利用mtDNA 16S rRNA序列差异鉴定江西青岚湖的河蚌物种
THE SPECIES IDENTIFICATION OF MUSSELS IN QING LAN LAKE (JIANGXI PROVINCE)BASED ON SEQUENCE DIFFERENCES OF MITOCHONDIRIAL 16S rRNA GENE
-
摘要: 对从青岚湖采集的10属14种河蚌的19个样本进行了16S rRNA的序列测定,并同GenBank中鄱阳湖流域相同物种河蚌的序列比较,分析了基于Kimura 2-parameter 模型参数得到的遗传距离,并构建了它们的UPGMA树.结果显示,所有用于比较的河蚌种间遗传距离变化范围在0.0274-0.2290,平均为0.132,种内遗传距离在0.0034-0.0068之间,平均为0.004,种间遗传距离远大于种内距离.表明以16S rRNA作为遗传标记,可以在物种水平上对青岚湖的河蚌进行准确的鉴定.Abstract: Generally,the identification of freshwater mollusk is mainly baesd on shell differences,but shells are greatly variable under different environmental conditions and hence usually result in taxonomic confusion.To clarify these problems,many studies on morphology,ecology,anatomy and physiology have been carried out in the past years,such as the researches on marsupia,glochidium,sensory hairs and spines.However,they have not shown any conclusive evidence for definite identification. In order to identify the mussel species correctly,we employed the molecular systematic method.The mitochondrial 16S ribosomal RNA gene has been used in the pyhlogenetic studies of mussels successfully. In the present study,the 16S rRNA gene fragments were sequenced to 19 individuals of mussels,which belong to 14 species of 10 genera,and were from the Qing Lan Lake (Jiangxi province,where the water quality is excellent and the species of fishes and mussels are abundant).The target fragments of 16S rRAN were isolated by PCR amplification to the samples.Thirty five cycles of PCR(94℃,1min denatruing,57℃,30s annealing,72℃,1min extension) were performed.Some sequenees of mussels from Poyang Lake were downloaded from GenBank for comparison.Alignments were manipulated using Clustal X and refined manually where necessary.Sequence analysis was performed using MEGA (Version 2.1)software.Genetic distances were calculated based on Kimura 2 parameter model.The largest intraspecific genetic distance is 0.0068,much lower than the average interspecific distance,which has a value of 0.1325,and still much lower than the shortest interspecific distance,0.0274,suggesting that 16S rRNA is a useful genetic marker for identification of mussel species clearly.
-
Keywords:
- Qing Lan Lake /
- Mussel /
- 16S rRNA /
- Species identification
-
-
[1] Liu Y Y,Wang Y X ,Zhang W Z.Economic Fauna of China.Freshwater Mollusk[M].Beijing:Science Press,1976,[刘月英,王跃先,张文珍.中国经济动物志--淡水软体动物.北京:科学出版社,1976][2] Wei Q S ,Fu C H,Wang Y F.Fu X F,Comparative studies on morphology of glochidia of six mussel species(Mollusca:Unionidae)[J].Acta Hydrobiologica Sinica,1994,18(4):303-309.[魏青山,傅彩红,王玉凤,傅小飞.珠蚌科六种蚌的钩介幼虫形态比较研究.水生生物学报,1994.18(4):303-309][3] Wu X P.Studies on freshwater mollusca in Mid-lower reaches of Yangtze River[D].Ph.D.thesis,Institute of Hydrobiology,CAS,1998.[吴小平.长江中下游淡水贝类的研究.博士学位论文,中科院水生生研究所,1998.][4] Lydeard C,Mulvey M,Davis G M.Molecular systemmatics and evolution of reproductive traits of North American freshwater unionacean mussels(Mollusca:bivalvia)as inferred from 16S rRNA gene sequences [J].Biologial Sciences,1996,351(1347):1593-1603[5] Huang Y Y,Liu H Z,Wu X P,Ouyang S.Testing the relationships of Chinese freshwater unionidae(Bivalvia) besed on analysis of partial mitochondrial 16S rRNA sequences[J].J.Moll.Stud,2002,68:359-363[6] Lan K,Morton B.Morphological,ecological and mitochondrial DNA 16S sequence distinctions between and within Saccostrea(Bivalvia:Ostreidae)populations in Hong Kong and Australia[J].Abstracts of World Congress of Malacology,2001,184[7] Liu S, Yang G,Zhou K Y,et al.Phylogeny of river dophins based on partial DNA sequence from Mitochondrial 16S rRNA gene[J].Journal of Nanjing Normal University(natrual science),2000,23(4):74-78.[刘珊,杨光,周开亚,等.淡水豚类mtDNA 16S rRNA 基因的系统发生.南京师大学报(自然科学版)2000,23(4):74-78][8] Thompson J D,Gibson T J,Plewniak F.The Clustal-X windows interface:flexible strategies for multiple sequences alignment aided by quality analysis tools [J].Nucl., Acids Res., 1997,25:4876-4882[9] Kumar S,Tamura K,Jakobsen I B,et al.MEGA: Molecular Evolutionary Genetics Analysis,Version 2.1[M].Arizona:Arizona State University,2001[10] Seberg O,Humphries C J,Knapp S,et al.Shortcuts in systematics?A commentary on DNA-based taxonomy[J].Trends in Ecology and Evobution,2003,18(2):63-65[11] Lipscomb D,Plantnik N,Wheeler Q.The intellectual comtent of taxonomy:a comment on DNA taxonomy[J].Trends in Ecology and Evolution,2003,18(2):65-66[12] Tautz D,Arctander P,Minelli A,et al.A plea for DNA taxonomy[J].Trends in Ecology and Evolution,2003,18(2):70-74 Liu Y Y,Wang Y X ,Zhang W Z.Economic Fauna of China.Freshwater Mollusk[M].Beijing:Science Press,1976,[刘月英,王跃先,张文珍.中国经济动物志--淡水软体动物.北京:科学出版社,1976][2] Wei Q S ,Fu C H,Wang Y F.Fu X F,Comparative studies on morphology of glochidia of six mussel species(Mollusca:Unionidae)[J].Acta Hydrobiologica Sinica,1994,18(4):303-309.[魏青山,傅彩红,王玉凤,傅小飞.珠蚌科六种蚌的钩介幼虫形态比较研究.水生生物学报,1994.18(4):303-309][3] Wu X P.Studies on freshwater mollusca in Mid-lower reaches of Yangtze River[D].Ph.D.thesis,Institute of Hydrobiology,CAS,1998.[吴小平.长江中下游淡水贝类的研究.博士学位论文,中科院水生生研究所,1998.][4] Lydeard C,Mulvey M,Davis G M.Molecular systemmatics and evolution of reproductive traits of North American freshwater unionacean mussels(Mollusca:bivalvia)as inferred from 16S rRNA gene sequences [J].Biologial Sciences,1996,351(1347):1593-1603[5] Huang Y Y,Liu H Z,Wu X P,Ouyang S.Testing the relationships of Chinese freshwater unionidae(Bivalvia) besed on analysis of partial mitochondrial 16S rRNA sequences[J].J.Moll.Stud,2002,68:359-363[6] Lan K,Morton B.Morphological,ecological and mitochondrial DNA 16S sequence distinctions between and within Saccostrea(Bivalvia:Ostreidae)populations in Hong Kong and Australia[J].Abstracts of World Congress of Malacology,2001,184[7] Liu S, Yang G,Zhou K Y,et al.Phylogeny of river dophins based on partial DNA sequence from Mitochondrial 16S rRNA gene[J].Journal of Nanjing Normal University(natrual science),2000,23(4):74-78.[刘珊,杨光,周开亚,等.淡水豚类mtDNA 16S rRNA 基因的系统发生.南京师大学报(自然科学版)2000,23(4):74-78][8] Thompson J D,Gibson T J,Plewniak F.The Clustal-X windows interface:flexible strategies for multiple sequences alignment aided by quality analysis tools [J].Nucl., Acids Res., 1997,25:4876-4882[9] Kumar S,Tamura K,Jakobsen I B,et al.MEGA: Molecular Evolutionary Genetics Analysis,Version 2.1[M].Arizona:Arizona State University,2001[10] Seberg O,Humphries C J,Knapp S,et al.Shortcuts in systematics?A commentary on DNA-based taxonomy[J].Trends in Ecology and Evobution,2003,18(2):63-65[11] Lipscomb D,Plantnik N,Wheeler Q.The intellectual comtent of taxonomy:a comment on DNA taxonomy[J].Trends in Ecology and Evolution,2003,18(2):65-66[12] Tautz D,Arctander P,Minelli A,et al.A plea for DNA taxonomy[J].Trends in Ecology and Evolution,2003,18(2):70-74
计量
- 文章访问数: 898
- HTML全文浏览量: 2
- PDF下载量: 458
下载: