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

G J McKay, L R Cooke

Research output: Contribution to journalArticle

42 Citations (Scopus)

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.
Original languageEnglish
Pages (from-to)371-8
Number of pages8
JournalFEMS Microbiology Letters
Volume152
Issue number2
Publication statusPublished - 15 Jul 1997

Keywords

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

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