中药材北沙参种子DNA条形码鉴定研究

Identification of the Seeds of Glehniae Radix with DNA Barcoding Method

目的:本研究旨在建立鉴定北沙参种子的新方法,确保药材生产种植中种质基原准确。方法:对北沙参不同产地种子经初步形态特征鉴定后,取单粒种子为一份样品,提取基因组DNA、PCR扩增、双向测序拼接获得ITS2序列,结合序列比对、变异位点分析,基于中药材DNA条形码鉴定系统(http://www.tcmbarcode.cn),系统地完成种子样品的分子鉴定。结果:研究表明66份种子样品中,64份与《中国药典中药材DNA条形码标准序列》中北沙参序列特征一致,2份样品序列中各有一处SNP位点,为北沙参暂未发现过的遗传多样性信息,鉴定结果显示为北沙参。结论:DNA条形码技术能准确鉴定北沙参种子,并为中药材种子鉴定提供新方法,同时SNP位点的发现为后续研究北沙参品种多样性问题提供参考。

In this study,a new technique was adopted to discriminate and ensure the pure sources of the seeds of Glehniae Radix.The morphological characteristics of the seeds from different habitats had been first surveyed.Then the DNA extraction,PCR amplification,DNA sequencing and assembly were carried out.Species identification for each sample was implemented through the DNA barcoding system of traditional Chinese medicine（http：//www.tcmbarcode.cn）.The results showed that sixty-four sequences were genuinely based on ＂Standard DNA Barcodes of Chinese Materia Medica in Chinese Pharmacopoeia＂,and two samples contained SNP loci.The identification results referred to Glehniae Radix of all sequences according to the DNA barcoding system.In conclusion,DNA barcoding technique was suitable for the identification of the seeds of Glehniae Radix,which provided a new method for identification of the seeds of Chinese herbal medicines.The finding of SNP loci also furnished with a reference for studies on the variety diversity of Glehniae Radix.