Screening of Oenological Yeasts for Volatile Phenol Release in Smoke-exposed Red Wine

Due to climate change, the higher incidence and severity of bushfires is a significant challenge for wine producers worldwide as an increase in smoke contamination negatively affects the physicochemical components that contribute to the lower quality of fresh produce and final products (smoke taint in wines). This reduces prices and consumer acceptability, impacting the producers and manufacturers. Current methods available to winemakers for assessing contamination in berries and wine consist of costly laboratory analyses that require skilled personnel and are time-consuming, cost prohibitive, and destructive. Therefore, novel, rapid, cost-effective, and reliable methods using digital technologies such as the use of near-infrared (NIR) spectroscopy, electronic nose (e-nose), and machine learning (ML) have been developed by our research group. Several ML models have been developed for smoke taint detection and quantification in berries and wine from different varieties using NIR absorbance values or e-nose raw data as inputs to predict glycoconjugates, volatile phenols, volatile aromatic compounds, smoke-taint amelioration techniques efficacy, and sensory descriptors, all models with >97% accuracy. These methods and models may be integrated and automated as digital twins to assess smoke contamination in berries and smoke taint in wine from the vineyard for early decision-making.

In recent years, the exposure of vineyards and grapes to smoke from bushfires and controlled burn events has in some instances resulted in wines described as smoke tainted. Such wines are characterised by undesirable sensory characters described as smoky, burnt, ash, smoky bacon, medicinal and ashtray. This review summarises the knowledge about the composition of smoke from forest and grass fires, describes relationships between smoke exposure of vineyards and smoke taint in wine, and outlines strategies for managing and reducing the risk to producing smoke-affected wines. The sensitivity of grapes and vines at different phenological stages to the uptake of contaminants from smoke, especially smoke-derived volatile phenols, is outlined, and the pathways for entry and metabolic transformation of volatile phenols are discussed. The potential for translocation of phenolic contaminants within the grapevine and the differences in uptake of smoke contaminants of different grape cultivars are also discussed, along with preliminary work on dose/response relationships regarding concentration and duration of exposure and subsequent expression of smoke taint in wine. The chemical basis of smoke taint in wine is described, and the relationship between volatile phenols from combustion of wood/lignin and their glycosides, and sensory panel ratings of smoke attributes in affected wines is discussed. This includes a summary of sensory reconstitution studies and measurement of flavour compounds released from nonvolatile glycosides in mouth and during in vitro model experiments, which corroborate the sensory contribution of glycosylated phenolic substances and their role as flavour precursors. Finally, the review also discusses analytical methods used to quantify free volatile phenolic substances and their associated glycosides, biomarkers for identifying smoke exposure of grapes, and strategies for assessing the risk of quality loss post-smoke exposure. Case studies are presented that outline the influence of harvesting and winemaking practices on the expression of smoke taint, and that describe oenological approaches to ameliorate smoke taint in wine.

Recent wildfires near vineyards in the Pacific United States have caused devastating financial losses due to smoke taint in wine. When wine grapes (Vitis vinifera) are exposed to wildfire smoke, their berries absorb volatile phenols derived from the lignin of burning plant material. Volatile phenols are released during the winemaking process giving the finished wine an unpleasant, smokey, and ashy taste known as smoke taint. Bacteria are capable of undergoing a wide variety of metabolic processes and therefore present great potential for bioremediation applications in many industries. In this study, we identify two strains of the same species that colonize the grape phyllosphere and are able to degrade guaiacol, a main volatile phenol responsible for smoke taint in wine. We identify the suite of genes that enable guaiacol degradation in Gordonia alkanivorans via RNAseq of cells growing on guaiacol as a sole carbon source. Additionally, we knockout guaA, a cytochrome P450 gene involved in the conversion of guaiacol to catechol; ΔguaA cells cannot catabolize guaiacol in vitro, providing evidence that GuaA is necessary for this process. Furthermore, we analyze the microbiome of berries and leaves exposed to smoke in the vineyard to investigate the impact of smoke on the grape microbial community. We found smoke has a significant but small effect on the microbial community, leading to an enrichment of several genera belonging to the Bacilli class. Collectively, this research shows that studying microbes and their enzymes has the potential to identify novel tools for alleviating smoke taint.

Smoke taint in wine is thought to be caused by smoke-derived volatile phenols (VPs) that are absorbed into grape tissues, trapped as conjugates that are imperceptible by smell, and subsequently released into wines as their free odor-active forms via metabolism by yeasts during fermentation. Blocking VP uptake into grapes would, therefore, be an effective way for vineyards to protect ripening grape crops exposed to smoke. Here, we re-evaluated a biofilm that had previously shown promise in pilot studies in reducing levels of smoke-derived VPs. A suite of nine free and acid-labile VPs were quantitated in Pinot Noir grapes that had been exposed to smoke after being coated with the biofilm one, seven or 14 days earlier. In contrast with earlier studies, our results demonstrated that in all cases, the biofilm treatments led to increased concentrations of both free and total VPs in smoke-exposed grapes, with earlier applications elevating concentrations of some VPs more than the later time points. Tracking VP concentrations through the grape ripening process demonstrated that some (phenol, p/m-cresol, and guaiacol) were not entirely sequestered in grapes as acid-labile conjugates, suggesting the presence of VP storage forms beyond simple glycosides. Free VPs in grapes, though a minor portion of the total, most clearly correlated with concentrations present in the resulting wines. Finally, red table grapes, available year round, were observed to replicate the effects of the biofilm treatments and were capable of transforming most VPs into acid-labile conjugates in under 24 h, indicating that they might be an effective model for rapidly assessing smoke-taint prophylactic products in the laboratory.

BACKGROUND AND AIMS: Fermentation of smoke-affected grapes can lead to wines that exhibit objectionable smoke-related sensory attributes, i.e. smoke taint. Fining agents are routinely used at different stages of the winemaking process to address constituents that are considered to adversely affect juice or wine quality. This study aimed to evaluate the efficacy of commercial fining agents in reducing the concentration of volatile phenols and the intensity of sensory attributes associated with smoke-tainted wine. METHODS AND RESULTS: Smoke-affected wines were treated with a range of fining agents, two of which, an activated carbon and a synthetic mineral, were found to appreciably ameliorate the taint. Treated wines contained a significantly lower level of smoke-derived volatile phenols and exhibited less intense ‘smoke’ and ‘cold ash’ aromas, ‘smoky’ flavour and ‘ashy’ aftertaste, compared with that of untreated (control) wines; with little or no impact on wine colour. CONCLUSIONS: Selected fining agents can ameliorate smoke taint in wine. Whereas most fining agents showed poor specificity towards the wine components responsible for smoke taint, some, an activated carbon in particular, were highly effective. SIGNIFICANCE OF THE STUDY: This research identifies a treatment that can be used to mitigate the impact of grapevine exposure to smoke on wine composition and sensory properties.

The frequency and intensity of wildfires have been increasing over the last 50 years and negatively impacted the wine industry. Previous methods of smoke mitigation during grape processing have shown little impact in reducing smoke taint in wines. Therefore, a novel method of using edible spray coatings for vineyard application was developed to help prevent volatile smoke phenol uptake in wine grapes. Four cellulose nanofiber-based coating suspensions incorporated with chitosan and/or β-cyclodextrin were evaluated. Films derived from the coating suspensions were exposed to volatile phenols found in wildfire smoke (guaiacol, 4-methyl guaiacol, m-cresol, o-cresol, p-cresol, syringol, and 4-methyl syringol) and evaluated with ultraviolet-visible spectroscopy where the results indicated that the coatings could uptake smoke phenols in varying degrees. The coatings were also applied in a vineyard at three different application times during grape growth: pea-sized, pre-bunch closure, and both at pea-sized and pre-bunch closure. The results showed that the application time did not have a significant (p < 0.05) effect on berry size, weight, °Brix, pH, or titratable acidity. The type of coating, time of application and washing were found to impact the number of volatile phenols in the grapes after a smoke event. Results from this study indicated that edible coatings could help mitigate smoke uptake in wine grapes without sacrificing the growth and key composition parameters of wine grapes. PRACTICAL APPLICATION: This research provides a novel spray coating that can be applied to wine grapes in the vineyard to potentially mitigate volatile smoke compounds in wine grapes without impacting fruit growth and key compositional parameters of wine grapes, thus maintaining high quality of wines for consumers. Results from this study can also be potentially applied to other agricultural commodities to solve the issues caused by the wildfire smoke.

When bushfires occur near grape growing regions, vineyards can be exposed to smoke, and depending on the timing and duration of grapevine smoke exposure, fruit can become tainted. Smoke-derived volatile compounds, including volatile phenols, can impart unpleasant smoky, ashy characters to wines made from smoke-affected grapes, leading to substantial revenue losses where wines are perceivably tainted. This study investigated the potential for post-harvest ozone treatment of smoke-affected grapes to mitigate the intensity of smoke taint in wine. Merlot grapevines were exposed to smoke at ~7 days post-veraison and at harvest grapes were treated with 1 or 3 ppm of gaseous ozone (for 24 or 12 h, respectively), prior to winemaking. The concentrations of smoke taint marker compounds (i.e., free and glycosylated volatile phenols) were measured in grapes and wines to determine to what extent ozonation could mitigate the effects of grapevine exposure to smoke. The 24 h 1 ppm ozone treatment not only gave significantly lower volatile phenol and volatile phenol glycoside concentrations but also diminished the sensory perception of smoke taint in wine. Post-harvest smoke and ozone treatment of grapes suggests that ozone works more effectively when smoke-derived volatile phenols are in their free (aglycone) form, rather than glycosylated forms. Nevertheless, the collective results demonstrate the efficacy of post-harvest ozone treatment as a strategy for mitigation of smoke taint in wine.

Wine made from grapes exposed to bushfire smoke can exhibit unpleasant smoky, ashy characters, which have been attributed to the presence of smoke-derived volatile phenols, in free or glycosylated forms. Here we report the uptake and glycosylation of volatile phenols by grapes following exposure of Cabernet Sauvignon vines to smoke, and their fate during winemaking. A significant delay was observed in the conversion of volatile phenols to their corresponding glycoconjugates, which suggests sequestration, the presence of intermediates within the glycosylation pathway and/or other volatile phenol storage forms. This finding has implications for industry in terms of detecting smoke-affected grapes following vineyard smoke exposure. The potential for an in-canopy sprinkler system to mitigate the uptake of smoke-derived volatile phenols by grapes, by spraying grapevines with water during smoke exposure, was also evaluated. While “misting” appeared to partially mitigate the uptake of volatile phenols by grapes during grapevine exposure to smoke, it did not readily influence the concentration of volatile phenols or the sensory perception of smoke taint in wine. Commercial sensors were used to monitor the concentration of smoke particulate matter (PM) during grapevine exposure to low and high density smoke. Similar PM profiles were observed, irrespective of smoke density, such that PM concentrations did not reflect the extent of smoke exposure by grapes or risk of taint in wine. The sensors could nevertheless be used to monitor the presence of smoke in vineyards during bushfires, and hence, the need for compositional analysis of grapes to quantify smoke taint marker compounds.

The number and intensity of wildfires are increasing worldwide, thereby raising the risk of smoke contamination of grapevine berries and the development of smoke taint in wine. This study aimed to develop five artificial neural network (ANN) models from berry, must, and wine samples obtained from grapevines exposed to different levels of smoke: (i) Control (C), i.e., no misting or smoke exposure; (ii) Control with misting (CM), i.e., in-canopy misting, but no smoke exposure; (iii) low-density smoke treatment (LS); (iv) high-density smoke treatment (HS) and (v) a high-density smoke treatment with misting (HSM). Models 1, 2, and 3 were developed using the absorbance values of near-infrared (NIR) berry spectra taken one day after smoke exposure to predict levels of 10 volatile phenols (VP) and 18 glycoconjugates in grapes at either one day after smoke exposure (Model 1: R = 0.98; R2 = 0.97; b = 1) or at harvest (Model 2: R = 0.98; R2 = 0.97; b = 0.97), as well as six VP and 17 glycoconjugates in the final wine (Model 3: R = 0.98; R2 = 0.95; b = 0.99). Models 4 and 5 were developed to predict the levels of six VP and 17 glycoconjugates in wine. Model 4 used must NIR absorbance spectra as inputs (R = 0.99; R2 = 0.99; b = 1.00), while Model 5 used wine NIR absorbance spectra (R = 0.99; R2 = 0.97; b = 0.97). All five models displayed high accuracies and could be used by grape growers and winemakers to non-destructively assess at near real-time the levels of smoke-related compounds in grapes and/or wine in order to make timely decisions about grape harvest and smoke taint mitigation techniques in the winemaking process.

Background and Aims: Wines made from grapes harvested from vineyards exposed to bushfire smoke can exhibit objectionable ‘smoky’, ‘cold ash’, ‘medicinal’ and ‘ashy’ aroma and flavour characters. This study evaluated a combined reverse osmosis and solid phase adsorption process as a potential amelioration method for the treatment of smoke-tainted wines. Methods and Results: Smoke-tainted wines were treated using either pilot or commercial scale reverse osmosis systems and the chemical composition and sensory properties of wine compared before and after treatment. The concentrations of smoke-derived volatile phenols, including marker compounds, guaiacol and 4-methylguaiacol, decreased significantly with treatment. As a consequence, diminished smoke-related sensory attributes enabled treated wines to be readily differentiated from untreated wines. However, the taint was found to slowly return with time, likely because of hydrolysis of glycoconjugate precursors, which were not removed during the treatment process. Conclusions: Reverse osmosis and solid phase adsorption reduced the concentration of smoked-derived volatile phenols and improved the sensory attributes of smoke-tainted wines. Significance of the Study: This is the first study to evaluate the amelioration of smoke taint in wine using reverse osmosis and solid phase adsorption.

Where vineyard exposure to bushfire smoke cannot be avoided or prevented, grape and wine producers need strategies to transform smoke-affected juice and wine into saleable product. This study evaluated the potential for spinning cone column (SCC) distillation to be used for the remediation of 'smoke taint'. Compositional analysis of 'stripped wine' and condensate collected during SCC treatment of two smoke-tainted red wines indicated limited, if any, removal of volatile phenols, while their non-volatile glycoconjugates were concentrated due to water and ethanol removal. Together with the removal of desirable volatile aroma compounds, this enhanced the perception of smoke-related sensory attributes; i.e., smoke taint intensified. Stripped wines also became increasingly sour and salty as ethanol (and water) were progressively removed. A preliminary juice remediation trial yielded more promising results. While clarification, heating, evaporation, deionization and fermentation processes applied to smoke-tainted white juice gave ≤3 µg/L changes in volatile phenol concentrations, SCC distillation of smoke-tainted red juice increased the volatile phenol content of condensate (in some cases by 3- to 4-fold). Deionization of the resulting condensate removed 75 µg/L of volatile phenols, but fermentation of reconstituted juice increased volatile phenol concentrations again, presumably due to yeast metabolism of glycoconjugate precursors. Research findings suggest SCC distillation alone cannot remediate smoke taint, but used in combination with adsorbents, SCC may offer a novel remediation strategy, especially for tainted juice.

Smoke taint in wine has become a critical issue in the wine industry due to its significant negative impact on wine quality. Data-driven approaches including univariate analysis and predictive modeling are applied to a data set containing concentrations of 20 VOCs in 48 grape samples and 56 corresponding wine samples with a taster-evaluated smoke taint index. The resulting models for predicting the smoke taint index of wines are highly predictive when using as inputs VOC concentrations after log conversion in both grapes and wines (Pearson Correlation Coefficient PCC = 0.82; R2 = 0.68) and less so when only grape VOCs are used (Pearson Correlation Coefficient PCC = 0.76; R2 = 0.56), and the classification models also show the capacity for detecting smoke-tainted wines using both wine and grape VOC concentrations (Recall = 0.76; Precision = 0.92; F1 = 0.82) or using only grape VOC concentrations (Recall = 0.74; Precision = 0.92; F1 = 0.80). The performance of the predictive model shows the possibility of predicting the smoke taint index of the wine and grape samples before fermentation. The corresponding code of data analysis and predictive modeling of smoke taint in wine is available in the Github repository (https://github.com/IBPA/smoke_taint_prediction).

Rapid headspace solid-phase microextraction sheets with direct analysis in real time mass spectrometry (SPMESH-DART-MS) of derivatized volatile phenols in grape juices and wines

Bushfire smoke can affect the composition and sensory properties of grapes and wines, in some cases leading to wines which exhibit undesirable “smoky”, “ashy” and “medicinal” characters. This study investigated the extent to which fruit maturity (i.e., ripeness) influences the perception of smoke taint in wine. Two white grape varieties (Chardonnay and Sauvignon Blanc) and two red grape varieties (Merlot and Shiraz) were exposed to smoke under experimental conditions, at approximately seven days post-veraison. Fruit was then harvested at two levels of maturity: Harvest A, when total soluble solids were 16–20 °Brix, i.e., the berry ripeness typically required for production of sparkling or light-bodied wines; and Harvest B, when total soluble solids were 22–25 °Brix, i.e., the berry ripeness typically required for production of full-bodied wines. The intensity of smoke taint in resulting wines was found to be influenced by fruit maturity, but differed between grape varieties. Smoke-related sensory attributes were apparent in Sauvignon Blanc wine made from early-harvested fruit and in Chardonnay wine made from late-harvested fruit, only; whereas Merlot and Shiraz wines exhibited smoke taint irrespective of fruit maturity. Smoke-derived volatile phenols, and various alcohols, esters and acids, were also quantified to determine the impact of smoke exposure and fruit maturity respectively, on wine composition.

Bushfires are becoming an increasing issue for the wine sector due to grape and vine losses and smoke taint in wine. Smoke affects vine physiology and the smoke's volatile phenols are absorbed by plants and berries, contaminating the wine. Our hypothesis was that, for the first time, unmanned aerial vehicle (UAV)-based visible images can be used to study the physiology of smoke-affected vines and to assess compromised vines. Procanico vines were exposed to two smoke treatments, a week apart. Gas exchanges and leaf biochemical traits were measured in the short term (30 min after smoke exposure) and in the long term (24 h after smoke exposure). Canopy damage was assessed with conventional vegetation indices (VIs) and by an innovative index derived by UAV-based visible images, the Canopy Area Health Index (CAHI). Gas exchange showed a reduction after the first smoke exposure, but the vines recovered within 24 h. The second smoke exposure led to an irreversible reduction in functional parameters. The VIs exhibited significant differences and CAHI presented a damage gradient related to bushfire nearby. The vineyard damage assessment by UAV-based visible images may represent a tool to study the physiological activity of smoke-affected vines and to quantify the loss of destroyed or damaged vines. © 2020 Society of Chemical Industry.