核农学报2026，Vol.40Issue(10)：1970-1979,10.DOI:10.11869/j.issn.1000-8551.2026.10.1970
葡萄灰霉病菌LFD-RPA检测方法的建立与评价
Establishment and Evaluation of LFD-RPA Detection Method for Botrytis cinerea
摘要
Abstract
Gray mold,caused by infection with Botrytis cinerea,is one of the major diseases in grape cultivation.This study developed a rapid and precise detection approach employing recombinase polymerase amplification in conjunction with a lateral flow dipstick(LFD-RPA),based on the Bc-hch gene sequence of B.cinerea(GenBank accession no.OQ411027.1)Specific primers(FRA-F/FRA-R)and a probe(LF-P)were designed,and reaction conditions were optimized.The method's sensitivity and accuracy were validated against quantitative real-time PCR(qRT-PCR)and conventional PCR.Results showed that FRA-F/FRA-R and LF-P exhibited high specificity.The LFD-RPA assay detected B.cinerea within 30 minutes at 37℃,producing visible bands on the dipstick.Its detection limit was 10-1ng·μL-1,similar to conventional PCR but slightly lower than qRT-PCR.Field sample testing showed 100％consistency between LFD-RPA,conventional PCR,and qRT-PCR,domastrating its accuracy.All three methods detected B.cinerea during the latent infection stage,as confirmed by tests at various post-inoculation time points.The LFD-RPA assay provides a reliable and rapid tool for monitoring gray mold in grapevines.关键词
葡萄灰霉病菌/LFD-RPA/qRT-PCR/潜育期检测/田间快速检测Key words
Botrytis cinerea/LFD-RPA/qRT-PCR/latent infection detection/rapid field detection分类
农业科技引用本文复制引用
袁麒,张强强,马思远,王兴哲,辛颖,顾沛雯..葡萄灰霉病菌LFD-RPA检测方法的建立与评价[J].核农学报,2026,40(10):1970-1979,10.基金项目
宁夏回族自治区重点研发计划项目"酿酒葡萄病虫害监测预警关键技术研究与产业化应用"(2024BBF02006),宁夏回族自治区重大科技成果转化项目"酿酒葡萄病虫害信息化监测预警技术应用"(2023CJE09038) （2024BBF02006）
