Abstract
The Fusarium genus causes devastating plant diseases worldwide, in which Fusarium oxysporum is the most serious crop pathogen. Disease monitoring is the basis of integrated pest management of any disease. The lack of rapid, accurate, and reliable device to detect and identify plant pathogens is one of the main limitations in integrated disease management. This study describes an efficient and quantifiable diagnosis method for the specific detection of F. oxysporum f. sp. cubense (Foc) race 4 in field-infected banana. With the optimized PCR parameters using the SCAR (sequence characterized amplified region) primers FocSc-1/FocSc-2 and a real-time PCR strategy, the developed method showed high reproducibility and was very sensitive to detect extremely low quantities of Foc genomic DNA (gDNA). We also found that Foc gDNA in severely symptomatic banana pseudostems and leaves were 6946-fold and 26.69-fold more than in those of mild-symptomatic banana, respectively.
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Acknowledgements
We are grateful to Drs H.-L. Wang (National Kaohsiung Normal University), Y.-S. Lin (National Chung Hsing University, NCHU), W.-H. Hsieh (NCHU), C.-H. Kuo (National Chiayi University), Taitung District Agricultural Research and Extension Station, Agricultural Research Institute and AVRDC-The World Vegetable Center for providing the tested microorganisms. We also thank Miss Y.-L. Wan and Miss. Y.-R. Chen for technical assistance. This research was supported in part by Council of Agriculture, Taiwan, R.O.C. under grant numbers 93AS-1.9.2-BQ-B1(1), 94AS-13.3.2-BQ-B1(6), 96AS-4.1.2-IC-I1(2), 97AS-4.1.2-ICI1(6), 98AS-4.1.1-IC-I1(1), and 99-AS-9.3.1-BQ-B2(6); by National Science Council, Taiwan, R.O.C. under grant numbers 98-2313-B-005-025-MY3, 99-2622-B-005-006-CC2, and 101-2313-B-005-028-MY3; by the Ministry of Education, Taiwan, R.O.C. under the ATU plan; and also by National Chung Hsing University, Taiwan, R.O.C. Dr. Ying-Hong Lin was supported with the postdoctoral fellowships provided by National Science Council, Taiwan, R.O.C. under grant numbers 098-2811-B-005-024, 099-2811-B-005-025, 100-2811-B-005-017, and 101-2811-B-005-011.
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Lin, YH., Su, CC., Chao, CP. et al. A molecular diagnosis method using real-time PCR for quantification and detection of Fusarium oxysporum f. sp. cubense race 4. Eur J Plant Pathol 135, 395–405 (2013). https://doi.org/10.1007/s10658-012-0096-0
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DOI: https://doi.org/10.1007/s10658-012-0096-0