滇重楼的psbA-trnH条形码分子鉴定研究

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

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psb A-trn H序列并进行双向测序,所得序列经Codon Code 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psb A-trn H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psb A-trn H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。

To explore a new method of identifying Paris polyphylla var. yunnanensis,the genomic DNA from P. polyphylla was extracted and the chloroplast psb A-trn H sequence was amplified by PCR. After spliced by Codon Code 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 psb A-trn H fragments found in intraspecific variation was small and the average K2 P genetic distance was 0. 007,while interspecific was 0. 025. The maximum interspecific genetic distance of P.polyphylla was significantly less than the minimum K2 P of intraspecific. The DNA barcoding comparison and NJ tree showed that psb A-trn H sequences can be used to distinguish P. polyphylla. The method was stable,accurate and will provide new technical means for clinical medication safety.