The Isolation And Identification Of Yeasts From The Course Of Spontaneuos Fermentation Of Vitis.amurensis Rupr Grape

Wine yeasts were the important microorganism during vinification, Wine yeasts diversity and identification are the foundation for further fine wine yeast selection. The yeasts which from Vitis.amurensis Rupr grape variety northeast region during the spontaneous fermentation can present variety characteristic, and region property, also can help brewaged corporation brewing original and typical Vitis.amurensis Rupr grape wine.Due to the low total sugar and high acidity Vitis.amurensis Rupr grape possessed and low temperature during fermentation season, So autochthonic yeast must be selected to adapt to these factors and also to solve the stuck fermentation because the ordinary wine yeast can not endure low pH and high acidity must condition. In allusion to this actuality, separated the fine wine yeast from composition of wine yeast populations during spontaneous fermentation and keeped the Vitis.amurensis Rupr wine smell and nutrition was studied.Five Vitis.amurensis Rupr grapes fermented respectively. 300 wine-related yeasts are isolated at random during spontaneous fermentation, preliminary assessment wine yeasts are carried out by Wallerstein Laboratory Nutrient Agar. At the same time, yeast species assessment was carried out by amplification and digestion of a region of the ribosomal RNA gene repeat unit. All of the Saccharomyces strains can provide a great deal of experimental base for using to select fine wine yeast which can endure low pH, high acidity must condition and develop grape berry feature etc.The results were as follows:1.300 wine related yeasts were isolated from the Spontaneous fermentation of V.amurensis Rupr. Yeast species assessment was carried out by WL medium. 4 unique colony morphologies were identified.2.By analysis of the restriction pattern of amplified 5.8S-ITS rRNA gene(RFLP)with the enzyme HinfⅠ, HaeⅢand HhaⅠ, respectively, 11 different genotypes were existed in fermentation. Thereinto, pattern 5,6,8,9 and 11 were respectively Candida sorbosa,Rhodotorula mucilaginosa,Issatchenkia terricola,Hanseniaspora uvarum,Saccharomyces cerevisiae。PCR product sizes vary from 450 to 880 bp.3.By analysis of the restriction pattern of amplified 5.8S-ITS region(RFLP)5 different hereditary clusterings were found.4.This work shows Wallerstein Laboratory Nutrient Agar(WL medium)was strengthened for using in wine fermentation to observe yeast populations and useful for predigesting in wine yeast molecular identification.After this study, the preliminary ideas of Vitis.amurensis Rupr grape wine yeast diversity in northeast China were established. And it was settled a base for the fine wine yeast which could endure low temperature and low pH.