摘要
为快速有效检测烟草靶斑病菌,以烟草靶斑病菌beta-tubulin基因序列保守区域为靶标位点,设计重组酶介导扩增技术(Recombinase aided amplification,RAA)引物,结合侧流层析试纸条(Lateral flow dipstick,LFD),建立烟草靶斑病菌快速检测体系并进行优化。结果表明,LFD-RAA检测体系在39℃恒温条件下反应30 min即可完成RAA扩增,并具有较高的特异性,其检测极限值为100 fg/μL,与PCR检测灵敏度相当。对检测体系优化确定检测扩增的最优时间为20~35 min、扩增温度为31~39℃。LFD-RAA检测体系可应用于烟草上烟草靶斑病菌的快速检测。
To rapidly and effectively detect the tobacco target spot pathogen,the conserved region of the beta-tubulin gene sequence of the pathogen was targeted and the recombinase aided amplification(RAA)primers were designed.Lateral flow chromatographic strips were used to establish and further optimize a rapid detection system.The results showed that the developed RAA amplification could be completed within 30 minutes at a constant temperature of 39°C and had high specificity with a detection limit of 100 fg/μL.Its sensitivity was comparable to that of conventional PCR detections.The optimal amplification time and temperature of the optimized detection system were 20-35 min and 31-39℃.This LFD-RAA detection system could be applied for rapid detection of the pathogen of tobacco target spot on tobacco plants.
作者
肖艳松
李思军
吴文信
刘天波
周向平
周陆苏
钟杰
XIAO Yansong;LI Sijun;WU Wenxin;LIU Tianbo;ZHOU Xiangping;ZHOU Lusu;ZHONG Jie(Chenzhou Branch of Hunan Provincial Tobacco Company,Chenzhou 423000,Hunan,China;Tobacco Science Institute of Hunan Province,Changsha 410004,China;Yongzhou Branch of Hunan Provincial Tobacco Company,Yongzhou 425000,Hunan,China;College of Plant Protection,Hunan Agricultural University,Changsha 410128,China)
出处
《烟草科技》
CAS
CSCD
北大核心
2024年第5期41-47,共7页
Tobacco Science & Technology
基金
湖南省郴州市烟草公司烟草农业科技创新项目“郴州烟区烟草靶斑病和野火病病原微生物组解析及早期流行诊断”(CZYC2022JS03)
“郴州雪茄烟霉变机制与绿色防控技术研究”(CZYC2023JS05)
中国烟草总公司湖南省公司科技重点项目“湖南烟草靶斑病成灾机制与绿色防控技术研究”(HN2021KJ01)。
关键词
烟草靶斑病
重组酶介导扩增技术
检测
Tobacco target spot
Recombinase aided amplification
Detection
