基于ITS2和SNP技术鉴定浙江铁皮石斛的初步研究

铁皮石斛以营养价值和保健功效备受消费者青睐,但不同品种和不同品质铁皮石斛的区分和鉴别存在困难,急需建立一种快捷、准确、高效的鉴定方法。以浙江省12个铁皮石斛和其他省的10个石斛为材料,选取DNA条形码ITS2,优化扩增体系,对扩增产物进行测序,经序列比对与分析,获取浙江铁皮石斛DNA条形码ITS2中的特异性SNP位点;针对位点设计特异性引物进行了PCR验证,同时建立了基于高分辨率熔解曲线(HRM)技术对SNP位点的快速鉴别方法。研究结果表明,DNA条形码ITS2能较好地区分铁皮石斛与其他石斛,而铁皮石斛的ITS2序列中存在SNP位点,基于SNP的快速检测实现对铁皮石斛的快速鉴别,为铁皮石斛的鉴别和质量控制提供了新的理念和技术路径。

Preliminary study on identification of Dendrobium officinale from Zhejiang based on ITS2 and SNP technology

Dendrobium officinale is favored by consumers for its nutritional value and health benefits. However, it is difficult to distinguish and identify different varieties and different qualities of D. officinale. It is urgent to establish a fast, accurate and efficient identification method. In this study, 12 D. officinale from Zhejiang Province and 10 other Dendrobium were used, the DNA barcode ITS2 was selected, the amplification system for ITS2 was optimized, then the amplified products were sequenced. After sequence comparison and analysis, the specific SNP sites were obtained from DNA barcode ITS2 of D. officinale; specific primers designed for the SNP sites were verified by PCR, and a rapid identification method for SNP sites based on high-resolution melting curve (HRM) technology was established. The results showed that DNA barcode ITS2 could distinguish D. officinale from other Dendrobium. The rapid detection based on SNP realized the rapid identification of D. officinale, which provided a new concept and technical path for identification and quality control of D. officinale.