云南、河南两省日本三角涡虫基于mtDNA COI基因片段的分子系统学研究
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摘要
本文利用PCR等技术对河南、云南两省二十一产地淡水多目日本三角涡虫线粒体DNA细胞色素C氧化酶I亚基(cytochrome C oxidase subunit I ,COI )基因进行PCR扩增、克隆和测序,通过生物信息学方法用Clustal X 1.80和Mega 4.0程序对其进行序列比对和系统树构建,利用UPGMA、ME、NJ、MP统计软件构建了支序图,并对其系统进化关系进行了初步探讨。
结果表明:UPGMA树、ME树、NJ树、MP树四种支序图显示的结果基本一致。其中,采自河南省大别山系商城县的两个种群XYshangcheng1和XYshangcheng2的COI片段序列同源性最高为一个聚类枝(II);采自云南省四个城市的LClancang1、DLnanjianwl1、YXchengjiang和DLnanjianwl2为一个大的聚类枝(I);而采自河南省信阳固始的XYgushi和新乡卫辉的XXweihui却与云南省保山的BSbanqiao和丽江的LJyulongxueshan为一个聚类枝(III);LClancang2、BSbanqiao2和XYxinxian为一个聚类枝(Ⅳ)。但聚枝的先后顺序在使用不同构树方法时有差异,聚类枝(I)和(II)都与地理分布相一致,而聚类枝(III)中的XXweihui和聚类枝(Ⅳ)中的XYxinxian与云南省其他种群聚为一枝。
结果提示:二十一产地日本三角涡虫大部分在支序图中的位置与其形态学资料反映出的系统演化关系一致,说明用线粒体DNA细胞色素C氧化酶亚基I基因可以构建一个相对准确的分子树,由比对序列获得的物种之间的遗传距离也基本可以反映种、属、科间的不同变异水平。河南省两产地涡虫的细胞色素C氧化酶亚基I基因在支序图中的位置与其形态学资料反映出的系统演化关系不完全一致,其基因序列的特殊性有待进一步研究。
In this paper, We had amplified ,cloned and sequenced freshwater planarians mitochondrial DNA (mtDNA) portional cytochrome C oxidase subunit I (COI) gene through PCR and other techniques ,which freshwater planarians Dugesia japonica from 21 different places. The sequences were aligned with Clustal X1.80 and the NJ tree was reconstructed with MEGA4.0. ,we eventually constructed phylogenetic tree utilizing UPGMA、ME、NJ、MP and ML statistical software and discussed their phylogenetic evolution relationship.
Sequences analysis results show that the five phylogenetic trees: UPGMA、ME、NJ、MP and ML trees are identical. the homology of XYshangcheng1 and XYshangcheng2 was 100%, the highest,share a branch(II).Freshwater planarians coming form LClancang1、DLnanjianwl1、YXchengjiang and DLnanjianwl2 share a branch(I);Freshwater planarians coming form XYgushi and XXweihui but with BSbanqiao and LJyulongxueshan share a branch(III); LClancang2、BSbanqiao2 and XYxinxian share a branch(Ⅳ); However, which two together wither each other firstly is not able to ascertain.This clustering pattern I and II is identical to geographical position.but XXweihui in the branch III and XYxinxian in the branchⅣshare a branch with other clusters in Yun Nan.
The results note us that position of 21 species freshwater planarians in phylogenetic tree is identical to the evolution relationship which is reflected by morpholoyical materials ,which indicates the mitochondrial portional cytochrome oxidase C subunit I gene could be utilized to construct a relatively accurate molecular tree,and the genetic distance getting from sequence comparions between different species,also could reflect the variance level of different species, genera and families. plsition of CoI of two places freshwater planarian in Henna Province in phylogenetic tree differs from the relationship reflected by morpholoyical materials.Its gene sequence specificity awaits our further study.
结果表明:UPGMA树、ME树、NJ树、MP树四种支序图显示的结果基本一致。其中,采自河南省大别山系商城县的两个种群XYshangcheng1和XYshangcheng2的COI片段序列同源性最高为一个聚类枝(II);采自云南省四个城市的LClancang1、DLnanjianwl1、YXchengjiang和DLnanjianwl2为一个大的聚类枝(I);而采自河南省信阳固始的XYgushi和新乡卫辉的XXweihui却与云南省保山的BSbanqiao和丽江的LJyulongxueshan为一个聚类枝(III);LClancang2、BSbanqiao2和XYxinxian为一个聚类枝(Ⅳ)。但聚枝的先后顺序在使用不同构树方法时有差异,聚类枝(I)和(II)都与地理分布相一致,而聚类枝(III)中的XXweihui和聚类枝(Ⅳ)中的XYxinxian与云南省其他种群聚为一枝。
结果提示:二十一产地日本三角涡虫大部分在支序图中的位置与其形态学资料反映出的系统演化关系一致,说明用线粒体DNA细胞色素C氧化酶亚基I基因可以构建一个相对准确的分子树,由比对序列获得的物种之间的遗传距离也基本可以反映种、属、科间的不同变异水平。河南省两产地涡虫的细胞色素C氧化酶亚基I基因在支序图中的位置与其形态学资料反映出的系统演化关系不完全一致,其基因序列的特殊性有待进一步研究。
In this paper, We had amplified ,cloned and sequenced freshwater planarians mitochondrial DNA (mtDNA) portional cytochrome C oxidase subunit I (COI) gene through PCR and other techniques ,which freshwater planarians Dugesia japonica from 21 different places. The sequences were aligned with Clustal X1.80 and the NJ tree was reconstructed with MEGA4.0. ,we eventually constructed phylogenetic tree utilizing UPGMA、ME、NJ、MP and ML statistical software and discussed their phylogenetic evolution relationship.
Sequences analysis results show that the five phylogenetic trees: UPGMA、ME、NJ、MP and ML trees are identical. the homology of XYshangcheng1 and XYshangcheng2 was 100%, the highest,share a branch(II).Freshwater planarians coming form LClancang1、DLnanjianwl1、YXchengjiang and DLnanjianwl2 share a branch(I);Freshwater planarians coming form XYgushi and XXweihui but with BSbanqiao and LJyulongxueshan share a branch(III); LClancang2、BSbanqiao2 and XYxinxian share a branch(Ⅳ); However, which two together wither each other firstly is not able to ascertain.This clustering pattern I and II is identical to geographical position.but XXweihui in the branch III and XYxinxian in the branchⅣshare a branch with other clusters in Yun Nan.
The results note us that position of 21 species freshwater planarians in phylogenetic tree is identical to the evolution relationship which is reflected by morpholoyical materials ,which indicates the mitochondrial portional cytochrome oxidase C subunit I gene could be utilized to construct a relatively accurate molecular tree,and the genetic distance getting from sequence comparions between different species,also could reflect the variance level of different species, genera and families. plsition of CoI of two places freshwater planarian in Henna Province in phylogenetic tree differs from the relationship reflected by morpholoyical materials.Its gene sequence specificity awaits our further study.
引文
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陈广文,吕九全,马金友,刘德增,2001a.中国淡水三肠目涡虫已知属种及其地理分布.动物学报,47(专刊):9-12.
陈广文,陈晓虹,刘德增,2001b.中国涡虫纲分类学研究进展.水生生物学报,25(4):406-412.
陈广文,李景奎,2000.我国的淡水涡虫.生物学通报,35(7):11-13.
陈丽梅,孔晓瑜,喻子牛,于珊珊,徐晖,2005.3种蛏类线粒体16S rRNA和COI基因片段的序列比较及其系统学初步研究.海洋科学,29(8):27—32.
董云伟,牛翠娟,2004.mtDNA COI基因在轮虫分子系统重建中的意义.海洋湖沼通报,3:35-41.
杜增瑞,1949.扁虫及其生殖.教育与科学,2(6):1-3.
杜增瑞,朴相根,1959.三角涡虫在中国及朝鲜的分布.动物学杂志,3(9):416-419.
高天祥,张秀梅,吉崎悟朗,等.中华绒鳌蟹和日本绒鳌蟹线粒体12S rRNA基因序列研究[J].中国海洋大学学报,2000,30(1):43-47.
黄浙等,1956.昆明三角涡虫的分布和生殖研究.山东大学学报,2(4): 104-118.
刘辰莹,江慧,2005.珞珈山东亚三角头涡虫群体遗传多样性贫乏的RAPD证据.氨基酸和生物资源.27( 3):9一14.
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