摘要
兰科药用植物形态分类困难,此研究采用DNA条形码分子鉴定法从分子水平验证兰科药用植物的传统形态分类。以matK,psbA-trnH和ITS2序列作为DNA条形码对已进行了形态鉴定的49属135种163份兰科药用植物样品进行分子鉴定,经DNA提取,PCR扩增、双向测序及校对拼接后,将得到的序列在GenBank中进行BLAST比对,然后运用MEGA 7.0软件中的Neighbor-joining(NJ)法构建物种系统进化树。结果表明,163份样品均能成功提取DNA;matK,psbA-trnH和ITS2序列的PCR扩增效率分别为100%,100%,98.77%;共获得487条序列,其中345条序列在GenBank数据库中比对到了相应物种的序列,142条为新增序列;运用NJ法构建的兰科药用植物系统进化树中,基于matK序列所构建的物种系统进化树要优于基于psbA-trnH和ITS2序列所构建的系统进化树。matK,psbA-trnH和ITS2序列在鉴定兰科药用植物过程中互为补充,DNA条形码分子鉴定法可用于辅助兰科药用植物的分类鉴定。
In this study, DNA barcoding was used to validate the traditional morphological classification of medicinal plants of Orchidaceae. The 163 samples of 135 species belong to 49 genera which have been confirmed by morphological identification were collected. Candidate sequences, including matK, psbA-trnH and ITS2 sequences, were amplified, bidirectionally sequenced, and assembled. All the sequences were blasted to GenBank database at NCBI, then analyzed using Neighbor-joining tree method by MEGA 7. O. The results showed that the DNAs of 163 samples were successfully extracted. The amplification efficiency of matK, psbA-tmH and ITS2 sequences were 100% , 100% and 98.77% , respectively. The 487 sequences were obtained, 345 sequences of which have matched corresponding sequences in the GenBank database and 142 sequences were new sequences. The topology of NJ tree which were constructed with the mark sequences was better than the trees of psbA-trnH and ITS2 sequences. In conclusion, the matK, psbA-tmH and ITS2 sequences were complementary and suitable for identification of medicinal plants of Orchidaceae. DNA barcoding can be used as an auxiliary means for identification of medicinal plants of Orehidaceae,
出处
《中国中药杂志》
CAS
CSCD
北大核心
2017年第11期2058-2067,共10页
China Journal of Chinese Materia Medica
基金
四川省科技计划项目(2013SZ0117)
关键词
兰科
DNA条形码
药用植物
分子鉴定
Orchidaceae
DNA barcoding
medicinal plants
molecular identification
