当前位置: X-MOL 学术Can. J. Plant Pathol. › 论文详情
Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)
Real-time and visual detection of Cercospora canescens based on the cytb gene using loop-mediated isothermal amplification (LAMP)
Canadian Journal of Plant Pathology ( IF 2 ) Pub Date : 2020-12-09
Zhen Liu, Xinchun Zhang, Di Chen, Jumei Hou, Tong Liu

Abstract

Cercospora canescens Ellis & Martin is an important fungal pathogen that causes leaf spot and defoliation in mungbean (Vigna radiata (Linn.) Wilczek.). Molecular methods for detection of this pathogen have not yet been developed. In this study, we developed a real-time and visual loop-mediated isothermal amplification (LAMP) assay based on the cytochrome b (cytb) gene to detect C. canescens in mungbean. No cross-reactions were observed in specificity tests conducted with 10 other plant pathogenic fungal species. The minimum detectable concentration was 100 pg/μl C. canescens genomic DNA, and the amplified product could be visualized with the addition of SYBR Green I fluorescent dye, which could make the assay more convenient for deployment in the field. The LAMP method presented provides a specific, rapid and visual diagnostic tool for the identification of C. canescens, and has the potential to be standardized as a detection method for this pathogen in the field.



中文翻译:

使用环介导的等温扩增(LAMP)基于cytb基因实时可视化检测Cescospora canescens

摘要

Cercospora canescens Ellis&Martin是一种重要的真菌病原体,会引起绿豆(Vigna radiata(Linn。)Wilczek。)的叶斑和落叶。尚未开发出检测这种病原体的分子方法。在这项研究中,我们开发了基于细胞色素b(cytb)基因的实时可视环介导的等温扩增(LAMP)分析法,以检测绿豆中的C. canescens。在与其他10种植物病原性真菌进行的特异性测试中未观察到交叉反应。最低可检测浓度为100 pg /μlC . canescens基因组DNA,并且可以通过添加SYBR Green I荧光染料来可视化扩增的产物,这可以使该测定法更便于在现场进行部署。提出的LAMP方法提供了一种特定的,快速的,可视化的诊断工具,可用于识别C. canescens,并有可能被标准化为该病原体的检测方法。

更新日期:2020-12-09
down
wechat
bug