Simultaneous Detection of Three Bacterial Seed-Borne Diseases in Rice Using Multiplex Polymerase Chain Reaction

Abstract

<i>Burkholderia glumae</i> (bacterial grain rot), <i>Xanthomonas oryzae</i> pv. <i>oryzae</i> (bacterial leaf blight), and <i>Acidovorax avenae</i> subsp. <i>avenae</i> (bacterial brown stripe) are major seedborne pathogens of rice. Based on the 16S and 23S rDNA sequences for <i>A. avenae</i> subsp. <i>avenae</i> and <i>B. glumae</i>, and transposase A gene sequence for <i>X. oryzae</i> pv. <i>oryzae</i>, three sets of primers had been designed to produce 402 bp for <i>B. glumae</i>, 490 bp for <i>X. oryzae</i>, and 290 bp for <i>A. avenae</i> subsp. <i>avenae</i> with the 63°C as an optimum annealing temperature. Samples collected from naturally infected fields were detected with two bacteria, <i>B. glumae</i> and <i>A. avenae</i> subsp. <i>avenae</i> but <i>X. oryzae</i> pv. <i>oryzae</i> was not detected. This assay can be used to identify pathogens directly from infected seeds, and will be an effective tool for the identification of the three pathogens in rice plants.