Rapid detection and quantification of yeast species during spontaneous wine fermentation by PCR-RFLP analysis of the rDNA ITS region

Abstract

L. GRANCHI, M. BOSCO, A. MESSINI and M. VINCENZINI.1999.PCR–RFLP analysis of the rDNA–ITS (internal transcribed spacer) region was applied to 174 yeast strains belonging to 30 species of oenological significance and including 27 type strains in order to define a rapid identification protocol for yeast colonies. DraI‐or HaeIII‐PCR–RFLP patterns were species‐specific with the exception of teleomorphic and anamorphic forms. An improved protocol taking about 30 h was used for the detection and quantification of yeast species occurring in the course of a spontaneous wine fermentation at industrial level. Wine samples were taken and plated daily on an agar medium and the developed colonies were analysed by PCR–RFLP after 24 h of incubation. A representative sample of these colonies was also identified by traditional methods. Both procedures gave identical results. However, PCR–RFLP analysis allowed a more precise enumeration of the yeast populations, proving to be a reliable and simple method for monitoring the development of the yeast community throughout wine fermentation.

Presence of Candida zemplinina in Sicilian Musts and Selection of a Strain for Wine Mixed Fermentations

“…We have conducted our analysis by considering 59 Candida isolates of the non-Saccharomyces IRVV collection. These were initially identified as representative of Candida stellata (Romancino et al, 2008), based on microbiological (Cavazza & Poznanski, 1998) and molecular analyses (Esteve-Zarzoso et al, 1999;Granchi et al, 1999). As new studies showed the prevalence of the C. zemplinina species in wine musts (Csoma & Sipiczki, 2008), we reanalysed our isolates and were able to establish that all of them belonged to the C. zemplinina species.…”

Isolation of a yeast strain able to produce a polygalacturonase with maceration activity of cassava roots

“…To achieve greater polymorphism, the amplified genes were treated with the restriction enzymes CfoI, HinfI, and HaeIII for the identification of yeasts at the species level (ESTEVE ZARZOSO et al, 1999;GRANCHI et al, 1999). The PCR amplified 5.8S ITS rRNA region was sequenced using the ABI Prism 3100 genetic analyzer.…”

Yeast biodiversity fromVitis viniferaL., subsp. sylvestris (Gmelin) Hegi to face up the oenological consequences of climate change

“…PCR products varying in length from 390 to 880 bp were digested with HinfI, HaeIII and DdeI enzymes, and the molecular mass of the restriction products obtained was compared with those described previously in the literature [10][11][12][13]. In general, we observed 18 different profiles of the ITS-5.8S rDNA region.…”

“…The approximate lengths were determined by the use of molecular weight ladder (100 bp, Roche). PCR and RFLP fragment lengths were used for identification of yeasts by comparison of the restriction bands with those available in literature [10][11][12][13]. Several groups appeared on the basis of their RFLP of the ITS region.…”