Vascular function in berries of Vitis vinifera (L) cv. Shiraz

Anatomical studies on the movement of a xylem tracer dye were combined with functional studies on changes in grape berry volume during final stages of berry ripening to gauge xylem effectiveness. Movement of a xylem tracer dye into pre-veraison fruit was compared with movement into post-veraison fruit by feeding a solution of acid fuchsin to excised shoots with bunches still attached, and then sectioning fruit for photo-microscopy. Those comparisons confirmed published studies showing an apparent blockage to dye movement along major vessels within the brush tissue of post-veraison fruit. However, our functional approach yielded a different impression of vascular activity. A continuation of xylem transport in ripening fruit was inferred from comparisons of berry volume where pedicels were either girdled (phloem interrupted, but xylem intact) or excised (both phloem and xylem interrupted). Volume changes in manipulated berries were compared with immediately adjacent intact control berries within the same bunch. Control fruit lost volume subsequent to 78 days after flowering (DAF) while manipulated fruit lost volume from the first day of treatment at 67 DAF. By harvest time at 95 DAF, both control fruit and girdled fruit had fallen to 91% and excised fruit to 46% of maximum volumes recorded around 78 DAF. Berry volume loss in girdled fruit was further enhanced by deficit irrigation. We conclude that xylem flow into those Shiraz berries must have continued beyond veraison despite dye evidence of a vessel blockage within the brush region of analogous post-veraison fruit.     

Dynamics of grape berry growth and physiology of ripening

Data from two experiments on development of grape berries is re-examined with emphasis on partitioning of berry weight into non-solutes per berry (largely water) and solutes per berry (largely sugar), using weight times juice °Brix. This approach is based on the thought that, since xylem flow is blocked after veraison, time curves of solutes per berry indicate the activity of phloem transport into the berry during ripening growth. Experiment 1: Measurements of Muscat Gordo Blanco berries from inflorescences with a spread of flowering times showed typical double-sigmoid volume/time curves but with divergent rates and amounts of volume increase. Despite this divergence, °Brix curves after veraison were almost coincident because, in each case, the rate of increase in solutes per berry was proportional to that of berry volume. These results indicate that sugar and water increments after veraison are linked and depend on the same source, namely, phloem sap. Experiment 2: An irrigation experiment on cv. Shiraz also showed divergent berry weight curves between treatments and years but with the difference that all berries shrank after a maximum berry weight was attained at 91 days after flowering (at about 20 °Brix). At this point, the curves of solutes per berry slowed then plateaued, indicating that inflow of phloem sap had become impeded. Prior to shrinkage these berries accumulated primary metabolites (mainly phloem sugar) but, during shrinkage, when berries were apparently isolated from vascular transport, non-anthocyanin glycosides accumulated. These results have implications for the study of berry flavour buildup and berry composition, and also for the understanding of sink competition within the vine, fresh and dried yield, and juice °Brix levels.     

Is weight loss in ripening grape berries cv. Shiraz caused by impeded phloem transport?

Berry shrinkage in ripening grapes cv. Shiraz is systemic within a given grapevine and coincident between grapevines in any given season. In this present study on weight loss in ripening berries, ripening curves of non-solutes per berry (largely water) were similar to curves for berry weight (as a function of time). Both sets of curves were equivalent with respect to timing of maximum weights and subsequent rates of weight loss. However, curves of solutes per berry (largely sugar) increased steeply up to the time of maximum berry weight, then slowed and plateau-ed. We suggest that phloem sap is the sole source for water and solutes that enter grape berries subsequent to veraison, and accumulate until maximum berry weight. We further suggest that phloem flow becomes impeded at maximum berry weight.
As berry ripening proceeds, continuation of berry transpiration leads to berry shrinkage and a concentration of solutes; i.e. any increase in juice Brix depends on shrinkage. One implication is that assimilates enter a berry up to the onset of shrinkage, whereafter accumulation of non-anthocyanin glycosides (including glycosides of flavour compounds) depends upon their synthesis in situ.     

Direct measurement of hydraulic properties in developing berries of Vitis vinifera L. cv Shiraz and Chardonnay

Berries of Vitis vinifera L. cv Shiraz can undergo weight loss during later stages of ripening. Existing published views on how weight loss occurs are based on changes in capacity of the vascular system to import water during development (McCarthy and Coombe, Australian Journal of Grape and Wine Research, 5, 17–21, 1999). One important element of these views is the proposed cessation of water flow through the xylem after veraison. We have now measured the water flow into berries of Shiraz and Chardonnay as they develop using the pressure probe and the high pressure flow meter (HPFM). The pressure probe connected to the pedicel of individual berries provided measurements of single berry hydraulic conductance. By systematic excision of tissue segments of the berry and pedicel we determined where in the pathway hydraulic conductance changed during development. The HPFM was used on whole bunches showing that berries (including pedicels) represent parallel high hydraulic resistances and that the hydraulic resistance of the bunch axis was rather small. The hydraulic conductance per berry could be determined from excision experiments. There was close agreement between the pressure probe and HPFM measurements. Both showed a ten‐fold reduction in hydraulic conductance of whole berries from veraison to full ripeness. Shiraz had hydraulic conductances that were 2‐ to 5‐fold higher than those for Chardonnay. Shiraz maintained a higher hydraulic conductance past 90 days after flowering than Chardonnay. The decrease in hydraulic conductance occurred in both the distal and proximal parts of the berry for both varieties. The pressure probe also provided measurements of the xylem pressure that non‐transpiring berries could develop. These pressures were –0.2 to –0.1 MPa until veraison and increased to zero when the juice osmotic potential reached about –3 MPa in Chardonnay and –4 MPa in Shiraz. The results suggest values of the reflection coefficient of the osmotic barrier around the xylem vessels of about 0.1–0.2 at veraison decreasing to 0 at harvest. It is suggested that in addition to changes in xylem anatomy, aquaporins in berry membranes may play a role in regulating hydraulic conductance. Water movement from the berry back to the parent vine via the xylem (backflow) may be an important component of berry weight loss in Shiraz, particularly if the phloem ceases functioning at high osmotic potentials near maximum weight. Backflow could account for a weight loss of 43 mg per day in Shiraz berries for a relatively small gradient of 0.1 MPa.     

Early post-veraison growth in grapes: evidence for a two-step mode of berry enlargement

An account is provided of berry softening plus cell wall loosening in flesh and skin tissues, that relates to changes in berry turgor and sugar accumulation around veraison in Golden Muscat ( Vitis vinifera × V. labrusca ). The onset of change in berry deformability was taken as a demarcation between pre-veraison and post-veraison growth phases. Our results confirmed that post-veraison berry enlargement was preceded by berry softening and sugar accumulation. Berry 'turgor' (as inferred from the rate of juice exudation from punctured fruit) increased abruptly and concurrently with berry softening and sugar accumulation, but dropped steadily with subsequent berry expansion. Elasticity of skin tissues decreased abruptly during early phases of expansion, while berry deformability increased. We suggest that cell wall loosening in flesh tissues precedes cell wall loosening in skin tissues, and that this loosening sequence results in a two step sequence where berry softening is later followed by berry expansion.     

Effects of manipulated grape berry transpiration on post-veraison sugar accumulation

A series of experiments were conducted on container-grown Pinot Noir and Sangiovese grapevines ( Vitis vinifera L.) to investigate whether changes in berry water loss at veraison influence the pattern of sugar accumulation. Berry transpiration was induced to vary either by changing the vapour pressure deficit (VPD) around bunches through temperature or relative humidity (RH) manipulations, or by applying drying-accelerating emulsion or a hydrophobic coating (vaseline) over the berry skin. In all the experiments, every berry of every bunch was rated prior to treatment as to softness and colour intensity. Transpiration rates were derived either from measurements of attached bunches using a custom-built open gas-exchange system or from weight loss calculated for single excised berries. Berry development and ripening were monitored throughout each experiment as deformability, fresh weight, and sugar concentration and content. Berries either did not respond to VPD-enhancement or showed reduced water loss when bunches were subjected to high temperature. Low berry transpiration in the latter treatment led to lower sugar content per berry up to harvest, and berry transpiration and net sugar intake were linearly correlated up to 0.20–0.25 mmol/m².s. A similar correlation was found also in the coating experiment within the first five days after treatments. When berry transpiration was restricted by applying vaseline, sugar accumulation was retarded. Low values of VPD, induced by raising the RH around the bunches, lowered sugar concentration but not sugar content per berry. The present study provides preliminary evidence that sugar accumulation in grapevine berries responds to changes in the evaporative demand around the bunch at veraison.     

Synthesis of flavonols and expression of flavonol synthase genes in the developing grape berries of Shiraz and Chardonnay (Vitis vinifera L.)

Flavonols were determined in Shiraz and Chardonnay grapes throughout berry development. The predominant flavonols were quercetin-3-glycosides with trace amounts of kaempferol-3-glycosides detected in Shiraz flowers but not in developing berries. Flavonols were present in the skin of ripening grapes but were not detected in seeds or flesh. Flavonols were also present in buds, tendrils, inflorescences, anthers and leaves. The concentration of flavonols in flowers (mg/g fresh weight) was high and decreased between flowering and berry set then remained relatively constant through berry development. The total amount of flavonols in berries (mg/berry) was low until pre-veraison then increased during berry development, particularly before veraison, the onset of ripening, in Chardonnay and during ripening in Shiraz. Two cDNA fragments with homology to genes encoding the enzyme flavonol synthase (FLS) were isolated from Shiraz flowers. In the overlapping region of the two cDNAs, they had 80% sequence identity at the nucleotide level and both had high homology to FLS genes from other plants. VvFLS1 was expressed in leaves, tendrils, pedicels, buds and inflorescences as well as in developing grapes. Expression was highest between flowering and fruit set then declined, increasing again during ripening coincident with the increase in flavonols per berry. Expression of VvFLS2 was much lower than for VvFLS1 and did not change during berry development. The results indicate that two distinct periods of flavonol synthesis occur in grapes, the first around flowering and the second during ripening of the developing berries.     

Proline and arginine accumulation in developing berries of Vitis vinifera L. in Australian vineyards: Influence of vine cultivar, berry maturity and tissue type

Using HPLC analysis, free amino acid profiles were obtained for ripe berries of six grapevine cultivars (Cabernet Sauvignon, Grenache, Muscat Gordo, Pinot Noir, Riesling and Sangiovese) grown under comparable conditions and harvested at similar maturities. Compositional differences were observed between cultivars, however proline and arginine were always the major amino acids. Mature berries of Cabernet Sauvignon contained a very high concentration of proline, but a much lower concentration of arginine. Those of other cultivars contained moderate levels of both arginine and proline.
Changes in free amino acid profiles during grape berry development were further investigated in four cultivars (Cabernet Sauvignon, Chardonnay, Gewurztraminer and Muscat Gordo). In all cases, most of the proline accumulation occurred late in ripening, around four weeks post-veraison. In contrast, arginine accumulation began before veraison and continued to full maturity, except for those cultivars in which a high concentration of proline accumulated, in which case the concentration of arginine reached a plateau relatively early in development. Accumulation of both proline and arginine appears to be develop-mentally regulated. These observations are discussed with reference to proline and arginine metabolism and possible links between them.
The distribution of free proline, arginine and other amino acids amongst pulp, skin and seed was examined using mature fruit of Cabernet Sauvignon and Riesling. Amino acid profiles were found to vary considerably between the berry components. The skin contained a greater ratio of arginine:proline compared with the pulp. This suggests that the yeast-assimilable nitrogen content of juices, and therefore their fermentability, could be enhanced in the presence of skins.     

Impacts of crop level,soil and irrigation management in grape berries of cv ‘Trincadeira’ (Vitis vinifera L.)

Berry size and crop yield are widely recognized as important factors that contribute to wine quality. The final berry size indirectly affects the phenolic concentration of the wine due to skin surface-to-berry volume ratio. The effects of different irrigation levels, soil management and plant crop level on growth of ‘Trincadeira’ berries were studied. In order to test the influence of different irrigation levels (rainfed, pre-veraison and post-veraison), different soil management (tillage and natural cover crops) and different plant crop levels (8 and 16 clusters per vine), leaf water potential, skin anthocyanin, polyphenols, berry skin and seed fresh weight were measured in fruits. The segregation of berries into three different berry classes: small, medium and large, allowed to identify different levels of contribution of soil management and irrigation level into berry, skin and seeds ratios. As expected, higher water availability due to irrigation and soil tillage management during berry development induced an increase in berry flesh weight and this was more evident in larger berries; however, berry skin and seed fresh weight remained unchanged. Also, anthocyanins did not show significant differences.     

The relationship between the expression of abscisic acid biosynthesis genes, accumulation of abscisic acid and the promotion of Vitis vinifera L. berry ripening by abscisic acid

Background and Aims:  Grapevines ( Vitis vinifera L.) are considered to have non-climacteric fruit, but the trigger initiating ripening (veraison) is poorly understood. This study aimed to further investigate the role of abscisic acid (ABA) during berry ripening.
Methods and Results:  In field-grown grapes over three seasons, free ABA levels increased at veraison then subsequently declined to low levels. Bound ABA increased as the free ABA level decreased after 10–11 weeks post-flowering (wpf), but ABA must also be degraded and/or exported. The absence of a large pool of bound ABA before veraison makes it unlikely that the increase in free ABA is due to the mobilization of conjugated ABA. The expression pattern of genes crucial for ABA synthesis, zeaxanthin epoxidase and two 9-cis-epoxycartenoid dioxygenases (NCEDs) indicates that berries may have the potential to synthesize ABA in situ . However, the expression profile of these genes did not correlate well with ABA levels indicating that ABA accumulation is under more complex control. The application of (+)-ABA advanced ripening as measured by colour formation, berry size increase and to a lesser extent sugar accumulation and altered the expression of one of the NCED genes.
Conclusions:  The changes in berry ABA levels around the time of veraison, which influence the timing of ripening, are under complex developmental control.
Significance of the Study:  The improved understanding of the control of berry ripening is vital to attempts to successfully manipulate this process.     

Deciphering the ability of different grapevine genotypes to accumulate sugar in berries

Background and Aims: To analyse the genetic determinants of berry sugar content in grapevine varieties, a method that combined measurements of total soluble solids (TSS) from berries sampled at a fixed stage after veraison with data on the yield and the leaf area was tested. Methods and Results: Over three seasons, differences in TSS at harvest for Riesling and Gewurztraminer grapes were explained by differences in the veraison dates and in the fruit‐to‐leaf ratios. TSS, yield and leaf area were also evaluated for 120 genotypes, which were progeny from a cross between Riesling and Gewurztraminer. Thermal timescales (degree‐days) proved relevant for describing sugar accumulation in the parent varieties, and when berries were sampled at a fixed heat sum after veraison in the progeny, the genotypic variability of sugar content was considerably reduced compared with the measurements on the same date at the end of the season. A negative relationship between the fruit‐to‐leaf ratio and the sugar content was also detected in the progeny. Conclusions: The evolution of TSS during berry ripening can be described using thermal timescales. Additionally, information on the yield and the leaf area needs to be integrated to analyse the behaviour of genotypes. Significance of the Study: A method to characterize the genotypic effect on fruit sugar accumulation after eliminating the contribution of veraison dates and crop load was proposed.     

Relative proportions of seed, skin and flesh, in ripe berries from Cabernet Sauvignon grapevines grown in a vineyard either well irrigated or under water deficit

Relationships between the fresh mass of seed, skin (exocarp), and flesh (mesocarp) in six different berry size categories, were assessed on ripe fruit from Cabernet Sauvignon grapevines exposed to either High (H), Control (C) or Low (L) water status during post‐veraison berry growth in a vineyard. Berries harvested from each treatment were segregated into six mass categories in order to distinguish between changes in fresh mass components associated with general variation in berry size (on well‐watered grapevines), and those associated with berry size differences due to water stress. Berry fresh mass across all treatments ranged from about 0.4 to 2.0 g. Fresh mass components for both H and C berries comprised approximately 5% seed, 15% skin, and 80% flesh, regardless of variation in berry size, although there was some increase in seed mass relative to whole‐berry fresh mass in larger berries. Berry growth (as inferred from fresh mass at harvest) was much less sensitive to water deficit than published reports for grapevine shoot growth. Midday leaf water potentials around –1.20 MPa (Control) were not sufficient to inhibit berry growth. However, midday water potentials around –1.50 MPa (Low water status) inhibited berry growth by 13–18% of that attained by grapevines grown at high water status (i.e. treatment H where midday leaf water potentials remained around –1.00 MPa). Inhibition of berry growth by water deficit was attributed almost exclusively to reduced growth of mesocarp tissues (for most berry size categories). Water deficit thus increased the proportion of whole‐berry fresh mass represented by seeds and skin (for most berry size categories). Changes in those proportions due to irrigation treatments exceeded differences associated with general (non‐stress) variation in whole‐berry fresh mass. Excluding adverse environmental impacts on whole‐berry fresh mass, our results point to a limited role for variation in berry size per se as a factor determining the solute concentration of juice or wine derived from different sized fruit. By implication, and for all categories of whole‐berry fresh mass represented here, late season water deficit can result in ripe fruit with more skin and seed tissues (relative to whole‐berry fresh mass) compared with well‐watered control fruit.     

Analysis of tannins in seeds and skins of Shiraz grapes throughout berry development

The flavan-3-ol and proanthocyanidin composition of both seeds and skin of Vitis vinifera L. cv. Shiraz grapes was determined by reversed-phase HPLC after acetone extraction and acid-catalysis in the presence of excess phloroglucinol. Samples were taken at weekly intervals from fruit-set until commercial harvest. The main period of proanthocyanidin accumulation in grape seeds occurred immediately after fruit-set with maximum levels observed around veraison. Over two seasons there was variation in both the timing and content of proanthocyanidins in seeds. In skin, proanthocyanidin accumulation occurred from fruit set until 1–2 weeks after veraison. Proanthocyanidin subunit composition was different in seeds and skin and changed during berry development but the mean degree of polymerisation of the tannin polymers in skins was higher than in the seeds at all stages of berry development. Proanthocyanidin levels in both seeds and skin decreased between veraison and harvest. Additional proanthocyanidin subunits were released when the residues remaining after acetone extraction were subjected to direct acid-catalysis in the presence of phloroglucinol. In the seeds, these accounted for much of the post-veraison decrease, but not in grape skin. At harvest, 75% of extractable berry proanthocyanidin was in the seeds. Accumulation of proanthocyanidins in the seeds appears to be independent of that in the skins, but in both tissues synthesis occurs early in berry development and maximum levels are reached around veraison.     

Influence of shriveling on berry composition and antioxidant activity of Cabernet Sauvignon grapes from Shanxi vineyards

BACKGROUND: Berry shrivel (BS), a berry development disorder, appears soon after veraison. It occurs worldwide and affects the quality of grape berries and wine. However, it had not been reported in China until recently. This study aimed to investigate the changes in berry composition and antioxidant activity of Cabernet Sauvignon grapes from Xiangning Valley, Shanxi Province, China, during BS. RESULTS: Shrinkage contributed to an increase in the concentration of basic grape ingredients such as sugar and acid. An appropriate degree of shrinkage was apparently helpful in improving the phenolic content and increasing the antioxidant activity, but the berries that continued to shrivel showed a low antioxidant activity. Further, the results indicated distinct differences between the berries harvested from the southern side of the canopy and those harvested from the northern side, presumably due to variations in sunlight exposure. CONCLUSION: Moderate BS was beneficial since it increased berry quality and antioxidant activity of Cabernet Sauvignon grapes from Shanxi vineyards. Copyright © 2011 Society of Chemical Industry     

ABA during reproductive development in non-irrigated grapevines (Vitis vinifera L. cv. Tempranillo)

In grapevines, stomatal aperture decreases after a mid-morning peak during summer days. Afternoon stomatal closure increases in non-irrigated plants as water limitation progresses, which suggests the involvement of abscisic acid (ABA) in the control of stomatal aperture. The objective of this work was to study the seasonal and diurnal time-courses of CO₂ assimilation rate, leaf conductance, leaf water potential +, and ABA concentration in xylem sap, leaves, flowers and berries in non-irrigated field-grown Tempranillo grapevines throughout reproductive development. Leaf decreased throughout fruit development because water availability decreased towards the end of the reproductive cycle. CO₂ assimilation rate, leaf conductance and xylem ABA concentration also decreased during the course of the growing season. Combining all measurements xylem ABA was either not correlated, or only slightly correlated, with leaf water status + and daily leaf conductance, respectively. This lack of relationship indicates that xylem ABA during fruit ripening had functions other than provision of a non-hydraulic signal. On a seasonal basis, xylem ABA concentration measured in non-irrigated grapevines was well related to berry ABA concentration, especially at the end of fruit development (veraison and harvest).     

Variation in Shiraz berry size originates before fruitset but harvest is a point of resynchronisation for berry development after flowering

Background and Aims:  Asynchronous berry development results in variation in berry size and berry composition. Variation in berry size affects vineyard yield, wine quantity and berry composition, while variation in berry composition affects fruit flavour and wine quality. The objective of this study is to identify when variation in berry size begins.
Methods and Results:  Shiraz bunches were sampled at seven stages throughout the growing season and the development of individual berries was characterised by measurements of weight, volume, surface area, deformability, seed number and seed weight. Coefficients of variation with respect to berry weight, volume and surface area were elevated throughout the post-flowering period but declined as the berries approached harvest ripeness.
Conclusions:  Variation in berry size originates prior to berry set, most probably during differentiation in the floral primordium at budburst. Harvest represents a point of resynchronisation in berry development when a number of physiological changes become synchronised between berries and within the bunch.
Significance of the Study:  Variation in berry size is presumed by the wine industry to have a negative impact on crop level, fruit composition and wine quality. Much of this presumption is scientifically unsubstantiated. The findings of this paper should encourage grapegrowers and winemakers to modify the timing of the harvest to coincide with a reduction in variation in berry size.     

Weight loss from ripening berries of Shiraz grapevines (Vitis vinifera L. cv. Shiraz)

Environmental and phenological correlatives associated with the onset of weight loss during ripening in Vitis vinifera cv. Shiraz berries were examined. An irrigation experiment concerned with transient water deficit effects on berry growth over four consecutive seasons provided source data. Timing of maximum berry weight was more closely correlated with the number of days after flowering than with temperature summation. Prior to maximum berry weight, Brix was more closely correlated with days after flowering than with temperature summation, while the number of days from flowering to 10 and 15Brix was remarkably uniform despite contrasting seasons. Driving variables for weight loss during advanced ripening remain a matter of conjecture, but loss of berry water under highly evaporative conditions of late summer is a likely contributor.     

Relationships between seed and berry development of Vitis Vinifera L. cv Shiraz: Developmental changes in seed morphology and phenolic composition

Berries were collected regularly from fruit set to berry maturity from irrigated (Shiraz) grapevines in a Barossa valley vineyard. Seeds were removed for detailed study of physical attributes (weight, moisture, colour) and phenolic composition (seed tannins). Three phases of seed growth and development were discerned: (1) a phase of seed growth characterised by a steady increase in both fresh weight and dry weight, biosynthesis and accumulation of flavan‐3‐ols and tannins, and green appearance; (2) a transition phase where seed fresh weight and dry weight reached a maximum, but with continuing enlargement of the basal end. Accumulation of flavan‐3‐ols and seed tannins also reached a maximum during phase 2, and was accompanied by an onset of tannin oxidation, and yellow appearance; and finally, (3) a phase of seed drying and maturation defined by a decrease in fresh weight due to water export, a sustained oxidation of tannins, and overall brown appearance. These phases in seed development correspond to particular stages in berry development. Seeds reached maximum fresh seed weight and full size at the beginning of berry colouring (veraison), while maximum dry seed weight coincided with maximum berry weight. Changes in seed phenolics were linked to berry development and maturation. Changes in seed coat colour were also related to developmental changes in berry anthocyanins and total skin phenolics, indicating that the external appearance and colour of the seed coat may be used as an additional indicator of overall berry ripeness. A graduated colour chart was developed to provide an objective index of seed coat colour and thus developmental status of seeds and berry.