Isolation and Molecular Identification (PCR-Delta and PCR-RFLP-ITS) of yeasts from the Black muscat grape cultived in El Malah (Wilaya of Ain Temouchent,Algeria).

Abstract

The grape is an exemplary product of microbial diversity, it is considered as home to many wild microorganisms (yeast). In this context, we explored in this study, the divergence of the indigenous yeast flora in the vineyards of the region El Malah (Wilaya of Ain Temouchent, Algeria) by collecting varietal grape samples (Black Muscat). A high molecular diversity of the flora has been demonstrated, using two molecular techniques, where one is interested in the study of variability of rDNA, more precisely the region 5,8S rRNA ITS1-ITS2 by PCR-RFLP-ITS and the region Delta by PCR-Delta is an additional study which helped sort the yeast Saccharomyces and non Saccharomyces.So 6 yeast species belonging to the 11 studied at 4 different types were characterized according to their molecular profiles. Thus the strains studied were characterized by two restriction enzymes (HinfI and HaeIII). Meanwhile, microscopic and macroscopic studies of classical identification of yeast were able to strengthen these results. Among the yeast species identified: Torulaspora delbrueckii, Torulaspora pretoriensis, Candida solani, Candida pseudointermedia and Saccharomyces cerevisiae. This variety is an excellent tank of non Saccharomyces yeasts as evidenced by the results.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.