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运用ITS2 DNA条形码技术鉴定中药材黄花紫堇、多刺绿绒蒿及其近缘物种,以保证药材质量,确保用药安全。文中对采自西藏拉萨的黄花紫堇、多刺绿绒蒿等13个药材样品进行了DNA提取、PCR扩增和测序;对实验序列和GenBank下载的71条切除两端的5.8S,28S区域,使用MEGA 5.0进行多重序列比对,并进行种内、种间遗传距离计算和进化树构建;同时预测了黄花紫堇、多刺绿绒蒿与其近缘种ITS2序列的二级结构。结果表明,作为DNA条形码,ITS2序列能够有效地对黄花紫堇、多刺绿绒蒿及其同属近缘物种进行分子鉴定。该研究不仅可为黄花紫堇、多刺绿绒蒿及其近缘物种、相关混伪品建立基于ITS2序列的DNA条形码的规范和安全的检测技术及鉴别方法,也为其质量控制、安全用药及合理开发利用提供理论依据及数据支撑。
ITS2 DNA barcoding using Chinese herbal medicine Cinnabarin, Prickly heat and its related species, in order to ensure the quality of medicinal herbs to ensure drug safety. In this paper, 13 medicinal herbs such as Corydalis, Prickly-Hope (Artemisia selengensis) collected from Lhasa, Tibet, were collected for DNA extraction, PCR amplification and sequencing. The experimental sequences and 5.8S, 28S regions of 71 excised ends downloaded from GenBank, Multiple sequence alignment (MEGA 5.0) was used to calculate intraspecific and interspecific genetic distance and phylogenetic tree construction. At the same time, the secondary structure of ITS2 sequence of Corydalis avenae, Prorocentrum amurense and its related species was predicted. The results showed that the ITS2 sequence could be used as a DNA barcode for molecular identification of Cineraria, Prickly-Burmese and their related species. This study not only established the standard and safety detection methods and identification methods of DNA barcodes based on ITS2 sequences for Corydalis pratensis, Prickly-Burmese and its related species, and related congealed products, but also for quality control, safe medication And provide reasonable theoretical basis for the development and utilization of data and support.