A PCR-based method to characterise and identify benzimidazole resistance in Helminthosporium solani

FEMS Microbiol Lett. 1997 Jul 15;152(2):371-8. doi: 10.1111/j.1574-6968.1997.tb10454.x.

Abstract

Control of Helminthosporium solani, the cause of silver scurf in potato tubers, has been impaired by selection of benzimidazole-resistant strains as a result of repeated use of the fungicide thiabendazole. Identification of thiabendazole-resistant strains of H. solani by conventional techniques takes several weeks. Primers designed from conserved regions of the fungal beta-tubulin gene were used to PCR amplify and sequence a portion of the gene. A point mutation was detected at codon 198 in thiabendazole-resistant isolates causing a change in the amino acid sequence from glutamic acid to alanine or glutamine. Species-specific PCR primers designed to amplify this region were used in conjunction with a restriction endonuclease to cause cleavage in sensitive isolates only and thus provide a rapid diagnostic test to differentiate field isolates.

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Benzimidazoles / pharmacology*
  • DNA Mutational Analysis
  • DNA, Fungal / genetics*
  • Drug Resistance, Microbial / genetics
  • Fungicides, Industrial / pharmacology*
  • Genes, Fungal / genetics
  • Helminthosporium / drug effects*
  • Molecular Sequence Data
  • Point Mutation / genetics
  • Polymerase Chain Reaction / methods*
  • Sensitivity and Specificity
  • Tubulin / genetics

Substances

  • Benzimidazoles
  • DNA, Fungal
  • Fungicides, Industrial
  • Tubulin
  • benzimidazole

Associated data

  • GENBANK/Y10670