液氮保藏菌种对平菇菌丝体和子实体的影响

Effects of liquid nitrogen preservation on mycelial and fruiting bodies of Pleurotus ostreatus

以皖平菇1号为试验材料,对菌种液氮保藏过程中的降温方式、保护剂、解冻3个关键环节设计三因素三水平正交试验。结果表明,正交试验9个不同的处理中对菌种萌发率影响最小的条件处理组合为不加保护剂直接放入液氮,38～40℃水浴解冻;对菌丝生长速度影响最小的处理组合为-70℃-液氮逐步降温,10%的二甲基亚砜作保护剂,38～40℃水浴解冻;对子实体平均产量影响最小的处理组合为4℃--20℃--70℃分别放置2 h的逐步降温法,10%的蔗糖水溶液作保护剂,38～40℃水浴解冻。保护剂和解冻方式的选择对菌种影响较大,结合成本等其他因素综合考虑,不加保护剂直接将菌种放入液氮,38～40℃水浴快速解冻的处理方式为最佳选择。

In this paper, ultra-low temperature preservation of Pleurotus ostreatus ＂Wanping No. 1＂ spawn was studied by designing an orthogonal experiment with three factors（cryoprotectant, pretreatment and thawing） at three levels. The results showed that in the 9 treatments, A3B3C3, with non-pretreatment, without a cryoprotectant into liquid nitrogen and thawing in a 38-40℃ water bath after taken out from liquid nitrogen is the best combination which bring the minimal influence on the germination phase and viability rate of mycelial; A2B1C3, with --70℃-liquid nitrogen cooling of spawn, 10%dimethylsulfoxide as cryoprotectant and thawing in a 38-40℃ water bath after taken out from liquid nitrogen is the best combination which bring the minimal influence on the mycelial growth rate and growth condition; A1B2C3, with 4℃ -- 20 ℃ -- 70 ℃-liquid nitrogen cooling of spawn, 10% sugar solution as cryoprotectant and thawing in a 38-40℃ water bath after taken out from liquid nitrogen is the best combination which bring the minimal influence on the yield of fruiting bodies. Cryoprotectant and thawing play important roles in the process of liquid nitrogen preservation ofPleurotus ostreatus. The best method for preservation of spawn in liquid nitrogen is the combination of non-pretreatment, without a cryoprotectant into liquid nitrogen and thawing in a 38-40℃ water bath after taken out from liquid nitrogen, taken cost into consideration.