PCR-based rapid assay for discriminative detection of latent infections of rice bacterial blight.

摘要

A triplex PCR method has been developed for the race-specific detection of Xanthomonas oryzae pv. oryzae (Xoo), the bacterial blight (BB) pathogen of rice. For this, three primer sets were designed: for specific internal regions of two genes (hpaA and XorII very-short-patch-repair endonuclease) and for a genomic locus derived from an amplified fragment length polymorphism (AFLP) fragment specific for the K3 and K5 races. The sizes of the PCR products when using XOOF/XOOR, XRMF/XRMR and XAF3F/XAF3R primer pairs were 327, 427bp and 1kb, respectively, when the assay was applied to detect the pathogen in solution and lesion exudates, and as a template. Amplicons were obtained without the need for any prior processing (e.g. DNA preparation from infected leaf or bacterial cell isolation from the lesion). Furthermore, the pathogen could be quickly detected in the asymptomatic rice leaf 3days after inoculation and at a distance of 6cm from the lesion site. This PCR-based simple and rapid assay will be a useful method for the detection and identification of Xoo as well as for disease forecasting in paddy fields.