探讨DNA条形码分子标记作为华溪蟹属(Sinopotamon)淡水蟹类辅助分类工具的可行性。选取分布于鄱阳湖流域的6种华溪蟹属淡水蟹类78个样本,采用线粒体COI和16SrRNA基因片段以及ITS2核基因片段,作为分子标记,结合GenBank数据库中同属序列片段,经DNA提取、引物筛选、PCR扩增、产物纯化测序及测序数据处理和分析,在计算得到的遗传距离和所构建系统发生树进行华溪蟹DNA条形码标准基因的选择。在本实验涉及的淡水蟹类个体及类群的识别上,16SrRNA和ITS2基因片段不适宜作为DNA条形码标准基因,而COI基因片段结合GenBank部分华溪蟹属序列数据分析,可区分本次研究所涉及的75%的物种。以线粒体COI基因片段为DNA条形码标准序列,可作为华溪蟹属淡水蟹类物种鉴定的辅助分类工具。
Freshwater crab genus Sinopotamon is the largest and specific group in China mainland,and a kind of precious natural resource of freshwater crustacean.Because of the morphological similarity among different species,there has long been one of difficulties of traditional morphological classification.To study DNA barcoding molecular markers as auxiliary classifying tool for taxonomic study,this research selects 78 samples covering 6 species of freshwater crab genus Sinopotamon distributing in Poyang lake basin,with u-sing mitochondrial COI and 16s rRNA genes and ITS2 nuclear genes,to carry on the preliminary discussion for DNA barcodes of genus Sinopotamon.The result indicates,in this experiment involving freshwater crabs individuals and groups recognition,the 16s rRNA and ITS2 genes are not appropriate for DNA bar-coding standard gene,however,combing COI gene fragment with some sequence data analysis of parts of genus Sinopotamon original from GenBank,there could distinguish 75% species of this research.Above it shows that COI as DNA barcodes standard sequence,can be used as the assistant classifying tool of fresh-water crab genus Sinopotamon;DNA barcoding shows great advantage with discriminating implicit biodi-versity of genus Sinopotamon,and experimental data indicates that inside the individual group of S.fuk-ienense,there is the most likely to find some hidden taxonomic unit based on supplementary samples collec-tion and morphological measurement;further more,experimental data reflects the phenomenon that the group S.lansi diffusing along with variety river systems,prompts that the water system interlink each oth-er may help freshwater crab groups diffusion rather than absolute geographic isolation on distribution.