PCR-RFLP Analysis of Amplified 28S Ribosomal DNA for Identification of Rhizoctonia spp., the Causal Agents of Sheath Diseases of Rice Plants

Abstract

Isolates of five Rhizoctonia spp., the causal agents of sheath disease of rice plants, were characterized by the restriction fragment length polymorphisms (RFLPs) analysis of a 28S ribosomal DNA (rDNA) gene amplified by the polymerase chain reaction (PCR). PCR-amplified products specific to AG 1-IA and AG 2-2 of R. solani, WAG-O of R. oryzae, AG-Ba of R. fumigata and AG-Bb of R. oryzae-sativae were selected and used for RFLP analysis. RFLP profiles obtained after digestion of 28S rDNA with the restriction enzymes were different depending on the four enzymes, HpaII, HhaI, Sau3AI and HaeIII, used. The RFLP profiles by the digestion with HpaII seemed to be useful for the benchmarks to discriminate Rhizoctonia spp.