Abstract Marssonina coronaria is a devastating plant pathogen that causes apple blotch on apple. Early diagnosis of M. coronaria is the key to control apple blotch disease effectively. In this study, we developed a loop-mediated isothermal amplification (LAMP) assay for rapid detection of M. coronaria. The ribosomal DNA internal transcribed spacer (rDNA-ITS) sequence of M. coronaria was selected as the target for primer design. The LAMP reaction was optimal at 60 °C for 70 min based on the color change and gel electrophoresis. In specificity tests, the LAMP assay was able to distinguish M. coronaria from other apple pathogenic fungi. In sensitivity tests, the detection limit of the LAMP assay was 100 fg μL−1 genomic DNA of M. coronaria. Furthermore, the LAMP assay was successfully applied to detect M. coronaria in diseased apple leaf samples from the field. Thus, our study provides a simple and efficient method for quick diagnosis of apple blotch caused by M. coronaria.

中文翻译：

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一种快速检测马氏冠状病毒引起的苹果斑病的LAMP方法的开发与应用

摘要 Marssonina coronaria 是一种破坏性的植物病原体，可导致苹果出现苹果斑病。冠状病毒分枝杆菌的早期诊断是有效防治苹果斑病的关键。在这项研究中，我们开发了一种环介导等温扩增 (LAMP) 检测方法，用于快速检测冠状病毒分枝杆菌。选择冠状病毒核糖体 DNA 内部转录间隔区 (rDNA-ITS) 序列作为引物设计的目标。根据颜色变化和凝胶电泳，LAMP 反应在 60 °C 下进行 70 分钟是最佳的。在特异性测试中，LAMP 检测能够将冠状病毒分枝杆菌与其他苹果病原真菌区分开来。在敏感性测试中，LAMP 检测的检测限为 100 fg μL−1 冠状病毒分枝杆菌的基因组 DNA。此外，LAMP 测定已成功应用于检测 M. 田间患病苹果叶样本中的冠状病毒。因此，我们的研究为快速诊断冠状病毒分枝杆菌引起的苹果斑病提供了一种简单有效的方法。