culture cassette bottom portion And the sterile water of appropriateness is added to soak cotton, then silkworm chrysalis is positioned on cotton, to keep humidity to be infected in favor of silkworm chrysalis；It puts It is placed in and cultivates at room temperature about 12-20 days, culture to silkworm chrysalis body surface grows white hypha；

Step 5: transplanting the silkworm chrysalis with white hypha in flowerpot, thickness of earth-fill cover is divided into the processing of at least two different depth, Each processing has identical quantity silkworm chrysalis, cultivates at room temperature, then records growing state, when culture to fructification grows simultaneously ripe, Cicada fungus can be harvested.