Detection of a mutation at codon 43 of the rpsL gene in Xanthomonas oryzae pv. oryzicola and X. oryzae pv. oryzae by PCR-RFLP

Genet Mol Res. 2015 Dec 29;14(4):18587-95. doi: 10.4238/2015.December.28.6.

Abstract

The aim of this study was to develop a method to detect a point mutation in the ribosomal S12 protein (rpsL) gene in streptomycin-resistant strains of Xanthomonas oryzae pv. oryzicola and X. oryzae pv. oryzae. The polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method was developed to detect a point mutation in codon 43 of the rpsL gene in X. oryzae pv. oryzicola and X. oryzae pv. oryzae. The 304-bp PCR product from the rpsL gene was digested by MboII to form two fragments (201 and 103 bp) if there was a mutation at codon 43, or three fragments (146, 103, and 55 bp) if there was no mutation. Compared with the results from nucleotide sequencing, the PCR-RFLP method was accurate in detecting the point mutation at codon 43 of the rpsL gene in streptomycin-resistant strains of X. oryzae pv. oryzicola and X. oryzae pv. oryzae. These results indicate that the PCR-RFLP is a simple, rapid and reliable method for detecting the point mutation at codon 43 of the rpsL gene.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Anti-Bacterial Agents / pharmacology
  • Bacterial Proteins / genetics*
  • Base Sequence
  • Codon*
  • Drug Resistance, Bacterial
  • Mutation*
  • Polymerase Chain Reaction
  • Polymorphism, Restriction Fragment Length
  • Ribosomal Proteins / genetics*
  • Sequence Analysis, DNA
  • Xanthomonas / drug effects
  • Xanthomonas / genetics*

Substances

  • Anti-Bacterial Agents
  • Bacterial Proteins
  • Codon
  • Ribosomal Proteins