食用菌病毒脱毒方法的比较

Comparisons of Different Methods for Virus-elimination of Edible Fungi

应用dsRNA技术从平菇天达300(Pleurotus ostreatus TD300)菌丝中提取到4条dsRNA条带,大小分别是8.2、2.5、2.1和1.1kb。采用菌丝尖端脱毒法、原生质体再生法、冷冻真空干燥法和单孢杂交法进行脱毒,各得到一株脱毒菌株,分别是HTC8、PR15、FL01和SSH11。脱毒菌株HTC8仅残留有低含量的8.2kbdsRNA,PR15和SSH11完全脱去dsRNA,而FL01仍残留有低浓度的8.2kb和2.5kbdsRNA。该4株脱毒菌株的菌丝生长速度和漆酶活力均比出发菌种有不同程度的提高,尤其是HTC8和PR15,菌丝生长速度比出发菌种分别提高22.73%和18.18%,漆酶活力比出发菌种分别提高145.83%和134.38%。综合比较此4种脱毒方法,菌丝尖端脱毒法和原生质体再生脱毒法比冷冻真空干燥法和单孢杂交法脱毒效果好,制备脱毒菌株用时短、效率高。

Four dsRNA bands were extracted from Pleurotus ostreatus TD300 by the dsRNA isolation technique with sizes of 8.2kb,2.5kb,2.1kb,and 1.1kb,respectively. Four virus-eliminated methods,i.e. hyphal tips cut （HTC）,protoplast regeneration （PR）,single spore hybridization （SSH）,and frozen and lyophilized （FL）,were applied to prepare virus-eliminated strains,and one virus-eliminated strain was selected for each virus-elimination method. The virus-eliminated strains were named as HTC8,PR15,FL01,and SSH11,respectively. There were low concentration of 8.2kb dsRNA remained in HTC8,as well as low concentration of 8.2kb and 2.5kb dsRNA remained in FL01. However,no dsRNA remained in PR15 and SSH11. The hyphal growth rate and laccase activity of the virus-eliminated strains increased,especially HTC8 and PR15,whose hyphal growth rate was higher by 22.73% and 18.18%,and laccase activities higher by 145.83% and 134.38% than that of the original strain,respectively. The conclusion is that hyphal tips cut and protoplast regeneration are suitable to prepare virus-eliminated strains of edible fungi.