侵染昆虫前后冬虫夏草菌类枯草杆菌蛋白酶基因表达研究
Expression of subtilisin-like protease gene from Hirsutella sinensis before and after infecting Hepialus gonggaensis
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摘要:
通过检测类枯草杆菌蛋白酶Pr1基因的表达变化, 了解Pr1基因在冬虫夏草菌侵染贡嘎蝠蛾过程中所起的作用。根据GenBank中已登录的Pr1基因的同源序列设计引物, 利用PCR扩增的方法得到采自四川康定的冬虫夏草菌菌株Pr1基因部分序列, 结合荧光定量PCR (qRT-PCR) 方法, 检测冬虫夏草菌在侵染贡嘎蝠蛾幼虫前后Pr1基因的表达变化。PCR扩增得到Pr1基因部分片段535 bp, GenBank登录号KC009680。qRT-PCR结果显示冬虫夏草菌侵染贡嘎蝠蛾后Pr1基因表达量显著上升, 侵染后的Pr1基因表达量是侵染前的2～3倍, Pr1基因在冬虫夏草菌侵染贡嘎蝠蛾的过程中可能起重要作用。本研究为筛选高致病力的冬虫夏草菌奠定了基础。
Abstract:To address the role of Pr1 gene in the process of Hirsutella sinensis infecting Hepialus gonggaensis, differential expression of subtilisin-like protease gene was detected. In the present study, Pr1 gene analogues from H. sinensis were obtained by PCR strategy using specific primers designed from conserved regions of Pr1 gene reported in the GenBank. Then we detected the changes in the expression of Pr1 gene before and after infecting H. gonggaensis using real-time quantitative PCR. We obtained the partial sequence of Pr1 gene with the length of 535 bp (GenBank accession: KC009680). Real-time PCR results showed that the expression level of Pr1 gene was significantly different among 8 samples (P < 0.01). Pr1 gene showed the obvious higher expression level (2−3 folds) after infecting the H. gonggaensis, suggesting that the Pr1 gene may play an important role in the process of H. sinensis infecting H. gonggaensis. The present study paves a way for further identification on infectivity assessment of H. sinensis.
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