双孢蘑菇菌种液氮保种方法的研究

Study on the Technology of Liquid Nitrogen Preservation of Agaricus bisporus Spawn Stocks

主要进行双孢蘑菇菌种液氮保藏中菌丝的培养基质、不同冷冻保护剂、不同降温速率及菌种解冻方式等因素对菌种活力的影响研究。结果表明生长3 d^5 d菌龄的PDA菌丝块,以10%甘油作为冷冻保护剂及聚丙烯吸管安瓿为容器,制备的菌种保藏管在液氮罐口缓慢降温至-40℃左右时,迅速浸入液氮保藏,提取的液氮保藏菌种以快速解冻方式进行复苏,双孢蘑菇菌种经过4年的液氮保藏,存活率达100%,栽培出菇的农艺性状仍保持原有菌株特性,研究建立的双孢蘑菇液氮保藏方法已成为我所蘑菇菌种长期保藏的主要方法之一。

To investigate the optimum conditions of liquid nitrogen preservation of Agaricus bisporus spawn stocks, four preservation factors, i.e. culture substrate of mycelium, cryoprotectants, cooling rate and thawing methods were discussed in this study. The results showed that PDA medium blocks after 3 d-5 d, 10% glycerol as cryoprotectant and Polypropylene-straw-ampoules as container were the best. In cooling process, strain tubes for preservation immersed into liquid nitrogen jar rapidly after cooling slowly to about -40℃ at the opening of the tank, and thawed rapidly for recovery when used. In this way, Agaricus bisporus strains had a 100% survival rate after 4 years＇ preservation and maintain their agronomic characters. This method had become a long-term preservation method of Agaricus bisporus spawn stocks in the author＇ s institute.