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Solid-liquid alternating culturing method for mycelia of amanita rubrovolvata imai

A kind of technology of Amanita hongtuo, solid-liquid alternation, applied in the fields of botanical equipment and methods, fertilizer mixture, gardening, etc., can solve the problems of easy disappearance or extinction, slow growth of mycelium, high price, etc. The effect of increasing growth rate and low production cost

Inactive Publication Date: 2014-04-02
YUNNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, the main bottlenecks that lead to the difficulty in the development and sustainable utilization of Amanita are: ① Amanita resources are rare, and its population is small, with few individuals, low yield, and extremely limited distribution. In addition, it is sensitive to habitat. Once the habitat is destroyed, it is easy to Disappeared or extinct, belonging to a special group of endangered organisms, which increases the difficulty of its further research and utilization; ②Amanita, most of which belong to ectomycorrhizal fungi, most of which are still difficult to cultivate in nature Or uncultivated microorganisms, difficult to artificial, semi-artificial cultivation
Therefore, there is no toxin product that can be developed on a large scale at home and abroad so far. Peptide toxins used as biochemical reagents are still expensive (about 100,000 US dollars per gram more than 10 years ago—Chen Zuohong et al., 1999), which is difficult to meet the needs of scientific research and application required
[0005] The pure culture of Amanita amanita is the premise or key to ensure the sustainable utilization of Amanita amanita resources, but it is still a problem that is difficult to overcome, and there are many blind spots. Difficulty in inducing filaments and very slow mycelial growth are important links or factors that restrict and affect pure culture

Method used

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Examples

Experimental program
Comparison scheme
Effect test

example 1

[0021] Harvest young fruiting bodies that grow healthy and have not opened umbrellas in the field; take the internal aseptic tissue block of about 0.3 cm at the junction of the cap and the stipe 2 Size, gently embedded in the induction medium, the medium components are: potato 200g / L, glucose 13g / L, yeast extract 1.20g / L, MgSO 4 1.00g / L, CaCl 2 1.00g / L, KH 2 PO 4 0.50g / L, 120ml of baume wort, 120ml of agar, 10.00g / L of agar, the pH value of the control medium is 5.8, and cultured in dark at 22-23°C for 46 days, fluffy white hyphae grow on the surface of the tissue block; The silk was subcultured and multiplied three times in the above-mentioned induction medium;

[0022] Then transfer the mycelium to a 9cm petri dish containing solid medium, and culture it in dark at 22-23°C for 70 days. When the mycelium grows to 3.5cm, transfer and cultivate for 4 generations; The medium components are: potato 200g / L, glucose 15g / L, yeast extract 1.20g / L, ZnSO 4 0.40 g / L, MgSO 4 0.40...

example 2

[0026] Harvest young fruiting bodies that grow healthy and have not opened umbrellas in the field; take the internal aseptic tissue block of about 0.3 cm at the junction of the cap and the stipe 2 Size, gently embedded in the induction medium, the medium components are: potato 200g / L, glucose 15g / L, yeast extract 1.40g / L, MgSO 4 1.00g / L, CaCl 21.00g / L, KH 2 PO 4 0.50g / L, 140ml of 16.0 Baume's wort, 10.00g / L of agar, and the pH value of the control medium was 6.0. After 55 days of dark culture at 22-23°C, fluffy white hyphae grew on the surface of the tissue block; The silk was subcultured twice in the induction medium mentioned above;

[0027] Then transfer the mycelium to a 9 cm petri dish containing solid medium, and culture it in dark at 22-23°C for 80 days. When the mycelium grows to 4.5 cm, transfer it and cultivate it for 6 generations; The medium components are: potato 200g / L, glucose 18g / L, yeast extract 1.40g / L, ZnSO 4 0.60 g / L, MgSO 4 0.60g / L, zeatin ZT1.20mg / ...

example 3

[0031] Harvest young fruiting bodies that grow healthy and have not opened umbrellas in the field; take the internal aseptic tissue block of about 0.3 cm at the junction of the cap and the stipe 2 Size, gently embedded in the induction medium, the medium components are: potato 200g / L, glucose 14g / L, yeast extract 1.30g / L, MgSO 4 1.00g / L, CaCl 2 1.00g / L, KH 2 PO 4 0.50g / L, 130ml of 16.0 Baume's wort, 10.00g / L of agar, and the pH value of the control medium is 5.8. After 50 days of dark culture at 22-23°C, fluffy white hyphae grow on the surface of the tissue block; The silk was subcultured and multiplied three times in the above-mentioned induction medium;

[0032] Then transfer the mycelium to a 9 cm petri dish containing solid medium, and culture it in dark at 22-23° C. for 75 days. When the mycelium grows to 4.0 cm, transfer it and cultivate it for 5 generations; The medium components are: potato 200g / L, glucose 17g / L, yeast extract 1.30g / L, ZnSO 4 0.50 g / L, MgSO 4 0...

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Abstract

The invention relates to a solid-liquid alternating culturing method for mycelia of amanita rubrovolvata imai and belongs to the technical field of culturing of macro fungi. The solid-liquid alternating culturing method for mycelia of amanita rubrovolvata imai is characterized by comprising the steps of: performing solid culturing on the induced and propagated mycelia for 4-6 generations and then performing the liquid culturing on the induced and propagated mycelia for 3-4 generations by using the solid-liquid alternating culturing method; then performing the solid culturing, wherein the growth speed of the mycelia is 8-10 times that of the mycelia before the liquid culturing, and the growth is speeded obviously. According to the invention, the propagation speed of the mycelia is speeded up by simply changing the culturing method, the operation is simple and easy to implement, the effect is obvious, and the production cost is low. The toxin of the amanita has potential application in the field of developing new specific medicines such as antineoplastic medicines, antibiosis and antiviral medicines, sedatives or narcotics, but is different to develop and apply due to the bottleneck problems such as the valuable and rare resource and the difficulty in artificial acclimatization so far. According to the invention, focused research is performed on the problems for limiting the pure culturing such as very slow growth of the mycelia of amanita rubrovolvata imai, and conditions are provided to the large-scale culturing of the mycelia, the artificial acclimatization cultivation in future and the like.

Description

technical field [0001] The invention relates to a method for solid-liquid alternate cultivation of Amanita amanita mycelium, which belongs to the field of biotechnology, and specifically belongs to the category of indoor cultivation of large poisonous fungi. Background technique [0002] Amanita ( Amanita ) belongs to Basidiomycotina, Phytomycetes, Agaricales, Amanitaceae ( Amanitaceae ), is a more special and valuable worldwide widespread genus among poisonous macrofungi, and its species diversity is very rich. Nearly 400 species have been reported in the world, and nearly 100 species (including subspecies, variants and variants) have been recorded in China. According to research, there are still many species in China that have not yet been studied and named. However, with the emergence of the global ecological crisis, many large fungal resources in our country are in danger, in a serious decline and endangered state, and some species are facing the risk of extinction b...

Claims

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Application Information

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IPC IPC(8): A01G1/04C05G3/00
Inventor 李宗菊李彪张曦予冯辽辽赵昱左奎
Owner YUNNAN UNIV
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