灰树花原生质体高温胁迫条件研究及其再生菌株的胞外酶活性分析  

Study on Heat Stress Condition of Grifola frondosa Protoplast and Analysis of Extracellular Enzyme Activity of Regenerated Strains

作　　者：王春晖 徐宁 吴芳 肖颖群 王小艳 姜性坚 张宇 胡汝晓 WANG Chun-hui;XU Ning;WU Fang;XIAO Ying-qun;WANG Xiao-yan;JIANG Xing-jian;ZHANG Yu;HU Ru-xiao(Hunan Edible Fungus Research Institute,Changsha 410016,China;Chaling County Xingnong Edible fungus Planting Farmer Professional Cooperative,Zhuzhou 412415,China)

机构地区：[1]湖南省食用菌研究所,湖南长沙410016 [2]茶陵县兴农食用菌种植农民专业合作社,湖南株洲412415

出　　处：《中国食用菌》2022年第6期41-45,51,共6页Edible Fungi of China

基　　金：湖南省自然科学基金面上项目(2020JJ4048);湖南省科技特派员创新创业项目(2020NK4135);湖南省农业种质资源普查保护利用项目(湘财农指[2021]37号)。

摘　　要：以胁迫温度和胁迫时间为参数设置梯度胁迫条件,对菌株灰树花2#原生质体进行高温胁迫,研究高温致死胁迫条件,分析不同胁迫条件下再生菌株的胞外酶活性。结果表明,灰树花2#原生质体高温胁迫的最适温度为48℃,对原生质体致死发生响应的最短时间为0.5 min,使原生质体全部致死的最长时间为7.5 min,胁迫条件为48℃、0.5 min~7.5 min。不同高温胁迫条件下的再生菌株之间、再生菌株与出发菌株之间的胞外酶活性存在个体差异。再生菌株T_(48)M_(7)-1、T_(48)M_(5)-19和T_(48)M_(3)-36的羧甲基纤维素酶活性及漆酶活性极显著高于出发菌株灰树花2#(P<0.01)。再生菌株T_(48)M_(7)-1的羧甲基纤维素酶活性及固体培养基漆酶活性显著高于T_(48)M_(5)-19和T_(48)M_(3)-36菌株(P<0.05)。研究结果可为灰树花及其他食用菌原生质体育种技术提供科学参考。Stress temperature and stress time were used as parameters to set the gradient heat stress amount,and the stress conditions were studied under heat stress on the protoplast of strain Grifola frondosa 2#,and the activity of extracellular enzymes of the regenerated strain was analyzed under different heat stress conditions.The results showed that the optimum temperature of G.frondosa protoplast under heat stress was 48℃,the shortest time in response to protoplast death and longest time to kill all protoplasts were 48℃,0.5 min and 7.5 min,respectively,and the range of heat stress condition was 48℃,0.5 min-7.5 min.There were individual differences in extracellular enzyme activities between the regenerated strains and the original strains and between the regenerated strains under different heat stress londitions.The activities of carboxymethyl cellulase and laccase of the regenerated strains T_(48)M_(7)-1,T_(48)M_(5)-19 and T_(48)M_(3)-36 were higher than those of the starting strain,and the activities of carboxymethyl cellulase and laccase activities represent extremely significantly difference(P<0.01).The carboxymethyl cellulase activity and solid medium laccase activity of the regenerated strain T_(48)M_(7)-1 was significantly higher than that of T_(48)M_(5)-19 and T_(48)M_(3)-36(P<0.05),the study results could provide a basis for protoplast breeding of G.frondosa and other edible fungi.

关 键 词：灰树花2# 原生质体 高温胁迫 再生菌株 胞外酶活性 

分 类 号：S646.2[农业科学—蔬菜学]