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近年来,国内外发表了不少利用培养在琼脂斜面上的菌丝块进行超低温冷冻保存报道,这种方法虽然是成功的,但要考虑的因素较多,比如保护剂的浓度、处理时间以及毒性等。我们试用草菇、香菇、蘑菇、凤尾菇、金针菇、黑木耳、毛木耳七个菌种的麦粒种,不用保护剂进行液氮保存,取得了很好的效果。材料与方法(一)制麦粒菌种选新鲜、饱满的麦粒,在流水中浸一夜,洗净。锅中蒸20分钟左右,麦粒熟透。但不开裂即可。在麦粒中加入1～1.5%碳酸钙,调节pH值到7.2,搅拌均匀。装入试管,在1.4kg/cm~2下灭菌2小时,冷却后接种,在24～26℃下培养。(二)冷冻处理将菌龄为14天的麦粒种,装在经高压灭菌的特制聚丙烯液氮管中,系上绳子,迅速投入液氮罐中保存1～4天。
In recent years, many domestic and international published reports on the use of mycelium cultured on agar slash for cryopreservation. Although this method is successful, there are many factors to be considered, such as the concentration of protective agent, the treatment time and Toxicity and so on. We tried straw, mushrooms, mushrooms, mushrooms, mushrooms, black fungus, hair fungus seven kinds of wheat germ, without preservative liquid nitrogen preservation, and achieved good results. Materials and methods (A) Saccharomyces cereal selection of fresh, full of wheat, soaked in running water overnight, washed. Steamed pot about 20 minutes, grains cooked. But not cracking can be. Add 1 to 1.5% calcium carbonate in the grains, adjust the pH value to 7.2 and stir well. The test tube was filled in, sterilized at 1.4 kg / cm ~ 2 for 2 hours, cooled and inoculated and cultured at 24-26 ° C. (B) Freezing treatment The 14-day-old wheat seeds were placed in a specially-made polypropylene liquid nitrogen tube autoclaved and tied to a rope and quickly put into a liquid nitrogen tank for 1 to 4 days.