期刊
CROP PROTECTION
卷 68, 期 -, 页码 27-35出版社
ELSEVIER SCI LTD
DOI: 10.1016/j.cropro.2014.11.004
关键词
Musa acuminate; Fusarium oxysporum f. sp cubense race 1 (FOC1) and race 4 (FOC4); TaqMan probes; Single-tube duplex real-time fluorescence; PCR; Rapid quantitative detection
类别
资金
- Guangdong Entry-Exit Inspection and Quarantine Bureau [2010GDK63]
- Ministry of Science and Technology Special Fund for the Public Interest in China [201410080]
- earmarked fund for Modern Agro-Industry Technology Research System in China [CAR-32-05]
- Construction funds of State Key Laboratory of Conservation and Utilization of Subtropical Agro-bioresources from Guangdong Province Science and technology funds
Banana fusarium wilt caused by Fusarium oxysporum f. sp. cubense race 1 (FOCI) and race 4 (FOC4) is a destructive disease that affects bananas in tropical and subtropical areas worldwide. A sensitive and specific detection method is the primary step to preventing spread of the disease. In this study, a real-time fluorescence PCR method was developed based on specific conserved primers from the markers of sequence characterized amplified region (SCAR) and TaqMan probes for detecting FOCI and FOC4. The results showed that real-time fluorescence PCR could be used to detect FOCI and FOC4 accurately and effectively within 90 min (not including DNA extraction). The developed method had high specificity and could therefore be used to distinguish E oxysporum f. sp. cubense from other allied species and forma, such as Fusarium verticillioides, Fusarium oxysporum f. sp. melonis, Fusarium oxysporum f. sp. momodicae, Fusarium oxysporum f. sp. benincasae, and Fusarium oxysporum f. sp. opuntiarurn, and other plant pathogens, such as Penicillium. This detection system also demonstrated high sensitivity, yielding a total copy number of 91,258, which was 100 times higher than that of endpoint PCR. We found that FOC1 had approximately 36 times higher abundance in banana 'Guangfen #1' pseudostem than FOC4 at 14 days after infection. In addition, banana `Guangfen #1' root tissues showed an approximately 23 times higher abundance of FOCI than corm tissues in field samples. In conclusion, the developed single-tube duplex real-time PCR method can sensitively distinguish FOCI and FOC4 with high specificity. This method can be utilized to assist in the implementation of quarantine measures for the prevention and control of banana fusarium wilt caused by E oxysporum f. sp. cubense. (C) 2014 Elsevier Ltd. All rights reserved.
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