1, medium formula. Corn flour120g, soybean flour 20g, sucrose 20g and water 1000 ml.
2. Production of culture medium. First, mix the weighed corn flour and soybean flour with appropriate amount of water, and then add enough water to boil. When it boils, start counting and keep it for 35 minutes. Pay attention to stirring from time to time during heating to avoid boiling, and pay attention to adding water to supplement the water lost during heating.
3. Add the required sucrose after cooking, subpackage it with a syringe or other device while it is hot, plug the cotton plug after subpackage, and place the test tube vertically. Because the culture medium is rich in nutrition, it is relatively difficult to completely sterilize corn flour, and the sterilization time is slightly longer than that of the conventional culture medium.
4. Generally, it can be sterilized at high pressure (1.5 kg/cm2) for 35 minutes to 40 minutes, and at normal pressure for 60 minutes to 90 minutes. Aft sterilization, that test tube is taken out when it is cool to be not hot, and the culture medium of Armillaria mellea mother seed placed vertically is generally made into a column, and the culture medium is completely solidify in the test tube after being completely cooled.
5. Inoculation, culture and preservation. Under aseptic conditions, each test tube is inoculated with a pea-sized strain block. The inoculated strain must be alive, the medium is brown, and the strain with severe bacteriocin keratinization should not be used.
6. After inoculation, culture in the dark at 25℃. Generally, after 7 days-10 days, the mycelium can be full. After the mycelium is full, put the strain in the refrigerator, which can generally be preserved for six months to one year. If long-term preservation is needed, the test tube mouth can be sealed with wax.
Extended data:
Has the following advantages
1, the raw materials for seed production are simple and easy to find, the seed production technology is simple and easy to operate, and the production cost is low. Convenient operation and easy completion.
2. Because the culture medium improves the water supply and is rich in nutrition, corn flour and soybean flour contain various growth factors, and the produced mother seed is thick, dense and white, which is not easy to brown and keratinize, and has strong anti-aging and anti-degradation ability, and also has strong rejuvenation ability for severely aged and degraded strains.
3. The mother seed made by this method can also greatly prolong the preservation time and reduce the number of generations in the preservation process. Can play a great role in inhibiting strain degradation.
4. The produced mother seed has strong vitality, and the strain blocks grafted on the mother seed, original seed or cultivated seed medium germinate quickly, and the growth speed is obviously faster than that of the mother seed produced by conventional methods.