选取秦岭山脉地区华溪蟹属(Sinopotamon)淡水蟹进行DNA条形码研究,探讨DNA条形码在华溪蟹分类中的适用性。经与模式标本比对,采集样本为光泽华溪蟹指名亚种(Sinopotamondavidi davidi)、尖叶华溪蟹(S. acutum)、长安华溪蟹(S.changanense)和陕西华溪蟹(S.shensiense);选择线粒体COI基因片段及核 ITS2序列片段作为DNA条形码,经DNA提取、PCR扩增、产物纯化测序及测序数据分析,发现 ITS2序列获取效率过低,不适宜作为DNA条形码;将实验获得62条 COI序列与 GenBank中的55条华溪蟹 COI序列共同组成数据集,构建的NJ、ML树可明确鉴别光泽华溪蟹指名亚种(S.davidi davidi )、兰氏华溪蟹(S.lansi)、长江华溪蟹指名亚种(S. yangtsekiense yangtsekiense )、凹肢华溪蟹(S.depressum)及福建华溪蟹(S.fukienense),但尖叶华溪蟹(S.acutum)与长安华溪蟹(S.changanense)、陕西华溪蟹(S.shensiense)与长江华溪蟹桐柏亚种(S.yangtsekiense tongbaiense )及陕县亚种(S.yangtsekiense shanxianense )均因遗传距离过小而无法鉴别;NJ 树显示长安华溪蟹(S. changanense)、尖叶华溪蟹(S.acutum)基本依据在秦岭两侧分布而分为两支,提示山脉对于华溪蟹分布起一定阻隔作用。
To study the application of DNA barcoding technology in Genus Sinopotaomon,we selected freshwater crabs from Qinling Mountains as the research obj ects.Compared with the type specimens, samples of Sinopotamondavidi davidi,S.acutum,S.changanense and S.shensiense were collected.Partial mitochondrial COI sequence and nuclear ITS2 sequence were used as DNA barcodes.After DNA extraction,the DNA samples were amplified,purified and sequenced.As a result,ITS2 fragments were found not appropriate for DNA barcoding standard gene.COI sequences from GenBank were combined with the experimental data to form the data set.The molecular phylogenetic trees constructed by NJ and ML method showed that S.davidi davidi,S.lansi,S.yangtsekiense yangtsekiense,S.depressum and 66S. fukienense can be identified clearly,but the relationship between S.acutum and S.changanense,S. shensiense and S.yangtsekiense remains confused.Furthermore,the clades of S.changanense and S.acutum from NJ tree reveal their geographical position,indicating that the Qinling Mountains may inhibit the freshwater crab diffusion.