收稿日期: 2024-04-19
修回日期: 2024-07-24
网络出版日期: 2024-10-30
基金资助
国家科技基础条件平台国家寄生虫资源库项目(NPRC-2019-194-30)
Phylogenetic and sequence analysis of mitochondrial nad1 of Metorchis orientalis strains from different geographic regions
Received date: 2024-04-19
Revised date: 2024-07-24
Online published: 2024-10-30
Supported by
National Parasitic Resources Center, the Ministry of Science and Technology Fund(NPRC-2019-194-30)
为研究不同地区东方次睾吸虫线粒体烟酰胺腺嘌呤二核苷酸脱氢酶亚单位Ⅰ基因（nad1）的部分序列（pnad1）遗传差异及种系发育情况，取福建、江西麦穗鱼中分离出的东方次睾吸虫囊蚴分别感染鹌鹑30 d后，收集成虫，各取3条成虫分别抽提线粒体基因组DNA，PCR扩增pnad1基因并测序。采用BioEdit软件将获得的序列与GenBank中来自黑龙江的东方次睾吸虫（GenBank：KY232061、KY232066、KY232071）和同属相关吸虫线粒体pnad1序列进行比对，分析其序列的种内差异和种间差异。采用MEGA 11.0软件计算不同地区序列遗传距离，用邻接法构建基于线粒体pnad1序列的后睾科吸虫系统进化树。结果显示，来自福建、江西的东方次睾吸虫样品均扩增出500 bp条带，测序结果为523 bp线粒体pnad1基因。序列比对分析结果显示，pnad1序列种内遗传变异率为0~0.8%，种间遗传变异率为 12.6%~13.9%。系统进化树结果显示，来自福建、江西与黑龙江株东方次睾吸虫聚于同一分支上，再与胆囊次睾吸虫（GenBank：KT740986）和黄体次睾吸虫（GenBank：KT40991）聚为一个分支。提示线粒体pnad1序列种内相对保守，种间差异较大，可作为种间遗传变异研究的标记。
关键词： 东方次睾吸虫; 线粒体DNA; 线粒体烟酰胺腺嘌呤二核苷酸脱氢酶亚单位Ⅰ基因; 序列分析; 种系发育
陈云虹 , 谢贤良 , 李燕榕 , 陈宝建 , 林耀莹 , 余文武 , 余海峰 . 不同地域株东方次睾吸虫线粒体nad1基因序列分析及种系发育研究[J]. 中国寄生虫学与寄生虫病杂志, 2024 , 42(5) : 668 -671 . DOI: 10.12140/j.issn.1000-7423.2024.05.017
To examine the genetic variation and phylogenetic relationship of nicotinamide adenine dinucleotide dehydrogenase subunits 1 (pnad1) of Metorchis orientalis from different geographic regions. The quail were infected with M. orientalis metacercaria isolated from Pseudorasbora parvain in Fujian Province and Jiangxi Province, respectively. Adult worms were collected after 30 days, and the mitochondrial genomic DNA was extracted from 3 adults in each region. The pnad1 was amplified from M. orientalis by PCR and sequenced. The obtained sequences were compared with the mitochondrial pnad1 sequences from Heilongjiang (GenBank: KY232061, KY232066, KY232071) and published sequences of the same genus using BioEdit software and the species-specific and inter-species differences were analyzed. The phylogenetic distance of different regions was calculated by MEGA 11.0 software, and the phylogenetic tree of opisthorchidae based on mitochondrial pnad1 sequence was constructed by the adjacency method. The results showed that 500-bp bands were amplified in the mitochondria of the M. orientalis from Fujian and Jiangxi, and the mitochondrial pnad1 gene was 523 bp. The results of sequence comparison showed that the intraspecific genetic variation rate of pnad1 sequence was 0-0.8%, and the interspecific genetic variation rate was 12.6%-13.9%. The constracted phylogenetic tree based on the pnad1 gene sequences showed that M. orientalis isolates from Fujian, Jiangxi and Heilongjiang were clustered together, M. xanthosomus (GenBank: KT740991) and M. bilis (GenBank: NC079698) were clustered in a branch. It was concluded that the mitochondrial pnad1 sequence is relatively conserved within species and varies greatly between species, which can be used as a marker for studying interspecific genetic variation.
Key words： Meotrchis orientalis; Mitochondrial DNA; nad1 gene; Sequence analyses; Phylogeny
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