Impact of agronomic practices on grape aroma composition: a review.

Aroma compounds are secondary metabolites that play a key role in grape quality for enological purposes. Terpenes, C13 -norisoprenoids, phenols, and non-terpenic alcohols are the most important aroma compounds in grapes and they can be found as free volatiles or glycoconjugated (bound) molecules. The non-volatile glycosylated group is the largest, and it is present in all varieties of Vitis vinifera (L.), the most widely used species for wine production. These aroma precursors represent the reserve of aroma molecules that can be released during winemaking. Their relative and absolute concentrations at fruit ripening determine the organoleptic value of the final product. A large range of biotic and abiotic factors can influence their biosynthesis in several ways. Agronomic practices such as irrigation, training systems, leaf removal, and bunch thinning can have an effect at plant level. The spraying of stimulatory compounds on fruit at different developmental stages has also been shown to modify metabolic pathways at fruit level with some impact on the aroma composition of the grapevine fruit. Viticulturists could act to promote aroma precursors to improve the aromatic profile of grapes and the wine ultimately produced. However, agronomic practices do not always have uniform results. The metabolic and physiological changes resulting from agronomic practices are unknown because there has not been sufficient research to date. This review presents the state of the art regarding the influences of vineyard agronomic management on the biosynthesis of grape aroma compounds. Although literature regarding the topic is abundant there are still many unknown biological mechanisms involved and/or that have been insufficiently studied. The aim of this work is therefore to find the gaps in scientific literature so that future investigations can focus on them. © 2018 Society of Chemical Industry.

The Microvine, a plant model to study the effect of vine-shoot extract on the accumulation of glycosylated aroma precursors in grapes.

BACKGROUND: The Microvine plant model displays unique reproductive organ behavior and is suitable for grapevine fruit physiological studies, allowing one to undertake studies up to five times more rapidly than the current situation with grapevines. Recently, vine-shoot aqueous extracts, which have an interesting phenolic and aroma composition, have been proposed as viticultural biostimulants, since their post-veraison foliar application to grapevines impacts the wine aroma profile. Using Microvines, the aim of this study was to determine the effect of vine-shoot extract foliar application on 21 stages of grape development. The application was carried out from BBCH 53 (inflorescences clearly visible) to BBCH 85 (softening of berries) to reveal stage-specific responses of the accumulation of glycosylated aroma precursors at BBCH 89 (berries ripe for harvest), the phenological stage selected to study the treatment effect. RESULTS: Microvine use made it possible to carry out 15 sampling time points during 86 days of the experiment, which were established by the cumulative degree days (CDD) parameter. The results confirmed that vine-shoot extract treatment had a positive impact on total glycosylated compounds, especially aglycones such as alcohols, terpenes and C13 -norisoprenoids, with a higher effect when the treatment was applied during ripening. CONCLUSION: Extrapolation of the results to grapevines suggests that vine-shoot extract treatment could modulate the synthesis of grape glycosylated aroma precursors. © 2017 Society of Chemical Industry.

Temperature desynchronizes sugar and organic acid metabolism in ripening grapevine fruits and remodels their transcriptome.

BACKGROUND: Fruit composition at harvest is strongly dependent on the temperature during the grapevine developmental cycle. This raises serious concerns regarding the sustainability of viticulture and the socio-economic repercussions of global warming for many regions where the most heat-tolerant varieties are already cultivated. Despite recent progress, the direct and indirect effects of temperature on fruit development are far from being understood. Experimental limitations such as fluctuating environmental conditions, intra-cluster heterogeneity and the annual reproductive cycle introduce unquantifiable biases for gene expression and physiological studies with grapevine. In the present study, DRCF grapevine mutants (microvine) were grown under several temperature regimes in duly-controlled environmental conditions. A singly berry selection increased the accuracy of fruit phenotyping and subsequent gene expression analyses. The physiological and transcriptomic responses of five key stages sampled simultaneously at day and nighttime were studied by RNA-seq analysis. RESULTS: A total of 674 millions reads were sequenced from all experiments. Analysis of differential expression yielded in a total of 10 788 transcripts modulated by temperature. An acceleration of green berry development under higher temperature was correlated with the induction of several candidate genes linked to cell expansion. High temperatures impaired tannin synthesis and degree of galloylation at the transcriptomic levels. The timing of malate breakdown was delayed to mid-ripening in transgressively cool conditions, revealing unsuspected plasticity of berry primary metabolism. Specific ATPases and malate transporters displayed development and temperature-dependent expression patterns, besides less marked but significant regulation of other genes in the malate pathway. CONCLUSION: The present study represents, to our knowledge the first abiotic stress study performed on a fleshy fruits model using RNA-seq for transcriptomic analysis. It confirms that a careful stage selection and a rigorous control of environmental conditions are needed to address the long-term plasticity of berry development with respect to temperature. Original results revealed temperature-dependent regulation of key metabolic processes in the elaboration of berry composition. Malate breakdown no longer appears as an integral part of the veraison program, but as possibly triggered by an imbalance in cytoplasmic sugar, when efficient vacuolar storage is set on with ripening, in usual temperature conditions. Furthermore, variations in heat shock responsive genes that will be very valuable for further research on temperature adaptation of plants have been evidenced.

Identification of stable QTLs for vegetative and reproductive traits in the microvine (Vitis vinifera L.) using the 18 K Infinium chip.

BACKGROUND: The increasing temperature associated with climate change impacts grapevine phenology and development with critical effects on grape yield and composition. Plant breeding has the potential to deliver new cultivars with stable yield and quality under warmer climate conditions, but this requires the identification of stable genetic determinants. This study tested the potentialities of the microvine to boost genetics in grapevine. A mapping population of 129 microvines derived from Picovine x Ugni Blanc flb, was genotyped with the Illumina® 18 K SNP (Single Nucleotide Polymorphism) chip. Forty-three vegetative and reproductive traits were phenotyped outdoors over four cropping cycles, and a subset of 22 traits over two cropping cycles in growth rooms with two contrasted temperatures, in order to map stable QTLs (Quantitative Trait Loci). RESULTS: Ten stable QTLs for berry development and quality or leaf area were identified on the parental maps. A new major QTL explaining up to 44 % of total variance of berry weight was identified on chromosome 7 in Ugni Blanc flb, and co-localized with QTLs for seed number (up to 76 % total variance), major berry acids at green lag phase (up to 35 %), and other yield components (up to 25 %). In addition, a minor QTL for leaf area was found on chromosome 4 of the same parent. In contrast, only minor QTLs for berry acidity and leaf area could be found as moderately stable in Picovine. None of the transporters recently identified as mutated in low acidity apples or Cucurbits were included in the several hundreds of candidate genes underlying the above berry QTLs, which could be reduced to a few dozen candidate genes when a priori pertinent biological functions and organ specific expression were considered. CONCLUSIONS: This study combining the use of microvine and a high throughput genotyping technology was innovative for grapevine genetics. It allowed the identification of 10 stable QTLs, including the first berry acidity QTLs reported so far in a Vitis vinifera intra-specific cross. Robustness of a set of QTLs was assessed with respect to temperature variation.

Mis-expression of a PISTILLATA-like MADS box gene prevents fruit development in grapevine.

The FLESHLESS BERRY (Flb) somatic variant identified in the grapevine cultivar Ugni Blanc develops grape berries without flesh, suggesting a role for the altered gene in differentiation of flesh cells. Here we describe identification of the molecular defect responsible for this phenotype. Using a combination of genetic and transcriptomic approaches, we detected the insertion of a miniature inverted-repeat transposable element in the promoter region of the PISTILLATA-like (VvPI) gene, the grapevine homologue of Arabidopsis PISTILLATA. The transposon insertion causes specific ectopic expression of the corresponding VvPI allele during early fruit development, causing expression of genes specific for petal and stamen development within the fruit. A causal relationship between the insertion and the phenotype was demonstrated by phenotypic and molecular analyses of somatic revertants showing that ectopic expression and mutant phenotype were always linked to the presence of the transposon insertion. The various phenotypic effects of the flb mutation on ovary morphology, fruit set and fruit development, depending on the cell lineage affected, are presented for each phenotype, offering new insights into floral and fleshly fruit development. The results highlight the importance of VvPI repression after fertilization to achieve normal fleshy fruit development, and the complex genetic, genomic and cellular interactions required for the flower to fruit transition in grapevine.

Genetic dissection of a TIR-NB-LRR locus from the wild North American grapevine species Muscadinia rotundifolia identifies paralogous genes conferring resistance to major fungal and oomycete pathogens in cultivated grapevine.

The most economically important diseases of grapevine cultivation worldwide are caused by the fungal pathogen powdery mildew (Erysiphe necator syn. Uncinula necator) and the oomycete pathogen downy mildew (Plasmopara viticola). Currently, grapegrowers rely heavily on the use of agrochemicals to minimize the potentially devastating impact of these pathogens on grape yield and quality. The wild North American grapevine species Muscadinia rotundifolia was recognized as early as 1889 to be resistant to both powdery and downy mildew. We have now mapped resistance to these two mildew pathogens in M. rotundifolia to a single locus on chromosome 12 that contains a family of seven TIR-NB-LRR genes. We further demonstrate that two highly homologous (86% amino acid identity) members of this gene family confer strong resistance to these unrelated pathogens following genetic transformation into susceptible Vitis vinifera winegrape cultivars. These two genes, designated resistance to Uncinula necator (MrRUN1) and resistance to Plasmopara viticola (MrRPV1) are the first resistance genes to be cloned from a grapevine species. Both MrRUN1 and MrRPV1 were found to confer resistance to multiple powdery and downy mildew isolates from France, North America and Australia; however, a single powdery mildew isolate collected from the south-eastern region of North America, to which M. rotundifolia is native, was capable of breaking MrRUN1-mediated resistance. Comparisons of gene organization and coding sequences between M. rotundifolia and the cultivated grapevine V. vinifera at the MrRUN1/MrRPV1 locus revealed a high level of synteny, suggesting that the TIR-NB-LRR genes at this locus share a common ancestor.

Day and night heat stress trigger different transcriptomic responses in green and ripening grapevine (vitis vinifera) fruit.

BACKGROUND: Global climate change will noticeably affect plant vegetative and reproductive development. The recent increase in temperatures has already impacted yields and composition of berries in many grapevine-growing regions. Physiological processes underlying temperature response and tolerance of the grapevine fruit have not been extensively investigated. To date, all studies investigating the molecular regulation of fleshly fruit response to abiotic stress were only conducted during the day, overlooking possible critical night-specific variations. The present study explores the night and day transcriptomic response of grapevine fruit to heat stress at several developmental stages. Short heat stresses (2 h) were applied at day and night to vines bearing clusters sequentially ordered according to the developmental stages along their vertical axes. The recently proposed microvine model (DRCF-Dwarf Rapid Cycling and Continuous Flowering) was grown in climatic chambers in order to circumvent common constraints and biases inevitable in field experiments with perennial macrovines. Post-véraison berry heterogeneity within clusters was avoided by constituting homogenous batches following organic acids and sugars measurements of individual berries. A whole genome transcriptomic approach was subsequently conducted using NimbleGen 090818 Vitis 12X (30 K) microarrays. RESULTS: Present work reveals significant differences in heat stress responsive pathways according to day or night treatment, in particular regarding genes associated with acidity and phenylpropanoid metabolism. Precise distinction of ripening stages led to stage-specific detection of malic acid and anthocyanin-related transcripts modulated by heat stress. Important changes in cell wall modification related processes as well as indications for heat-induced delay of ripening and sugar accumulation were observed at véraison, an effect that was reversed at later stages. CONCLUSIONS: This first day - night study on heat stress adaption of the grapevine berry shows that the transcriptome of fleshy fruits is differentially affected by abiotic stress at night. The present results emphasize the necessity of including different developmental stages and especially several daytime points in transcriptomic studies.

Haplotype diversity of VvTFL1A gene and association with cluster traits in grapevine (V. vinifera).

BACKGROUND: Interaction between TERMINAL FLOWER 1 (TFL1) and LEAFY (LFY) seem to determine the inflorescence architecture in Arabidopsis. In a parallel way, overexpression of VvTFL1A, a grapevine TFL1 homolog, causes delayed flowering and production of a ramose cluster in the reiterated reproductive meristem (RRM) somatic variant of cultivar Carignan. To analyze the possible contribution of this gene to cluster phenotypic variation in a diversity panel of cultivated grapevine (Vitis vinifera L. subsp. vinifera) its nucleotide diversity was characterized and association analyses among detected sequence polymorphisms and phenology and cluster traits was carried out. RESULTS: A total of 3.6 kb of the VvTFL1A gene, including its promoter, was sequenced in a core collection of 140 individuals designed to maximize phenotypic variation at agronomical relevant traits. Nucleotide variation for VvTFL1A within this collection was higher in the promoter and intron sequences than in the exon regions; where few polymorphisms were located in agreement with a high conservation of coding sequence. Characterization of the VvTFL1A haplotype network identified three major haplogroups, consistent with the geographic origins and the use of the cultivars that could correspond to three major ancestral alleles or evolutionary branches, based on the existence of mutations in linkage disequilibrium. Genetic association studies with cluster traits revealed the presence of major INDEL polymorphisms, explaining 16%, 13% and 25% of flowering time, cluster width and berry weight, respectively, and also structuring the three haplogroups. CONCLUSIONS: At least three major VvTFL1A haplogroups are present in cultivated grapevines, which are defined by the presence of three main polymorphism LD blocks and associated to characteristic phenotypic values for flowering time, cluster width and berry size. Phenotypic differences between haplogroups are consistent with differences observed between Eastern and Western grapevine cultivars and could result from the use of different genetic pools in the domestication process as well as different selection pressures on the development of table and wine cultivars, respectively. Altogether, these results are coherent with previous classifications of grapevine phenotypic diversity mainly based on cluster and berry morphotypes as well as with recent results on the structure of genetic diversity in cultivated grapevine.

A negative MYB regulator of proanthocyanidin accumulation, identified through expression quantitative locus mapping in the grape berry.

Flavonoids are secondary metabolites with multiple functions. In grape (Vitis vinifera), the most abundant flavonoids are proanthocyanidins (PAs), major quality determinants for fruit and wine. However, knowledge about the regulation of PA composition is sparse. Thus, we aimed to identify novel genomic regions involved in this mechanism. Expression quantitative trait locus (eQTL) mapping was performed on the transcript abundance of five downstream PA synthesis genes (dihydroflavonol reductase (VvDFR), leucoanthocyanidin dioxygenase (VvLDOX), leucoanthocyanidin reductase (VvLAR1), VvLAR2 and anthocyanidin reductase (VvANR)) measured by real-time quantitative PCR on a pseudo F1 population in two growing seasons. Twenty-one eQTLs were identified; 17 of them did not overlap with known candidate transcription factors or cis-regulatory sequences. These novel loci and the presence of digenic epistasis support the previous hypothesis of a polygenic regulatory mechanism for PA biosynthesis. In a genomic region co-locating eQTLs for VvDFR, VvLDOX and VvLAR1, gene annotation and a transcriptomic survey suggested that VvMYBC2-L1, a gene coding for an R2R3-MYB protein, is involved in regulating PA synthesis. Phylogenetic analysis showed its high similarity to characterized negative MYB factors. Its spatiotemporal expression profile in grape coincided with PA synthesis. Its functional characterization via overexpression in grapevine hairy roots demonstrated its ability to reduce the amount of PA and to down-regulate expression of PA genes.

Water deficit differentially modulates leaf photosynthesis and transpiration of fungus-tolerant Muscadinia x Vitis hybrids.

Screening for drought performance among novel fungi-tolerant grapevine genotypes is a key point to consider in semiarid regions where water scarcity is a common problem during fruit ripening period. It is therefore important to evaluate the genotypes' responses at the level of carbon metabolism and water demand, under water deficit conditions. This study aimed to characterize leaf and plant water use efficiency (respectively named WUEi and WUEpl) of novel INRAE fungi-tolerant genotypes (including LowSugarBerry (LSB) genotypes), under mild and high-water deficit (WD) and to decipher the photosynthetic parameters leading to higher WUEi. For this purpose, experiments were conducted on potted plants during one season using a phenotyping platform. Two stabilized soil moisture capacity (SMC) conditions, corresponding to mild (SMC 0.6) and high (SMC 0.3) WD, were imposed from the onset of berry ripening until the physiological ripeness stage, which was defined as the point at which fruits reach their maximum solutes and water content. At the whole plant level, all genotypes increased WUEpl under high WD. The highest WUEpl was reached for 3176N, which displayed both a high rate of non-structural carbon accumulation in fruits due to high fruit-to-leaf ratio and low plant transpiration because of low total leaf area. However, when normalizing the fruit-to-leaf ratio among the genotypes, G14 reached the highest normalized WUEpl_n under high WD. At the leaf level, WUEi also increased under high WD, with the highest value attained for G14 and 3176N and the lowest value for Syrah. The higher WUEi values for all genotypes compared to Syrah were associated to higher levels of photosynthesis and changes in light-harvesting efficiency parameters (ΦCO2, qP and qN), while no clear trend was apparent when considering the photosynthetic biochemical parameters (Vcmax, Jmax). Finally, a positive correlation between leaf and plant WUE was observed regardless of genotypes. This study allowed us to classify grapevine genotypes based on their grapes primary metabolite accumulation and water consumption during the critical sugar-loading period. Additionally, the study highlighted the potential drought adaptation mechanism of the LSB genotypes.

Carbon trade-offs in the fruits of fungus-tolerant Muscadinia × Vitis hybrids exposed to water deficit.

Adopting disease-tolerant grapevines is an efficient option to implement a smarter management strategy limiting the environmental impacts linked to pesticide use. However, little is known on their production of fruit metabolites regarding expected future climate fluctuations, such as increased water shortage. Moreover, previous studies about how water deficit impacts grape composition, lack accuracy due to imprecise timing of fruit sampling. In this study, we phenotyped six new fungus-tolerant genotypes exposed to varying water status in field-grown conditions. The accumulation of water, main cations, primary and secondary metabolites were precisely monitored at the arrest of phloem unloading in fruits, which was targeted at the whole cluster level. The goal was to decipher the effects of both genotype and water deficit on the allocation of carbon into soluble sugars, organic acids, amino acids and anthocyanins. The results revealed that the effect of decreased water availability was specific to each berry component. While fruit sugar concentration remained relatively unaffected, the malic/tartaric acid balance varied based on differences among genotypes. Despite showing contrasted strategies on carbon allocation into berry metabolites, all genotypes reduced fruit yield and the amount of compounds of interest per plant under water deficit, with the extent of reduction being genotype-dependent and correlated with the response of berry volume to plant water status. This first set of data provides information to help reasoning the adaptation of these varieties according to the expected risks of drought and the possibilities of mitigating them through irrigation.

New Fungus-Resistant Grapevine Vitis and V. vinifera L. × M. rotundifolia Derivative Hybrids Display a Drought-Independent Response in Thiol Precursor Levels.

The use of new disease-resistant grapevine varieties is a long-term but promising solution to reduce chemical inputs in viticulture. However, little is known about water deficit effects on these varieties, notably regarding berry composition. The aim of this study was to characterize the primary metabolites and thiol precursors levels of 6 fungi-resistant varieties and Syrah. Vines were grown under field conditions and under different water supply levels, and harvested at the phloem unloading arrest. A great variability among varieties regarding the levels of thiol precursors was observed, with the highest concentration, of 539 µg/kg, being observed in 3176-N, a hybrid displaying red fruits. Water deficit negatively and equally impacted the accumulation of sugars, organic acids, and thiol precursors per berry and per plant, with minor effects on their concentration. The observed losses of metabolites per cultivation area suggest that water deficits can lead to significant economic losses for the producer.

miR156b-targeted VvSBP8/13 functions downstream of the abscisic acid signal to regulate anthocyanins biosynthesis in grapevine fruit under drought.

Anthocyanins are the primary color components of grapevine berries and wines. In cultivation practices, a moderate water deficit can promote anthocyanin accumulation in red grape skins. Our previous study showed that abscisic acid (ABA) plays a key role in this process. Herein, we identified a microRNA, vv-miR156b, that is generated in grapevine berries in response to drought stress, along with increasing anthocyanin content and biosynthetic structural gene transcripts. In contrast, vv-miR156b short tandem target mimic (STTM) function-loss callus exhibits the opposite phenotype. Results from in vivo and in vitro experiments revealed that the ABA-signaling-regulated transcription factor VvAREB2 binds directly to the ABA-responsive element (ABRE) of the MIR156b promoter and activates miR156b expression. Furthermore, two miR156b downstream targets, VvSBP8 and VvSBP13, exhibited reduced grape anthocyanin content in their overexpressors but there was a contrary result in their CRISPR-edited lines, the decrease in anthocyanin content was rescued in miR156b and SBP8/13 double overexpressors. We further demonstrated that both VvSBP8 and VvSBP13, encoding transcriptional repressors, displayed sufficient ability to interact with VvMYC1 and VvMYBA1, thereby interfering with MYB-bHLH-WD (MBW) repeat transcriptional complex formation, resulting in the repression of anthocyanin biosynthesis. Our findings demonstrate a direct functional relationship between ABA signaling and the miR156-SBP-MBW complex regulatory module in driving drought-induced anthocyanin accumulation in grape berries.

In vivo grapevine anthocyanin transport involves vesicle-mediated trafficking and the contribution of anthoMATE transporters and GST.

In cells, anthocyanin pigments are synthesized at the cytoplasmic surface of the endoplasmic reticulum, and are then transported and finally accumulated inside the vacuole. In Vitis vinifera (grapevine), two kinds of molecular actors are putatively associated with the vacuolar sequestration of anthocyanins: a glutathione-S-transferase (GST) and two MATE-type transporters, named anthoMATEs. However, the sequence of events by which anthocyanins are imported into the vacuole remains unclear. We used MYBA1 transformed hairy roots as a grapevine model tissue producing anthocyanins, and took advantage of the unique autofluorescence of anthocyanins to study their cellular trafficking. In these tissues, anthocyanins were not only visible in the largest vacuoles, but were also present at higher concentrations in several vesicles of different sizes. In the cell, small vesicles actively moved alongside the tonoplast, suggesting a vesicular trafficking to the vacuole. Subcellular localization assays revealed that anthoMATE transporters were closely related with these small vesicles, whereas GST was localized in the cytoplasm around the nucleus, suggesting an association with the endoplasmic reticulum. Furthermore, cells in hairy roots expressing anthoMATE antisense did not display small vesicles filled with anthocyanins, whereas in hairy roots expressing GST antisense, anthocyanins were accumulated in vesicles but not in the vacuole. This suggests that in grapevine, anthoMATE transporters and GST are involved in different anthocyanin transport mechanisms.

Transposon-induced gene activation as a mechanism generating cluster shape somatic variation in grapevine.

We have characterized the genetic and molecular origin of the reiterated reproductive meristem (RRM) somatic variant phenotype of grapevine cultivar Carignan. Here, we show that the extreme cluster proliferation and delayed anthesis observed in this somatic variant is caused by a single dominant mutation. Transcriptional profiling of Carignan and RRM plants during early stages of inflorescence development demonstrated the overexpression of a few regulatory genes, including VvTFL1A, a close TFL1 Arabidopsis homolog, in RRM inflorescences. Genetic and molecular analyses correlated the insertion of a class-II transposable element, Hatvine1-rrm, in the VvTFL1A promoter, with upregulation of the corresponding VvTFL1A allele in reproductive and vegetative organs of the shoot apex. These results suggest a role for this TFL1 grapevine homolog in the determination of inflorescence structure, with a critical effect on the size and branching pattern of grapevine fruit clusters. Our results demonstrate the existence of spontaneous cis-activation processes caused by class-II transposable elements in grapevine plants, and point to their possible role as a mechanism to generate somatic cell variation in perennial plants. This mechanism is expected to generate dominant phenotypes in chimeric sectors that can be readily exposed to natural selection.

The grape microvine - a model system for rapid forward and reverse genetics of grapevines.

A grapevine model system is described that is suitable for rapid forward and reverse genetic studies in small controlled environments. It is based on the Vvgai1 mutant allele that confers a dwarf stature, short generation cycles and continuous flowering ('microvine'). Black and white berry microvine genotypes were developed that can be transformed by Agrobacterium tumefaciens. Near-homozygous lines were created for efficient bi-allelic single nucleotide polymorphism (SNP) marker mapping and mutagenesis studies. A genetic mapping strategy based on picovine-derived microvine progeny populations was used to rapidly phenotype and map the flower sex and fleshless berry loci and identify a new lethal recessive locus, Vvlrl1. The microvine provides a unique model system for rapid genetic studies of grapevine by changing the perennial long life cycle of the plant to one with features and advantages similar to an annual plant.

A grapevine Shaker inward K(+) channel activated by the calcineurin B-like calcium sensor 1-protein kinase CIPK23 network is expressed in grape berries under drought stress conditions.

Grapevine (Vitis vinifera), the genome sequence of which has recently been reported, is considered as a model species to study fleshy fruit development and acid fruit physiology. Grape berry acidity is quantitatively and qualitatively affected upon increased K(+) accumulation, resulting in deleterious effects on fruit (and wine) quality. Aiming at identifying molecular determinants of K(+) transport in grapevine, we have identified a K(+) channel, named VvK1.1, from the Shaker family. In silico analyses indicated that VvK1.1 is the grapevine counterpart of the Arabidopsis AKT1 channel, known to dominate the plasma membrane inward conductance to K(+) in root periphery cells, and to play a major role in K(+) uptake from the soil solution. VvK1.1 shares common functional properties with AKT1, such as inward rectification (resulting from voltage sensitivity) or regulation by calcineurin B-like (CBL)-interacting protein kinase (CIPK) and Ca(2+)-sensing CBL partners (shown upon heterologous expression in Xenopus oocytes). It also displays distinctive features such as activation at much more negative membrane voltages or expression strongly sensitive to drought stress and ABA (upregulation in aerial parts, downregulation in roots). In roots, VvK1.1 is mainly expressed in cortical cells, like AKT1. In aerial parts, VvK1.1 transcripts were detected in most organs, with expression levels being the highest in the berries. VvK1.1 expression in the berry is localized in the phloem vasculature and pip teguments, and displays strong upregulation upon drought stress, by about 10-fold.VvK1.1 could thus play a major role in K(+) loading into berry tissues, especially upon drought stress.

Transcripts switched off at the stop of phloem unloading highlight the energy efficiency of sugar import in the ripening V. vinifera fruit.

Transcriptomic changes at the cessation of sugar accumulation in the pericarp of Vitis vinifera were addressed on single berries re-synchronised according to their individual growth patterns. The net rates of water, sugars and K+ accumulation inferred from individual growth and solute concentration confirmed that these inflows stopped simultaneously in the ripe berry, while the small amount of malic acid remaining at this stage was still being oxidised at low rate. Re-synchronised individual berries displayed negligible variations in gene expression among triplicates. RNA-seq studies revealed sharp reprogramming of cell-wall enzymes and structural proteins at the stop of phloem unloading, associated with an 80% repression of multiple sugar transporters and aquaporins on the plasma or tonoplast membranes, with the noticeable exception of H+/sugar symporters, which were rather weakly and constitutively expressed. This was verified in three genotypes placed in contrasted thermo-hydric conditions. The prevalence of SWEET suggests that electrogenic transporters would play a minor role on the plasma membranes of SE/CC complex and the one of the flesh, while sucrose/H+ exchangers dominate on its tonoplast. Cis-regulatory elements present in their promoters allowed to sort these transporters in different groups, also including specific TIPs and PIPs paralogs, and cohorts of cell wall-related genes. Together with simple thermodynamic considerations, these results lead to propose that H+/sugar exchangers at the tonoplast, associated with a considerably acidic vacuolar pH, may exhaust cytosolic sugars in the flesh and alleviate the need for supplementary energisation of sugar transport at the plasma membrane.

The reduction of plant sink/source does not systematically improve the metabolic composition of Vitis vinifera white fruit.

The quality of wine grapes depends on the balance between primary and secondary metabolites. Unlike many perennial crops that accumulate starch in the fruits before ripening, the non-climacteric grapes ripe with no previous carbon reserves. Based on the assumption that fruit carbon sink is limiting metabolite accumulation in grapes, bunch thinning is performed to limit plant Sink/Source (S/S). We studied the effects of severe bunch thinning on the accumulation of primary metabolites and on four families of glycosylated aroma precursors (GAPs) at the arrest of fruit phloem unloading of two white grape Vitis vinifera cvs. At plant level, crop reduction resulted in significant losses of metabolites to be accumulated in the fruits: i.e. up to 72% for sugars, 75% for organic acids and GAPs. Nevertheless, S/S manipulation could not modify the balance between GAPs and primary metabolites or increase the concentration in GAPs in the physiologically ripe grape.