THE BIOGEOGRAPHY OF FERMENTATIVE YEAST POPULATIONS FROM THE VINEYARDS OF THE AZORES ARCHIPELAGO

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THE BIOGEOGRAPHY OF FERMENTATIVE YEAST POPULATIONS FROM THE VINEYARDS OF THE AZORES ARCHIPELAGO J. Drumonde-Neves a b, M. T. Lima a, D. Schuller b University of Azores Department of Agricultural Sciences a Research Center for Agricultural Technology of Azores University of Minho Department of Biology b Molecular and Environmental Research Centre

AIMS Evaluate the biogeography of the fermentative yeast flora from the vineyards of the Azores Archipelago To build a Saccharomyces cerevisiae strain collection for - Preservation of yeast genetic diversity - Equitable sharing of genetic data - Selection and improvement of wine strains

INTRODUCTION Azores Archipelago Corvo Flores Graciosa Terceira Faial Pico S. Jorge São Miguel 600 km Santa Maria

INTRODUCTION Azores Archipelago Corvo Flores Graciosa Terceira - Biscoitos Faial Pico S. Jorge São Miguel Santa Maria

INTRODUCTION Non-abandonned Abandonned vineyards Appellationsvineyards of origin / non-appellations of origin Traditional varieties Lagidos (Pico Island) Verdelho Arinto Terrantez Hybrid varieties (American varieties) S. Lourenço (Santa Maria Island)

MATERIAL AND METHODS SAMPLING LOCATIONS Santa Maria São Miguel Terceira 3 Baía do Tagarete 2 Baía de S. Lourenço S. Vicente 10 Biscoitos 14 4 Sul 1 Baía da Maia 5 P. Delgada 6 Caloura 7 Vila Franca 8 Ponta Garça 9 Faial da Terra 13 Pesqueiro 12 Salga 11 Porto Martins Graciosa Pico 15 Vitória 16 Santa Cruz da Graciosa 18 Lagoa Outeiro 22 Lagidos 19 Candelária 23 21 Canada das Almas Appellations of origin No appellations of origin Abandonned vineyards 17 Luz S. Mateus 25 24 Terra do Pão 20 Manhenha Faial S. Jorge Flores Norte Pequeno 27 28 29 Capelinhos 26 Varadouro Praia do Norte 35 Urzelina Fajã do Ouvidor 34 Fajã da Ribeira 32 da Areia Fajã dos Vimes 33 Fajã de S. João 31 30 Ponta do Topo 36 Santa Cruz das Flores

MATERIAL AND METHODS Weight loss (g/l) Microvinification Isolation Species identification -70 g/ L ITS-RFLP 0-10 -20-30 -40-50 -60-70 Yeast collection from the vineyards of Azores -80-90 -100 1 6 11 16 21 26 Day Interdelta Sequences S. cerevisiae strains identification

MATERIAL AND METHODS Santa Maria São Miguel Terceira Graiosa Pico Faial São Jorge Flores

MATERIAL AND METHODS Santa Maria 8 São Miguel Terceira Graciosa 9 12 14 27 Pico 6 Faial 11 São Jorge Flores 1

MATERIAL AND METHODS Traditional varieties Terrantez Arinto Hybrid varieties Verdelho Non-abandonned vineyards Abandonned vineyards 8 Santa Maria 9 São Miguel 12 Terceira 14 Graciosa 27 Pico 6 Faial 11 São Jorge 1 Flores 4 13 13 34 24

RESULTS FERMENTATION Fermentation onset (day) Fermentation duration (day) Traditional varieties 2,3 18,6 Hybrid varieties in non- abandonned vineyards 2,4 29,3 Hybrid varieties in abandonned vineyards 2,0 25,9

RESULTS Summary of all spontaneous fermentations SAMPLES OF TRADITIONAL VARIETIES 16,7 SAMPLES OF HYBRID VARIETIES FROM NON-ABANDONED VINEYARDS 52,9 SAMPLES OF HYBRID VARIETIES FROM ABANDONED VINEYARDS Unfinished fermentations 66,7 Samples from other winemaking regions Samples from appellations of origin

RESULTS Summary of all spontaneous fermentations SAMPLES OF TRADITIONAL VARIETIES 56,0 SAMPLES OF HYBRID VARIETIES FROM NON-ABANDONED VINEYARDS 56,3 SAMPLES OF HYBRID VARIETIES FROM ABANDONED VINEYARDS 75,0 Fermentations completed by S. cerevisiae Samples from other winemaking regions Samples from appellations of origin

RESULTS Summary of all spontaneous fermentations SAMPLES OF TRADITIONAL VARIETIES 44,0 SAMPLES OF HYBRID VARIETIES FROM NON-ABANDONED VINEYARDS 43,7 SAMPLES OF HYBRID VARIETIES FROM ABANDONED VINEYARDS 25,0 Fermentations completed by non-saccharomyces species Samples from other winemaking regions Samples from appellations of origin

RESULTS Summary of all spontaneous fermentations SAMPLES OF TRADITIONAL VARIETIES 80 Pichia guilliermondii 44,0 20 Hanseniaspora vineae SAMPLES OF HYBRID VARIETIES FROM NON-ABANDONED VINEYARDS 50 Candida railenensis 33 43,7 Pichia guilliermondii 17 Fungal sp. SAMPLES OF HYBRID VARIETIES FROM ABANDONED VINEYARDS 50 Candida railenensis 25,0 50 Candida zemplinina

RESULTS Number of S. cerevisiae strains involved in spontaneous fermentations SAMPLES OF TRADITIONAL VARIETIES 4 3 1 7 1 2 2 23 19 2 5 7 3 1 1-23 80 SAMPLES OF HYBRID VARIETIES FROM NON-ABANDONED VINEYARDS 14 1 1 3 9 21 1 1 1 52 1-21 SAMPLES OF HYBRID VARIETIES FROM ABANDONED VINEYARDS 3-11 6 4 11 3 9 3 36 Fermentations completed by S. cerevisiae Samples from other winemaking regions Samples from appellations of origin

RESULTS Number of S. cerevisiae strains from each island SAMPLES OF TRADITIONAL VARIETIES 4 3 1 7 1 2 2 23 19 2 5 7 3 1 SAMPLES OF HYBRID VARIETIES FROM NON-ABANDONED VINEYARDS 14 1 1 3 9 21 1 1 1 St.ª Maria SAMPLES OF HYBRID VARIETIES FROM ABANDONED VINEYARDS S. Miguel Terceira Graciosa Pico 6 4 11 3 9 3 Faial S. Jorge Flores

RESULTS Number of S. cerevisiae strains from each island SAMPLES OF TRADITIONAL VARIETIES 15 3 46 16 4 3 1 7 1 2 2 23 19 2 5 7 3 1 SAMPLES OF HYBRID VARIETIES FROM NON-ABANDONED VINEYARDS 16 3 9 22 14 1 1 3 9 21 1 1 1 1 1 St.ª Maria S. Miguel Terceira SAMPLES OF HYBRID VARIETIES FROM ABANDONED VINEYARDS Graciosa Pico 23 6 6 4 11 233 9 3 Faial S. Jorge Flores

CONCLUSIONS The vineyards of the Azores islands harbor a high genetic diversity of S. cerevisiae strains; In abandoned vineyards, this species occurred with a lower genetic diversity, but a greater abundance or capacity to predominate in spontaneous must fermentations; The ocurrence of non-saccharomyces species depended on the type of vineyard and the grape variety (traditional or hybrid); Vineyard ecosystems from the Graciosa island harbor the highest number of S. cerevisiae strains, probably associated with the large extension of vineyards on this island;

ACKNOWLEDGMENTS This work was financially supported by the DRCT, the programs POCI 2010 (FEDER/FCT, POCI/AGR/56102/2004), PTDC (AGR-ALI/103392/2008) and the EC s Seventh Framework Programme (FP7/2007-2013) under grant agreement nº 232454. Serviços de Desenvolvimento Agrário dos Açores Numerous wine producers UNIÃO EUROPEIA FUNDO ESTRUTURAIS