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。
Identification accuracy of traditional Chinese medicine is crucial for the traditional Chinese medicine research, produc- tion and application. DNA barcoding based on the mitochondria] gene coding for eytoehrome e oxidase subunit I （ CO I ）, are more and more used for identification of traditional Chinese medicine. Using universal barcoding primers to sequence, we discussed the feasi- bility 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 intraspeeies genetic dis- tance of Trimeresurus albolabris, Ptyas dhumnades and Lycodon rufozonatus was greater than 2%. Further phylogenetic relationship re- sults 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 intraspeeific 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 med- icine.
出处
《中国中药杂志》
CAS
CSCD
北大核心
2015年第5期868-874,共7页
China Journal of Chinese Materia Medica
基金
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)
关键词
COⅠ
遗传距离
系统分类
乌梢蛇
CO I
genetic distance
phylogenetic systematics
Ptyas dhumnades
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
黄璐琦，E-mail：huangluqi01@126．com
李力，E-mail：liboshil963@vip.163．com林葵，E-mail：linkui8910@163．com
