SIMPLE, RAPID AND ACCURATE PCR-BASED DETECTION OF PANTOEA ANANATIS IN MAIZE, SORGHUM AND DIGITARIA sp.

Abstract

SUMMARY Detection of Pantoea ananatis in the early growing season is important for disease prediction and management. We developed a simple molecular marker based on PCR for conclusive diagnosis of P. ananatis in maize, sorghum and Digitaria sp. A pair of primers was used for amplifying only one of the seven internal transcribed spacer (ITS) regions of P. ananatis 16S-23S rRNA genes. Sixty-one strains of P. ananatis from diverse ecogeographical origins; total DNA of pool of maize white spot (MWS) lesions and MWS-like lesions from sorghum and Digitaria sp.; reference strains of P. agglomerans, P. ananatis, P. stewartii, P. allii, P. vagans, P. anthophila, P. eucalypti, P. deleyi, P. rodasii, P. rwandensis and P. wallisii were used for testing the specificity of the primers. A single amplicon per sample, 361 bp or 389 bp in size, was obtained from P. ananatis from different sources and P. allii isolated from Allium cepa and the identity of all amplicons was confirmed by DNA sequencing. The present results provide a rapid and reliable tool for the accurate identification of P. ananatis isolates and for direct PCR-based diagnosis of P. ananatis associated with maize, sorghum and Digitaria sp.