目的 基于 DNA 条形码及荧光定量 PCR（qPCR）检查法建立防风药材混淆品水防风的快速检查方法。 方法 建立基于 ITS2 序列防风、水防风 DNA 条形码，寻找防风和水防风的特异性位点，以邻接法构建防风及 其伪品水防风的系统发育树；根据水防风特异性位点设计特异性引物（SFF、SFR），建立水防风荧光定量 PCR 方法；考察水防风 qPCR 检查法的掺伪检出限及灵敏度，并对市场收集的多批防风及其混淆品水防风样品进行 快速检测。结果 防风及其伪品水防风 DNA 条形码系统发育树中 31 批防风样品与 6 批水防风样品呈现不同分 支。qPCR 结果显示，水防风 Ct 值明显小于防风，其中水防风检出限为 0.655 ng·mL-1 ；在防风 DNA 中掺杂 1％水防风 DNA 时仍可检出。37 批药材样品中，6 批 Ct 值均＜26 为水防风，30 批 Ct 值均＞33 为防风，1 批 Ct 值为 30.68，为防风掺水防风，qPCR 检查结果与 DNA 条形码、电泳检测结果一致。结论 本研究建立的防 风混淆品水防风 qPCR 检测方法准确、灵敏度高，操作简便快捷，可应用于防风混淆品水防风的快速鉴别。

Objective To establish a rapid detection method for the adulterant Libonotis laticalycina Shan et shen in the medicinal material Saposhnikovia divaricata（Trucz.） Schischk based on DNA barcoding and quantitative PCR （qPCR）. Methods DNA barcoding was established for Libonotis laticalycina and Saposhnikovia divaricata based on the ITS2 sequence. Specific nucleotide sites distinguishing the two species were identified， and a phylogenetic tree of Saposhnikovia divaricata and its adulterant Libonotis laticalycina was constructed using the neighbor-joining method. Specific primers （SFF and SFR） were designed based on the unique sites of Libonotis laticalycina to establish a qPCR method for its detection. The detection limit and sensitivity of the qPCR method for detecting adulteration were investigated. This method was then applied for rapid detection of multiple batches of Saposhnikovia divaricata and its adulterant Libonotis laticalycina collected from the market. Results In the DNA barcoding phylogenetic tree，31 batches of Saposhnikovia divaricata samples and 6 batches of Libonotis laticalycina samples formed distinct branches. qPCR results showed that the Ct value for Libonotis laticalycina was significantly lower than that for Saposhnikovia divaricata. The detection limit for Libonotis laticalycina was 0.655 ng·mL-1 ，and the method could detect as low as 1% Libonotis laticalycina DNA mixed with Saposhnikovia divaricata DNA. Among 37 batches of samples，6 batches with Ct values all < 26 were identified as Libonotis laticalycina， 30 batches with Ct values all > 33 were identified as genuine Saposhnikovia divaricata，and 1 batch with a Ct value of 30.68 was identified as Saposhnikovia divaricata adulterated with Libonotis laticalycina. These qPCR detection results were consistent with those from DNA barcoding and electrophoresis. Conclusion The established qPCR detection method for the adulterant Libonotis laticalycina in Saposhnikovia divaricata is accurate，highly sensitive，simple and rapid to operate，making it suitable for the detection of Libonotis laticalycina in Saposhnikovia divaricata samples.

R284.1

广东省基础与应用基础研究基金项目（2024A1515011249）。