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为探讨冬虫夏草寄主蝠蛾种属间的系统进化关系,采用CTAB法分别从18个冬虫夏草居群的虫草虫体头部和2种寄主蝠蛾中提取基因组DNA,PCR扩增获得冬虫夏草寄主线粒体Cytb基因片段序列.结合GenBank中的蝙蝠蛾科24种冬虫夏草寄主的同源序列,构建NJ系统树.结果表明,双栉蝠蛾属(Bipectilus)先于蝠蛾属(Hepialus)和类蝠蛾属(Hepialiscus)分化;蝠蛾属的冬虫夏草寄主种类众多形态各异,且均有特定的地理分布,蝠蛾属的种间遗传分化也较明显,蝠蛾属可能是多系起源;Cytb基因序列在冬虫夏草寄主蝠蛾种属间存在较丰富的变异,在蝠蛾属中除草地蝠蛾和金沙蝠蛾的该段序列完全一致外,其他种类蝠蛾种间变异率为0.23%—9.24%,Cytb基因序列可用于冬虫夏草寄主昆虫种的鉴定,但有必要获取更多种类的寄主昆虫来进一步验证.通过对冬虫夏草虫体头部提取的DNA进行PCR扩增获得其寄主蝠蛾Cytb基因序列的方法准确有效,有助于进一步获取更多的基因序列对更多的冬虫夏草居群和寄主种类进行系统发育、居群遗传结构及菌虫相互关系等研究.
In order to investigate the phylogenetic relationship among the host species of C. sinensis, the CTAB method was used to extract the genomic DNA from the head of Cordyceps militaris and the two kinds of host moths from 18 populations of Cordyceps sinensis respectively. The Cytb gene of the host Cordyceps sinensis was amplified by PCR Fragment sequence.The NJ phylogenetic tree was constructed based on the homologous sequences of 24 species of Cordyceps sinensis host in the GenBank.The results showed that the species Bipectilus preceded Hepialus and Hepialiscus ). The host species of C. sinensis had many morphological variations, and all had special geographical distribution. The genetic differentiation of the two species was also obvious, and the genus Cytb was probably multi-lineage. The Cytb gene sequence was highly expressed in the host of Cordyceps sinensis There were abundant variation among the moths. The sequences of the moths were different from those of the other two moths. The variation rate of Cytb gene was 0.23% -9.24% Can be used for the identification of Cordyceps sinensis host insects, but it is necessary to obtain more kinds of host insects for further verification by Cordyceps sinensis head of the DNA extracted by PCR amplification of its host bat moth Cytb Due to the sequence of accurate and effective method to help further more gene sequences of more Cordyceps sinensis populations and host species phylogenetic study population genetic structure of bacteria and insects mutual relations.