Storage method of edible fungus strain and mastery of topdressing method of edible fungus

1. In the method of periodic transplantation and storage at low temperature, the bacteria that need to be stored are inoculated on the suitable slope medium and cultured at the right temperature. When the mycelium grows all over the slope, the mycelium is taken out and stored in a low temperature dry place of 3 ℃-5 ℃ or in a 4 ℃ refrigerator or freezer. The bacteria are transferred every 4-6 months, which should be determined according to the characteristics of the bacteria. Note that the ambient temperature should not be too high during storage to prevent mold from entering the tube through the cotton stopper. Therefore, if the cotton plug is used, the cotton plug can be wrapped with clean sulfuric acid paper or Kraft paper, which can not only reduce the chance of pollution, but also prevent the culture medium from drying. Except volvariella volvacea, other edible fungi can be stored by this method.

two。 Liquid paraffin storage method take chemically pure liquid paraffin (no moisture, no mildew) and put it in a triangle bottle with cotton stopper and wrapped paper, sterilize under the pressure of 1 kg / square centimeter for 1 hour, and then put it in a 40 ℃ thermostat for several days to evaporate the moisture until the paraffin oil is completely transparent. The treated paraffin oil was transferred to the blank slope and cultured at 28 ℃-30 ℃ for 2-3 days. Then the liquid paraffin is injected into the inclined tube to be stored by aseptic operation. The appropriate amount of injection is 1-1.5 cm higher than the slope of the culture medium, plug it with a rubber plug, seal it with solid paraffin, and store it upright in a low temperature and dry place. The storage time is more than 1 year, and the storage time can be prolonged at low temperature.

3. In the sand tube storage method, the river sand was soaked and washed with water for several times, the coarse grains were removed through a sieve of 60 mesh, then soaked in 10% hydrochloric acid for 2-4 hours, the organic matter was removed, then washed with water until the pH value reached neutral, and dried. At the same time, soak the lean ridge soil or vegetable garden soil in water to make it neutral, remove the supernatant after precipitation, dry and grind it fine, sift it with a 100-mesh sieve, mix the treated sand with soil in a ratio of (2-4): 1, absorb the iron with a magnet, then pack it into small test tubes or ampoules, pack 0.5-2 grams per tube, plug cotton, and wrap sterilization with paper (1.5 kg / square cm, 1 hour). Then dry heat sterilization (160℃, 2-3 hours) 1-2 times, aseptic test, qualified use. The bacteria that have formed spores are injected into 3-5 ml of aseptic water under aseptic conditions, and the bacterial suspension is made by scraping the stalks, and then the bacterial solution is dripped into the sand pipe with a sterile straw to soak through the sand. Put the inoculated sand pipe into a vacuum dryer filled with desiccant and connect it with a vacuum pump for several hours until the sand is dry. Vacuum drying should be completed within 48 hours after spore insertion to avoid spore germination. The prepared sand pipe is sealed with paraffin and can be preserved for 2-10 years at low temperature.

4. Filter paper storage method: take white (collect dark spores) or black (collect white spores) filter paper, cut into small strips of 4 cm × 0.8 cm, lay them in a petri dish and wrap them with paper for sterilization (1 kg / square cm, 30 minutes). The spores were collected by hook suspension method, and the spores fell on the filter paper. Put the filter paper strip containing spores into the storage test tube, and then put the storage test tube into the dryer for 1-2 days, remove the moisture of the filter paper, make the moisture content of the filter paper reach about 2%, and then store it at a low temperature.

5. ① wheat grain storage method. Rinse the wheat without grained grains and impurities, soak for 12-15 hours, boil with water for 15 minutes, continue hot soaking for 15 minutes, make the wheat bulge without breaking, drain and spread out to dry, so that the water content of the wheat is about 25%. Mix calcium carbonate and gypsum into cooked wheat grains (10 kg: 133 g: 33 g), mix evenly, put them into test tubes, 2-3 grams per tube, then clean the test tubes, plug cotton plugs, sterilize (1.5 kg / sq cm, 2 hours), set aside after aseptic inspection, inoculate the test tube matrix after cooling, culture at the right temperature, and coat the cotton plugs with paraffin when the mycelia are full. Store at low temperature. Transfer every two years or so. ② bran koji preservation method. Take fresh wheat bran and sift through 60 mesh to remove coarse grains. Mix the bran and tap water at 1:1, put it into a small test tube, each tube is packed with a cotton stopper, wrap it with paper, sterilize it under high pressure (1.5kg / square centimeter, 30 minutes), and set aside after passing the sterility test. transfer the robust bacteria growing on the inclined medium to the aseptic bran koji tube, and try to tamp the bran in the small test tube as far as possible to make it loose. Culture at the appropriate temperature until the mycelium is full of bran, put the bran koji tubule in the dryer and store at low temperature or suitable temperature.

6. 0.7-0.9 grams of pure sodium chloride was taken by physiological saline storage method, put into 100% ml distilled water, stirred and packed evenly in test tubes, 5-10 ml per tube, sterilized (1 kg / square centimeter, 30 minutes). After passing the sterility test, the bacteria to be preserved were put into the potato glucose liquid medium and cultured at suitable temperature for 5-7 days. Aseptic operation absorbs a few cultured bacteria, injects them into qualified saline test tubes, plugs them with aseptic rubber plugs, smears them with paraffin, and stores them at room or low temperature.

7. The cultured and abundant bacteria or spores were suspended on the bacterial suspension made of sterilized serum, egg white and skim milk by freeze vacuum drying, and the suspension was packed aseptically in sterilized glass ampoules of 0.3-0.5ml each tube, then connected with the freeze-drying device with a pressure rubber tube. The ampoules were placed in a freezing tank for rapid freezing at-30 ℃ ~-40 ℃, and then emptied and dried in the frozen state. The ampoules were melted and sealed in vacuum and stored at-20 ℃. Generally, the ampoules could be preserved for more than 10 years, but the cost was high.

8. Liquid nitrogen cryopreservation method: first of all, the bacteria to be preserved are prepared into bacterial suspensions, and then ampoules are prepared, 0.8 ml cryoprotectant 10% (volume ratio) glycerin distilled water solution is added to each bottle, and cotton plugs are sterilized (1 kg / square centimeter, 5 minutes). After aseptic examination, put in the bacteria to be stored, fuse the mouth of the bottle and check whether there is air leakage. Put the sealed ampule bottle in the freezer and slowly cool it at a rate of 1 ℃ per minute, so that the collection is gradually and evenly frozen until-35 ℃. After that, the freezing speed does not need to be controlled. Ampoules are immediately put into liquid nitrogen tanks and preserved at-150 ℃ ~-196 ℃.