基于线粒体COⅡ基因和D-loop区序列对江西安南光唇鱼(Acrossocheilus)物种鉴定及系统发育位置分析
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摘要
以采自江西赣州安南的野生鱼为实验材料,分别设计引物扩增其线粒体DNA COⅡ和D-loop序列并进行序列比对分析,经扩增得到的COⅡ序列全长为691 bp,D-loop序列的全长为937～944 bp,其中COⅡ序列中A、T、G、C的平均占比分别为31. 00%、25. 80%、15. 30%和27. 90%,而D-loop序列的占比则分别为35. 60%、32. 30%、12. 00%和20. 00%,两序列都是A+T含量高于G+C,G的含量在4种碱基中最低。COⅡ序列中仅存在6个碱基位点的转换,D-loop则有多个碱基位点存在转换、颠换、缺失或插入,群体内遗传距离分别为0. 0075、0. 0050。用最大似然法构建的COⅡ和D-loop系统发育树显示赣州安南光唇鱼与半刺光唇鱼(Acrossocheilus hemispinus)亲缘性最近,序列相似性分别为98. 70%、98. 69%,遗传距离最近分别为0. 0130、0. 0260,未达到种间分化的水平(0. 05)。2个序列分析结果一致,结合外部形态特征观察,可以断定赣州安南光唇鱼与半刺光唇鱼为同一种,研究为江西赣州安南光唇鱼的资源利用提供了资料。
Using Acrossocheilus fasciatus from Annnan in Jiangxi province,China,as experimental materials,we analyzed the variability of mitochondrial cytochrome oxidase unit Ⅱ( COⅡ) gene and D-loop region,and discussed the phylogenetic positon and species identification of two kinds of A. fasciatus. The results showed that the COⅡ gene was 691 bp in length,and the D-loop gene was931-944 bp in length. Regardless of COII,or D-loop,the content of bases A + T was higher than that of bases G + C. Only 6 base sites were transformed in COⅡ sequence,while several base sites in D-loop sequence were transformed,transversed,and insertions/deletions. The genetic distances within the population were 0. 0075 and 0. 0050,respectively. The COⅡ and D-loop phylogenetic tree constructed by the maximum likelihood method showed the closest relationship between JXAnnan and Arossocheilus hemispinus. The sequence similarities were 98. 70% and 98. 69%,respectively. The genetic distances were 0. 0130 and 0. 0260,respectively,indicating that the genetic distance did not reach the level of interspecific divergence( 0. 05). The results demonstrated that JXAnnan is a geographical group of A. hemispinus. The results of this study can provide information for taxonomic study of Acrossocheilus.
Using Acrossocheilus fasciatus from Annnan in Jiangxi province,China,as experimental materials,we analyzed the variability of mitochondrial cytochrome oxidase unit Ⅱ( COⅡ) gene and D-loop region,and discussed the phylogenetic positon and species identification of two kinds of A. fasciatus. The results showed that the COⅡ gene was 691 bp in length,and the D-loop gene was931-944 bp in length. Regardless of COII,or D-loop,the content of bases A + T was higher than that of bases G + C. Only 6 base sites were transformed in COⅡ sequence,while several base sites in D-loop sequence were transformed,transversed,and insertions/deletions. The genetic distances within the population were 0. 0075 and 0. 0050,respectively. The COⅡ and D-loop phylogenetic tree constructed by the maximum likelihood method showed the closest relationship between JXAnnan and Arossocheilus hemispinus. The sequence similarities were 98. 70% and 98. 69%,respectively. The genetic distances were 0. 0130 and 0. 0260,respectively,indicating that the genetic distance did not reach the level of interspecific divergence( 0. 05). The results demonstrated that JXAnnan is a geographical group of A. hemispinus. The results of this study can provide information for taxonomic study of Acrossocheilus.
引文
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[19]伍献文.中国鲤科鱼类志[M].上海:上海科技出版社,1977:274-277.
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[21]单乡红,林人端,乐佩琦,等.中国动物志[M].北京:科学出版社. 2000:52-84.
[22]王莉,王绪祯,何舜平.基于线粒体ND4基因序列的七种条纹光唇鱼系统发育及对半刺光唇鱼分类地位的质疑[J].水生生物学报,2010,34(6):1218-1222.
[23]翁朝红,谢仰杰,肖志群,等.线粒体COⅠ和16S rRNA片段确定近江蛏和缢蛏属的分类地位[J].水生生物学报,2013,37(4):684-690.
[24]任巍,牛艳东,周毅,等.挂榜山小鲵线粒体DNA COⅠ基因及D-loop区序列研究[J].生命科学研究,2010,14(4):327-330.
[25]ZHANG P,CHEN Y Q,ZHOU H,et al. Phylogeny,evolution,and biogeography of Asiatic salamanders(HynobⅡdae)[J]. Proceedings of the National Academy of Sciences,2006,103(19):7360-7365.
[26]NEI M. Molecular evolutionary genetics[M]. New York:Columbia University Press,1987:448-512.
[2]张玉明,闫家强.光唇鱼胚胎发育的研究[J].绍兴文理学院学报,2010,30(9):44-48.
[3]袁乐洋.中国光唇鱼属鱼类分类整理[D].南昌:南昌大学,2005.
[4]姜建湖,戴海平,竺俊全,等.养殖光唇鱼(Acrossocheilus fasciatus)卵巢发育的组织学观察[J].海洋与湖沼,2013,44(2):348-354.
[5]潘娜,苗亮,李明云,等.浙江新昌光唇鱼(Acrossocheilus)COⅡ和D-loop基因克隆及系统发育分析[J].海洋与湖沼,2015,46(2):381-388.
[6]ROSEL P E,DIZON A E,HAYGOOD M G. Variability of the mitochondrial control region in populations of the harbour porpoise,Phocoena,on interoceanic and regional scales[J]. Can J Fish Aquat Sci,2011,52(6):1210-1219.
[7]肖武汉,张亚平.鱼类线粒体DNA的遗传与进化[J].水生生物学报,2000,24(4):384-391.
[8]LIU H,BECKENBACH A T. Evolution of the mitochondrial cytochrome oxidase II gene among 10 orders of insects[J]. Molecular Phylogenetics&Evolution,1992,1(1):41-52.
[9]LYDEARD C,ROE K J. The phylogenetic utility of the mitochondrial cytochrome b gene for inferring relationships among Actinopterygian fishes[M]. Salt Lake City:American Academic Press,1997:285-303.
[10]邵爱华.暗纹东方鲀(Takifugu fasciatus)线粒体DNA的分离纯化及其几个重要基因的克隆与序列分析[D].苏州:苏州大学,2005.
[11]凌去非,李思发.鲤科25种鱼类线粒体COⅡ基因序列差异及其系统进化关系[J].水产学报,2006,30(6):747-752.
[12]董徐辉,李明云,陈炯,等.日本鬼鲉(Inimicus japonicus)线粒体Cytb和COⅡ基因的克隆及形目系统发育分析[J].海洋与湖沼,2011,42(1):47-54.
[13]NIELSEN J L,ANN C,THOMAS W K. Differences in genetic diversity for mitochondrial DNA between hatchery and wild populations of Oncorhychus[J]. Canadian Journal of Fisheries and Aquatic Sciences,1994,51(51):290-297.
[14]SAMBROOK J,FFITSCH E F,MANIATIS T. Molecular cloning:a laboratory manual,2nd ed[C]. New York:Cold Spring Harbor Laboratory Press,1989.
[15]黄志坚,徐晓鹏,唐晶晶,等.鱼类线粒体DNA控制区扩增引物及其设计方法和应用[P]. 2010-8-25.
[16]THOMPSON J D,GIBSON T J,PLEWNIAK F,et al. The clustal_X windows interface:flexible strategies for multiple sequence alignment aided by quality analysis tools[J]. Nucleic Acids Research,1997,25(25):4876-4882.
[17]TAMURA K,DUDLEY J,NEI M,et al. MEGA4:molecular evolutionary genetics analysis(MEAN)software version 4. 0[J]. Molecular Biology and Evolution,2007,24(8):1596-1599.
[18]KOTTELAT M. Diagnoses of a new genus and 64 new species of fishes from Laos(Teleostei:Cyprinidae,Balitoridae,Bagridae,SyngnathiJdae,ChauhurⅡdae and Tetraodontidae)[J]. Journal of South Asian Natural History,2000,5(1):37-82.
[19]伍献文.中国鲤科鱼类志[M].上海:上海科技出版社,1977:274-277.
[20]毛节荣,徐寿山.浙江动物志:鱼类[M].杭州:浙江科学技术出版社,1991:106-107.
[21]单乡红,林人端,乐佩琦,等.中国动物志[M].北京:科学出版社. 2000:52-84.
[22]王莉,王绪祯,何舜平.基于线粒体ND4基因序列的七种条纹光唇鱼系统发育及对半刺光唇鱼分类地位的质疑[J].水生生物学报,2010,34(6):1218-1222.
[23]翁朝红,谢仰杰,肖志群,等.线粒体COⅠ和16S rRNA片段确定近江蛏和缢蛏属的分类地位[J].水生生物学报,2013,37(4):684-690.
[24]任巍,牛艳东,周毅,等.挂榜山小鲵线粒体DNA COⅠ基因及D-loop区序列研究[J].生命科学研究,2010,14(4):327-330.
[25]ZHANG P,CHEN Y Q,ZHOU H,et al. Phylogeny,evolution,and biogeography of Asiatic salamanders(HynobⅡdae)[J]. Proceedings of the National Academy of Sciences,2006,103(19):7360-7365.
[26]NEI M. Molecular evolutionary genetics[M]. New York:Columbia University Press,1987:448-512.