Establishment of a Rapid LAMP Detection Assay for Fusarium oxysporum f. sp. niveum
YANG Ke1,2, XIAO Ji-Ling2, ZHANG Yi2, WEI Lin3, LIANG Zhi-Huai1,2,*
1 Longping Branch, Graduate School of Hunan University, Changsha 410125;
2 Hunan Agricultural Biotechnology Research Institute, Changsha 410125;
3 Hunan Institute of Plant Protection, Changsha 410125
AbstractFusarium wilt of watermelon caused by Fusarium oxysporum f. sp. niveum (FON) is the main disease of watermelon across all major watermelon-growing regions in China. Therefore, the establishment of a rapid detection method of FON is quite important for the diagnosis, prevention and control of Fusarium wilt of watermelon. In this study, using the specific DNA sequences of FON as the target, a set of sensitive and specific loop-mediated isothermal amplification (LAMP) primers were designed, and combined with the rapid DNA extraction method, a rapid LAMP detection assay for FON was established. The results of primer specificity experiments showed that only FON-0, FON-1 and FON-2 showed yellow-green color for LAMP detection, and the agarose gel electrophoresis results of LAMP products showed trapezoidal bands, indicating that the primers had high specificity. The sensitivity detection results showed that the lower limit of detection of the LAMP assay for FON-1 genomic DNA was 10 pg/μL, and for FON-1 content in plants was 5×102 spores/g. The results of potting experiments showed that the assay could make a correct judgment on whether watermelon seedlings were infested by FON, using root tissues as samples. In this assay, from the beginning of DNA extraction to the detection results, only about 90 min was needed, and the pathogenic mycelia, the tissues of the plants could be used as detection materials. The results of LAMP reaction were discernible to the naked eye, with simple operation, strong specificity, high sensitivity, stability and wide applicability. It provides technical support for rapid molecular diagnosis of FON in the field.
YANG Ke,XIAO Ji-Ling,ZHANG Yi, et al. Establishment of a Rapid LAMP Detection Assay for Fusarium oxysporum f. sp. niveum[J]. 农业生物技术学报, 2021, 29(9): 1836-1844.
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