Isolation and Identification of the Indigenous Yeast Population during Spontaneous Fermentation of Isabella (Vitis labrusca L.) Grape Must.

María L. Raymond Eder,Cristina Reynoso,Santiago C. Lauret,Alberto L. Rosa

doi:10.3389/fmicb.2017.00532

María L. Raymond Eder, Cristina Reynoso + Show 2 more

Open Access

https://doi.org/10.3389/fmicb.2017.00532

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Abstract

Grape must harbors a complex community of yeast species responsible for spontaneous alcoholic fermentation. Although there are detailed studies on the microbiota of Vitis vinifera L. grapes, less is known about the diversity and behavior of yeast communities present on fermenting grape must from other species of Vitis. In this work, we used a culture-dependent method to study the identity and dynamics of the indigenous yeast population present during the spontaneous fermentation of Isabella (Vitis labrusca L.) grape must. Alcoholic fermentation was conducted using standard enological practices, and the associated non-Saccharomyces and S. cerevisiae yeast community was analyzed using selective growth media and 5.8-ITS DNA sequencing. Candida californica, Candida hellenica, Starmerella bacillaris (synonym Candida zemplinina), Hanseniaspora uvarum, and Hanseniaspora vineae were the main non-Saccharomyces species identified on Isabella fermenting must. Issatchenkia hanoiensis, a yeast species rarely found on Vitis vinifera L. grapes, was also recognized on Isabella grape must. Candida azymoides, Candida californica and Pichia cecembensis, identified in this work on Isabella fermenting must, have not previously been found on Vitis vinifera L. grape must. Interestingly, C. azymoides, I. hanoiensis and P. cecembensis have recently been isolated from the surface of Vitis labrusca L. grapes from vineyards in the Azores archipelago, suggesting that specific Vitis-yeast species associations are formed independently of geographic origin. We suggest that C. azymoides, C. californica, and P. cecembensis are yeast species preferentially associated with Vitis labrusca L. grapes. Specific biological interactions between grapevines and yeast species may underlie the assembly of differential Vitis-microbial communities.

Highlights

The surface of grapes lodges the microbiota responsible for spontaneous fermentation of grape must into wine (Mortimer and Polsinelli, 1999; Barata et al, 2012)
We found C. azymoides, C. californica and P. cecembensis on Isabella grape must, which have not been recognized on Vitis vinifera L. grapes
The study of DrumondeNeves et al (2016) and the results reported here strongly suggest that at least C. azymoides and P. cecembensis are preferentially associated with the microbiota of Vitis labrusca L. grapes

Summary

Introduction

The surface of grapes lodges the microbiota responsible for spontaneous fermentation of grape must into wine (Mortimer and Polsinelli, 1999; Barata et al, 2012). The complexity of indigenous non-Saccharomyces and Saccharomyces yeast microbiota present during spontaneous grape must fermentation has a major impact on the organoleptic and sensory properties of the final wines (Jolly et al, 2013; Padilla et al, 2016b; Varela and Borneman, 2016). Detailed studies on the microbiota of grapes and grape musts may enable valuable indigenous yeast strains that contribute to the regional character of wines to be identified (Knight et al, 2015; Capece et al, 2016; Padilla et al, 2016b; Varela and Borneman, 2016). The sequencing of rDNA regions D1–D2 (James et al, 1997; Kurtzman and Robnett, 1998), RFLP analyses (Guillamón et al, 1998) and/or the sequencing of ribosomal 5.8S-ITS are frequently used for rapid identification of indigenous isolated yeasts (Barata et al, 2012)

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