蒿属外来药用资源中亚苦蒿的特异性PCR鉴别

Specific PCR Identification of Artemisia absinthium,A New Foreign Medicinal Resource of Artemisia

目的:基于特异性聚合酶链式反应(PCR)建立了一种中亚苦蒿药材的鉴别方法,可以准确、便捷地鉴别中亚苦蒿及其近缘种。方法:利用Chloroplast Genome Information Resource(CGIR)数据库查找中亚苦蒿及其近源种叶绿体基因组序列,并筛选获得中亚苦蒿特异性单核苷酸多态性(SNP)位点,根据SNP位点设计一对中亚苦蒿特异性鉴别引物zykh1-F和zykh1-R。采集中亚苦蒿及其常见近缘种的原植物样本,建立中亚苦蒿特异性PCR鉴别方法并优化反应体系,对该方法进行耐受性和适用性考察。利用该方法对从新疆药材市场购买中亚苦蒿药材样本进行鉴别。结果:中亚苦蒿特异性PCR鉴别方法采用鉴别引物zykh1-F和zykh1-R,退火温度为54℃、循环次数为33次。经PCR扩增和凝胶电泳后中亚苦蒿在210 bp处可观察到单一明亮条带,其余近缘种如艾、黄花蒿、白叶蒿、野艾蒿均无条带。结论:该研究所建立的中亚苦蒿特异性PCR鉴别方法可准确鉴别中亚苦蒿及其常见近缘种,具有高度特异性,且该方法节约时间和成本,为中亚苦蒿资源引种和利用提供一种方便快捷的物种鉴别方法。

Objective:To establish a specific polymerase chain reaction(PCR)method for the identification of Artemisia absinthium to allow accurate and convenient identification of A.absinthium and its related species.Method:The chloroplast genome sequences of A.absinthium and its related species were searched from Chloroplast Genome Information Resource(CGIR),and the specific single nucleotide polymorphism(SNP)sites of A.absinthium were screened out.A pair of specific identification primers(zykh1-F and zykh1-R)of A.absinthium was designed.The original plant samples of A.absinthium and its related species were collected.The specific PCR method was established and optimized,and the tolerance and feasibility of this method were investigated and verified.The method was used to identify A.absinthium samples purchased from Xinjiang medicinal materials market.Result:A 210 bp bright band was obtained from A.absinthium after PCR amplification and gel electrophoresis under the following conditions:specific primers zykh1-F and zykh1-R,annealing temperature of 54℃,and the number of cycles of 33.No such band was observed from its relative species,such as A.argyi,A.annua,A.leucophylla,and A.lavandulaefolia.Conclusion:The specific PCR identification method of established in this study can accurately identify A.absinthium and its common related species with high specificity.The method can save time and cost and allows a convenient and fast species identification for the introduction and utilization of A.absinthium resources.