Microbiological and Molecular Identification (PCR-RFLP-ITS) of the yeast from Cinsault grape cultivated in Abdelmalek Ramdane (Wilaya of Mostaganem, Algeria)

Abstract

The grape is an exemplary fruit of the microbial diversity, it is considered as the habitat of multiple wild microorganisms (yeasts). In this study, we explored the divergence of the indigenous yeast flora in the vineyards of the Abdelmalek Ramdane region (Wilaya de Mostaganem, Algeria) by collecting grape samples (Cinsault). Extraction of DNA as well as a PCR-ITS-RFLP has been developed for the ITS1-ADNR 5,8S-ITS2 region; it is sensitive enough to detect the biodiversity of the different species of isolated yeasts. Then 4 species of yeast out of the 11 studied belonging to 4 different genera were characterized according to their molecular profiles. Thus, the strains studied were characterized with two restriction enzymes (Hinf I and HaeIII). Meanwhile, classical microscopic yeast identification studies were able to enhance the results. Among the yeast species identified: Pichia fermentans, Torulaspora delbrueckii, Zygosaccharomyces microellipsoide and Saccharomyces cerevisiae. According to the results obtained, the Cinsault variety presents an excellent reservoir of yeasts Non saccharomyces. However, the tools of molecular biology have brought a notorious revolution in precise yeast identification tests. The PCR-RFLP-ITS is one of the most widely used methods of identification.