Effect of fermentation temperature on microbial population evolution using culture-independent and dependent techniques

Abstract The population dynamics of micro-organisms during grape-must fermentation has been thoroughly studied. However, the main approach has relied on microbiological methods based on plating. This approach may overlook micro-organisms that (i) grow slowly or do not grow well on artificial media or (ii) whose population size is small enough to be detected by regular sampling. Culture-independent methods have been used and compared with the traditional plating method during wine fermentations performed at two different temperatures (13 °C and 25 °C). These methods include a qualitative technique, the DGGE; a semi-quantitative technique, the direct cloning of amplified DNA; and a quantitative technique, the QPCR. The biodiversity observed in the must and at the beginning of fermentation was much higher when DGGE or direct cloning were used. Quantification of the most frequent non- Saccharomyces yeast, Hanseniaspora uvarum and Candida zemplinina, showed that they survived throughout the fermentation process and, specifically, it revealed the quantitatively relevant presence of C. zemplinina until the end of fermentation.