Introduction

One of the most important and difficult tasks for a viticulturist or winemaker is predicting/assessing harvest dates that will ensure the production of wines with preferred styles. More than ever, within a context of global warming, wine producers need objective indicators of grape maturity in order to make informed decisions about when to harvest. The progressive development of grape-derived aromas and flavour precursors during berry ripening has garnered the attention of researchers over recent years (Kontoudakis et al., 2010, Bindon et al., 2013; Cramer et al., 2014; Pascual et al., 2016; Rabot et al., 2017; Böttcher et al., 2018; Boss et al., 2018; Allamy et al., 2018). The decision to harvest grapes for wine production can be determined by measuring grape berry sugar concentration as total soluble solids and expressed as °Brix (technological maturity). Brix degree may be the sole criteria for harvest decision or considered in combination with basic analytical parameters related to berry acidity (pH, titratable acidity) and colour for red cultivars; phenolic maturity is generally assessed by performing analysis of anthocyanins and tannins content in grape skin and seed, and their level of extractability. Wine aroma is one of the most important components of wine quality, but none of the mentioned attributes gives objective information about the aromatic potential of a grape or the resulting wine aromatic profile (Deloire, 2013; Calderon-Orellana et al., 2014). Up to now, berry tasting is the main method used in the field to assess aromatic maturity. While berry tasting is relevant, it is also highly subjective as the perception of flavours depends on the taster's personal ability, experience and training (Rabot et al., 2017). Therefore, it would be useful for growers and winemakers to be able to objectively predict wine aromatic profile evolution during ripening by measuring simple physiological grape parameters, such as berry sugar accumulation which is dependent on vineyard growing conditions (soil × climate × cultural practices).

Empirically, the aroma of red wines made from grapes from early to later harvests (i.e., grapes of increasing maturity levels) are often described as green/herbaceous, fresh fruit, mature/ripe fruit and jammy/stewed fruit. Sensory studies have demonstrated such aromatic evolution in Merlot, CabernetSauvignon, Petit Verdot and Shiraz wines through sequential harvest (Casassa et al., 2013; Bindon et al., 2013; Longo et al., 2018a; Longo et al., 2018b). However, in all these studies the choice of harvest dates were based on grape sugar concentration, with harvests being performed from 19 °Brix onwards. A lack of correlation between berry sugar content and aroma development has recently been found in Shiraz wines made from grapes harvested from end of veraison onwards (Boss et al., 2018; Šuklje et al., 2019a). The aromatic profile of red wines made from sequential harvested grapes post plateau of berry sugar accumulation evolves from fresh fruit to mature and jammy fruit; however, in this period, sugar concentration (°Brix) is directly related to berry volume evolution and does not seem to be the most appropriate indicator of aromatic maturity. On the other hand, a recent study using an untargeted analysis was able to demonstrate a coherent evolution of wine volatome in Shiraz wines made from sequential harvested grapes based on the berry sugar accumulation method (Šuklje et al., 2019a). Therefore, the objective of the present study was to demonstrate the existence of reproducible aromatic wine styles by carrying out sequential harvests based on the measurement of grape berry sugar accumulation (mg/berry and not sugar concentration). Wines were made from Shiraz and Cabernet-Sauvignon grapes sourced during two consecutive vintages from nine vineyards located in three different mesoclimate areas. It was hypothesised that genotype wine influences aromatic evolution during berry ripening more than environment; therefore, a variety of vineyards and mesoclimates was accordingly chosen. The evolution of nexuses in grape composition, wine composition and the wine sensory profile were also investigated.

Materials and Method

2. Harvest and winemaking

Harvest dates were chosen using the Sugar Accumulation method as described by Deloire (2013), and they are delineated in Figure 1. A representative 10 bunches were sampled in triplicate per block on a weekly basis from veraison onwards. In the laboratory, 100 berries per replicate were carefully excised with the pedicel to avoid juice loss. Berries were weighed and then crushed for measurements of juice total soluble solids (TSS) content (expressed as Brix degree) and sugar per berry (mg/berry) using sugar concentration (expressed in g/L; 1 °Brix = 10 g/L) and berry fresh mass. Harvest dates linked to potential final wine styles were determined for Shiraz and Cabernet-Sauvignon using the proposed sugar accumulation and wine style models as shown in Figure 1. Two and three successive harvests (H1, H2 and H3) were conducted during the 2014 and 2015 vintages respectively at 12, 18 and 24 days post sugar accumulation plateau for Shiraz and 20, 30 and 40 days post sugar accumulation plateau for CabernetSauvignon. The timing of sequential harvests for both varieties was based on previous studies that suggested there is a relationship between the time after plateau of sugar accumulation and the style of the corresponding wines assessed by sensory evaluation (www.vivelys.com). Harvests did not deviate from these models by more than 2 days. The first harvest (H1), termed Fresh Fruit (FF), was thought to be linked to fresh red fruit notes in wines, while the third harvest (H3), termed Mature Fruit (MF), was associated with a mature dark fruit style. An intermediate harvest date (H2) was introduced during the 2015 vintage. Dates for sugar accumulation plateaus and harvests are presented in Table 2. Wines were made from 60 kg of grapes in triplicate as described by Šuklje et al. (2019a).

Figure 1. Proposed method for determining optimum harvest dates in relation to the potential wine style based on sugar accumulation per berry for Shiraz and CabernetSauvignon.

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Table 2. Dates of sugar accumulation plateau, TSS at plateau date and harvest dates expressed in number of days after sugar accumulation plateau for Shiraz and Cabernet-Sauvignon vineyards.

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Vintage	Vineyard	Plateau date	TSS at plateau date (°Brix)	Plateau-harvest duration for H1 (number of days)	Plateau-harvest duration for H2 (number of days)	Plateau-harvest duration for H3 (number of days)
2014	G1_SH	17/01	21.7	11	No harvest	24
	G2_SH	17/01	21.6	11	No harvest	24
	G2_CS	28/01	19	23	No harvest	42
2015	G1_SH	03/02	20.9	12	17	24
	G2_SH	10/02	20.3	12	19	24
	G3_SH	05/02	19.3	12	18	24
	G4_SH	10/02	19.8	12	19	24
	G2_CS	10/02	20.6	19	30	40
	O1_SH	02/02	20.3	11	17	24
	O1_CS	05/02	19.8	21	32	39
	O2_SH	26/02	20.7	11	18	25

Results and discussion

1. Berry Sugar accumulation measurements to predict harvest date

Berry sugar accumulation and fresh mass curves were determined on a population of berries from each of the experimental plots in the 2014 and 2015 vintages (Figure 2 and Supplementary Data Figure S4). Irrespective of variety, the plateaus of berry sugar accumulation were evident for most of the vineyards in both regions, excluding O1 site in 2014 (Figure 2A, 2C, 2D and Supplementary Data Figure S4). Differences were observed between vineyards and vintages in terms of the dates when the berry population reached the plateau of sugar accumulation; this reflects the variations in berry ripening dynamics and the asynchrony of berry development (Shahood et al., 2015) linked to climatic conditions, viticultural practices and varieties. The average TSS at plateau was 20.1 ± 0.8 °Brix (Table 2). In contrast, Shahood et al. (2020) recently proposed a model of berry ripening based on measurements carried out on thousands of individual berries, whereby the plateau of berry sugar accumulation was reached at 1M sugar (18 °Brix), and further sugar concentration increases were due to berry water loss only. Climatic conditions and vine physiology during flowering usually alter ovule fecundation and the resulting seed number per berry, which is at the origin of the asynchrony of berry development and can also impact berry heterogeneity (Shahood, 2017). This phenomenon is usually observed in commercial vineyards and has been well-described in the literature (Friend et al., 2009; Gouthu and Deluc, 2015). The present work also confirmed that the evolution of sugar per berry in a population of berries is a relevant physiological indicator which can be used to determine the plateau of berry sugar accumulation and, in turn, the harvest date as previously suggested (Deloire, 2013; Šuklje et al., 2019a). The plateau is generally reached at approximately 20 ± 1 °Brix, and any further increases in sugar concentration are most likely due to berry water loss and ultimately shrivelling (Deloire et al., 2021).

In the 2014 vintage, no clear plateau of sugar accumulation was found in either Shiraz or Cabernet-Sauvignon from the O1 vineyards (650 m) (Figure 2B and Supplementary Data Figure S4). Instead, a slow and unsteady accumulation of berry sugar was observed throughout the ripening period, which was attributed to severe water restrictions as determined by vineyard inspections. The 2014 vintage in O1_SH and O_CS vineyards was characterised by severe water stress during the veraison period, followed by several rain events around time of harvest. A severe vine water deficit inhibited leaf photosynthesis, which in turn inhibited the ripening process by stopping or slowing down berry sugar accumulation (Wang et al., 2003a). Rain events temporarily supplied water in the late maturation stage and may have reactivated the photosynthetic mechanism, resulting in uneven berry sugar accumulation. During the 2015 season, berry sugar accumulation for both cultivars and all vineyards in the Orange region closely followed the predicted model curve (Figure 2D and Supplementary Data Figure S4). The observed plateau of berry sugar accumulation in the 2015 O1 vintage (Figure 2D), in contrast to the 2014 vintage (Figure 2B), can be explained by higher water availability and milder climatic conditions during the veraison and post-veraison periods. Therefore, berry fresh mass and sugar accumulation per berry could be used as relevant morphological and physiological indicators to diagnose vineyards issues such as water shortage (Wang et al., 2003b; Ojeda et al., 2001; Rossouw et al., 2017), in combination with other methods such as δ¹³C measurement (van Leeuwen et al., 2001).

Figure 2. Berry fresh mass (grey) and sugar per berry (black) evolution for selected vineyards in Griffith and Orange.

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A: G2_SH_2014; B: O1_CS_2014; C: G2_SH_2015; D: O1_CS_2015. The black arrow indicates the date of the plateau of sugar accumulation.

2. Sensory analysis

A similar aromatic evolution was observed for Shiraz and Cabernet-Sauvignon wines from Griffith for the 2014 vintage. Wines from H1 were associated with red fruit and herbaceous attributes reminiscent of tomato leaf, green olive and, specifically, capsicum for Cabernet-Sauvignon (Figure 3). Shiraz wines were perceived as being more acidic, but not Cabernet-Sauvignon wines (Figure 4). On the other hand, wines from H3, were characterised by dark fruit, plum sensorial attributes, higher astringency and were perceived as being more alcoholic. In 2015, wines from two other Shiraz vineyards in Griffith (G3 and G4) and from two different climates in the Orange region, as well as an H2, were also evaluated. Once again, H1 wines (FF) were correlated with red fruit and the perception of acidity, whilst H3 wines (MF) were associated with dark fruit, plum, black cherry, astringency and alcohol perception regardless of the vineyard (Figures 5A and 5B). H2 wines were closely grouped with the corresponding “Mature Fruit” wines from the Orange region, but this grouping was not so evident for the H2 wines from Griffith.

H1 (FF) and H3 (MF) wines showed a similar pattern of aromatic evolution irrespective of the variety, site and climatic conditions across regions and vintages. Descriptors such as red fruit, dark fruit and plum consistently discriminated “Fresh Fruit” (H1) from “Mature Fruit” (H3) stages. In 2015, no selected descriptors could consistently discriminate the intermediate stage (H2) from FF and MF stages (Figure 5A and 5B). H2 wines were perceived differently to wines from H1(FF) and H3(MF) (Figures 5A and 5B), but these differences depended on site, climate and variety (data not shown). While the wine aromatic profile was well-defined for H1 and H3 wines, H2 wines style clearly depended on the vineyard. Therefore, a sequential harvest based solely on grape sugar concentration and maturation time will not guarantee a given aromatic profile of a final wine. Conversely, the number of days after the plateau of berry sugar accumulation (mg/berry) seems to be synchronised with the aromatic maturity perceived in the resulting wines.

Figure 3. Principal Component Analysis biplot of sensory data for 2014 G1 and G2 Shiraz wines

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Figure 4. Odour comparison profiles performed on the means of the perceived aromatic intensity of each selected sensory descriptor for 2014 Cabernet-Sauvignon wines.

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Significant differences are indicated with stars: (*), significant at p < 0.05, (**), significant at p < 0.01, (***) significant at p < 0.001

Figure 5. Mean rating ±SD of sensory attributes intensity showing significant differences assessed in 2015 Shiraz wines (A: n = 18 per harvest date) and Cabernet-Sauvignon wines (B: n = 6 per harvest date) between harvest dates (H1, H2, H3).

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One-way ANOVA was used to compare data. Different letters above bars indicate significance at p < 0.05 (Fischer’s LSD). All quoted uncertainty is the standard deviation of 18 Shiraz (A) and 6 Cabernet-Sauvignon (B) wines respectively.

Conclusions

The present study has demonstrated the existence of a predictable aromatic sequence during grape ripening in Australian Shiraz and Cabernet-Sauvignon from different meso-climates. Two distinct maturity stages were identified and characterised: i) Fresh Fruit distinguished by fresh/red fruits attributes, and ii) Mature Fruit associated with dark fruit and plum character. Berry sugar accumulation (mg/berry) is characterised by two phases between veraison and harvest: a) pre-plateau of berry sugar accumulation, and b) post plateau of berry sugar accumulation, in which the increase in sugar concentration (°Brix) is mainly due to berry water loss. After the plateau of berry sugar accumulation, the aromatic and phenolic maturity of the wine was separate from technological maturity. Compositional analyses revealed that wine grape and yeast-derived compounds were affected by grape maturity. A few individual compounds can be considered as maturity markers irrespective of grape genotype. The contribution of DMS to the dark fruit aroma specific to Mature Fruit wines was the most important marker for discriminating the Fresh Fruit stages from the Mature Fruit stages. Specific cultivar markers which potentially contribute to aromas were also identified; for example, (Z)‐3‐hexenol may contribute to fresh fruit aroma in Shiraz wines. While terpenoids were not generally affected by the post plateau ripening process, the composition of esters was significantly altered with a marked varietal effect. The results of the study have contributed to knowledge regarding the nexuses between grape composition, wine composition and subsequent sensory characteristics; however, further research would be required to gain an even greater understanding of these complex relationships.

Acknowledgements

The authors would like to thank Campbell Meeks for making the experimental wines for this study and to Australia’s grape growers and winemakers for their financial support through their investment body, Wine Australia. The authors would also like to thank the wine producers in Griffith and Orange for allowing us access to their vineyards and for their grape donations.

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