Preparation of secondary strain of volvariella volvacea

The secondary species is a strain made by transferring the primary species into the culture material with secondary species. Second-class species can be directly used for production. In general, secondary species are transferred to the same culture material for expanded culture to become productive species. Secondary species are generally packed in 750 ml mushroom bottles or plastic bags.

(1) preparation of culture materials

Formula one: 90% rice straw, 9% wheat bran (rice bran), 1% gypsum powder. The preparation method is to select the golden and mildew-free dried straw, cut it into 3 cm long, soak it in clean water for about 12 hours, make it fully absorb water (only the rice straw that absorbs water is easy to be sterilized thoroughly), after picking it up, squeeze out the excess water, add wheat bran and gypsum powder in proportion, mix well, and the humidity should be dripped continuously with water droplets in hand. Then put it in a bottle, compacted with a stick on the one hand, loose and tight evenly and moderately, generally 75g of hay in 750ml bottle. The culture material should be packed about 5 cm away from the mouth of the bottle and should not be too full. Then use a sharp stick to make a small hole in the middle of the bottle to facilitate vaccination. Wash the culture medium attached inside and outside the bottle mouth, dry the inner wall of the bottle mouth, plug with cotton stopper, and sterilize.

Formula 2: cottonseed hull 94%, wheat bran 5%, gypsum powder 1%, lime 6%. Preparation method: pre-wet the cottonseed shell and add auxiliary materials, and the water content is about 60%. It can be bottled (bag).

Formula 3: 98.5% wheat grain, 1% gypsum powder, 0.5% calcium carbonate. Preparation method: soak the wheat grain in 12%-15% lime water for about 10 hours, heat and boil for 15 minutes, then filter, let the wheat grain dry slightly, cool, mix in gypsum powder and calcium carbonate, and bottle immediately. When bottling, a thin layer of rice chaff with a humidity of 30% to 40% is placed at the bottom of the bottle, and a thin layer of cottonseed shell or rice chaff is placed at the shoulder of the bottle as sealing material to reduce miscellaneous bacteria pollution. The bottling method of other culture materials is the same as that of formula one. This method should be used as a secondary species, such as a productive species, which is easy to cause pollution because of good nutrition.

(2) sterilized bottles (bags) filled with culture materials should be sterilized on the same day to avoid fermentation deterioration. The sterilization pressure is 137 ~ 147 KPA and the sterilization time is 1.5 hours. If the atmospheric pressure sterilization stove is used, the water needs to be boiled for 8 hours and stuffy overnight, which can also achieve the purpose of sterilization.

(3) immediately after inoculation and sterilization, take out the strain bottle (bag) and send it to the inoculation box or super clean table. After cooling, it can be inoculated. Generally, each first-class seed is connected with 4-8 bottles of second-class species, and each bottle of second-class species is connected with about 60 bottles to produce seeds.

(4) after culture and inoculation, all the bacteria were transferred to the incubator or incubator. The suitable room temperature is 3032 ℃, and it can be checked after 24 hours. It is best to check the mycelium once a day before sealing, and pick out the contaminated bacteria in time. The optimum bacterial age is 3-5 days after the strain grows in the bottle, and the age of the 750 ml mushroom bottle is 25-30 days. In particular, it should be pointed out that the training room should be clean and ventilated, and the outdoor environment should also be clean.