Alternative splicing regulation appears to play a crucial role in grape berry development and is also potentially involved in adaptation responses to the environment

Background Alternative splicing (AS) produces transcript variants playing potential roles in proteome diversification and gene expression regulation. AS modulation is thus essential to respond to developmental and environmental stimuli. In grapevine, a better understanding of berry development is crucial for implementing breeding and viticultural strategies allowing adaptation to climate changes. Although profound changes in gene transcription have been shown to occur in the course of berry ripening, no detailed study on splicing modifications during this period has been published so far. We report here on the regulation of gene AS in developing berries of two grapevine (Vitis vinifera L.) varieties, Gewurztraminer (Gw) and Riesling (Ri), showing distinctive phenotypic characteristics. Using the software rMATS, the transcriptomes of berries at four developmental steps, from the green stage to mid-ripening, were analysed in pairwise comparisons between stages and varieties. Results A total of 305 differential AS (DAS) events, affecting 258 genes, were identified. Interestingly, 22% of these AS events had not been reported before. Among the 80 genes that underwent the most significant variations during ripening, 22 showed a similar splicing profile in Gw and Ri, which suggests their involvement in berry development. Conversely, 23 genes were subjected to splicing regulation in only one variety. In addition, the ratios of alternative isoforms were different in Gw and Ri for 35 other genes, without any change during ripening. This last result indicates substantial AS differences between the two varieties. Remarkably, 8 AS events were specific to one variety, due to the lack of a splice site in the other variety. Furthermore, the transcription rates of the genes affected by stage-dependent splicing regulation were mostly unchanged, identifying AS modulation as an independent way of shaping the transcriptome. Conclusions The analysis of AS profiles in grapevine varieties with contrasting phenotypes revealed some similarity in the regulation of several genes with developmental functions, suggesting their involvement in berry ripening. Additionally, many splicing differences were discovered between the two varieties, that could be linked to phenotypic specificities and distinct adaptive capacities. Together, these findings open perspectives for a better understanding of berry development and for the selection of grapevine genotypes adapted to climate change.

Transcriptomic analysis of temporal shifts in berry development between two grapevine cultivars of the Pinot family reveals potential genes controlling ripening time

Background Grapevine cultivars of the Pinot family represent clonally propagated mutants with major phenotypic and physiological differences, such as different colour or shifted ripening time, as well as changes in important viticultural traits. Specifically, the cultivars ‘Pinot Noir’ (PN) and ‘Pinot Noir Precoce’ (PNP, early ripening) flower at the same time, but vary in the beginning of berry ripening (veraison) and, consequently, harvest time. In addition to genotype, seasonal climatic conditions (i.e. high temperatures) also affect ripening times. To reveal possible regulatory genes that affect the timing of veraison onset, we investigated differences in gene expression profiles between PN and PNP throughout berry development with a closely meshed time series and over two separate years. Results The difference in the duration of berry formation between PN and PNP was quantified to be approximately two weeks under the growth conditions applied, using plant material with a proven PN and PNP clonal relationship. Clusters of co-expressed genes and differentially expressed genes (DEGs) were detected which reflect the shift in the timing of veraison onset. Functional annotation of these DEGs fit to observed phenotypic and physiological changes during berry development. In total, we observed 3,342 DEGs in 2014 and 2,745 DEGs in 2017 between PN and PNP, with 1,923 DEGs across both years. Among these, 388 DEGs were identified as veraison-specific and 12 were considered as berry ripening time regulatory candidates. The expression profiles revealed two candidate genes for ripening time control which we designated VviRTIC1 and VviRTIC2 (VIT_210s0071g01145 and VIT_200s0366g00020, respectively). These genes likely contribute the phenotypic differences observed between PN and PNP. Conclusions Many of the 1,923 DEGs show highly similar expression profiles in both cultivars if the patterns are aligned according to developmental stage. In our work, putative genes differentially expressed between PNP and PN which could control ripening time as well as veraison-specific genes were identified. We point out connections of these genes to molecular events during berry development and discuss potential candidate genes which may control ripening time. Two of these candidates were observed to be differentially expressed in the early berry development phase. Several down-regulated genes during berry ripening are annotated as auxin response factors / ARFs. Conceivably, general changes in auxin signaling may cause the earlier ripening phenotype of PNP.

Influence of Berry Ripening Stages over Phenolics and Volatile Compounds in Aged Aglianico Wine

The harvest time of grapes is a major determinant of berry composition and of the wine quality, and it is usually established through empirical testing of main biochemical parameters of the berry. In this work, we studied how the ripening stage of Aglianico grapes modulates key secondary metabolites of wines, phenolics and volatile compounds. Specifically, we analyzed and compared four berry ripening stages corresponding to total soluble solids of 18, 20, 22, and 25 °Brix and related aged wines. Wine color intensity, anthocyanins level and total trans-resveratrol (free + glycosidic form) increased with grape maturity degree. Wines obtained from late-harvested grapes significantly differed from the others for a higher content of aliphatic alcohols, esters, acetates, a-terpineol and benzyl alcohol. The content of glycosidic terpene compounds, such as nerol, geraniol and a-terpineol, was higher in wines obtained with grapes harvested at 25 °Brix compared to the earlier harvests. Our work indicated that the maturity of the grape is a determining factor in phenolic and volatile compounds of red Aglianico wines. Moreover, extending grape ripening to a sugar concentration higher than 22 °Brix improves the biochemical profile of aged wine in terms of aroma compounds and of phytochemicals with known health-related benefits.

Kaolin impacts on hormonal balance, polyphenolic composition and oenological parameters in red grapevine berries during ripening

BACKGROUND: Kaolin particle-film application is a well-known strategy to avoid fruit damage. However, its putative role in balancing berry ripening under a changing climate remains poorly explored. OBJECTIVE: We assessed kaolin treatment effect on several ripening berry components, hormonal balance and oenological parameters of the field-grown Touriga-Franca (TF) and Touriga–Nacional (TN) grapevine varieties at veraison (EL35) and ripening (EL38) during two growing seasons (2017 and 2018). RESULTS: Under the adverse summer conditions (two heatwave events) of 2017, kaolin application increased 211.2 %and 51.4 %the salicylic acid (SA) and abscisic acid (ABA) levels in TF berries at EL38, while no significant differences were observed in TN. Conversely, TF, and TN kaolin treated berries showed lower SA and ABA accumulation in 2018, respectively. Tartaric acid content increased about 17.2 %, and 24.2 %in TF and TN treated berries at stage EL35 in the 2017 growing season. Though kaolin treatment had no consistent effect on anthocyanins accumulation, flavonoids, ortho-diphenols and tannins increased in kaolin treated grapevines in 2017. CONCLUSIONS: This study highlights the key role of climate in triggering ripening related processes and fruit quality potential. Nevertheless, kaolin treated grapevines displayed an improved response to oxidative stress signals by increasing secondary metabolites accumulation in warm vintages. Kaolin application promoted different varietal responses, with a possible ripening delaying effect in TF, reinforcing its efficiency in alleviating severe summer stress impacts.

Compositional Changes in Grapes and Leaves as a Consequence of Smoke Exposure of Vineyards from Multiple Bushfires across a Ripening Season

The negative effects of smoke exposure of grapes in vineyards that are close to harvest are well documented. Volatile phenols in smoke from forest and grass fires can contaminate berries and, upon uptake, are readily converted into a range of glycosylated grape metabolites. These phenolic glycosides and corresponding volatile phenols are extracted into the must and carried through the winemaking process, leading to wines with overtly smoky aromas and flavours. As a result, smoke exposure of grapes can cause significant quality defects in wine, and may render grapes and wine unfit for sale, with substantial negative economic impacts. Until now, however, very little has been known about the impact on grape composition of smoke exposure very early in the season, when grapes are small, hard and green, as occurred with many fires in the 2019–20 Australian grapegrowing season. This research summarises the compositional consequences of cumulative bushfire smoke exposure of grapes and leaves, it establishes detailed profiles of volatile phenols and phenolic glycosides in samples from six commercial Chardonnay and Shiraz blocks throughout berry ripening and examines the observed effects in the context of vineyard location and timing of smoke exposure. In addition, we demonstrate the potential of some phenolic glycosides in leaves to serve as additional biomarkers for smoke exposure of vineyards.