Department of Plant Pathology, Zhejiang Agriculture and Forest University, Hangzhou 311300, China.
Research Institute for the Agriculture Science of Hangzhou, Hangzhou 310013, China.
Plant Dis. 2023 Sep;107(9):2665-2672. doi: 10.1094/PDIS-12-22-2841-RE. Epub 2023 Sep 12.
causes crown rot, wilt, root rot, and many other major plant diseases worldwide. During the progression of strawberry crown rot disease, the pathogen is transmitted from the mother plant to the seedling through the stolon, with obvious characteristics of latent infection. Therefore, rapid and timely detection of . is important for efficient disease management. In this study, a recombinase polymerase amplification-lateral flow dipstick (RPA-LFD) detection technique was developed for the rapid detection of . on strawberry plants by targeting the gene, which is unique to spp. Because this RPA-LFD detection technique was highly specific to . , other and non fungi were not detected. The optimal reaction temperature and time for this technique were 39°C and 8 min, respectively. The detection limit was 1 pg of . genomic DNA in a 50-μl reaction system. A total of 46 strawberry plants with or without crown rot symptoms collected from Jiande, Changxing, and Haining in Zhejiang Province were further assessed for infection using both RPA-LFD and traditional tissue isolation techniques. The RPA-LFD test showed that 32 of the 46 strawberry plants tested were positive for , while in the traditional isolation technique, was isolated from 30 of the 46 strawberry plants. These results suggest that our established RPA-LFD method is rapid, sensitive, and highly specific in detecting . infection in strawberry plants.
引起草莓冠腐病、枯萎病、根腐病等多种重大植物病害。在草莓冠腐病的发展过程中，病原体通过匍匐茎从母株传播到幼苗，具有明显的潜伏感染特征。因此，快速、及时地检测. 对于有效的疾病管理非常重要。本研究利用针对. spp. 特有的基因，开发了重组酶聚合酶扩增-侧流层析试纸条（RPA-LFD）检测技术，用于快速检测草莓植株上的. 。由于该 RPA-LFD 检测技术对. 具有高度特异性，其他. 和非真菌均未被检测到。该技术的最佳反应温度和时间分别为 39°C 和 8 分钟。在 50μl 反应体系中，该技术的检测限为 1pg 的. 基因组 DNA。共检测了来自浙江建德、长兴和海宁的 46 株具有或不具有冠腐症状的草莓植株，使用 RPA-LFD 和传统组织分离技术对. 感染进行了进一步评估。RPA-LFD 检测结果显示，46 株草莓植株中有 32 株为. 阳性，而在传统分离技术中，从 46 株草莓植株中有 30 株分离出了. 。这些结果表明，我们建立的 RPA-LFD 方法在检测草莓植株中的. 感染方面具有快速、敏感和高度特异性。
