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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。
Taking Wanping mushroom No.1 as test material, orthogonal experiment of three factors and three levels was designed for the three key steps of temperature reduction, protective agent and thawing during the process of liquid nitrogen storage. The results showed that in the nine orthogonal experiments, the germicidal rate had the least effect on the germination rate. The combination of treatment with no protective agent was directly placed in liquid nitrogen and thawed at 38-40 ℃ in water bath. The treatment with the least influence on mycelial growth rate The combination was -70 ℃ -the liquid nitrogen was gradually cooled down, 10% dimethylsulfoxide was used as the protective agent and the water bath was thawed at 38-40 ℃. The treatment combination with the least effect on the average fruiting body yield was 4 ℃ -20 ℃ -70 ℃ were placed 2 h gradual cooling method, 10% sucrose as a protective agent, 38 ~ 40 ℃ water bath thawing. The selection of protective agent and thawing method has a great impact on the strain, taking the combination of cost and other factors into consideration, the strain is directly placed in liquid nitrogen without protective agent, and the best way is to quickly thaw in the water bath at 38-40 ℃.