Strain typing of Zygosaccharomyces yeast species using a single molecular method based on polymorphism of the intergenic spacer region (IGS)

Abstract

Unlike previously reported methods that need a combination of several typing techniques, we have 22 developed a single method for strain typing of the Zygosaccharomyces bailii, Z. mellis and Z. rouxii spoilage 23 species. Strains belonging to other species have also been included for comparison. We have demonstrated 24 that the IGS-PCR RFLP method has a high discriminative power. Considering the three endonucleases used in 25 this work, we have obtained a variability of 100% for Z. mellis and Z. rouxii strains and up to 70% for Z.bailii. 26 We have also detected two misidentified Z. mellis strains (CBS 711 and CBS 7412) which have RFLP patterns 27 with a set of bands characteristic of Z. rouxii strains. Sequencing of 26S rDNA D1/D2 domains and the 5.8-ITS 28 rDNA region confirmed these strains as Z. rouxii. The method also groups three certified hybrid strains of 29 Zygosaccharomyces in a separate cluster.