A culture-independent PCR-based method for the detection of Lachancea thermotolerans in wine

When inoculated in association with Saccharomyces cerevisiae, the yeast Lachancea thermotolerans determines a reduction of volatile acidity and an increase in the production of glycerol, 2-phenylethanol, and polysaccharides. Moreover, L. thermotolerans is a natural L-lactic acid producer, thus it contributes to wine acidification and microbiological stabilization. In view of its utilization in winemaking, a culture-independent PCR-based method was developed for the detection of L. thermotolerans during wine fermentations. This method, which utilizes species-specific PCR primer pairs that anneal to intron 2 of the mitochondrial COX1 gene, is rapid and reliable, and detects L. thermotolerans in wine at 10(4) cells/ml and with a S. cerevisiae/L. termotholerans ratio of 1,000/1.