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标题：Development and Evaluation of a Loop-mediated Isothermal Amplification Assay for the Rapid Detection and Identification of Pectobacterium carotovorum on Celery in the Field
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作者：Yanxia Shi ; Zhiwen Jin ; Xianglong Meng 等
期刊名称：Horticultural Plant Journal
印刷版ISSN：2468-0141
出版年度：2020
卷号：6
期号：5
页码：313-320
DOI：10.1016/j.hpj.2020.07.004
语种：English
出版社：Elsevier
摘要：AbstractPectobacterium carotovorumis the causal agent of bacterial soft rot in a wide range of vegetable host species. OnceP. carotovoruminfects the plant, the spread of the disease is difficult to control. In this study, a rapid and sensitive method based on loop-mediated isothermal amplification (LAMP) was developed for detectingP. carotovorumin celery with soft rot using a primer set designed from thepmrAconserved sequence ofP. carotovorum. The specificity of the LAMP primer set forP. carotovorumwas extensively validated on bothP. carotovorumstrains and non-target strains. The sensitivity was 1 pg ofP. carotovorumgenomic DNA, which demonstrated 10 times more sensitive than the conventional PCR assay. LAMP was also used to detectP. carotovorumin bacterial suspension. The lowest detection concentration was 104CFU · mL−1. In addition, a LAMP assay, in conjunction with a crude DNA extraction method, was successfully performed onP. carotovorum-infected samples derived from both artificially and naturally infected plants. In summary, the LAMP assay established in this study constitutes a simple, sensitive, and rapid method for the detection ofP. carotovorum, and has potential application in the control of celery soft rot disease through early detection.
关键词：KeywordsBacterial soft rotP. carotovorumLoop-mediated isothermal amplification (LAMP)celerypmrAgenefield detection