Development of Rapid Detection Methods for Fusarium oysporum f. sp. melonis in Melon Seeds
Journal
International Journal of Molecular Sciences
Journal Volume
25
Journal Issue
10
Start Page
5371
ISSN
1422-0067
Date Issued
2024-05-14
Author(s)
Abstract
Melon (Cucumis melo L.) is a global commercial crop that is sensitive to seed-borne wilt infections caused by Fusarium oxysporum f. sp. melonis (Fom). To address the challenge of detecting Fom contamination, we designed a probe-based real-time PCR method, TDCP2, in combination with rapid or column-based DNA extraction protocols to develop reliable molecular detection methods. Utilizing TDCP2, the detection rate reached 100% for both artificially Fom-inoculated (0.25–25%) and pod-inoculated melon seeds in conjunction with DNA samples from either the rapid or column-based extraction protocol. We performed analyses of precision, recall, and F1 scores, achieving a maximum F1 score of 1 with TDCP2, which highlights the robustness of the method. Additionally, intraday and interday assays were performed, which revealed the high reproducibility and stability of column-based DNA extraction protocols combined with TDCP2. These metrics confirm the reliability of our developed protocols, setting a foundation for future enhancements in seed pathology diagnostics and potentially broadening their applicability across various Fom infection levels. In the future, we hope that these methods will reduce food loss by improving the control and management of melon diseases.
Subjects
disease management
polymerase chain reaction (PCR)
Probe-qPCR
rapid DNA extraction method
Publisher
MDPI AG
Description
Article number: 5371
Type
journal article