Abstract:Seedborne bacteria, including Pseudomonas viridiflava (Pv), P. syringae pv. lachrymans (Psl), P. syringae pv. syringae (Pss), Acidovorax citrulli (Ac), Xanthomonas cucurbitae (Xc) and X. melonis (Xm), are important diseases in melon crops production. By using three sets of multiplex PCR and a chromogenic macroarray system, it was allowed us to detect multiple target bacterial species in bacterial suspension, naturally infected or artificially infected seed samples simultaneously without the use of agarose gel electrophoresis. The average qualitative detection limit could be reach N×103~N×104 CFU/mL with pure bacteria culture. Target bacteria could be detected from artificial infected watermelon(Citrullus vulgaris), melon(Euphorbia meloformis) and cucumber(Cucumis sativus) seeds and naturally infected A. citrulli watermelon seeds. The new method is efficient, high-throughput and practical, and gets rid of the dependence of fluorescence scanner. This study provides a method for the simultaneous detection of plant seedborne pathogens.