Abstract: Fusarium acuminatum（FA）is one of the important pathogens causing root rot of Astragalus membranaceus var.mongholicus（AMM）.To rapidly detect and accurately quantify FA in AMM plants and soil，a quantitative real⁃time PCR（qPCR）detection method was established and its practicability in detecting the pathogen in AMM plants and soil samples was verified. The results showed that primer pair Fae F4/Fae R4 designed based on the elongation factor⁃1α gene（EF‑1α）sequence of FA was highly specific. For the FA in AMM plants and soil，the sensitivities of qPCR detection method were 2.56×10^⁃3 ng/μL of DNA concentration and 1×10² conidia/g，respectively；the correlation coefficients of the constructed standard curves were 0.999 7 and 0.983 8；the amplification efficiencies were 1.07 and 0.87.Repeatability evaluation found that the method was stable and reliable. With the established method，the pathogen FA could be detected in inoculated⁃AMM samples as early as 1 hour after inoculation，and the contents increased with time.The detection rate of FA was 100% in suspected diseased AMM samples.FA was tested positive in all soil samples and the number of conidia in diseased soil samples was significantly greater than that in healthy soil at the same cultivation years.In conclusion，the qPCR detection method established in this study could be used to rapidly and quantitatively detect FA in AMM plants and soil，providing a reliable technical support for the accurate detection and timely prevention and control of AMM root rot.

Key words: Astragalus membranaceus var.mongholicus, Root rot, Fusarium acuminatum, Quantitative Real?time PCR, Detection

摘要： 锐顶镰刀菌（Fusarium acuminatum）是黄芪根腐病的重要病原菌之一。为实现锐顶镰刀菌的快速检测和准确定量，建立了一种实时荧光定量PCR（qPCR）检测方法，并对其在黄芪植株和土壤样本检测中的实用性进行验证。结果表明，基于锐顶镰刀菌EF-1α序列设计的引物Fae F4/Fae R4特异性强，所建方法对黄芪和土壤中锐顶镰刀菌的检测灵敏度分别为DNA 质量浓度2.56×10^-3 ng/μL和分生孢子1×10²个/g，标准曲线的相关系数分别为0.999 7和0.983 8，扩增效率分别为1.07和0.87。重复性评价显示方法可靠、稳定。利用该方法检测发现，黄芪样本接种锐顶镰刀菌1 h后即可检测到病菌存在，且侵染量随时间逐渐上升；田间疑似根腐发病样本的锐顶镰刀菌阳性检出率为100%；田间发病和未发病土壤样本中，病菌阳性检出率也为100%，且相同种植年限的发病土带菌量显著高于未发病土。因此，所建qPCR检测方法可用于黄芪植株和土壤中锐顶镰刀菌的检测，为黄芪根腐病的精准监测和及时防控提供可靠的技术支撑。

关键词: 黄芪, 根腐病, 锐顶镰刀菌, 实时荧光定量PCR, 检测