斗鱼属鱼类亲缘关系的Cyt b基因序列和RAPD分析
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广东淡水鱼类种质资源库建设项目(No.2006B60101029)


Cytochrome b Complete Sequences and RAPD Analysis of the Phylogenetic Relationships among the Fishes of the Genus Macropodus
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    摘要:

    采用mtDNACytb基因序列分析和RAPD两种分子标记技术,研究了中国分布的叉尾斗鱼(Macropodusopercularis)、圆尾斗鱼(M.chinensis)和香港斗鱼(M.hongkongensis)以及越南的红鳍斗鱼(M.erythropterus)4种鱼类之间的亲缘关系。获得4种斗鱼14条Cytb基因全序列(1155bp),结合GenBank中搜索到的近缘物种同源序列进行分析。从133条随机引物中筛选到36条引物,在优化的反应条件下,10个群体96个个体共扩增出清晰稳定的条带749条,构建矩阵进行分析和聚类。基于Cytb全序列以邻接法和最小进化法构建的系统树及RAPD数据UPGMA聚类分析的结果都显示,香港斗鱼和红鳍斗鱼先聚为一分支,再与叉尾斗鱼聚类,圆尾斗鱼处于外缘。本研究结果反映了圆尾斗鱼与其他斗鱼的亲缘关系较远,种间遗传距离为0.1845~0.2253(Cytb)和0.6536~0.7465(RAPD),两者为同一单系群中两个独立演化的自然类群;香港斗鱼与叉尾斗鱼间遗传分化明显,Cytb碱基差异为11.00%,RAPD遗传距离达0.5777,支持其为独立物种的观点,且香港斗鱼群体间遗传差异较大,Cytb碱基差异为3.12%,RAPD遗传距离0.0601;叉尾斗鱼群体间Nei's基因多样度和Shannon信息指数分别为0.0582和0.0869,而各群体内的数值分别为0.0161~0.0317和0.0235~0.0467,表明遗传差异主要来自群体间,并按分布流域分别聚类。

    Abstract:

    In this study,mitochondrial cytochrome b(Cyt b) gene sequencing and random amplified polymorphic DNA(RAPD) technique were applied to study the phylogenetic relationships of four species of genus Macropodus: M.opercularis,M.chinensis,M.hongkongensis and M.erythropterus.The complete sequences of the mitochondrial Cyt b gene(1 155 bp) of 4 species from 14 individuals were sequenced.Combined these sequences with homologous sequences from GenBank Database,M.opercularis and M.concolor were used as ingroup;Pseudosphromenus cupanus,P.dayi,Malpulutta kretseri and Batta splendens were used as outgroup.Moreover,36 random primers were selected from 133 primers,and 749 reproducible bands were amplified in 96 individuals from 10 populations under optimized reaction conditions.Molecular phylogenetic trees based on Cyt b gene sequences were constructed by neighbor-joining(NJ) and minimum evolution(ME) methods,respectively.The results indicated that M.hongkongensis was grouped with M.erythropterus firstly,then clustered with M.opercularis,and M.chinensis was in the outer edge.The UPGMA cluster analysis based on RAPD data was identical to the molecular phylogenetic trees based on Cyt b.The results reflect that: M.chinensis has a distant relationship with other fishes of genus Macropodus;genetic distances between species' were 0.184 5-0.225 3(Cyt b) and 0.653 6-0.746 5(RAPD);M.chinensis and other Macropodus represent two independently evolving natural groups of the same monophyletic group.Genetic differentiation among M.hongkongensis and M.opercularis was obvious,Cyt b sequence divergence between species' was 11.00%,genetic distance between species' was 0.577 7(RAPD),which agrees with the view that M.hongkongensis is a valid species.The genetic diversity between different M.hongkongensis populations was significant,Cyt b sequence divergence was 3.12%,genetic distance was 0.060 1(RAPD).Nei's gene diversity(HE) and Shannon's Information index(I) between M.opercularis populations were 0.058 2 and 0.086 9,and among populations were 0.016 1-0.031 7 and 0.023 5-0.046 7,respectively,which show that the genetic diversity of M.opercularis mainly between populations too,and the individuals cluster respectively according to their distribution of river basin.

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王培欣,罗建仁,白俊杰,叶星,胡隐昌,汪学杰.2009.斗鱼属鱼类亲缘关系的Cyt b基因序列和RAPD分析.动物学杂志,44(5):14-23.

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  • 收稿日期:2009-02-09
  • 最后修改日期:2009-04-22
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