湖南和河南两省来源的褐黄血蜱遗传多样性及遗传进化关系分析

Genetic diversity and genetic evolutionary relationship analysis of Haemaphysalis flava collected from Hunan and Henan Provinces

为了探究湖南和河南两省来源的褐黄血蜱遗传多样性及遗传进化关系,试验采用PCR方法扩增来自于湖南、河南两省的20只褐黄血蜱的核糖体ITS-1及线粒体cox1基因序列,通过DNAStar、MegAlign、ClustalX、PhyML 3.0及MEGA 5.0软件分析其遗传多样性及遗传进化关系。结果表明:褐黄血蜱ITS-1和cox1基因序列的长度分别为1500 bp和850 bp,经校准、对齐后的长度分别为1302 bp和800 bp;来自于湖南省褐黄血蜱ITS-1及cox1基因序列的种内差异分别为0~1.3%、0~0.4%;来自于河南省褐黄血蜱ITS-1、cox1基因序列的种内差异分别为0.1%~1.0%、0~0.1%,两省褐黄血蜱ITS-1、cox1基因序列的种内差异分别为0~1.3%、0~0.4%,种间差异分别为50.66%~52.77%、2.5%~14.3%;ITS-1基因序列的A+T和G+C含量分别为46.1%~46.3%、53.7%~53.9%;cox1基因序列的A+T和G+C含量分别为60.6%~62.6%、37.4%~39.4%;在构建的2个遗传进化树中,来自于湖南与河南两省的褐黄血蜱均处于一个分支上,两者亲缘关系较近。说明尽管来自于两省的褐黄血蜱之间均存在遗传变异现象,但亲缘关系较近,未出现遗传分化。

In order to explore the genetic diversity and genetic evolutionary relationship of Haemaphysalis flava collected from Hunan and Henan Provinces, in the experiment, PCR was used to amplify the ribosomal ITS-1 and mitochondrial cox1 gene sequences of 20 individuals of Haemaphysalis flava collected from Hunan and Henan Provinces;DNAStar, MegAlign, ClustalX,PhyML 3.0 and MEGA 5.0 softwares were used to analyze the genetic diversity and genetic evolutionary relationship. The results showed that the lengths of the ITS-1 and cox1 gene sequences of Haemaphysalis flava were 1 500 bp and 850 bp respectively, and the calibrated and aligned lengths were 1 302 bp and 800 bp, respectively. The intra-species differences in the ITS-1 and cox1 gene sequences of Haemaphysalis flava from Hunan Province were 0-1.3% and 0-0.4%,respectively;the intra-species differences in the ITS-1 and cox1 gene sequences of Haemaphysalis flava from Henan Province were 0.1%-1.0% and 0-0.1%,respectively. The intra-species differences in the ITS-1 and cox1 gene sequences of Haemaphysalis flava between the two provinces were 0-1.3% and 0-0.4%,respectively;the inter-species differences were 50.66%-52.77% and 2.5%-14.3%,respectively. The A+T and G+C contents of ITS-1 gene sequences were 46.1%-46.3% and 53.7%-53.9%,respectively;the A+T and G+C contents of cox1 gene sequences were 60.6%-62.6% and 37.4%-39.4%,respectively. In the two phylogenetic trees constructed, the Haemaphysalis flava isolates from Hunan and Henan Provinces were on the same branch, which were closely related. The results suggested that although there were genetic variations between Haemaphysalis flava individuals from the two provinces, the genetic relationship was relatively close and there was no genetic differentiation.