Development and Application of a New PCR Method for Detection of Blumeria graminis f. sp. tritici

J Mol Microbiol Biotechnol. 2018;28(3):137-146. doi: 10.1159/000494432. Epub 2018 Dec 6.

Abstract

We developed new PCR assays that target beta-tubulin (TUB2) and 14 alpha-demethylase (CYP51) genes and used them for the species-specific detection of Blumeria graminis f. sp. tritici (Bgt). Based on fungi DNA sequences available in the NCBI (National Center for Biotechnology Information) GenBank database we developed simplex and duplex PCR assays. The specificities of the primer sets were evaluated using environmental samples of wheat leaves collected during the 2015/2016 growing season across Poland. Primer sets LidBg17/18 and LidBg21/22 strongly amplified fragments of the expected length for all 67 tested samples. Primer specificity was confirmed using field samples of Zymoseptoria tri-tici, Puccinia triticina (syn. P. recondita f. sp. tritici), P. striiformis f. sp. tritici, and Pyrenophora tritici-repentis.

Keywords: Blumeria graminis; CYP51; Molecular identification; PCR assay; Phytopathology; TUB2.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Ascomycota / enzymology
  • Ascomycota / genetics*
  • Ascomycota / isolation & purification*
  • Basidiomycota / genetics
  • Cytochrome P450 Family 51 / genetics
  • DNA Primers
  • DNA, Fungal / genetics
  • DNA, Fungal / isolation & purification
  • Databases, Nucleic Acid
  • Genes, Fungal / genetics
  • Limit of Detection
  • Plant Diseases / microbiology
  • Plant Leaves / microbiology
  • Polymerase Chain Reaction / methods*
  • Saccharomyces cerevisiae Proteins
  • Sequence Alignment
  • Sequence Analysis, DNA / methods
  • Species Specificity
  • Triticum / microbiology
  • Tubulin / genetics

Substances

  • DNA Primers
  • DNA, Fungal
  • Saccharomyces cerevisiae Proteins
  • Tub2 protein, S cerevisiae
  • Tubulin
  • Cytochrome P450 Family 51