Optimizing protocol for a rapid and cost effective DNA isolation for Marker Assisted Selection pipeline

Domesticated grapes are assumed to have originated in the Middle East. However, a considerable number of species are native in East Asian countries such as China, Korea and Japan as well. Evidence suggests that a total of seven species and eight varieties have been found to be native to Japan. A wide level variation in morphology, genetic and fruit composition exist in wild grape native to Japan.

Excessive fertilization of the vineyard leads to low quality grapes, increased costs and a negative impact on the environment. In order to establish an integrated management system aimed at a sustainable fertilization of the vineyards, nutritional reference levels were established. For this purpose, 30 representative vineyards of the Albariño variety were studied, in which soil and petiole analyses were carried out for two years and grape yield and quality at harvest were measured. In both years of study, soil pH, calcium, sodium and cation exchange capacity were positively correlated with calcium content and negatively correlated with manganese in grapes. Irrigated vineyards had higher levels of aluminium in soil and lower levels of calcium in petiole. Climatic conditions were very different in the years of the study. The year 2019 was colder than usual, in 2020 there was a marked water stress with high summer temperatures. This resulted in medium-high acidity in grapes in 2019 and low acidity in 2020, with sugar levels being similar both years. A very marked decrease in must amino nitrogen was observed in 2020, with ammonia nitrogen remaining stable. The correlation of acidity and sugar values in grapes with soil and petiole analysis data made it possible to establish reference levels for the nutritional diagnosis of the Albariño variety in this region. Based on these results, an easy-to-use TIC application is currently being created for grapegrowers, aimed at improving the sustainability of the vineyard through reasoned fertilization. This study has now been extended to other Galician vine varieties.

Alcoholic fermentation is the main step for winemaking, mainly performed by the yeast Saccharomyces cerevisiae. But other wine yeasts called non-Saccharomyces may contribute to alcoholic fermentation and modulate the wine aroma complexity. The recurrent problem with the use of these non-Saccharomyces yeasts is their trend to die off prematurely during alcoholic fermentation, leading to a lack of their interesting aromatic properties searched in the desired wine. This phenomenon appears to be mainly due to interactions with S. cerevisiae. These interactions are often negatives but remain unclear because of the species and strain specific response. Among the non-Saccharomyces yeasts, Lachancea thermotolerans is a wine yeast naturally found in grape must and well known as a great L-lactic acid producer and an aromatic molecules enhancer, but its behavior during alcoholic fermentation can be completely different in co-fermentation with S. cerevisiae in function of strain used.

Grape downy mildew, caused by plasmopara viticola, is one of the main diseases affecting viticulture worldwide and its control usually relies on frequent sprays with agrochemicals. Grapevine varieties resistant to p. Viticola represent an effective solution to control downy mildew and reduce the environmental impact of viticulture. Loci of resistance to p. Viticola (Rpv) have been introgressed from wild vitis species and some of them, like Rpv1, Rpv3.1 and Rpv10, are currently the most utilized genetic resources in grape breeding.

Wine contains secondary metabolites derived from aromatic amino acids (AADC), which can determine quality, stability and bioactivity. Several yeast species, as well as some lactic acid bacteria (LAB), can contribute in the production of these aromatic compounds. Winemaking should be studied as a series of microbial interactions, that work as an interconnected network, and can determine the metabolic and analytical profiles of wine. The aim of this work was to select microorganisms (yeast and LAB) based on their potential to produce AADC compounds, such as tyrosol and hydroxytyrosol, and design a microbial consortium that could increase the production of these AADC compounds in wines.