Establishment of the recombinase polymerase amplification-lateral flow dipstick (RPA-LFD) detection technique for Fusarium oxysporum

• Published in: Plant disease
• Main Authors: Hu, Shuodan, Yan, Chenyi, Yu, Hong, Zhang, Yu, Zhang, Chuan-Qing
• Format: Journal Article
• Language: English
• Published: United States 01.09.2023
• Subjects: Fungi; Fruit; Crop Type; Pathogen detection; small fruits; Causal Agent; Subject Areas
• ISSN: 0191-2917
• DOI: 10.1094/PDIS-12-22-2841-RE

Fusarium oxysporum can cause crown rot, wilt, root rot, and many other important plant diseases worldwide. During the development of strawberry crown rot disease, it is transmitted from the mother plant to the strawberry seedling via the stolon with obvious characteristics of latent infection. Therefore, the rapid and timely detection of F. oxysporum can enable efficient disease management. In this study, a recombinase polymerase amplification-lateral flow dipstick (RPA-LFD) detection technique was developed for the rapid detection of F. oxysporum on strawberry targeting on the CYP51C gene, which is unique to Fusarium spp. This RPA-LFD detection technique was highly specific to F. oxysporum, other Fusarium and non-Fusarium fungi cannot be detected. The optimal reaction conditions for this detection were 39°C for 8 min. The limit of detection was 1 pg of F. oxysporum genomic DNA in a 50 µL reaction. A total of 46 strawberry plants with or without crown rot symptoms from Jiande, Changxing, and Haining in Zhejiang province were further assessed for infection of F. oxysporum using both RPA-LFD and traditional tissue isolation. The RPA-LFD test showed that 32 of the 46 strawberry plants were tested were positive, means 32 strawberry plants were infected with F. oxysporum. While F. oxysporum was isolated from 30 of the 46 tested strawberry plants These results suggest that our established RPA-LFD method can quickly, sensitively, and specifically detect F. oxysporum infection in strawberry.