Journal of Taiwan Agricultural Research

Author:Chia-Hsin Tsai、Ya-Ting Liu、Ssu-Yu Lin、Ching-Shan Tseng、 Cheng-Ping Kuan*

Abstract:

Bacterial fruit blotch (BFB), caused by Paracidovorax citrulli, threatens cucurbit production
worldwide and often results in severe economic losses. In this study, we developed a detection assay
for sensitive detection of P. citrulli. The assays employ polymerase chain reaction (PCR) with specific
co-amplification of a plant gene as an internal control from total nucleic acids, enabling parallel
detection and minimizing the risk of false-negative PCR results during routine testing and eliminating
the need to remove contaminating plant DNA from extracts. The assay exhibited high specificity, as
no cross-reactions were observed with non-target bacterial species or with nucleic acid extracted from
healthy cucurbit tissues. Sensitivity testing demonstrated reliable detection of low concentrations
of pathogen DNA, and the method was validated using artificially inoculated seedlings and field
samples. Compared with conventional PCR, the multiplex detection assay provides higher throughput
and quantitative readouts based on mean fluorescence intensity. This platform offers a reliable tool for
seed health testing, quarantine surveillance, and integrated management of BFB in cucurbit crops.

Key words:Paracidovorax citrulli, Bacterial fruit blotch, Cucurbits, Diagnosis

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