(1) medium formula: I: PDA (potato 200g, glucose 18g, agar 18g, water 1000m 1). Ⅱ: PDA+magnesium sulfate 1.5g+ potassium dihydrogen phosphate 3g+ peptone 2g, water 1000ml. ⅲ: potato 100g, sawdust 100g, glucose 20g, agar 20g, water 1000ml.
Sterilize the culture medium with the above formula under high pressure for 20 minutes, make it into a test tube inclined plane, and use it up within 3 days.
(2) Production method: Inoculate the selected stropharia rugoso-annulata tissue block to the culture medium (or inoculate the preserved species to be rejuvenated) through tissue separation technology in NoI. After culturing at 24-28℃ for 10- 15 days, when the mycelium grows to 1: 3- 1: 4 in the test tube, select the thick part of the front end of the mycelium and transfer it to No.2 or No.3 medium, and adopt two-point inoculation method to make the mycelium grow in the medium (generally 65438+)
2. Production of original seeds and production seeds
(1) Formula: I: wheat or barley 88%, rice bran or wheat bran 10%, gypsum or calcium carbonate 1%, lime 0.5%, water content 65% (adding rice bran or wheat bran is mainly loose culture medium to increase air permeability and make hyphae move well). No.2: wheat 25%, straw 64%, wheat bran 10%, gypsum powder 1%, water content 65%-70%. ⅲ: sawdust 42%, shavings 42%, rice bran or wheat bran 15%, magnesium sulfate 0.2%, gypsum or calcium carbonate 1%, and water content 70%.
The above formula was sterilized under the pressure of 1.3- 1.5 kg/m2 for 2-2.5 hours and used up within 3 days.
(2) Selection and preparation of raw materials: Practice has proved that the main raw material of the culture medium made by the original and production species of Pleurotus ostreatus is wheat, followed by sawdust and shavings, and straw.
(3) Selection of wheat grains: It is best to choose wheat grains with dark yellow or dark red skins, which are tough and not easy to break when soaked. In terms of water absorption, barley is superior to wheat.
(4) Soaking: Soaking can be divided into two methods: soaking in lime water and adding lime after precooking with clear water. The former is soaked in 1% lime water, the pH value is 9 ~ 10, wheat is soaked for 8 ~ 9 hours, barley is soaked for 9 ~ 10 hours, the temperature is 25℃, and the time should be shortened when the water temperature is high in summer. Soaked wheat grains, pinched with fingernails, are very elastic, and nail prints should disappear soon. The latter first soaked the wheat with clear water for the same time as on the table. Wash the soaked wheat grains with clear water, drain them, put them in a pot until they just boil, open them after cooling, mix them with paving materials, adjust the pH to pH6.5 with 0.5% lime, and the water content is 65%.
(5) Bottle disinfection: select a pale white high-temperature glass bottle with a volume of 700ml and an inner diameter of about 3cm (wide-mouth bottles are not suitable to avoid the invasion of miscellaneous bacteria), put the evenly mixed culture materials into bottles, each bottle is 200-300 g, and disinfect for 2-2.5 hours under the pressure of 1.3 ~ 1.5kg/m2.
(4) Inoculation culture: multi-point inoculation is adopted, that is, 3-4 points are inoculated around the culture material in the bottle at a proper distance, and then a small piece of strain is inoculated on the material surface, and it is cultured in the dark at 24-28℃ for 7- 10 days. When the hyphae germinate and grow to a plaque with a diameter of about 4cm, the hyphae at the inoculation point are stirred manually immediately, and the hyphae at the stirring point are broken.
3. Production of production species:
(1) Formula, culture medium making, bottle selection and disinfection are the same as the original species.
(2) Inoculation: Inoculate a full bottle of original seeds on the surface of the production medium, and spread the material surface as far as possible to avoid contamination by miscellaneous bacteria. Inoculate no more than 30 bottles of seed for production per bottle of original seed. After 7- 10 days of culture at 24-28℃, when the hyphae germinate and grow to be white and thick, the hyphae are artificially stirred to break and stimulate their rapid growth, usually for 20 days.