A catalogue of cultivable yeasts from the microbiota of grape berries cv. Vinhão and Loureiro

Abstract

The relevance of non-Saccharomyces native yeasts for improving the complexity of wine aroma is increasingly recognised, besides their use in a range of biotechnological applications. Previously, we used a metagenomic approach to assess the effects of vineyard calcium sprays on the microbiota of grape berries cv. Vinhão and Loureiro. In this study, we aimed at assembling a catalogue of the cultivable yeasts from grape berries of these cultivars and to investigate the direct effect of calcium and other elicitors on the isolated taxa. Seventeen unique colony morphologies were identified, and the sequencing of the region spanning the rDNA internal transcribed spacers or the rDNA D1/D2 domain revealed that they are distributed into 13 different genera. Although the cultivar and vintage influenced the taxa abundance and diversity, Aureobasidium pullulans and Hanseniaspora uvarum were the most abundant species, ubiquitously populating the fruits together with Pichia terricola, Rhodotorula glutinis, Starmerella bacillaris and Vishniacozyma heimaeyensis. The rarest genera comprised Candida, Trichosporon and Zygoascus. Analysis of yeast growth in the presence of specific elicitors revealed that Pichia spp. are the most sensitive to exogenous calcium, while Sporobolomyces roseus is the most sensitive to copper exposure. Moreover, A. pullulans, H. uvarum and S. bacillaris displayed the highest sensitivity to osmotic stress. In turn, Wickerhamomyces anomalus, Metschnikowia pulcherrima, P. fermentans, P. kluyveri and P. kudriavzevii showed mild resistance to ethanol, unlike the remaining non-Saccharomyces yeasts. Correlation networks expressing the overall yeast performance showed that H. uvarum and S. bacillaris share the strongest positive interspecies correlation, followed by the pairs P. fermentans–P. kluyveri, B. albus–R. glutinis and A. pullulans–V. heimaeyensis. The strongest negative correlations were found for the pairs P. kluyveri–S. roseus, P. terricola–W. anomalus, P. kudriavzevii–V. heimayensis, P. kluyveri–V. heimayensis, and A. pullulans–P. kluyveri. This study exposed microbial niches with a predictably common response to abiotic stress.

Non-Saccharomyces Yeast Strains, Aromatic Compounds and Sensory Analysis of Italy and Negra Criolla Pisco from the Moquegua Region of Peru

“…Similarly, V. heimaeyensis has been found in the microbiota of grape berries from cv. Vinhão and Loureiro grapes, although its technological relevance has not been reported [49]. Moreover, A. pullulans, V. carnescens, and V. heimaeyensis were also able to metabolize saccharose and glucose, reducing the pH but without gas production (Figure 1B).…”

Identificación de levaduras nativas en café Geisha (Coffea arabica) y su efecto como cultivo iniciador en las características sensoriales y químicas de café lavado

“…Además, se realizaron análisis microscópicos para caracterizar las células por forma, tamaño, color, ornamentación, gemación y presencia de pseudohifas. De esta manera y consultando fuentes de literatura, las cepas fueron identificadas hasta nivel de géneros (Martins et al 2022).…”

Identification of grape berry indigenous epiphytic yeasts with <i>in vitro</i> and <i>in vivo</i> antagonistic activity towards pathogenic fungi

“…The native yeast strains used in this study had previously been isolated from the microbiota of grape berries (Martins et al, 2022) during two harvest seasons in two vineyards located in the DOC regions of Vinhos Verdes and Douro (Northern Portugal). Briefly, the isolation methodology consisted of platting freshly extracted grape juice in a solid Wallerstein differential medium, and isolating approximately 200 unique colonies (per vineyard) in a fresh medium after 5 days of incubation at 24 °C.…”

“…Briefly, the isolation methodology consisted of platting freshly extracted grape juice in a solid Wallerstein differential medium, and isolating approximately 200 unique colonies (per vineyard) in a fresh medium after 5 days of incubation at 24 °C. Species identification was performed following Sanger sequencing of the rDNA internal transcribed spacers (ITS) and the rDNA D1/D2 domain region applying previously optimised methods (Martins et al, 2022). The sequences were queried onto BLASTn (nucleotide BLAST -basic local alignment search tool) of NCBI (National Center for Biotechnology Information) and strain identification was based on the result with the highest score (sequence of the best match, determined through values of query cover, percentage identity and E value).…”

“…The six filamentous fungi were A. niger, B. cinerea, C. acutatum, C. nymphaea, P. expansum and P. glabrum. In our previous study, preliminary assays showed the ability of A. pullulans and M. pulcherrima to inhibit the growth of A. niger in vitro (Martins et al, 2022). Extensive studies that involve a vast array of indigenous strains are thus necessary in order to be able to design an effective foliar formulation to ensure sustainable agriculture.…”