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浙江新昌光唇鱼(Acrossocheilus)COⅡ和D-loop基因克隆及系统发育分析*
潘娜1, 苗亮2, 李明云2, 郭晓飞2, 赵亮2, 陈炯2, 张玉明3, 吕益龙3
1.宁波大学 应用海洋生物技术教育部重点实验室;2.宁波大学,应用海洋生物技术教育部重点实验室;3.浙江新昌水利水电局
摘要:
采用扩增COⅡ、D-loop序列、进行序列比对和系统树分析并结合形态特征观察的方法,研究了浙江新昌光唇鱼(Acrossocheilus)的系统分类和种类鉴定。形态特征结果表明,新昌光唇鱼具有光唇鱼属鱼类的基本特征;扩增得到COⅡ序列全长691bp,D-loop序列全长931bp-944bp, A T含量两基因序列都高于G C(A T的含量分别为57.30%-57.80%和65.30%-65.90%),G的含量在四种碱基中最低;COⅡ序列有6个碱基位点存在转换,而D-loop则存在多个碱基的转换、颠换、缺失或插入位点。浙江新昌光唇鱼群体内COⅡ、D-loop基因的平均遗传距离分别为0.003、0.005;基于COⅡ和D-loop序列构建的NJ法系统树均显示新昌光唇鱼与温州光唇鱼(Acrossocheilus wenchowensis)形成一个紧密的簇,序列相似性分别为99.94%和99.36%,遗传距离分别为0.000、0.011,未达到种间分化水平(遗传距离大于0.05),表明新昌光唇鱼和温州光唇鱼(A. wenchowensis)为同一种,且根据形态特征可以断定为二个不同的地理群体,而D-loop序列因进化速度较快更适于光唇鱼属种内及近缘物种种间的亲缘关系的研究。研究结果既有助于新昌光唇鱼资源的开发利用,又可为研究光唇鱼类的分类提供参考资料。
关键词:  光唇鱼  COⅡ基因  D-loop基因  序列分析  系统发育分析
DOI:10.11693/hyhz20140900235
分类号:
基金项目:国家星火计划项目(2011GA701001);浙江省科技厅重大科技专项遗传育种项目(2012C12907-8)
ClONING MITOCHONDRIAL COⅡ and D-loop AND ANALYSIS THE PHYLOGENTIC POSITION OF A FISH IN ACROSSOCHEILUS FROM XINCHANG IN ZHEJIANG PROVINCE
Pan Na,Miao Liang,Li Mingyun,Guo Ciaofei,Zhao Liang,Chen Jiong,Zhang Yuming,LV Yilong
1.Key Laboratory of Applied Marine Biotechnology, Ministry of Education, Ningbo University, Ningbo, Zhejiang 315211;2.The Water Conservancy Bureau of Xinchang County, Xinchang 312500
Abstract:
In order to study the taxonomy and species identification of a Acrossocheilus fish (denoted by ZJxinchang) caputred from Xinchang County of Zhejiang Province, we amplified and analysis mitochondrial COⅡ and D-loop gene, constructed phylogenetic trees, and combined with morphological observation. Morphological features’ finding that ZJxinchang was similar to Acrossocheilus in external morphology. The entire COⅡand D-loop sequences of ZJxinchang were 691bp and 931bp-944bp, respectively. The content of A T was higher than G C in both COⅡ and D-loop. The proportion of A T were 57.30%-57.80% and 65.30%-65.90%, and the content of G was the lowest in the four bases. There were only six transition sites in COⅡ gene, and there were several variation sites (transition, transversion, deletion or insertion) in D-loop. The average genetic distances of COⅡ and D-loop genes among ZJxinchang group were respectively 0.003 and 0.005. Phylogenetic tree (NJ method) based on COⅡ and D-loop both showed that ZJxinchang and Acrossocheilus wenchowensis formed a tight cluster, the sequence similarity between ZJxinchang and A. wenchowensis were 99.94% and 99.36% for COII and D-loop, respectively. The genetic distance of COII and D-loop ZJxinchang and A. wenchowensis were 0.000, 0.011, respectively, both were below species" divergence (0.05). The results demonstrated that ZJxinchang is a geographical group of A. wenchowensis, the morphological variation between ZJxinchang and the type species of A. wenchowensis may due to diferent living envrionment. D-loop gene evolved faster than COⅡ, it"s more suitable for exploring the intra- and inter-specific comparability of genus Acrossocheilus. The resluts of this study is useful to the development and utilization of Xingchang A. wenchowensis, and can provide information for taxonomic study of Acrossocheilus.
Key words:  Acrossocheilus  COⅡ  D-loop  sequence analysis  phylogenetic analysis
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