Development of a TaqMan probe-based insulated isothermal PCR (TiiPCR) for the detection of Acidovorax citrulli, the bacterial pathogen of watermelon fruit blotch

in Sciences Citation Index(SCI), 科學引文索引資料庫(SCI)
標題Development of a TaqMan probe-based insulated isothermal PCR (TiiPCR) for the detection of Acidovorax citrulli, the bacterial pathogen of watermelon fruit blotch
出版類型SCI(Sciences Citation Index)
出版年度2016
AuthorsPei-Yi Wu, 吳佩宜, Lien-Chun Ho 何蓮君, Jun-Jie Chang 張俊傑, Kuo-Ching Tzeng 曾國欽, Wen-Ling Deng 鄧文玲, & Yi-Hsien Lin 林宜賢
出版日期2016 / 10
其他編號MOST 104-2311-B-020-001
中文摘要

Watermelon (Citrullus lanatus) is an important crop of the Cucurbitaceae family in fruit production worldwide. During its production, bacterial fruit blotch (BFB) caused by Acidovorax citrulli (Acidovorax avenae subsp. citrulli) is an important limiting factor on the volume and value of crops. This pathogen is known as a seed-borne pathogen, and the infested seeds can be a primary source of inoculum. Hence, a rapid and sensitive method for detecting A. citrulli on seeds would be an important tool in the management of BFB. In this study, we sought to develop a method to detect A. citrulli bacterial cells based on a TaqMan probe-based insulated isothermal PCR (TiiPCR) assay. Firstly, the specific primers and probe were designed based on a specific DNA fragment from the genome of A. citrulli. Then, PCR amplification was performed with the plasmid DNA to adjust the components of the PCR reagents, such as the concentrations of primers, magnesium chloride, and Taq DNA polymerase. Results revealed that 10 copies of plasmid DNA were detectable within the modified reagents by TiiPCR. Moreover, 10 bacterial cells in each reaction tube were detectable at a 100 % detection rate in this condition with a fluorescent signal intensification over 1.8. Based on these results, we concluded that a specific, rapid, and sensitive method based on TiiPCR had been successfully developed to detect bacterial cells of A. citrulli.

網址http://link.springer.com/article/10.1007/s10658-016-1054-z
期刊名稱European Journal of Plant Pathology
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