Microbiome dynamics during spontaneous fermentations of sound grapes in comparison with sour rot and Botrytis infected grapes.

The main losses in viticulture around the world are normally associated with rotten grapes affecting both the chemical composition and the grape microbiota that later might affect the alcoholic fermentation. We analyzed the population in musts obtained from sour rotten, botrytized and healthy Macabeo grapes and the population dynamics during the spontaneous alcoholic fermentation by culture dependent and various culture independent methods including, for the first time, qPCR and massive sequencing. Grape health state affected the fermentation kinetics and also the microbial diversity and composition. Unexpectedly, the fermentation proceeded the fastest in the rotten must followed by the healthy and the botrytized grapes. As in previous studies, plate cell counts and qPCR results confirmed the increase in the number of both bacteria and fungi in the musts from damaged grapes. Massive sequencing detected higher biodiversity than the other techniques at each stage, with Saccharomyces and Oenococcus found already in the grape must. Hanseniaspora osmophila replaced to Hanseniaspora uvarum as the predominant yeast during the mid-fermentation stage for both damaged grapes. Furthermore, musts and beginning of fermentation from rotten and botrytized grapes consistently had a higher presence of the fungi Zygosaccharomyces, Penicillium and Aspergillus while high abundance of Botrytis were observed just for botrytized grapes. As expected, the acetic acid bacteria number increased in musts from rotten and botrytized grapes, mostly due to changes in proportion of the genus Gluconoacetobacter which remained more abundant during damaged grapes fermentation than during healthy ones. Interestingly, the presence of Oenococcus oeni at the end of the alcoholic fermentation was strongly affected by the health status of the grapes.