Alternative solid-liquid culture method of Amanita hongtuo mycelium

Abstract

The invention relates to a method for alternating solid-liquid culture of Amanita amanita mycelium, belonging to the technical field of large fungus culture. It is characterized in that: the induced and multiplied mycelia are cultured alternately with solid and liquid, that is, after 4 to 6 generations of solid culture, 3 to 4 generations of liquid culture, and then solid culture, the growth rate of the mycelium is 8 to 10 times before the liquid culture, the growth was significantly accelerated. The invention speeds up the propagation speed of the mycelium by simply changing the culture mode, and the operation is simple and easy to realize, the benefit is obvious, and the production cost is low. Amanita toxins have potential applications in the development of new specific drugs such as antitumor, antibacterial and antiviral, sedative or anesthetic drugs, but so far it has been difficult to develop and apply due to bottlenecks such as rare resources and difficulty in artificial domestication. The present invention focuses on the problem of restricting pure culture such as the very slow growth of Amanita hongtuo mycelia, and provides conditions for large-scale cultivation of mycelia and artificial domestication cultivation in the future.

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. But now with the emergence of the global ecological crisis, many large fungal resources in my country are in danger, in a serious decline and endangered state, and some species are facing the risk of extinction bef...

Examples

example 1

[0021] Pick young fruiting bodies that grow vigorously and have not opened umbrellas in the field; take the internal aseptic tissue block of about 0.3cm 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 0.40g / L, MgSO 4 0.40g / L,...

example 2

[0026] Pick young fruiting bodies that grow vigorously and have not opened umbrellas in the field; take the internal aseptic tissue block of about 0.3cm 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 2 1.00g / L, KH 2PO 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 0.60g / L, MgSO 4 0.60g / L, zeatin ZT1.20mg / L, ...

example 3

[0031] Pick young fruiting bodies that grow vigorously and have not opened umbrellas in the field; take the internal aseptic tissue block of about 0.3cm 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 0.50g / L, MgSO 4 0.50g...

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. But now with the emergence of the global ecological crisis, many large fungal resources in my country are in danger, in a serious decline and endangered state, and some species are facing the risk of extinction bef...

Examples

example 1

[0021] Pick young fruiting bodies that grow vigorously and have not opened umbrellas in the field; take the internal aseptic tissue block of about 0.3cm 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 0.40g / L, MgSO 4 0.40g / L,...

example 2

[0026] Pick young fruiting bodies that grow vigorously and have not opened umbrellas in the field; take the internal aseptic tissue block of about 0.3cm 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 2 1.00g / L, KH 2PO 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 0.60g / L, MgSO 4 0.60g / L, zeatin ZT1.20mg / L, ...

example 3

[0031] Pick young fruiting bodies that grow vigorously and have not opened umbrellas in the field; take the internal aseptic tissue block of about 0.3cm 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 0.50g / L, MgSO 4 0.50g...