Development of a TaqMan probe-based insulated isothermal PCR (TiiPCR) in seed detection of watermelon fruit blotch
Wu Pei-Yi: National Pingtung University of Science and Technology
<div>Watermelon is an important crop of the Cucurbitaceae family in fruit production worldwide. During its production, <em>Acidovorax citrulli</em> is the causal agent of bacterial fruit blotch (BFB). BFB is an important international quarantine disease with zero tolerance and the infested seeds can be a primary source of inoculum in the field. Hence, a rapid and sensitive method for detecting <em>A. citrulli</em> contaminated seeds would play an important role in management of BFB. In this study, we sought to develop a method to detect contaminated seeds with <em>A. citrulli </em>based on a TaqMan probe-based insulated isothermal PCR (TiiPCR). First, specific primers and probes were designed based on the DNA fragment of <em>A. citrulli</em> genome. In our reacting conditions, results revealed the detection rate could reach 100% under 10 cells/tube. In addition, the minimal fluorescent signal intensification in this condition was over 1.8. Furthermore, detection carried out with 10% or 2% contaminated seeds still revealed 100% detection rate by TiiPCR. Therefore, we demonstrated that a rapid and sensitive detection method of BFB on seeds were established based on TiiPCR in this study.</div>
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