基于ITS2和psbA-trnH区域对马鞭草科药用植物进行DNA条形码研究

[DNA bacording of Verbenaceae medicinal plant by using ITS2 and psbA-trnH region].

作者信息

Chen Qian, Liu Yimei, Liu Zhen, Chen Shilin, Chen Keli

机构信息

Key Laboratory of Ministry of Education on Traditional Chinese Medicine Resource and Compound Prescription & Hubei University of Chinese Medicine, Wuhan 430065, China.

出版信息

Zhongguo Zhong Yao Za Zhi. 2012 Apr;37(8):1107-13.

PMID:22779359

Abstract

OBJECTIVE

To evaluate the ability of several hotspot candidate sequence of DNA barcodes for identifying medicinal plant species in Verbenaceae.

METHOD

Using universal primers, three chloroplast sequences, psbA-trnH, rbcL, matK, two nuclear ribosomal DNA ITS2 and ITS were amplified and sequenced. PCR amplification and sequencing efficiency, intra- and inter-specific variation, barcoding gap and identification efficiency (with BLAST 1 and Nearest Distance methods) were used to evaluate these loci.

RESULT

The rate of successful amplification using matK was too low to further analyze, and the rate of successful amplification and sequencing using psbA-trnH, ITS, and ITS2 and rbcL sequence was 83.6%, 83.6%, 96.4%, 98.2%, respectively. The rate of successful identification using psbA-trnH, ITS, and ITS2 was 100% at the species level except that rbcL was 77.8%, 75.9% for 55 samples belonging to 32 species, but ITS2 did better in intra- and inter-specific variation, barcoding gap than the other loci. The rate of successful identification of ITS2 was 89.5%, 87.6% even when joining the date of 165 samples from GenBank.

CONCLUSION

It proposes that the combination of ITS2 and psbA-trnH senquence is promising for the identification of the species in Verbenaceae.

摘要

目的

评估DNA条形码的几个热点候选序列对马鞭草科药用植物物种的鉴定能力。

方法

使用通用引物扩增并测序三个叶绿体序列psbA-trnH、rbcL、matK，两个核糖体DNA ITS2和ITS。采用PCR扩增和测序效率、种内和种间变异、条形码间隙和鉴定效率（使用BLAST 1和最近距离法）来评估这些位点。

结果

matK的成功扩增率过低无法进一步分析，psbA-trnH、ITS、ITS2和rbcL序列的成功扩增和测序率分别为83.6%、83.6%、96.4%、98.2%。对于属于32个物种的55个样本，psbA-trnH、ITS和ITS2在物种水平的成功鉴定率为100%，rbcL为77.8%、75.9%，但ITS2在种内和种间变异、条形码间隙方面比其他位点表现更好。即使加入来自GenBank的165个样本的数据，ITS2的成功鉴定率仍为89.5%、87.6%。