Identification of yeasts by RFLP analysis of the 5.8S rRNA gene and the two ribosomal internal transcribed spacers

Abstract

The identification and classification of yeasts have traditionally been based on morphological, physiological and biochemical traits. Various kits have been developed as rapid systems for yeast identification, but mostly for clinical diagnosis. In recent years, different molecular biology techniques have been developed for yeast identification, but there is no available database to identify a large number of species. In the present study, the restriction patterns generated from the region spanning the internal transcribed spacers (ITS1 and ITS2) and the 5.8s rRNA gene were used to identify a total of 132 yeast species belonging to 25 different genera, including teleomorphic and anamorphic ascomycetous and basidiomycetous yeasts. In many cases, the size of the PCR products and the restriction patterns obtained with endonucleases Cfol. Haelll and Hinfl yielded a unique profile for each species. Accordingly, the use of this molecular approach is proposed as a new rapid and easy method of routine yeast identification.

Differentiation of Brewing and Related Yeasts Based on PCR Amplification and Restriction Fragment Length Polymorphism of Ribosomal DNA

“…Here an overlap between the Saccharomyces sensu stricto and sensu lato groups was evident. As reported by Esteve-Zarzoso et al 3 , the Saccharomyces sensu stricto group could also not be separated using this digestion enzyme.…”

“…In recent times, there have been a number of reports describing the application of molecular genetic tools for the differentiation and characterisation of yeasts used in industrial fermentation 2,3,4,7,8,9,11 . Data from these studies show that RFLP patterns allow recognition of individual species as well as individual strains within a species.…”

“…paradoxus and Sacch. pastorianus 3 . Here the size of the PCR products and the restriction patterns obtained by using endonucleases Cfo I, Hae III and Hin fI produced a unique pattern for many species.…”

An analysis of inter- and intraspecific genetic variabilities in the Kluyveromyces marxianus group of yeast species for the reconsideration of the K. lactis taxon

“…The alignment of the 5.8S-ITS region sequences yielded 648 nucleotide positions, of which 109 (17%) were variable and, of them, 80 (12%) were phylogenetically informative. The alignment of the 5.8S-ITS sequences confirmed the existence of nucleotide substitutions involving restriction site gains or losses in the strains belonging to these closely related species, which have been very useful to formulate the strategy for discrimination among these species described in Table 3 (Belloch et al, 1998a;Esteve-Zarzoso et al, 1999). Table 3.…”

Detection of Metarhizium anisopliae var. anisopliae within infected sugarcane borer Diatraea saccharalis (Lepidoptera, Pyralidae) using specific primers

“…The rDNA unit presents sequence variations which may be used in systematic studies at different taxonomic levels (Fouly et al, 1997;Argentina, 1999). The rDNA 18S and 28S regions are the most conserved units, and may be used in differentiating genera and species, while the ITS and IGS spacer regions have accumulated more variability and are better used to differentiate species or strains within the same species (Esteve-Zarzoso et al, 1999).…”