Zeolite application and irrigation during ripening reduced berry sunburn damage and yield loss in cv. Sangiovese (Vitis vinifera L.).

Climate change poses significant challenges to the grapevine cultivation for wine production. In the last years, the occurrence of extreme weather events such as intense heat waves and prolonged period of drought increased sunburn damage with negative consequences to yield and berry composition. Short-term adaptation strategies are urgently needed to mitigate these effects. In this light, our study aimed to evaluate the efficacy of zeolite application on the canopy and irrigation during ripening on sunburn damage, yield, and berry composition in cv. Sangiovese (Vitis vinifera L.). Over 3 years (2021-2023), canopy management and irrigation treatments were arranged in a strip-plot design. Canopy management treatments included leaf removal on the cluster zone, with and without zeolite application, and no defoliated control; irrigation treatments comprised irrigation from berry softening and no irrigation. Both zeolite application and irrigation reduced sunburn damage, thus mitigating yield loss. Regarding berry composition, zeolite application did not alter the main parameters whereas irrigation led to reductions in sugar and anthocyanin concentrations. These findings suggest that zeolite application and irrigation during ripening represent promising and cost-effective solutions for grape growers facing climate change-induced challenges. However, further studies are necessary to optimize timing of irrigation to avoid detrimental effects on anthocyanin accumulation.

Use of an automatic fruit-zone cooling system to cope with multiple summer stresses in Sangiovese and Montepulciano grapes.

Grapevines are frequently subjected to heatwaves and limited water availability during ripening. These conditions can have consequences for the physiological health of the vines. Moreover, the situation is often exacerbated by intense solar radiation, resulting in reduced yield due to sunburn and a decline in quality. In light of these challenges, our study aimed to develop a fruit-zone cooling system designed to mitigate grape sunburn damage and improve the microclimate conditions within the vineyard. The system comprises a network of proximal sensors that collect microclimate data from the vineyard and an actuator that activates nebulizers when the temperature exceeds the threshold of 35°C. The research was conducted over two years (2022 and 2023) in Bologna (Italy) using potted Sangiovese and Montepulciano vines. These two vintages were characterized by high temperatures, with varying amounts of rainfall during the test period, significantly impacting the evaporative demand, which was notably higher in 2023. Starting from the veraison stage we compared three treatments: Irrigated control vines (WW); Control vines subjected to 50% water restriction during the month of August (WS); WS vines treated with nebulized water in the bunch area during the stress period (WS+FOG). The application of nebulized water effectively reduced the temperature of both the air around the clusters and the clusters themselves. As we expected, Montepulciano showed better single leaf assimilation rate and stomatal conductance under non-limiting water conditions than Sangiovese while their behavior was unaffected under water-scarce conditions. Importantly, for the first time, we demonstrated that nebulized water positively affected gas exchange in both grape varieties. In addition to this, the vines treated with the misting system exhibited higher productivity compared to WS vines without affecting technological maturity. In the 2023 vintage, the activation of the system prevented the ripening blockage that occurred in Montepulciano under water stress. Regarding the concentration of total anthocyanins, a significant increase in color was observed in WS+FOG treatment, suggesting a predominant role of microclimate on anthocyanin biosynthesis and reduction of oxidative phenomena. In conclusion, the fruit-zone cooling system proved to be an invaluable tool for mitigating the adverse effects of multiple summer stresses.

When Cannabis sativa L. Turns Purple: Biosynthesis and Accumulation of Anthocyanins.

Environmental cues elicit anthocyanin synthesis in plant vegetative and reproductive tissues. Their accumulation in different organs accounts for their diverse biological functions, mainly related to their antioxidant properties, and it depends on a temporally and spatially regulated mechanism controlled by the action of a well-known multi-transcription factor complex. Despite the highly recognizable value of Cannabis sativa L. as a natural biorefinery of phytochemicals, very little information is known on anthocyanin pigmentation in this species. In this work, a targeted quantification of anthocyanins via HPLC-MS/MS, combined with the transcriptional profile via RT-qPCR of genes encoding for structural and decorating enzymes and regulatory transcription factors in different C. sativa tissues, help gain insights into the anthocyanin pathway in this species. To the best of our knowledge, this is the first report on the identification of cyanidin-3-rutinoside (keracyanin) as the major anthocyanin in C. sativa vegetative and floral tissues. Keracyanin amounts were higher than in small berries, suggesting that Cannabis biomass is a valuable source of colored antioxidants to be exploited in diverse applications. Furthermore, a gene putatively encoding for an anthocyanin DTX35 type transporter and CsTTG1 were identified in silico and their transcriptional levels were assessed via RT-qPCR. The results allow us to provide the first model of anthocyanin regulation in C. sativa, opening a new research scenario in this species for both breeding purposes and phytochemical exploitation.

Post-veraison trimming slow down sugar accumulation without modifying phenolic ripening in Sangiovese vines.

BACKGROUND: Vineyard strategies have recently been developed to combat the effects of global warming on grapevines, which is causing grapes to ripen quickly, excessive sugar accumulation in berries, and high alcohol levels in wines. We evaluate the effectiveness of post-veraison trimming as a means of slowing down sugar accumulation, without modifying phenolic ripening, in Sangiovese vines grown in highly fertile and well watered soil. RESULTS: The removal of about two-thirds of the leaf area by shoot trimming after veraison led to a reduction in sugar content without affecting yield. Total and extractable anthocyanins, skin, and seed tannins showed no significant variation at harvest after the treatment during the three-year trial, while the replenishment of carbohydrates in canes at the end of the trial was negatively affected. CONCLUSIONS: These results indicate that, in highly fertile and well-watered soil, post-veraison trimming may represent a powerful tool for decreasing sugar concentration during harvest without affecting yield or total and extractable phenolic compounds. However, the reduction in starch reserves compared to the control serves as a warning about repeated trimming over the years. © 2018 Society of Chemical Industry.

Low night temperature at veraison enhances the accumulation of anthocyanins in Corvina grapes (Vitis Vinifera L.).

Climate change is a major concern in grape production worldwide. Nights have been warming much faster than the days, raising attention on the effect of night temperatures on grape and wine composition. In this study we evaluated the effect of night temperatures on grape coloration in the cv. Corvina (Vitis vinifera L.). In 2015 and 2016 potted plants were cooled overnight (10-11 °C) during two berry ripening phases, veraison (TV) or post-veraison (TPV), and compared to control vines (C) grown at ambient night temperature (15-20 °C on average). Cooling treatment around veraison (TV) hastened berry anthocyanin accumulation, while the same treatment applied after veraison (TPV) was ineffective. Molecular analysis revealed an increased transcription of four key genes in anthocyanin biosynthesis (CHS3, F3H1, MYBA1 and UFGT) in TV treatment. These results suggest that the anthocyanin biosynthesis capacity was enhanced by cool nights during veraison. However, since the gene expression was not always temporally correlated to the increase in anthocyanin concentration, we speculate on the presence of mechanisms, such as enzymatic regulation or anthocyanin transport, which may contribute in determining the anthocyanin accumulation under low night temperatures.

Whole Plant Temperature Manipulation Affects Flavonoid Metabolism and the Transcriptome of Grapevine Berries.

Among environmental factors, temperature is the one that poses serious threats to viticulture in the present and future scenarios of global climate change. In this work, we evaluated the effects on berry ripening of two thermal regimes, imposed from veraison to harvest. Potted vines were grown in two air-conditioned greenhouses with High Temperature (HT) and Low Temperature (LT) regimes characterized by 26 and 21°C as average and 42 and 35°C as maximum air daily temperature, respectively. We conducted analyses of the main berry compositional parameters, berry skin flavonoids and berry skin transcriptome on HT and LT berries sampled during ripening. The two thermal conditions strongly differentiated the berries. HT regime increased sugar accumulation at the beginning of ripening, but not at harvest, when HT treatment contributed to a slight total acidity reduction and pH increase. Conversely, growing temperatures greatly impacted on anthocyanin and flavonol concentrations, which resulted as strongly reduced, while no effects were found on skin tannins accumulation. Berry transcriptome was analyzed with several approaches in order to identify genes with different expression profile in berries ripened under HT or LT conditions. The analysis of whole transcriptome showed that the main differences emerging from this approach appeared to be more due to a shift in the ripening process, rather than to a strong rearrangement at transcriptional level, revealing that the LT temperature regime could delay berry ripening, at least in the early stages. Moreover, the results of the in-depth screening of genes differentially expressed in HT and LT did not highlight differences in the expression of transcripts involved in the biosynthesis of flavonoids (with the exception of PAL and STS) despite the enzymatic activities of PALs and UFGT being significantly higher in LT than HT. This suggests only a partial correlation between molecular and biochemical data in our conditions and the putative existence of post-transcriptional and post-translational mechanisms playing significant roles in the regulation of flavonoid metabolic pathways and in particular of anthocyanins.

Transcriptional Responses to Pre-flowering Leaf Defoliation in Grapevine Berry from Different Growing Sites, Years, and Genotypes.

Leaf removal is a grapevine canopy management technique widely used to modify the source-sink balance and/or microclimate around berry clusters to optimize fruit composition. In general, the removal of basal leaves before flowering reduces fruit set, hence achieving looser clusters, and improves grape composition since yield is generally curtailed more than proportionally to leaf area itself. Albeit responses to this practice seem quite consistent, overall vine performance is affected by genotype, environmental conditions, and severity of treatment. The physiological responses of grape varieties to defoliation practices have been widely investigated, and just recently a whole genome transcriptomic approach was exploited showing an extensive transcriptome rearrangement in berries defoliated before flowering. Nevertheless, the extent to which these transcriptomic reactions could be manifested by different genotypes and growing environments is entirely unexplored. To highlight general responses to defoliation vs. different locations, we analyzed the transcriptome of cv. Sangiovese berries sampled at four development stages from pre-flowering defoliated vines in two different geographical areas of Italy. We obtained and validated five markers of the early defoliation treatment in Sangiovese, an ATP-binding cassette transporter, an auxin response factor, a cinnamyl alcohol dehydrogenase, a flavonoid 3-O-glucosyltransferase and an indole-3-acetate beta-glucosyltransferase. Candidate molecular markers were also obtained in another three grapevine genotypes (Nero d'Avola, Ortrugo, and Ciliegiolo), subjected to the same level of selective pre-flowering defoliation (PFD) over two consecutive years in their different areas of cultivation. The flavonol synthase was identified as a marker in the pre-veraison phase, the jasmonate methyltransferase during the transition phase and the abscisic acid receptor PYL4 in the ripening phase. The characterization of transcriptome changes in Sangiovese berry after PFD highlights, on one hand, the stronger effect of environment than treatment on the whole berry transcriptome rearrangement during development and, on the other, expands existing knowledge of the main molecular and biochemical modifications occurring in defoliated vines. Moreover, the identification of candidate genes associated with PFD in different genotypes and environments provides new insights into the applicability and repeatability of this crop practice, as well as its possible agricultural and qualitative outcomes across genetic and environmental variability.

Influence of berry ripeness on accumulation, composition and extractability of skin and seed flavonoids in cv. Sangiovese (Vitis vinifera L.).

BACKGROUND: The anthocyanin and tannin concentration and composition of Vitis vinifera L. cv. Sangiovese berries were investigated from post-veraison to harvest. Exhaustive extraction with methanol and acetone was performed to determine the total flavonoid concentration, while a model hydroalcoholic solution was used to prepare extracts representing the winemaking process. The aim of this study was to improve the knowledge of the phenolic maturity of Sangiovese grape. RESULTS: The total anthocyanin concentration increased during ripening, but the quantity of extractable anthocyanins increased more rapidly than the total. The total skin tannin concentration declined from post-veraison to harvest, whereas the extractable portion increased, with little difference in the composition of the fractions. Both the total and extractable seed tannin concentration diminished rapidly just after veraison, and only small fluctuations were detected until harvest. CONCLUSION: These results indicate that the extractability of anthocyanins and skin tannins increases during ripening, whereas there is no clear trend for seed tannins during the same period. This is the first survey to study the behavior of phenolic compounds during different steps of ripening of Sangiovese grape. © 2016 Society of Chemical Industry.

The grapevine VviPrx31 peroxidase as a candidate gene involved in anthocyanin degradation in ripening berries under high temperature.

Anthocyanin levels decline in some red grape berry varieties ripened under high-temperature conditions, but the underlying mechanism is not yet clear. Here we studied the effects of two different temperature regimes, representing actual Sangiovese (Vitis vinifera L.) viticulture regions, on the accumulation of mRNAs and enzymes controlling berry skin anthocyanins. Potted uniform plants of Sangiovese were kept from veraison to harvest, in two plastic greenhouses with different temperature conditions. The low temperature (LT) conditions featured average and maximum daily air temperatures of 20 and 29 °C, respectively, whereas the corresponding high temperature (HT) conditions were 22 and 36 °C, respectively. The anthocyanin concentration at harvest was much lower in HT berries than LT berries although their profile was similar under both conditions. Under HT conditions, the biosynthesis of anthocyanins was suppressed at both the transcriptional and enzymatic levels, but peroxidase activity was higher. This suggests that the low anthocyanin content of HT berries reflects the combined impact of reduced biosynthesis and increased degradation, particularly the direct role of peroxidases in anthocyanin catabolism. Overexpression of VviPrx31 decreased anthocyanin contents in Petunia hybrida petals under heat stress condition. These data suggest that high temperature can stimulate peroxidase activity thus anthocyanin degradation in ripening grape berries.

Selective defoliation affects plant growth, fruit transcriptional ripening program and flavonoid metabolism in grapevine.

BACKGROUND: The selective removal of grapevine leaves around berry clusters can improve the quality of ripening fruits by influencing parameters such as the berry sugar and anthocyanin content at harvest. The outcome depends strongly on the timing of defoliation, which influences the source-sink balance and the modified microclimate surrounding the berries. We removed the basal leaves from Vitis vinifera L. cv Sangiovese shoots at the pre-bloom and veraison stages, and investigated responses such as shoot growth, fruit morphology and composition compared to untreated controls. Moreover, we performed a genome-wide expression analysis to explore the impact of these defoliation treatments on berry transcriptome. RESULTS: We found that pre-bloom defoliation improved berry quality traits such as sugar and anthocyanin content, whereas defoliation at veraison had a detrimental effect, e.g. less anthocyanin and higher incidence of sunburn damage. Genome-wide expression analysis during berry ripening revealed that defoliation at either stage resulted in major transcriptome reprogramming, which slightly delayed the onset of ripening. However, a closer investigation of individual gene expression profiles identified genes that were specifically modulated by defoliation at each stage, reflecting the uncoupling of metabolic processes such as flavonoid biosynthesis, cell wall and stress metabolism, from the general ripening program. CONCLUSIONS: The specific transcriptional modifications we observed following defoliation at different time points allow the identification of the developmental or metabolic processes affected in berries thus deepening the knowledge of the mechanisms by which these agronomical practices impact the final berry ripening traits.

Increasing the source/sink ratio in Vitis vinifera (cv Sangiovese) induces extensive transcriptome reprogramming and modifies berry ripening.

BACKGROUND: Cluster thinning is an agronomic practice in which a proportion of berry clusters are removed from the vine to increase the source/sink ratio and improve the quality of the remaining berries. Until now no transcriptomic data have been reported describing the mechanisms that underlie the agronomic and biochemical effects of thinning. RESULTS: We profiled the transcriptome of Vitis vinifera cv. Sangiovese berries before and after thinning at veraison using a genome-wide microarray representing all grapevine genes listed in the latest V1 gene prediction. Thinning increased the source/sink ratio from 0.6 to 1.2 m2 leaf area per kg of berries and boosted the sugar and anthocyanin content at harvest. Extensive transcriptome remodeling was observed in thinned vines 2 weeks after thinning and at ripening. This included the enhanced modulation of genes that are normally regulated during berry development and the induction of a large set of genes that are not usually expressed. CONCLUSION: Cluster thinning has a profound effect on several important cellular processes and metabolic pathways including carbohydrate metabolism and the synthesis and transport of secondary products. The integrated agronomic, biochemical and transcriptomic data revealed that the positive impact of cluster thinning on final berry composition reflects a much more complex outcome than simply enhancing the normal ripening process.