基于响应面法对香菇原生质体制备条件的优化Optimization of Lentinula edodes protoplast producing conditions based on response surface methodology
孙佳星;杨红;孙晓东;郝然;张伟东;吕国忠;
摘要(Abstract)：
香菇Lentinula edodes是我国食用菌年产量最大的单品，优化香菇原生质体的制备条件，获得高质量、高活力的原生质体可以为香菇育种和遗传多样性分析提供技术保障。本研究利用单因素试验对稳渗剂浓度、酶解液类型、酶解时间、酶解温度4个因素的最优作用条件和相关性进行检验和筛选；依据单因素试验的结果，对酶解时间、酶解温度、酶解液种类进行响应面法3因素3水平试验优化分析。单因素试验结果表明，使用2%的溶壁酶+蜗牛酶+纤维素酶混合酶制剂为酶解液、0.6mol/L甘露醇为稳渗剂的效果显著高于其他处理(P<0.05)。经Box-Behnken设计得出香菇原生质体最佳制备条件为酶解时间3.25h，酶解液浓度1.7%，酶解温度30.49℃，获得原生质体产量为14.02×106 CFU/mL，与理论产量(13.862×106 CFU/mL，与理论产量(13.862×106 CFU/mL)偏差小，可为后续原生质体融合育种和多样性分析提供重要基础。
关键词(KeyWords)： 原生质体;单因素试验;多酶协同酶解;Box-Behnken设计
基金项目(Foundation): 辽宁省科技厅民族创新联合基金项目(2020-MZLH-14)~~
作者(Authors): 孙佳星;杨红;孙晓东;郝然;张伟东;吕国忠;
DOI: 10.13346/j.mycosystema.210295
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