Journal of Henan Agricultural Sciences ›› 2026, Vol. 55 ›› Issue (3): 107-117.DOI: 10.15933/j.cnki.1004-3268.2026.03.011

• Plant Protection • Previous Articles     Next Articles

A Quantitative Real⁃time PCR Method for Rapid Detection of Astragalus membranaceus var.mongholicus Root Rot Pathogen Fusarium acuminatum

ZU Weixi1,ZHAO Limei2,ZHAO Xuejiao2,WANG Xue2,GAO Fen3   

  1. (1.Department of Hospitality Management,Taiyuan Tourism College,Taiyuan 030032,China;2.Modern Research Center for Traditional Chinese Medicine,Shanxi University,Taiyuan 030006,China;3.Institute of Applied Chemistry,Shanxi University,Taiyuan 030006,China)
  • Received:2025-05-16 Accepted:2025-07-04 Published:2026-03-15 Online:2026-03-27

黄芪根腐病菌锐顶镰刀菌的实时荧光定量PCR快速检测方法

祖未希1,赵丽梅2,赵雪姣2,王雪2,高芬3   

  1. (1.太原旅游职业学院 酒店管理系,山西 太原 030032;2.山西大学 中医药现代研究中心,山西 太原 030006;3.山西大学 应用化学研究所,山西 太原 030006)
  • 通讯作者: 高芬,副教授,博士,主要从事药用植物病害及其生物防治研究。E-mail:gaofen@sxu.edu.cn
  • 作者简介:祖未希,副教授,硕士,主要从事食品营养与卫生、基于药食同源植物的健康产品研发等研究。E-mail:zuweixi@163.com
  • 基金资助:
    山西省基础研究计划资助项目(202103021224029);山西省回国留学人员科研资助项目(2022-023)

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

CLC Number: