DNA条形码技术在常见中药材蛇类鉴别中的应用(1)
[摘要] 中药材的准确鉴别是中药材研究、生产和应用至关重要的一步。DNA 条形码技术以线粒体细胞色素氧化酶Ⅰ亚基(COⅠ)基因序列作为标记，在中药材鉴别中的应用日益增多。研究应用DNA 条形码通用引物扩增测序，探讨DNA条形码技术在常见中药材蛇类(6科15属19种共109个样品)鉴别的可行性，采用邻接法构建该类群分子系统发育树。结果表明，该片段的G+C平均量为43.9%，低于A+T平均量(56.1%)。基于Kimura 双参数模型计算，白唇竹叶青、乌梢蛇和赤练蛇的种内平均遗传距离均大于2%。通过构建系统发育关系后进一步分析发现，白唇竹叶青一样品鉴别有误。乌梢蛇中的某些样品也可能存在鉴别有误，即原本是灰鼠蛇的可能被错误鉴定为乌梢蛇。造成赤练蛇种内遗传距离差异大的因素需进一步分析。DNA条形码用于常见中药材蛇类种间的鉴别是可行的，极大程度地弥补统形态学分类方法的缺陷，值得加大推广应用和进一步深入研究。
[关键词] COⅠ;遗传距离;系统分类;乌梢蛇
[收稿日期] 2014-08-28
, http://www.100md.com
[基金项目] 国家自然科学基金项目(31460559);广西自然科学基金项目(2014GXNSFBA118113);广西科技厅项目(桂科重12118005-2-5);南宁市科技攻关项目(20123238);广西分析测试研究中心基本业务费项目(2012ACZ07)
[通信作者] 李力，E-mail： liboshi1963@vip.163.com;林葵，E-mail： linkui8910@163.com
[作者简介] 黄勇，博士，主要从事药用动物系统分类和分子进化研究，E-mail： huangykiz@163.com
DNA barcoding and its utility in commonly-used medicinal snakes
HUANG Yong， ZHANG Yue-yun， ZHAO Cheng-jian， XU Yong-li， GU Ying-le， HUANG Wen-qi， LIN Kui， LI Li
, 百拇医药
(1. Guangxi Botanical Garden of Medicinal Plants， Nanning 530023， China;
2. Guangxi Research Center of Analysis， Nanning 530022， China)
[Abstract] Identification accuracy of traditional Chinese medicine is crucial for the traditional Chinese medicine research， production and application. DNA barcoding based on the mitochondrial gene coding for cytochrome c oxidase subunitⅠ(COⅠ)， are more and more used for identification of traditional Chinese medicine. Using universal barcoding primers to sequence， we discussed the feasibility of DNA barcoding method for identification commonly-used medicinal snakes (a total of 109 samples belonging to 19 species 15 genera 6 families). The phylogenetic trees using Neighbor-joining were constructed. The results indicated that the mean content of G+C(46.5%) was lower than that of A+T(53.5%). As calculated by Kimera-2-parameter model， the mean intraspecies genetic distance of Trimeresurus albolabris， Ptyas dhumnades and Lycodon rufozonatus was greater than 2%. Further phylogenetic relationship results suggested that identification of one sample of T. albolabris was erroneous. The identification of some samples of P. dhumnades was also not correct， namely originally P. korros was identified as P. dhumnades. Factors influence on intraspecific genetic distance difference of L. rufozonatus need to be studied further. Therefore， DNA barcoding for identification of medicinal snakes is feasible， and greatly complements the morphological classification method. It is necessary to further study in identification of traditional Chinese medicine., 百拇医药(黄勇 张月云 赵成坚 徐永莉 谷颖乐 黄文琦 林葵 李力)
[关键词] COⅠ;遗传距离;系统分类;乌梢蛇
[收稿日期] 2014-08-28
, http://www.100md.com
[基金项目] 国家自然科学基金项目(31460559);广西自然科学基金项目(2014GXNSFBA118113);广西科技厅项目(桂科重12118005-2-5);南宁市科技攻关项目(20123238);广西分析测试研究中心基本业务费项目(2012ACZ07)
[通信作者] 李力，E-mail： liboshi1963@vip.163.com;林葵，E-mail： linkui8910@163.com
[作者简介] 黄勇，博士，主要从事药用动物系统分类和分子进化研究，E-mail： huangykiz@163.com
DNA barcoding and its utility in commonly-used medicinal snakes
HUANG Yong， ZHANG Yue-yun， ZHAO Cheng-jian， XU Yong-li， GU Ying-le， HUANG Wen-qi， LIN Kui， LI Li
, 百拇医药
(1. Guangxi Botanical Garden of Medicinal Plants， Nanning 530023， China;
2. Guangxi Research Center of Analysis， Nanning 530022， China)
[Abstract] Identification accuracy of traditional Chinese medicine is crucial for the traditional Chinese medicine research， production and application. DNA barcoding based on the mitochondrial gene coding for cytochrome c oxidase subunitⅠ(COⅠ)， are more and more used for identification of traditional Chinese medicine. Using universal barcoding primers to sequence， we discussed the feasibility of DNA barcoding method for identification commonly-used medicinal snakes (a total of 109 samples belonging to 19 species 15 genera 6 families). The phylogenetic trees using Neighbor-joining were constructed. The results indicated that the mean content of G+C(46.5%) was lower than that of A+T(53.5%). As calculated by Kimera-2-parameter model， the mean intraspecies genetic distance of Trimeresurus albolabris， Ptyas dhumnades and Lycodon rufozonatus was greater than 2%. Further phylogenetic relationship results suggested that identification of one sample of T. albolabris was erroneous. The identification of some samples of P. dhumnades was also not correct， namely originally P. korros was identified as P. dhumnades. Factors influence on intraspecific genetic distance difference of L. rufozonatus need to be studied further. Therefore， DNA barcoding for identification of medicinal snakes is feasible， and greatly complements the morphological classification method. It is necessary to further study in identification of traditional Chinese medicine., 百拇医药(黄勇 张月云 赵成坚 徐永莉 谷颖乐 黄文琦 林葵 李力)