Comprehensive analysis of colloid formation, distribution, and properties of monovarietal red wines using asymmetrical flow field-flow fractionation with online multidetection.

Red wine colloids, crucial in determining wine quality and stability, are understudied due to inadequate techniques for studying them effectively in the natural wine environment. Recently, Asymmetrical Flow Field-flow Fractionation (AF4) with online multidetection has emerged as a novel analytical tool for quantifying, fractionating, and characterizing red wine colloids in their native state. This study aimed to characterize the colloidal composition of 24 monovarietal Italian wines produced without filtration, oak contact, fining treatments, malolactic fermentation, macerating enzymes or ageing on yeast lees. AF4 analysis allowed quantification and characterization of wine colloids based on light scattering signal (MALS; gyration radius - Rg), size (hydrodynamic radius - Rh) and absorbance (A280 & A520 nm). The results showed that each wine contained up to five distinct colloids' populations, varying in size and gyration radii. Despite possessing very similar Rh, most colloids exhibited great differences in compactness, as indicated by their varying Rg values. Comparing the A280 signal of whole wines to those of wines containing only species larger than 5 kDa (considered colloids) allowed to calculate the percentage of molecules involved in colloidal particles assembly, ranging from 1 to 44 % of the total A280 absorbing compounds, reflecting the diversity among wines. The A520 signal indicated the presence of polymeric pigments in the colloidal fraction. Notably, colored colloids all had Rg > 20 nm, indicating their association with other colloidal-forming compounds. This observation led to the conclusion that, apart from free anthocyanins and polymeric pigments, the color of red wines is also due to colloidal particles formed by the latter bound to proteins, with their quantity being highly variable across wines of different origin. These findings, which highlight the fundamental role of proteins in shaping the colloidal status of red wines, were utilized to propose an updated hypothetical model for colloidal aggregation in red wine.

Dimethyl Sulfide (DMS) in Amarone Wines: Influence of Aging, Withering, Grape Variety, and Geographical Origin.

Occurrence of dimethyl sulfide (DMS), a potent aroma compound accumulating during aging, was investigated in commercial and experimental Amarone wines. In commercial Amarone, DMS was observed in concentrations ranging from 2.9 to 64.3 µg/L. Model aging studies on experimental wines indicated that DMS in Amarone is strongly associated with aging and that wines from different vineyards can vary significantly in their ability to accumulate DMS during aging. The capacity of certain vineyards to give wines with higher DMS-forming potential was consistent across three consecutive vintages, representing a true terroir factor to be expressed with aging. Wine content of primary amino acids (PAN), a commonly analyzed enological parameter of grape must, was shown to be positively correlated with DMS accumulation during aging. Grape withering also increased DMS-forming potential mostly due to increased PAN resulting from concentration due to water loss. Increased pH due to withering also contributed to a higher DMS content of withered wines, but to a lower extent. In certain vineyard sites, an influence of vintage conditions on DMS-forming potential was also observed.

Multivariate prediction of Saliva Precipitation Index for relating selected chemical parameters of red wines to the sensory perception of astringency.

Astringency is an essential sensory attribute of red wine closely related to the saliva precipitation upon contact with the wine. In this study a data matrix of 52 physico-chemical parameters was used to predict the Saliva Precipitation Index (SPI) in 110 Italian mono-varietal red wines using partial least squares regression (PLSr) with variable selection by Variable Importance for Projection (VIP) and the significance of regression coefficients. The final PLSr model, evaluated using a test data set, had 3 components and yielded an R2test of 0.630 and an RMSEtest of 0.994, with 19 independent variables whose regression coefficients were all significant at p < 0.05. Variables selected in the final model according to the decreasing magnitude of their absolute regression coefficient include the following: Procyanidin B1, Epicatechin terminal unit, Total aldehydes, Protein content, Vanillin assay, 520 nm, Polysaccharide content, Epigallocatechin PHL, Tartaric acid, Volatile acidity, Titratable acidity, Catechin terminal unit, Proanthocyanidin assay, pH, Tannin-Fe/Anthocyanin, Buffer capacity, Epigallocatechin PHL gallate, Catechin + epicatechin PHL, and Tannin-Fe. These results can be used to better understand the physico-chemical relationship underlying astringency in red wine.

Contribution of terpenes, methanethiol, and fermentative esters to sparkling wine aroma in relation to production technology, vintage, and aging: a case study on Durello wines.

BACKGROUND: The present study aimed to assess the chemical and sensory variability existing across sparkling white wines of a single appellation in relation to production technique and aging, identifying the existence of specific aroma chemical and sensory (odor) signatures. For this purpose, we selected Durello, an emerging sparkling white wine from the Veneto region produced using either Charmat or Classico methods. RESULTS: Comparison of Durello varietal volatiles profile with other Italian sparkling and still wines highlighted specific features of Durello, such as high content of 1,4-cineole and non-megastigmane norisoprenoids. The production technique (Charmat or Champenoise) impacted both the volatile chemical profile, influencing the content of esters, fatty acids, cyclic terpenoids, C6 alcohols, ß-damascenone and methanethiol. With regard to wine age and vintage, an influence on esters, p-menthane-1,8-diol, cineoles and methyl salicylate was observed. CONCLUSION: Sensory evaluation by means of a sorting task approach indicated the existence of two main types of odor profile, one clearly associated with Classico method. The main drivers of such aroma diversity were esters, 1,4-cineole and methanethiol. © 2023 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Reconstruction of Simplified Microbial Consortia to Modulate Sensory Quality of Kombucha Tea.

Kombucha is a fermented tea with a long history of production and consumption. It has been gaining popularity thanks to its refreshing taste and assumed beneficial properties. The microbial community responsible for tea fermentation-acetic acid bacteria (AAB), yeasts, and lactic acid bacteria (LAB)-is mainly found embedded in an extracellular cellulosic matrix located at the liquid-air interphase. To optimize the production process and investigate the contribution of individual strains, a collection of 26 unique strains was established from an artisanal-scale kombucha production; it included 13 AAB, 12 yeasts, and one LAB. Among these, distinctive strains, namely Novacetimonas hansenii T7SS-4G1, Brettanomyces bruxellensis T7SB-5W6, and Zygosaccharomyces parabailii T7SS-4W1, were used in mono- and co-culture fermentations. The monocultures highlighted important species-specific differences in the metabolism of sugars and organic acids, while binary co-cultures demonstrated the roles played by bacteria and yeasts in the production of cellulose and typical volatile acidity. Aroma complexity and sensory perception were comparable between reconstructed (with the three strains) and native microbial consortia. This study provided a broad picture of the strains' metabolic signatures, facilitating the standardization of kombucha production in order to obtain a product with desired characteristics by modulating strains presence or abundance.

Volatile Compounds in Monovarietal Wines of Two Amarone Della Valpolicella Terroirs: Chemical and Sensory Impact of Grape Variety and Origin, Yeast Strain and Spontaneous Fermentation.

Aroma profiles of withered Corvina and Corvinone wines from two different Valpolicella terroirs were investigated in relationship to yeast strain and use of spontaneous fermentation. The results indicated that volatile chemical differences between wines were mainly driven by grape origin, which was associated with distinctive compositional profiles. Wine content in terpenes, norisoprenoids, benzenoids and C6 alcohols, as well as some fermentative esters, were indeed significantly affected by grape origin. Conversely, yeast strain influence was mainly associated with fermentation-derived esters. Sensory analysis, besides confirming the major role of grape origin as driver of wine differentiation, indicated that spontaneous fermentations reduced the sensory differences associated with grape origin and variety, mainly due to high content of acetic acid and ethyl acetate.

Diversity of Italian red wines: A study by enological parameters, color, and phenolic indices.

An extensive survey was conducted on 110 Italian monovarietal red wines from a single vintage to determine their standard compositional, color, and phenolic characteristics, analysing more than 35 parameters evaluated through methods commonly used in the wine industry. 'Primitivo' achieved the highest average alcohol strength (15.4% v/v) and dry extract values, while 'Cannonau' showed the lowest total acidity. 'Corvina' had the lowest phenolic content (1065 mg/L by Folin-Ciocalteu assay), remarkably different from the highest found in 'Sagrantino' (3578 mg/L), the latter being also the richest variety in both proanthocyanidins and vanillin-reactive flavanols. 'Teroldego' wines were the richest in both total and monomeric anthocyanins (702 and 315 mg/L, respectively), followed by 'Aglianico' and 'Raboso Piave', while 'Corvina', 'Nebbiolo', and 'Nerello Mascalese' were the poorest. 'Montepulciano' and 'Sangiovese' showed intermediate values for the majority of the parameters analyzed. A multivariate PCA-DA approach allowed achieving both a classification of the different wines as well as the discrimination of 'Sangiovese' wines produced in two regions (Emilia Romagna and Toscana) that returned a 42-66% success rate depending on the zone considered. Taking into account the number and diversity of the wines analyzed, a correlation study helped in better understanding the underlying relations between the most common and widespread analytical techniques for phenolic and color determinations.

Two Sides to One Story-Aroma Chemical and Sensory Signature of Lugana and Verdicchio Wines.

Lugana and Verdicchio are two Italian white wines with a Protected Designation of Origin (PDO) label. These two wine types are produced in different regions using the same grape variety. The aim of this work is to investigate the existence of volatile chemical markers that could help to elucidate differences between Lugana and Verdicchio wines both at chemical and sensory levels. Thirteen commercial wine samples were analyzed by Gas Chromatography-Mass Spectrometry (GC-MS), and 76 volatile compounds were identified and quantified. Verdicchio and Lugana had been differentiated on the basis of 19 free and glycosidically bound compounds belonging to the chemical classes of terpenes, benzenoids, higher alcohols, C6 alcohols and norisoprenoids. Samples were assessed by means of a sorting task sensory analysis, resulting in two clusters formed. These results suggested the existence of 2 product types with specific sensory spaces that can be related, to a good extend, to Verdicchio and Lugana wines. Cluster 1 was composed of six wines, 4 of which were Lugana, while Cluster 2 was formed of 7 wines, 5 of which were Verdicchio. The first cluster was described as "fruity", and "fresh/minty", while the second as "fermentative" and "spicy". An attempt was made to relate analytical and sensory data, the results showed that damascenone and the sum of 3 of esters the ethyl hexanoate, ethyl octanoate and isoamyl acetate, was characterizing Cluster 1. These results highlighted the primary importance of geographical origin to the volatile composition and perceived aroma of Lugana and Verdicchio wines.

Influence of Different Modalities of Grape Withering on Volatile Compounds of Young and Aged Corvina Wines.

Withering is a practice traditionally used in various regions to produce sweet or dry wines. During withering there is an increase in sugar content but also a modification in volatile compound profiles. Controlling metabolic changes through the dehydration process to obtain wines with desired characteristics is therefore a challenging opportunity. The effects of two different withering technologies, post-harvest or on-vine with blocked sap vessel flow, on the volatile profile of young and aged Corvina red wines was investigated. The results showed that modulation of wine aroma due to the withering process is associated with fermentative metabolites, such as esters, higher alcohols, and acids, as well as grape-related compounds such as C6 alcohols, terpenes and norisoprenoids. Significant differences were also found by comparing the two withering techniques. Post-harvest in a traditional "fruttaio" warehouse wines showed higher content of ethyl acetate, ethyl butanoate, ß-citronellol and 3-oxo-α-ionol, whereas post-harvest withering on-vine increased ß-damascenone in wines. The type of withering technique has an influence on the evolution of some aroma compounds during the aging of wine, among them linalool, (E)-1-(2,3,6-trimethylphenyl)buta-1,3-diene (TPB), n-hexyl acetate, ethyl acetate, ethyl 3-methylbutanoate, 3-oxo-α-ionol and ß-damascenone.

Oxygen and SO2 Consumption of Different Enological Tannins in Relationship to Their Chemical and Electrochemical Characteristics.

The oxidative behavior of five commercial enological tannins of different sources (tea, grape marc, grape seed, untoasted oak, and toasted oak) was investigated in model wine solutions in the presence or absence of SO2. Solutions of the tannins were also analyzed for total phenolics, methyl cellulose precipitable tannins, high-performance liquid chromatography, and linear sweep voltammetry. Tea and oak-derived tannin solutions were characterized by the highest oxygen consumption rates, with oak-derived tannins exhibiting the highest oxygen consumption rates per milligram of phenolic material present. Linear sweep and derivative voltammetry parameters were well-correlated with oxygen consumption rates, whereas total phenolics or total tannins were not. All tannins were associated with consumption of SO2 upon reaction with oxygen, with the lowest rate of SO2 lost per milligram of O2 reacted being observed for oak tannins.

Fate of Grape-Derived Terpenoids in Model Systems Containing Active Yeast Cells.

Terpenes are important contributors to wine aroma. Free and glycosidically bound terpenes are primarily formed in grapes. During fermentation, they undergo important transformation catalyzed by yeast, so that the terpene profile of grape is substantially different from that of the corresponding wine. The present paper assessed the ability of a Saccharomyces cerevisiae strain to transform 17 different terpenes. Biotransformation was performed by placing target compounds in incubation with resting cells. Volatile compounds produced were extracted by solid-phase extraction and analyzed by gas chromatography-mass spectrometry. Geranyl acetate, neryl acetate, citronellyl acetate, and menthyl acetate were formed from the corresponding terpene alcohols. ß-Citronellol was the main product of geraniol transformation; geranial, an intermediate of this pathway, has also been detected. Limonene was hydroxylated by yeast to form carveol, trans-2,8-menthadien-1-ol, and cis-2,8-menthadien-1-ol. Moreover, yeast cells were found to be able to adsorb a significant portion of the terpenes present in the reaction batches, with the extent of this phenomenon being linked to terpene hydrophobicity.

Addition of volatile sulfur compounds to yeast at the early stages of fermentation reveals distinct biological and chemical pathways for aroma formation.

Volatile sulfur compounds (VSCs) greatly influence the sensory properties and quality of wine and arise via both biological and chemical mechanisms. VSCs formed can also act as precursors for further downstream VSCs, thus elucidating the pathways leading to their formation is paramount. Short-term additions of exogenous hydrogen sulfide (H2S), ethanethiol (EtSH), S-ethylthio acetate (ETA), methanethiol (MeSH) and S-methylthio acetate (MTA) were made to exponentially growing fermentations of synthetic grape medium. The VSC profiles produced from live yeast cells were compared with those from dead cells and no cells. Interestingly, this experiment allowed the identification of specific biochemical and/or chemical pathways; e.g. most of the conversion of H2S to EtSH, and the further step from EtSH to ETA, required the presence of live yeast cells, as did the conversion of MeSH to MTA. In contrast, the reaction from MTA to MeSH and ETA to EtSH was due primarily to chemical degradation. Ultimately, this research unravelled some of the complex interactions and interconversions between VSCs, pinpointing the key biochemical and chemical nodes. These pathways are highly interconnected and showcase the complexity of both the sulfur pathways in yeast and the reactive chemistry of sulfur-containing compounds.

Volatile organic compounds from Starmerella bacillaris to control gray mold on apples and modulate cider aroma profile.

Gray mold caused by Botrytis cinerea is a fungal disease that can determine significant economic losses of apple during the storage phase. An alternative to reduce the use of traditional synthetic fungicides is to employ the yeast Starmerella bacillaris as biological control agent (BCA), also with positive effect on apple juice fermentation for the production of cider. Thus, we aimed to evaluate the safety of 16 S. bacillaris strains and their ability to control B. cinerea. In addition, the fermentation performances in apple juice and the volatile organic compounds (VOCs) profile were assessed, both in single-strain and in sequential fermentations with Saccharomyces cerevisiae. The in vitro assays showed that all S. bacillaris strains can be considered safe from the analyzed virulence factors, and were able to significantly constrain the growth of B. cinerea, reducing mycelial growth of 50% in dual-culture and of 90% through VOCs. Moreover, in vivo antagonistic assays revealed a visible decrease of gray mold rot symptoms on apples confirming the potential of S. bacillaris as BCA. GC-MS analysis of the ciders obtained showed increased concentrations in the sequential fermentation of some higher alcohols and terpenes, positively correlated with the cider aromatic quality, and suggested the involvement of benzyl alcohol, known for its antimicrobial action, in the biocontrol efficacy.

Fermentative and post-fermentative oxygenation of Corvina red wine: influence on phenolic and volatile composition, colour and wine oxidative response.

BACKGROUND: During the production of red wine, moderate uptake of oxygen in the post-fermentative phase helps the stabilization of colour and the decrease of astringent tannins. However, the influence of oxygen uptake during the fermentative phase in must has not been completely investigated. In this study we evaluated the effect of controlled oxygen supply during fermentation-maceration of Corvina grapes on colour characteristics, tannins, volatile compounds, acetaldehyde production and oxidative stability of the finished wine. RESULTS: Oxygen supply during fermentation improved the formations of stable pigments of Corvina wines due to the higher production of acetaldehyde. However, in wines treated with oxygen a lower production of fruity esters by yeasts was observed. Wines obtained from higher oxygen exposure during fermentation-maceration showed reduced ability to react with oxygen during storage. CONCLUSIONS: Fermentative and post-fermentative oxygenation should be considered as a technological approach for modifying colour composition and stability, as well as oxidative behaviour of wine during aging. © 2020 Society of Chemical Industry.

Contribution of non-Saccharomyces yeasts to wine volatile and sensory diversity: A study on Lachancea thermotolerans, Metschnikowia spp. and Starmerella bacillaris strains isolated in Italy.

Saccharomyces cerevisiae starter cultures are largely used in winemaking to repress the wild microorganisms and achieve more predictable and desired outcomes. Notwithstanding, alternative microbial resources received increasing attention for their potential to produce wines with more distinctive and typical features. Our previous survey revealed a great inter- and intra-species diversity in an extensive collection of non-Saccharomyces wine yeasts from multiple regions of Italy. This study aimed to explore the detected biodiversity evaluating the quality of wines obtained by sequential inoculation of specific selected strains of the collection (Lachancea thermotolerans or Metschnikowia spp. or Starmerella bacillaris), and S. cerevisiae EC 1118. Fermentations of natural grape must at laboratory scale were followed by microbiological, chemical and sensorial analysis of the wines. The results indicated that each yeast species and strain exerted a distinctive impact on the wine, giving final products clearly separated with Principal Component Analysis. In particular, L. thermotolerans contributed producing relevant amounts of lactic acid and had the highest potential to reduce ethanol content; the presence of S. bacillaris increased the level of glycerol, and, remarkably, reduced acetaldehyde and total SO2; Metschnikowia spp. promoted the formation of higher alcohols and esters, and reduced volatile phenols. The sensory analysis based on the orthonasal aroma confirmed the separation between the wines obtained with the sequential fermentations and the control with single inoculation of EC 1118, although the three non-Saccharomyces species used could not be clearly distinguishable by the panelists. This study indicates that the use of selected native non-Saccharomyces strains in conjunction with S. cerevisiae positively modulates some relevant chemical parameters, and improves the aromatic intensity of wine, therefore justifying investments in non-conventional yeasts as co-starter cultures.

Volatile terpenoids, norisoprenoids and benzenoids as markers of fine scale vineyard segmentation for Corvina grapes and wines.

In this study, the diversity existing at the very small scale of single vineyard parcels in volatile composition of grapes and wines from a single estate in the Valpolicella wine region has been studied. Corvina grapes from eight contiguous vineyards were used for the study and vinified with the same protocol. The compounds analyzed by GC-MS were representative of the terpenoid, norisoprenoid and benzenoid chemical families. Free and bound compounds analysis showed that differences between parcels were relatively small on grapes samples, whereas after fermentation larger differences between wine samples were highlighted. Multivariate statistical analysis of wine volatiles highlighted the existence of similarities between wine volatile profiles, which reflected to a good extent the geographical location of the corresponding vineyard parcels. The main drivers of this diversity were the monoterpene alcohols linalool, α-terpineol, linalool oxide; the benzenoids vanillin, ethyl vanillate and methyl vanillate; and the norisoprenoid ß-damascenone. Wine from one vineyard parcel was not correctly classified, possibly due to the influence of the peculiar training system applied to this parcel. With aging the vineyard parcel geographical diversity was still reflected by the chemical diversity of wines, even if the separation was less fine. As many reactions occurred, some drivers of the diversity were changed after aging. They were benzenoids: ethyl vanillate, methyl vanillate and vanillin; the norisoprenoid 3-oxo-α-ionol; the terpenes linalool oxide, linalool, p-methane-1,8-diol, α-terpineol, and the precursors of nerol, geraniol, linalool.

Changes in chemical and sensory properties of Amarone wine produced by Penicillium infected grapes.

There is very little information on effects of Penicillium on aroma of passito wine. This study analyzed chemical composition and sensory properties of Amarone wines produced from withered grapes artificially contaminated by P. expansum or P. crustosum. Changes in properties of the two wines were evident by comparing wines obtained from healthy and Botrytis cinerea infected grapes used as controls. Penicillium infection affected primary and volatile composition of Amarone wine. Sensory profiles of these wines, obtained by descriptive analysis, resulted in clear differences in the wines between themselves and the control wines. Partial least square regression analysis explained only partially the relationship between molecules and sensory descriptors, and showed the existence of complex interactions of compounds mainly involved in specific aroma attributes. GC-olfactive analysis showed a greater number of odour regions in P. crustosum wine compared to control wines. Useful insight was provided into understanding how Penicillium rotten grapes affect Amarone wine properties.

Norisoprenoids, Sesquiterpenes and Terpenoids Content of Valpolicella Wines During Aging: Investigating Aroma Potential in Relationship to Evolution of Tobacco and Balsamic Aroma in Aged Wine.

During wine aging, tobacco and balsamic aroma notes appear. In this paper, volatile compounds directly or potentially related to those aromas have been investigated in Corvina and Corvinone wines during aging. Corvina and Corvinone are two northern-Italy autochthonous red grape varieties, used to produce Valpolicella Classico and Amarone wines, both characterized by tobacco and balsamic aroma notes. Wines were analyzed shortly after bottling or following model aging at 60°C for 48, 72, and 168 h. Volatile compounds were analyzed by HS-SPME-GC-MS. Results showed that compounds related to tobacco aroma [ß-damascenone, 3-oxo-α-ionol, (E)-1-(2,3,6-Trimethylphenyl)-buta-1,3-diene (TPB), and megastigmatrienones] increased in relationship to storage time with different patterns. ß-Damascenone and 3-oxo-α-ionol rapidly increased to reach a plateau in the first 48-72 h of model aging. Instead, TPB and megastigmatrienones concentration showed a linear correlation with aging time. During model aging, several cyclic terpenes tended to increase. Among them 1,8-cineole and 1,4-cineole, previously reported to contribute to red wine eucalyptus notes increased proportionally to storage time, and this behavior was clearly associated with reactions involving α-terpineol, limonene, and terpinolene, as confirmed by studies with model wine solutions. Among other relevant volatile compounds, sesquiterpenes appear to contribute potentially balsamic and spicy aroma notes. In this study, linear sesquiterpenes (nerolidol, farnesol) underwent acid hydrolysis during long wine aging, while cyclic sesquiterpenes seemed to increase with time. The chemical pathways associated with evolution of some of the compounds investigated have been studied in model wine.

Facile gas chromatography-tandem mass spectrometry stable isotope dilution method for the quantification of sesquiterpenes in grape.

Sesquiterpenes are a widespread class of compounds of increasing interest found in grapes and wines, amongst many other natural sources. Due to a lack of standards and the complexity of the mass spectra fragmentation, accurate quantification of these low concentration compounds had not previously been accomplished. The current paper presents a new method for the concurrent quantification of several sesquiterpenes. The multivariate method optimisation is presented. Synthesised isotopic standards were utilised in conjunction with solid phase microextraction (SPME) and gas chromatography-tandem mass spectrometry (GC-MS/MS) to perform a standard isotope dilution assay (SIDA). The method was successfully applied to several grape must samples of four different cultivar. To the best of our knowledge this was the first time some of these sesquiterpenes were quantified in grape.

Induction of Terpene Biosynthesis in Berries of Microvine Transformed with VvDXS1 Alleles.

Terpenoids, especially monoterpenes, are major aroma-impact compounds in grape and wine. Previous studies highlighted a key regulatory role for grapevine 1-deoxy-D-xylulose 5-phosphate synthase 1 (VvDXS1), the first enzyme of the methylerythritol phosphate pathway for isoprenoid precursor biosynthesis. Here, the parallel analysis of VvDXS1 genotype and terpene concentration in a germplasm collection demonstrated that VvDXS1 sequence has a very high predictive value for the accumulation of monoterpenes and also has an influence on sesquiterpene levels. A metabolic engineering approach was applied by expressing distinct VvDXS1 alleles in the grapevine model system "microvine" and assessing the effects on downstream pathways at transcriptional and metabolic level in different organs and fruit developmental stages. The underlying goal was to investigate two potential perturbation mechanisms, the former based on a significant over-expression of the wild-type (neutral) VvDXS1 allele and the latter on the ex-novo expression of an enzyme with increased catalytic efficiency from the mutated (muscat) VvDXS1 allele. The integration of the two VvDXS1 alleles in distinct microvine lines was found to alter the expression of several terpenoid biosynthetic genes, as assayed through an ad hoc developed TaqMan array based on cDNA libraries of four aromatic cultivars. In particular, enhanced transcription of monoterpene, sesquiterpene and carotenoid pathway genes was observed. The accumulation of monoterpenes in ripe berries was higher in the transformed microvines compared to control plants. This effect is predominantly attributed to the improved activity of the VvDXS1 enzyme coded by the muscat allele, whereas the up-regulation of VvDXS1 plays a secondary role in the increase of monoterpenes.