Development of a PCR-based assay for specific and sensitive detection of Fusarium buharicum from infected okra plant.

Autor: Paul SK; Laboratory of Plant Pathology, Faculty of Life and Environmental Sciences, Shimane University, Shimane, Japan.; Department of Agronomy, Bangladesh Agricultural University, Mymensingh, Bangladesh., Gupta DR; Laboratory of Plant Pathology, Faculty of Life and Environmental Sciences, Shimane University, Shimane, Japan.; Institute of Biotechnology and Genetic Engineering, Bangabandhu Sheikh Mujibur Rahman Agricultural University, Gazipur, Bangladesh., Ino M; Laboratory of Plant Pathology, Faculty of Life and Environmental Sciences, Shimane University, Shimane, Japan., Ueno M; Laboratory of Plant Pathology, Faculty of Life and Environmental Sciences, Shimane University, Shimane, Japan.
Jazyk: angličtina
Zdroj: PloS one [PLoS One] 2024 Apr 16; Vol. 19 (4), pp. e0302256. Date of Electronic Publication: 2024 Apr 16 (Print Publication: 2024).
DOI: 10.1371/journal.pone.0302256
Abstrakt: Fusarium wilt, caused by the fungus Fusarium buharicum, is an emerging disease of okra in Japan. The disease was first reported in Japan in 2015, causing significant damage to okra seedlings. Due to the potential threat in okra cultivation, the development of an accurate detection method for F. buharicum is needed for the surveillance and management of the disease. In this study, we designed a primer set and developed conventional and nested PCR assays for the specific detection of F. buharicum in infected okra plants and contaminated soil, respectively. We compared the diversity of the translation elongation factor 1 alpha (EF-1α) gene of F. buharicum with 103 other fungal species/isolates to design a species-specific primer. This primer pair successfully amplified approximately 400 bp of PCR product that was only detected in the F. buharicum isolate, not in the other fungal isolates. The developed nested PCR method was highly sensitive and could detect the fungus from a 0.01 fg DNA sample. The primer successfully detected the pathogen in artificially infected plants and soil by conventional and nested PCR, respectively. This is the first report of the development of the F. buharicum-specific primer set and detection assays, which can be used for the specific and sensitive detection of F. buharicum in field samples and for taking early control measures.
Competing Interests: The authors declare that they have no competing interests.
(Copyright: © 2024 Paul et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)
Databáze: MEDLINE
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