Impact of light on protective fractions of Cu in white wine: Influence of oxygen and bottle colour.

Cu(II)-organic acid (fraction I) and Cu(I)-thiol (fraction II) complexes can suppress sulfhydryl off-aromas in wine. This study investigated the impact of light exposure on the protective fractions of Cu of bottled white wine. Fluorescent light-exposed Chardonnay with two initial concentrations of dissolved oxygen (0.5 and 10 mg/L) was stored in different coloured bottles and concentrations of Cu fractions and riboflavin, a photo-initiator at 370-440 nm, were measured during 110 days storage. Light-exposed wines with lower oxygen concentrations resulted in a 100-fold decrease in the Cu fraction I half-life, and a 60-fold decrease for Cu fractions I and II combined. The half-life for Cu fraction I decay during light exposure was extended 30-fold with the use of brown compared to flint glass. Light exposure can rapidly exhaust the protective Cu fractions in wine, and bottles with less light transmission below 440 nm can slow this loss.

Validation of Raman spectroscopic models to verify the origin of Australian beef grown under different production systems.

Verification of beef production systems and authentication of origin is becoming increasingly important as consumers base purchase decisions on a greater number of perceived values including the healthiness and environmental impact of products. Previously Raman spectroscopy has been explored as a tool to classify carcases from grass and grain fed cattle. Thus, the aim of the current study was to validate Partial Least Squares Discriminant Analysis (PLS-DA) models created using independent samples from carcases sampled from northern and southern Australian production systems in 2019, 2020 and 2021. Validation of the robustness of discrimination models was undertaken using spectral measures of fat from 585 carcases which were measured in 2022 using a Raman handheld device with a sample excised for fatty acid analysis. PLS-DA models were constructed and then employed to classify samples as either grass or grain fed in a two-class model. Overall, predictions were high with accuracies of up to 95.7% however, variation in the predictive ability was noted with models created for southern cattle yielding an accuracy of 73.2%. While some variation in fatty acids and therefore models can be attributed to differences in genetics, management and diet, the impact of duration of feeding is currently unknown and thus further work is warranted.

Rapid In-Field Volatile Sampling for Detection of Botrytis cinerea Infection in Wine Grapes.

Fungal infection of grape berries (Vitis vinifera) by Botrytis cinerea frequently coincides with harvest, impacting both the yield and quality of grape and wine products. A rapid and non-destructive method for identifying B. cinerea infection in grapes at an early stage prior to harvest is critical to manage loss. In this study, zeolitic imidazolate framework-8 (ZIF-8) crystal was applied as an absorbent material for volatile extraction from B. cinerea infected and healthy grapes in a vineyard, followed by thermal desorption gas chromatography-mass spectrometry. The performance of ZIF-8 in regard to absorbing and trapping the targeted volatiles was evaluated with a standard solution of compounds and with a whole bunch of grapes enclosed in a glass container to maintain standard sampling conditions. The results from the sampling methods were then correlated to B. cinerea infection in grapes, as measured and determined by genus-specific antigen quantification. Trace levels of targeted compounds reported as markers of grape B. cinerea infection were successfully detected with in-field sampling. The peak area counts for volatiles 3-octanone, 1-octen-3-one, 3-octanol, and 1-octen-3-ol extracted using ZIF-8 were significantly higher than values achieved using Tenax®-TA from field testing and demonstrated good correlation with B. cinerea infection severities determined by B. cinerea antigen detection.

Detection and prediction of Botrytis cinerea infection levels in wine grapes using volatile analysis.

Infection of grape berries (Vitis vinifera) by the fungus Botrytis cinerea (grey mould) frequently occurs in vineyards, resulting in off-flavours and other odours in wine and potential yield losses. In this study, volatile profiles of four naturally infected grape cultivars, and laboratory-infected grapes were analysed to identify potential markers for B. cinerea infection. Selected volatile organic compounds (VOCs) were highly correlated with two independent measures of B. cinerea infection levels, demonstrating that ergosterol measurements provide accurate quantification of lab-inoculated samples, while B. cinerea antigen detection is more suitable for naturally infected grapes. Excellent predictive models of infection level were confirmed (Q2Y of 0.784-0.959) using selected VOCs. A time course experiment confirmed that selected VOCs 1,5-dimethyltetralin, 1,5-dimethylnaphthalene, phenylethyl alcohol and 3-octanol are good markers for B. cinerea quantification and 2-octen-1-ol could be considered as an early marker of the infection.

Characterisation of internal oxygen concentration of strawberry (Fragaria × ananassa) and blueberry (Vaccinium corymbosum).

Gas exchange mechanisms play crucial roles in maintaining fruit post-harvest quality in perishable fruit such as strawberry (Fragaria×ananassa Duch.) and blueberry (Vaccinium corymbosum L.). The internal oxygen concentration ([O2 ]) of strawberry and blueberry were measured using Clark-type oxygen sensing electrodes. The volume of intercellular voids in strawberry was obtained by micro-computed tomography (micro-CT). In both berries, internal [O2 ] was consistent and relatively high across measured tissues. The overall [O2 ] was well above the Michaelis constant (K m ) for cytochrome c oxidase in both fruit and different from previously examined grape (Vitis vinifera L.) berry mesocarp with near zero minimum [O2 ]. In strawberry and blueberry, cell vitality was also maintained at full maturity in the mesocarp. Higher storage temperature (i.e. 20 vs 4°C) reduced internal [O2 ] of strawberry. Pedicel detachment in blueberry was associated with greater fruit dehydration and lower internal [O2 ] after short-term storage of 12h. The results suggest that the intercellular voids of the fruit's mesocarp provide an efficient gas exchange route for maintaining high fruit internal [O2 ] post-harvest.

Assessing chemometric models developed using Raman spectroscopy and fatty acid data for Northern and Southern Australian beef production systems.

A total of 960 beef carcases from northern and southern Australian production regions were assessed by examining the subcutaneous fat. Carcases from four different production systems within each region were assessed, by Raman spectroscopy and the fatty acid composition determined to develop models that best classified the various production systems. As a result, 12 Partial Least Square Discriminant Analysis models were developed. A two-class model based on fatty acid composition was able to correctly classify 99% of grass and grain fed animals. The best Raman spectroscopic model correctly classified 94% of grass vs grain carcases produced in the northern region. For the southern production region, the models had the following classification accuracies; southern long-term grain fed (98%), southern short-term grain fed (95%), southern grass (96%), southern grass supplemented (97%), and the southern model classified grass vs grain (97%). Raman spectroscopy is considered a useful rapid method for classification of beef carcases based upon production system.

Vascular Connections Into the Grape Berry: The Link of Structural Investment to Seededness.

Vascular bundles in the grape pedicel and berry contain the conduits, phloem and xylem, for transport of water, sugar, nutrients and signals into and through the grape berry and play a critical role in berry growth and composition. Here, we assess the vascular anatomy within the proximal region of the berry. Guided using a 3D berry model generated by micro-CT, differential staining of transverse sections of berries and receptacles was followed by fluorescent microscopy. Morphometric and vascular characteristics were analyzed within the central proximal region (brush zone, a fibrous extension from the pedicel vascular system into the berry) of the seeded cultivars Shiraz and Sauvignon Blanc, as well as the stenospermocarpic cultivars Ruby Seedless and Flame Seedless. Observations revealed a change in vascular arrangement from the receptacle into the berry brush zone and differences in xylem element size as well as xylem and phloem area relationships. Xylem anatomical and derived hydraulic parameters, as well as total tissue area of xylem and phloem varied between cultivars and in receptacle and berry components. Variation in vascular growth between grape pedicels and berries was independent of seededness. Differences in receptacle xylem vessel size and distribution could contribute to cultivar-dependent xylem backflow constraint.

Elucidating the interaction of carbon, nitrogen, and temperature on the biosynthesis of Aureobasidium pullulans antifungal volatiles.

The combined biochemical impact of carbon, nitrogen and temperature on the biosynthesis of the antifungal volatile organic compounds (VOCs): ethanol, 2-methyl-1-propanol, 3-methyl-1-butanol and 2-phenylethanol produced by Aureobasidium pullulans A1 and A3 was investigated using a Box-Behnken experimental design and response surface methodology (RSM). Normalized peak areas derived from solid phase micro extraction-gas chromatography-mass spectrometry (SPME-GC-MS) analysis, indicated that initial carbon content had a significant influence on the biosynthesis of ethanol and alcohols with greater than three carbon atoms. This result suggests a dominant activity of the A. pullulans anabolic pathway to biosynthesize three higher alcohols via de novo biosynthesis of amino acids from sugar metabolism. Low concentrations of carbon (3-13 g l-1 ) with nitrogen as both ammonium and amino acids in the growth medium resulted in a higher number of significant linear and quadratic relationships. Nitrogen availability and growth temperature had significant negative linear and quadratic correlations with VOCs biosynthesis in most instances. Isolate-dependant metabolic response was evident for all abiotic parameters tested on alcohol production. The findings of this study offer new perspectives to improve the production of key antifungal compounds by antagonists in biological control systems.

Secondary Metabolites Coordinately Protect Grapes from Excessive Light and Sunburn Damage during Development.

Sunburn is a physiological disorder that reduces grape quality and vineyard yield. It is the result of excessive sunlight and high temperatures. As climate change continues to increase air temperatures, reports of sunburn damage in vineyards worldwide are becoming more frequent. Grapes produce secondary metabolites (carotenoids, polyphenols and aroma compounds) to counter photooxidative stress and acclimate to higher radiation environments. This study evaluated changes in these compounds in during ripening when grapes were exposed post-flowering (ED) and at véraison (LD), and compared them to a nondefoliated control (ND). ND contained more α-terpineol and violaxanthin, and the defoliated treatments contained more zeaxanthin, ß-carotene, C6 compounds and flavonoids. ED berries adapted better to higher-light environments, displayed larger changes in secondary metabolite concentrations and lower levels of sunburn damage than LD berries did. The composition of berries with increasing sunburn damage was evaluated for the first time. Berries with no damage had the lowest concentrations of flavonoids and oxidized glutathione, and the highest concentrations of chlorophyll and α-terpineol. As damage increased, destruction of photosynthetic pigments, increase in polyphenols and loss of aroma compounds were evidenced. A significant effect of temperature and developmental stage on grape composition was also observed. This study provides a holistic overview of changes in secondary metabolites experienced by grape berries when exposed to excessive light, how these vary along development and how they affect sunburn incidence.

Volatile organic compounds produced by Aureobasidium pullulans induce electrolyte loss and oxidative stress in Botrytis cinerea and Alternaria alternata.

Aureobasidium pullulans is a yeast-like fungus that produces volatile organic compounds (VOCs) with antifungal properties. VOCs have the potential to trigger the production of intracellular reactive oxygen species (ROS), lipid peroxidation and electrolyte loss in microorganisms. The relationship among A. pullulans VOCs, induced ROS accumulation and electrolyte leakage was investigated in Botrytis cinerea and Alternaria alternata in vitro. Exposure to a mixture of A. pullulans VOCs: ethanol, 2-methyl-1-propanol, 3-methyl-1-butanol and 2-phenylethanol, resulted in electrolyte leakage in both B. cinerea and A. alternata. Fluorescence microscopy using 2',7'-dichlorofluorescein diacetate indicated triggered ROS accumulation in exposed fungal mycelia and the presence of the superoxide radical was evident by intense red fluorescence with dihydroethidium. Partial inhibition of enzymes of the mitochondrial respiratory chain complex I of B. cinerea and A. alternata by pre-treatment with rotenone reduced ROS accumulation in hypha exposed to A. pullulans VOCs and reversed the VOCs inhibition of fungal growth. Scanning electron micrographs revealed that B. cinerea and A. alternata hypha exposed to A. pullulans VOCs had altered cell wall structures. Our findings give insights into the potential mechanisms involved in the antifungal properties of A. pullulans in the suppression of B. cinerea and A. alternata growth in vitro.

Nanosecond Pulsed Dielectric Barrier Discharge Ionization Mass Spectrometry.

Dielectric barrier discharge ionization (DBDI) is an emerging technique for ionizing volatile molecules directly from complex mixtures for sensitive detection by mass spectrometry (MS). In conventional DBDI, a high frequency and high voltage waveform with pulse widths of ∼50 µs (and ∼50 µs between pulses) is applied across a dielectric barrier and a gas to generate "low temperature plasma." Although such a source has the advantages of being compact, economical, robust, and sensitive, background ions from the ambient environment can be formed in high abundances, which limits performance. Here, we demonstrate that high voltage pulse widths as narrow as 100 ns with a pulse-to-pulse delay of ∼900 µs can significantly reduce background chemical noise and increase ion signal. Compared to microsecond pulses, ∼800 ns pulses can be used to increase the signal-to-noise and signal-to-background chemical noise ratios in DBDI-MS by up to 172% and 1300% for six analytes, including dimethyl methylphosphonate (DMMP), 3-octanone, and perfluorooctanoic acid. Using nanosecond pulses, the detection limit for DMMP and PFOA in human blood plasma can be lowered by more than a factor of 2 in comparison to microsecond pulses. In "nanopulsed" plasma ionization, the extent of internal energy deposition is as low as or lower than in electrospray ionization and micropulsed plasma ionization based on thermometer ion measurements. Overall, nanosecond high-voltage pulsing can be used to significantly improve the performance of DBDI-MS and potentially other ion sources involving high voltage waveforms.

Changes in Red Wine Composition during Bottle Aging: Impacts of Grape Variety, Vineyard Location, Maturity, and Oxygen Availability during Aging.

This work investigated the influence of grape variety, vineyard location, and grape harvest maturity, combined with different oxygen availability treatments, on red wine composition during bottle aging. Chemometric analysis of wine compositional data (i.e., wine color parameters, SO2, metals, and volatile compounds) demonstrated that the wine samples could be differentiated according to the different viticultural or bottle-aging factors. Grape variety, vineyard location, and grape maturity showed greater influence on wine composition than bottle-aging conditions. For most measured wine compositional variables, the evolution patterns adopted from the viticultural factors were not altered by oxygen availability treatment. However, contrasting evolution patterns for some variables were observed according to specific viticultural factors, with examples including dimethyl sulfide, phenylacetaldehyde, maltol, and ß-damascenone for vineyard locations, 2-methylbutanal, 1,4-cineole, and linalool for grape variety, and methanethiol, methional, and homofuraneol for grape maturity.

Geographical Origin Has a Greater Impact on Grape Berry Fungal Community than Grape Variety and Maturation State.

We used barcoded sequencing to analyze the eukaryotic population in the grape berries at different ripening states in four Australian vineyards. Furthermore, we used an innovative compositional data analysis for assessing the diversity of microbiome communities. The novelty was the introduction of log-ratio balances between the detected genera. Altogether, our results suggest that fungal communities were more impacted by the geographical origin of the Australian vineyards than grape variety and harvest time. Even if the most abundant genera were Aureobasidium and Mycosphaerella, they were ubiquitous to all samples and were not discriminative. In fact, the balances and the fungal community structure seemed to be greatly affected by changes of the genera Penicillium, Colletotrichum, Aspergillus, Rhodotorula, and Botrytis. These results were not evident from the comparison of relative abundance based on OTU counts alone, remarking the importance of the balance analysis for microbiome studies.

Regional Discrimination of Australian Shiraz Wine Volatome by Two-Dimensional Gas Chromatography Coupled to Time-of-Flight Mass Spectrometry.

Shiraz wine volatomes from two Australian geographical indications (GIs), that is, Orange and Riverina, were compared using comprehensive two-dimensional gas chromatography coupled to time-of-flight mass spectrometry. Shiraz wines were made in triplicate from grapes harvested at two harvest dates from six vineyards in the two GIs. A total of 133 compounds showed a significant trend between wines from the cooler Orange GI and warmer Riverina. Compounds associated with wines from the cooler climate were grape-derived volatiles, such as monoterpenes, sesquiterpenes, green leaf volatiles, and some norisoprenoids. Fermentation-derived compounds, such as esters and S-containing compounds, showed no specific trend related to grape origin. In addition, wines could be also clearly separated according to the harvest date, irrespective of the climate, with C6 compounds, higher alcohol acetates, and other esters contributing utmost to the differentiation of samples, whereas terpenoids and norisoprenoids did not have an influence. This study demonstrated the plasticity of wine volatome related to grape origin and also the maturity level (harvest date), irrespective of climate.

A GC-MS untargeted metabolomics approach for the classification of chemical differences in grape juices based on fungal pathogen.

Fungal bunch rot of grapes leads to production of detrimental flavour compounds, some of which are well characterised but others remain unidentified. The current study uses an untargeted metabolomics approach to classify volatile profiles of grape juices based on the presence of different fungal pathogens. Individual grape berries were inoculated with Botrytis cinerea, Penicillium expansum, Aspergillus niger or A. carbonarius. Grape bunches were inoculated and blended with healthy fruit, to provide 10% (w/w) infected juice. Juices from the above sample batches were analysed by GC/MS. PLS-DA of the normalised summed mass ions indicated sample classification according to pathogen. Compounds identified from those mass ion matrices that had high discriminative value for classification included 1,5-dimethylnaphthalene and several unidentified sesquiterpenes that were relatively higher in B. cinerea infected samples. A. niger and A. carbonarius samples were relatively higher in 2-(4-hexyl-2,5-dioxo-2,5-dihydrofuran-3-yl)acetic acid, while P. expansum samples were higher in γ-nonalactone and m-cresol.

Analytical strategies for the measurement of different forms of Cu and Fe in wine: Comparison between approaches in relation to wine composition.

The speciation of Cu and Fe in wine was assessed by a number of methodologies and those with superior performance were utilised on 49 wines and compared to compositional data. The adopted analytical strategies were stripping potentiometry, HPLC and an extraction (solid-phase or liquid-liquid) followed by atomic absorption or optical emission spectroscopic measurement. Stripping potentiometry was specific for sulfide-bound Cu in wines, and showed that this was the predominant form of Cu. A solid phase extraction technique provided a hydrophobic fraction of Cu that was specific for sulfide-bound Cu in white wine but red wine matrix effects hindered isolation of sulfide-bound Cu. The other methods assessed either perturbed the sulfide-bound Cu or were not sufficiently sensitive. The solid phase extraction method enabled routine fractionation of Fe in red and white wines whilst the additional techniques surveyed either measured total Fe or suffered from background contamination.

Unravelling wine volatile evolution during Shiraz grape ripening by untargeted HS-SPME-GC × GC-TOFMS.

The influence of grape maturity on wine volatome was investigated using HS-SPME-GC × GC-TOFMS. Shiraz wines were made from grapes harvested from four different vineyards from two berry maturity levels. A total of 1276 putative compounds were detected in at least one of the wine samples and 175 showed significant trends related to grape maturity. The first two dimensions of the Principal component analysis accounted for 57% of the variation and separated the samples according to the harvest date. Wines from the first harvest date were characterised by an abundance of lipoxygenase derived compounds, norisoprenoids and sulfur-containing compounds whereas a significant increase in some acetate esters was observed in wines produced from the more mature grapes. This study demonstrated a common evolution of grape volatiles for Shiraz inside the same mesoclimate. During the late ripening stage of the grape, a direct nexus between sugar concentration and wine volatile evolution was not observed.

Late-Season Shiraz Berry Dehydration That Alters Composition and Sensory Traits of Wine.

Late-season berry dehydration (LSD) is a common occurrence in Shiraz grapes, particularly those grown in hot climates. LSD results in significant yield reductions; however, the effects on wine composition and sensory characteristics are not well-documented. Wines made of 100% nonshriveled clusters (control) were related to red fruit flavors by the sensory panel, whereas wines made of 80% shriveled clusters (S-VCT) were perceived as more alcoholic and associated with dark fruit and dead/stewed fruit characters. The latter wines also resulted in higher concentrations of massoia lactone and γ-nonalactone, compounds known to contribute to prune and stewed-fruit aromas. Wines made of shriveled grapes were also characterized by an increase in C6-alcohols and a decrease in esters, whereas wine terpenoids were altered compound-specific. An increase in orange pigments and wine chemical age in S-VCT wines indicated faster oxidative aging compared to the control. LSD appeared to alter final wine composition directly but also appeared to influence yeast metabolism, potentially due to an alteration of the composition of lipids in the grape juice. This study emphasized the relevance of sorting shriveled and nonshriveled berries for final wine chemical composition and wine style.

Volatile and sensory profiling of Shiraz wine in response to alcohol management: comparison of harvest timing versus technological approaches.

The aim of this study was to compare the volatile and sensory profiles of Australian Shiraz red wines produced by several methods to achieve alcohol concentrations of 10.5 and 13.5% v/v. These levels were considerably lower contents than the commercial wine (16-17% v/v) that was produced from this vineyard site. Wines were produced by: (i) harvest timing (19.3, 24 and 29.3 Brix); (ii) blending equal proportions of early harvest (19.3 Brix) and late harvest wines (29.3 Brix); and (iii) dealcoholization using reverse osmosis followed by a membrane contactor. Dealcoholization caused a significant loss of volatile compounds, particularly esters, while the blending treatment had an averaging effect on most analytes. Sensory descriptive analysis of treatments with 10.5% v/v alcohol showed that the perception of the herbaceous attribute was more intense in the early harvest wines in comparison to the dealcoholized wines, while those of dark fruit, raisin/prune, astringency and alcohol were lower. No sensory differences were found amongst the 13.5% v/v wines, except for alcohol. Sensory and compositional data were modelled by means of Common Dimension (ComDim) multi-block analysis and indicated which chemical components are important to the perceived wine sensory properties. Insights from this study will provide knowledge that may be applied to control or moderate both unripe sensory attributes in addition to a deficiency of ripe fruit aromas or mouthfeel characteristics in reduced-alcohol red wines.

A comparative study of partial dealcoholisation versus early harvest: Effects on wine volatile and sensory profiles.

Two Verdelho and Petit Verdot wines were produced from sequential harvests of grapes. The alcohol concentration of early harvest (EH) and late harvest (LH) wines were respectively 9% and 13.5% v/v for Verdelho, and 10.5% and 13% v/v for Petit Verdot. LH wines were dealcoholised to match the same alcohol level of EH samples using a combined reverse osmosis-evaporative perstraction process. In dealcoholised wines, there was a decrease in volatile compounds (esters particularly) compared to LH treatments. For both varieties, the sensory attribute ratings for overall aroma intensity and alcohol mouthfeel also decreased following dealcoholisation. Dealcoholised wines were distinctively different from both LH and EH wines even though these wines had similar alcohol level to EH wines. When dealcoholisation is considered for high-alcohol wines, it is important to consider that membrane effects can significantly change depending on the wine non-volatile matrix composition and the level of alcohol reduction required.