环介导等温扩增检测技术快速检测向日葵茎基溃疡病菌

Loop-Mediated Isothermal Amplification Assay for Rapid Detection of , the Causal Agent of Sunflower Black Stem.

机构信息

College of Plant Protection, Shenyang Agricultural University, Shenyang 110866, China.

Liaoning Key Laboratory of Plant Pathology, Shenyang Agricultural University, Shenyang 110866, China.

出版信息

Plant Dis. 2022 Jan;106(1):260-265. doi: 10.1094/PDIS-07-21-1409-RE. Epub 2022 Jan 21.

DOI:10.1094/PDIS-07-21-1409-RE

PMID:34402633

Abstract

, the causal agent of sunflower black stem, severely affects sunflower yield and quality. A rapid and sensitive detection method is necessary for diagnosis of this disease. In this study, a loop-mediated isothermal amplification (LAMP) assay was developed for rapid detection of the pathogen from diseased sunflower tissues. The LAMP primers were designed to target the rDNA region of the fungus. The reaction condition was optimized to 60°C water baths for 45 min. The detection limit of the LAMP assay was 100 fg DNA or 10 conidia/g seeds. The LAMP assay was validated by detecting from infected sunflower tissue samples, including leaves, stems, and seeds, and applying to seed samples randomly collected from sunflower fields. This LAMP assay will be useful for estimating disease prevalence and implementing sustainable management of sunflower black stem.

摘要

向日葵黑茎病菌是一种严重影响向日葵产量和品质的病原菌。因此，有必要建立一种快速、灵敏的检测方法用于该病害的诊断。本研究针对病原菌的 rDNA 区域设计了环介导等温扩增（LAMP）检测引物，建立了检测该病原菌的 LAMP 方法。最佳反应条件为 60°C 恒温水浴 45 min。该方法的检测下限为 100 fg 基因组 DNA 或 10 个分生孢子/g 种子。应用该方法对来自叶片、茎和种子的感病组织样品进行检测，并对向日葵田间采集的种子样品进行检测，结果均为阳性，表明该方法具有良好的特异性。因此，该 LAMP 方法可用于估计病害的流行程度，并实施向日葵黑茎病的可持续管理。