农业生物技术学报2026，Vol.34Issue(2)：420-430,11.DOI:10.3969/j.issn.1674-7968.2026.02.015
甘薯丛枝病植原体TaqMan探针qPCR检测方法的建立与应用
Establishment and Application of TaqMan Probe qPCR Method for Detecting Sweet Potato Witches'-broom Phytoplasma Disease
摘要
Abstract
Phytoplasma-induced sweet potato infection is a significant disease that impacts sweet potato(Ipomoea batatas)production in China,particularly in southeastern coastal regions.This study developed specific primers and TaqMan probes based on the secY sequence of the sweet potato witches'-broom phytoplasma transporter protein gene for sensitive and rapid detection of the sweet potato witches'-broom phytoplasma and its presence in propagation media.A qPCR detection method was established using TaqMan,and was applied to test collected sweet potato samples,insect vectors,and various sweet potato tissues infected with phytoplasma.The results demonstrated that this TaqMan qPCR detection method could detect sweet potato witches'-broom phytoplasma sensitively,rapidly,and accurately,with a detection limit of 1.71×10¹ copies/μL.It could also specifically distinguish between Melia azedarach witches'-broom phytoplasma(16SrⅠ-B),Areca catechu yellowing phytoplasma(16SrⅠ-B),Catharanthus roseus periwinkle little leaf phytoplasma(16SrⅠ-A),the causal agent of sweet potato bacterial wilt(Ralstonia solanacearum),and black rot pathogen of sweet potato(Dickeya dadantii).This method was used to quantitatively detect the pathogen content in different tissues of sweet potatoes infected with phytoplasma,revealing a pathogen content range of 3.98×10⁴～6.03×105 copies/μL,and identifying the sweet potato stems and the stem base as having the highest phytoplasma content.The insect vectors in the field were also quantitatively tested using this detection method,confirming Orosius orientalis as the transmission medium of sweet potato witches'-broom phytoplasma.In conclusion,the qPCR detection method developed in this study demonstrated good sensitivity,specificity,and repeatability.It not only enables rapid detection of sweet potato witches'-broom phytoplasma,but also provides a reference for grading the disease severity of sweet potato witches'-broom disease according to pathogen quantity.关键词
甘薯/甘薯丛枝病/植原体/TaqMan探针实时荧光定量PCRKey words
Sweet potato/Sweet potato witches'-broom/Phytoplasma/TaqMan probe real-time fluorescence quantitative PCR分类
农业科技引用本文复制引用
李华伟,许泳清,李国良,张鸿,崔纪超,林赵淼,邱永祥,汤浩,邱思鑫..甘薯丛枝病植原体TaqMan探针qPCR检测方法的建立与应用[J].农业生物技术学报,2026,34(2):420-430,11.基金项目
国家重点研发计划(2024YFD1401200) （2024YFD1401200）
福建省属公益类科研院所基本科研专项(2025R10290010) （2025R10290010）
国家现代农业产业技术体系(CARS-10-B14) （CARS-10-B14）
福建省省政府5511协同创新工程(XTCXGC2021005) （XTCXGC2021005）
福建省农业科学院自由探索科技创新项目(ZYTS202407) （ZYTS202407）
