黑龙江省不同地区东方次睾吸虫线粒体cox1和nad1基因序列分析及种系发育研究

Phylogenetic analysis and sequence analysis of mitochondrial cox1 and nad1 for Metorchis orientalis isolates from different regions in Heilongjiang province

为了阐明黑龙江省不同地区的东方次睾吸虫线粒体细胞色素C氧化酶第I亚基(cox1)和烟酰胺腺嘌呤二核苷酸脱氢酶亚单位I(nad1)的遗传变异及种系发育情况,本研究利用PCR方法扩增黑龙江省3个地区的东方次睾吸虫线粒体cox1和nad1部分基因,对其序列分析并构建种系发生树,探讨其遗传进化关系。结果显示pcox1和pnad1基因片段长度分别为699 bp和492 bp,且基因长度均一致。pcox1基因种内变异率在0~0.7%,种间变异率在12.2%~15.8%;pnad1基因种内变异率在0~0.2%,种间变异率在12.7%~23.3%,种间变异要大于种内变异。分子进化树结果显示,3个地区的东方次睾吸虫聚集在一起,次睾属的另两个成员聚集在同一分支,且系统发生树中的Bootstrap值均较高。本研究表明线粒体cox1基因和nad1基因可以用于东方次睾吸虫的种系发育分析,本研究为东方次睾吸虫进一步的分子流行病学和数量遗传学研究奠定了实验基础。

In order to clarify the genetic variation and phylogenetic relationship of cytochrome c oxidase subunit 1 （coxl） and NADH dehydrogenase subunits 1 （nadl） of Metorchis orientalis in different geographical regions of Heilongiiang province, a part of the coxl （pcoxl） and nadl （pnadl） were amplified separately from individual specimen by PCR and sequenced to determine sequence variations and examine their phylogenetic relationships. The results showed that the length of pcoxl and pnadl were 699 bp and 492 bp, respectively. Herein, the intra-specific sequence variations were 0-0.7% for pcoxl, 0-0.2% for pnadl. In contrast, the inter-specific sequence differences among members of the Opisthorchiidae were significantly higher, being 12.2%-15.8% for pcoxl, 12.7%-23.3% for pnadl. The phylogenetic relationship results indicated that M.orientalis isolates from three different regions in Heilongjiang province were cluster together, M.xanthosomus and M.bilis were clustered in a branch, and the phylogenetic tree showed a high Bootstrap values. These findings had important implications for studying systematics, molecular epidemiology and population genetics of M.orientalis.