Development and evaluation of a loop-mediated isothermal amplification assay for rapid detection of bacterial blight pathogen (Xanthomonas axonopodis pv. dieffenbachiae) in anthurium
文献类型: 外文期刊
作者: Niu Jun-hai 1 ; Gao Yue-rong 1 ; Yin Jun-mei 1 ; Leng Qing-yun 1 ; Yang Guang-sui 1 ; Wang Cun 1 ; Ren Yu 1 ;
作者机构: 1.Trop Crops Genet Resources Inst, Minist Agr, CATAS Key Lab Crop Gene Resources & Germplasm Enh, Hainandanzhou 571737, Peoples R China
关键词: Anthurium;Bacterial blight;Xanthomonas axonopodis pv;Dieffenbachiae;Loop-mediated isothermal amplification (LAMP);Molecular diagnosis
期刊名称:EUROPEAN JOURNAL OF PLANT PATHOLOGY ( 影响因子:1.907; 五年影响因子:2.022 )
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收录情况: SCI
摘要: Anthurium bacterial blight caused by Xanthomonas axonopodis pv. dieffenbachiae (Xad) is an important and destructive disease worldwide, and no effective technique has been developed for its control. Detection of infection (latent) in anthurium plants is critical to evaluate disease progress and strengthening management to avoid a serious epidemic in the fields. In this paper, we presented a novel molecular method to detect Xad in anthurium using loop-mediated isothermal amplification (LAMP) technique (Xad-LAMP). The Xad-LAMP reaction could be finished by incubating at 61-65 A degrees C for 1 h, and the amplificons were confirmed through gel electrophoresis, HpaII enzyme analysis, and visually inspected using SYBR Green I/calcein stain and lateral flow dipstick (LFD) assay. The specificity of Xad-LAMP primers set was widely validated on Xad and nontarget strains. In sensitivity testing, Xad-LAMP allowed detection as low as 1-10 f. pure genome DNA or 10(4) CFU/ml cells, 10-100 times more sensitive than conventional PCR. In addition, combining with the optimized DNA extraction, Xad-LAMP detections were successfully performed on both latent and disease samples, derived from artificially and naturally infected plants respectively. In all, this study provided a promising and practical molecular tool for Xad detecting, will facilitate the forecasting and control of anthurium blight disease.
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