天然产物研究与开发 ›› 2015, Vol. 27 ›› Issue (5): 758-762.doi: 10.16333/j.1001-6880.2015.05.002

• 研究论文 • 上一篇    下一篇

滇重楼的psbA-trnH条形码分子鉴定研究

刘涛1,赵英良2,杨莹2,王玲1,李玛1,王欣林2,周博2,杨生超1*   

  1. 1 云南农业大学;2 云南中烟工业有限责任公司,昆明 650201
  • 出版日期:2015-05-30 发布日期:2015-06-05

Molecular Identification of Paris polyphylla var. yunnanensis using psbA-trnH DNA Barcoding

LIU Tao1, ZHAO Ying-liang2, YANG Ying2, WANG Ling1, LI Ma1, WANG Xin-lin2, ZHOU Bo2, YANG Sheng-chao1*   

  1. 1 Yunnan Agriculture University;2 China Tobacco Yunnan Industrial Co.,Ltd,Kunming 650201,China
  • Online:2015-05-30 Published:2015-06-05

摘要: 为探讨滇重楼药材鉴定新方法,本研究通过对重楼样品提取基因组DNA,PCR扩增叶绿体psbA-trnH序列并进行双向测序,所得序列经CodonCode Aligner拼接后,采用MEGA5.0软件进行序列比对,计算种内和种间遗传距离(K2P),构建邻接数(neighbor-joining tree)进行结果分析。研究表明,叶绿体psbA-trnH片段在滇重楼种内变异较小,平均K2P遗传距离为0.007,而滇重楼与其它重楼种间平均K2P遗传距离为0.025,滇重楼种间最大K2P遗传距离明显小于滇重楼与重楼属其它种内的最小K2P遗传距离。中药材DNA条形码鉴定系统比对和NJ树鉴定结果均表明psbA-trnH序列可区分滇重楼及其同属物种,且具有较好的稳定性和准确性,为保障临床安全用药提供了新的技术手段。

关键词: 滇重楼, DNA条形码, psbA-trnH, 分子鉴定

Abstract: To explore a new method of identifying Paris polyphylla var. yunnanensis, the genomic DNA from P.polyphylla was extracted and the chloroplast psbA-trnH sequence was amplified by PCR.After spliced by CodonCode Aligne,the intraspecific and interspecific genetic distance (K2P) and neighbor-joining (NJ) tree were built by MEGA5.0 software.The result showed that the chloroplast psbA-trnH  fragments found in intraspecific variation was small and the average K2P genetic distance was 0.007,while interspecific was 0.025.The maximum interspecific genetic distance of P.polyphylla was significantly less than the minimum K2P of intraspecific.The DNA barcoding comparison and NJ tree showed that psbA-trnH sequences can be used to distinguish P.polyphylla. The method was stable,accurate and will provide new technical means for clinical medication safety.

Key words: Paris polyphylla var. yunnanensis, DNA barcoding, psbA-trnH, molecular identification

中图分类号: 

S184