Pilar Santamaría

Ecology of spontaneous fermentation in one winery during 5 consecutive years

1999.The ecology of spontaneous fermentation in a new winery in La Rioja (Spain) was studied during 5 consecutive years using mitochondrial DNA restriction analysis. The number of different strains detected for each vintage and their appearance frequency varied from one year to another. A small number of strains were present in consecutive years, but the presence of each one varied in function of the specific year. Only one strain was present in all the 5 years studied. For the 1997 vintage, an unusual dominance of non‐Sacharomyces yeasts in the vigorous fermentation was detected; this may explain the abnormal analytical data for the wines of that year.

Improvement of grape and wine phenolic content by foliar application to grapevine of three different elicitors: Methyl jasmonate, chitosan, and yeast extract.

Phenolic compounds play a key role in grape and wine organoleptic properties, being therefore a key parameter in wine quality. Elicitor application constitutes an interesting field of research since it is indirectly involved in the accumulation of phenolic compounds. The aim of this study was to compare the effect of the application of three different elicitors on both grape and wine phenolic content. Methyl jasmonate, chitosan, and a commercial yeast extract were applied to the canopy at veraison and one week later. Results showed that foliar treatments carried out with methyl jasmonate and yeast extract achieved the best results, increasing grape and wine anthocyanin content when compared to the control. Moreover, the application of the yeast elicitor also enhanced grape stilbene content. In contrast, the chitosan treatment did not have a substantial impact on the phenolic compounds. The results of this study indicate that methyl jasmonate and yeast extract applications could be a simple practice to increase grape and wine phenolic content.

Methyl Jasmonate Foliar Application to Tempranillo Vineyard Improved Grape and Wine Phenolic Content

The importance of phenolic compounds for grape and wine quality has drawn attention to studying different practices with the aim of increasing their content. Cluster application of elicitors is a viticultural practice that has shown promising results in recent years. However, cluster application requires a previous defoliation, which is time-consuming and expensive. In the present study, methyl jasmonate was foliar applied to Tempranillo grapevines in order to study its effect on grape and wine phenolic composition. Methyl jasmonate foliar application increased anthocyanin and stilbene content in both grape and wine, besides enhancing wine flavonol content. This treatment induced the synthesis of 3-O-glucosides of petunidin and peonidin and trans-p-coumaroyl derivatives of cyanidin and peonidin. For stilbenes, trans-piceid content was considerably increased in both grape and wine. The results obtained suggest that methyl jasmonate foliar application could be a simple and accessible practice to enhance grape and wine quality.

Pulsed electric field treatment enhanced stilbene content in Graciano, Tempranillo and Grenache grape varieties

The purpose of this paper was to study the effect of pulsed electric fields (PEF) on the stilbene content of three grape varieties. For this purpose, four different PEF treatments were applied using a continuous system over three varieties, Graciano, Tempranillo and Grenache, destemmed and crushed. In addition, the influence of PEF on their physicochemical composition was studied. PEF treatments did not affect the pH or total acidity of Graciano, however, musts from Tempranillo and Grenache had higher pH values and lower total acidity. In the three varieties, all treatments resulted in an increase of potassium content, deeper colour intensity and total polyphenol index and lower tonality, more pronounced in the treatments with higher time and energy. The stilbene content of the must significantly increased with respect to the control. This increase depended on the variety and the treatment applied. Tempranillo increased up 200% the total stilbene concentration, Grenache 60% and Graciano 50%. For the three varieties, the treatment with the highest time and energy was the most effective on the total stilbene extraction. These results indicate that PEF could be a suitable technology for obtaining musts with deeper colour and higher phenolic content, including resveratrol and piceid.

Phenylalanine and urea foliar applications to grapevine: effect on wine phenolic content.

Phenolic compounds play a key role in the organoleptic properties of wines. Viticultural practices may influence grape and wine phenolic content, thus determining their quality. The objective of this study was to evaluate the effect of foliar applications of phenylalanine and urea, at two different doses, on wine phenolic composition. Grapes were harvested at their optimal technological maturity and their respective wines were elaborated at small scale. Wine detailed phenolic composition was determined. Results revealed that the content of several anthocyanins and flavonols was enhanced by the application of both phenylalanine doses and by the application of the low dose of urea. In contrast, flavanols and non-flavonoid compounds were less affected by the foliar treatments. The findings seem to be related to the time of application, since anthocyanins and flavonols are accumulated after veraison. In conclusion, nitrogen foliar fertilization increased the phenolic content of Tempranillo wines. This could be of interest since anthocyanins and flavonols are associated with wine quality, especially with its color.

Genomic diversity of Oenococcus oeni populations from Castilla La Mancha and La Rioja Tempranillo red wines.

The Oenococcus oeni populations of Tempranillo wines from Castilla La Mancha and La Rioja winemaking regions were analysed from one to three years and up to ten wineries. The objective was to evaluate the genetic variability and the O. oeni population structure. For this purpose a MLST scheme based on four loci (gyrB, purK, pgm and recP genes) and PFGE with SfiI restriction enzyme were developed for later combination. The results showed an O. oeni population completely adapted to winemaking regions. A purifying selection influenced the genes evolution, especially recP that along with purK were the most interesting loci to analyse the genetic variability of the isolates. In this way linkage disequilibrium and intergenic and intragenic recombination were determined between isolates. PFGE typing with UPGMA data were not coincident with the phylograms assessed for MLST by Maximum likelihood and combination of both techniques differentiated all the isolates as strains. Those results led the research to conclude that O. oeni population from CM and LR was a panmictic population with a slight clonal evolution, so subpopulations could not be described. A broader study including more winemaking regions with different grape varieties and distinct ways of elaborating would be interesting to complete the knowledge about O. oeni populations.

Ecology of Indigenous Lactic Acid Bacteria along Different Winemaking Processes of Tempranillo Red Wine from La Rioja (Spain)

Ecology of the lactic acid bacteria (LAB) during alcoholic fermentation (AF) and spontaneous malolactic fermentation (MLF) of Tempranillo wines from four wineries of La Rioja has been studied analyzing the influence of the winemaking method, processing conditions, and geographical origin. Five different LAB species were isolated during AF, while, during MLF, only Oenococcus oeni was detected. Although the clonal diversity of O. oeni strains was moderate, mixed populations were observed, becoming at least one strain with distinct PFGE profile the main responsible for MLF. Neither the winemaking method nor the cellar situation was correlated with the LAB diversity. However, processing conditions influenced the total number of isolates and the percentage of each isolated species and strains. The winemaking method could cause that genotypes found in semicarbonic maceration did not appear in other wineries. Four genotypes of O. oeni were isolated in more than one of the rest wineries. These four together with other dominant strains might be included in a future selection process.

Molecular analysis of Oenococcus oeni and the relationships among and between commercial and autochthonous strains

The presence and distribution of genotypes from malolactic starter cultures between the autochthonous microbiota in fermenting Rioja wines have been studied in this paper. The commercial cultures characterization allowed to identify different species and common pulsed field gel electrophoresis and randomly amplified polymorphic DNA genotypes in several brands of Oenococcus oeni starter cultures. Four indistinguishable genotypes were found between the commercial and the autochthonous O. oeni strains. These four genotypes appeared during different vintages in seven of the 10 sampled wineries despite bacterial starter cultures had never been used. Therefore, the detection of commercial LAB genotypes indistinguishable from the autochthonous ones involved their removal from a selection process. These genotypes represented the 17.7% of autochthonous O. oeni isolated in this region during three consecutive years. This fact demonstrated that performing similar studies as previous selection criteria is advisable to avoid large work and the proposal of one commercialized bacteria.

Effect of methyl jasmonate application to grapevine leaves on grape amino acid content

Over the last few years, considerable attention has been paid to the application of elicitors to vineyard. However, research about the effect of elicitors on grape amino acid content is scarce. Therefore, the aim of this study was to evaluate the influence of foliar application of methyl jasmonate on must amino acid content. Results revealed that total amino acid content was not modified by the application of methyl jasmonate. However, the individual content of certain amino acids was increased as consequence of methyl jasmonate foliar application, i.e., histidine, serine, tryptophan, phenylalanine, tyrosine, asparagine, methionine, and lysine. Among them, phenylalanine content was considerably increased; this amino acid is precursor of phenolic and aromatic compounds. In conclusion, foliar application of methyl jasmonate improved must nitrogen composition. This finding suggests that methyl jasmonate treatment might be conducive to obtain wines of higher quality since must amino acid composition could affect the wine volatile composition and the fermentation kinetics.

Wine aromatic compound production and fermentative behaviour within different non-Saccharomyces species and clones.

Twenty‐five enological yeasts belonging to nine different species (Candida zeylanoides, Cryptococcus uzbekistanensis, Debaryomyces hansenii, Lachancea thermotolerans, Metschnikowia pulcherrima, Torulaspora delbrueckii, Williopsis pratensis, Zygosaccharomyces bailii and Saccharomyces cerevisiae) were screened for aroma formation and fermentative behaviour as part of a non‐Saccharomyces yeast selection programme.