This study aimed to assess the response of four red grapevine (Vitis vinifera L.) varieties to elevated temperature, drought and their combination, focusing on the concentration and profile of grape flavonoids. Fruit-bearing cuttings of Tempranillo, Cabernet Sauvignon, Merlot and Grenache grew in greenhouses under, either ambient temperature (T) or ambient temperature + 4 °C (T+4). Plants also received either full irrigation (FI, substrate field capacity) or deficit irrigation (DI, 50 % substrate field capacity). In general, T+4 decreased the concentration of anthocyanins, but DI mitigated this effect. T+4 and DI increased the abundance of methylated anthocyanins and flavonols with additive effects. Grapes under T+4 had higher abundance of acylated anthocyanins, while DI increased the proportion of tri-hydroxylated anthocyanins and flavonols. The impact of interacting elevated temperature and drought on grape composition was genotype dependent. In terms of anthocyanin concentration and profile, Tempranillo was the most affected variety, whereas Grenache was less sensitive.

In Mediterranean regions, severe summers are becoming more common, leading to restrictions to vine productivity and yield quality, requiring sustainable practices to support this sector. We assessed the behaviour of three red grapevine varieties from the Douro Region to examine their tolerance to summer climate stress from the perspective that the less common varieties may have potential for increased use in a climate change scenario. Leaf and fruit biochemical profile, antioxidant activity and fruit colorimetric parameters were assessed at different phenological stages in Aragonez (AR), Tinto Cão (TC) and Touriga Nacional (TN) grape varieties. All three varieties exhibit significant variability in phenological timing, influenced by genetic and environmental factors. Photosynthetic pigment strategies differed among varieties. Chlorophyll content in AR was high to cope with high radiation, while TN displaying a balanced approach, and TC had lower pigment levels, with higher levels of phenolics, antioxidants, and soluble sugars, particularly during stress. The variations in berry biochemical profile highlight the distinct characteristics of the varieties. TC and TN show potential for coping with climate change, having elevated total acidity, while AR has larger and heavier berries with distinct coloration. These findings reinforce the need to study the behaviour of different varieties in each Terroir, to understand their diverse strategies to deal with summer climate stress. This will help in selecting the most suitable variety for these conditions under vineyard management in the Douro Region.

Corinthian currants are dried fruits produced from Vitis vinifera L. var. Apyrena grape. This study investigated the distribution of phenolic compounds in male Wistar rat livers following two distinct Corinthian currant long-term dietary intake protocols (3 and 10% w/w). Method optimization, comparing fresh and lyophilized tissues, achieved satisfactory recoveries (>70%) for most analytes. Enzymatic hydrolysis conditions (37 °C, pH 5.0) minimally affected phenolics, but enzyme addition showed diverse effects. Hydrolyzed lyophilized liver tissue from rats consuming Corinthian currants (3 and 10% w/w) exhibited elevated levels of isorhamnetin (20.62 ± 2.27 ng/g tissue and 33.80 ± 1.38 ng/g tissue, respectively), along with similar effects for kaempferol, quercetin, and chrysin after prolonged Corinthian currant intake. This suggests their presence as phase II metabolites in the fasting-state liver. This study is the first to explore phenolic accumulation in rat liver, simulating real conditions of dried fruit consumption, as seen herein with Corinthian currant.

The main bottleneck in the application of biotechnological breeding methods to woody species is due to the in vitro regeneration recalcitrance shown by several genotypes. On the other side, woody species, especially grapevine (Vitis vinifera L.), use most of the pesticides and other expensive inputs in agriculture, making the development of efficient approaches of genetic improvement absolutely urgent. Genome editing is an extremely promising technique particularly for wine grape genotypes, as it allows to modify the desired gene in a single step, preserving all the quality traits selected and appreciated in elite varieties. A genome editing and regeneration protocol for the production of transgene-free grapevine plants, exploiting the lipofectamine-mediated direct delivery of CRISPR-Cas9 ribonucleoproteins (RNPs) to target the phytoene desaturase gene, is reported. We focused on Nebbiolo (V. vinifera), an extremely in vitro recalcitrant wine genotype used to produce outstanding wines, such as Barolo and Barbaresco. The use of the PEG-mediated editing method available in literature and employed for highly embryogenic grapevine genotypes did not allow the proper embryo development in the recalcitrant Nebbiolo. Lipofectamines, on the contrary, did not have a negative impact on protoplast viability and plant regeneration, leading to the obtainment of fully developed edited plants after about 5 months from the transfection. Our work represents one of the first examples of lipofectamine use for delivering editing reagents in plant protoplasts. The important result achieved for the wine grape genotype breeding could be extended to other important wine grape varieties and recalcitrant woody species.

Mounting evidence recognizes structural variations (SVs) and repetitive DNA sequences as crucial players in shaping the existing grape phenotypic diversity at intra- and inter-species levels. To deepen our understanding on the abundance, diversity, and distribution of SVs and repetitive DNAs, including transposable elements (TEs) and tandemly repeated satellite DNA (satDNAs), we re-sequenced the genomes of the ancient grapes Aglianico and Falanghina. The analysis of large copy number variants (CNVs) detected candidate polymorphic genes that are involved in the enological features of these varieties. In a comparative analysis of Aglianico and Falanghina sequences with 21 publicly available genomes of cultivated grapes, we provided a genome-wide annotation of grape TEs at the lineage level. We disclosed that at least two main clusters of grape cultivars could be identified based on the TEs content. Multiple TEs families appeared either significantly enriched or depleted. In addition, in silico and cytological analyses provided evidence for a diverse chromosomal distribution of several satellite repeats between Aglianico, Falanghina, and other grapes. Overall, our data further improved our understanding of the intricate grape diversity held by two Italian traditional varieties, unveiling a pool of unique candidate genes never so far exploited in breeding for improved fruit quality.

The present study aimed: i) to evaluate the impact of the changes in temperature and relative humidity (RH), projected by the year 2100, on grape ripening, and ii) to assess if bunch transpiration is a key physiological process involved in the advancement in grape development under future climate conditions. Fruit-bearing cuttings of Vitis vinifera L. cv. \'Tempranillo\' were grown, from fruit set to maturity, in glasshouses under two conditions: 24°C/14°C and 55%/70% RH (day/night) (T) vs 28°C/18°C and 43%/58% RH (T+4). To elucidate the role of bunch transpiration in grape development in a future climate scenario, the bunches of half of the plants in the T+4 glasshouse were sprayed with an antitranspirant (AT+4). T+4 increased bunch transpiration, hastened the ripening process, increasing the rate of total soluble solid (TSS) accumulation and malic acid degradation, and reduced the concentration of total anthocyanins. The application of antitranspirant partially alleviated the effects of combined high temperature and low RH on maturation times, through lower TSS accumulation rates. Berries in AT+4 had the lowest concentrations of anthocyanins and color, likely related to a reduction in light transmittance by the antitranspirant film and to higher anthocyanin degradation due to the longer exposure to elevated temperatures. The results show a negative impact of elevated temperature and low RH on grape composition. The increased bunch transpiration under these conditions played an important role in the changes observed in phenology and sugar accumulation.

Plant growth-promoting rhizobacteria (PGPRs) that produce 1-aminocyclopropane-1-carboxylate (ACC) deaminase are capable of reducing limits to plant growth due to water-deficient conditions. Here, seven PGPR strains that can produce ACC deaminase were successfully obtained from the rhizosphere soil of grapevine (Vitis vinifera L.) in arid regions of China. The strains belonged to three different genera: Pseudomonas, Enterobacter, and Achromobacter, according to their 16S rDNA sequencing analysis. A drought tolerance experiment revealed two PGPR strains (DR3 and DR6) with exceptionally high phosphate solubilization, nitrogen fixation, indoleacetic acid (IAA), and exopolysaccharides secretion potential. Both strains were selected for use in a pot experiment to evaluate their growth-promoting effects on grapevines under drought conditions. Each of these two PGPRs and their mixed inoculation into grapevines were expected to alleviate the comprehensive growth inhibition of grapevines caused by drought stress. The mixed inoculation was hypothesized to elicit the best growth-promoting effects. Inoculation with the PGPRs not only enhanced the root-adhering soil/root tissue ratios and soil aggregate stability, but it also increased the nitrogen and phosphorus levels in the soil and plant leaves. Further, inoculation with PGPRs significantly altered the plant height, biomass of shoot and root organs, relative water contents, and net photosynthetic rate of leaves, enabling grapevines to better cope with drought. Moreover, the contents of IAA, abscisic acid, and malondialdehyde in these grapevines under drought stress were significantly changed by PGPRs. They indirectly affected biochemical and physiological properties of grapevines to alleviate their drought stress. Taken together, these results demonstrate that the DR3 and DR6 PGPRs might be useful for effectively weakening the growth inhibition caused by drought in grapevines. The strains might also be applied as effective bioinoculants to maintain the quality of wine grapes.

The aim of the study was to analyse selected aspects of the natural variability of selected varieties of Vitis vinifera. Grapevine is one of the most popular and desirable crops in the world due to the great tradition of wine production, but grape extracts also have a wide range of pharmaceutical effects on the human body. It is important to identify different varieties for the conservation of genetic resources, but also for commercial and cultivation purposes. The variability of conserved DNA-derived polymorphism profiles, as well as microbial characteristics, were analysed in this study. Six different varieties of Vitis vinifera L. were used in the study: Cabernet Savignon, Chardonney, Welschriesling, Weisser Riesling, Gewurztramines and Gruner Veltliner. Genetic polymorphism was analysed by CDDP markers for WRKY genes. Polymorphic amplicon profiles were generated by all primer combinations used in the study. Gruner Veltliner and Welschriesling were the most similar, with a similarity value at 0.778. Microbiological quality of grape and antimicrobial activity against Gram-positive and Gram-negative bacteria and yeasts were analysed further. The plate diluting method for microbial quality and the disc diffusion method for antimicrobial activity were evaluated. The number of total count of bacteria ranged between 3.12 in Cabernet Sauvignon to 3.62 log cfu/g in Grűner Veltliner. The best antimicrobial activity showed Gewurztramines against Salmonella enterica, Yersinia enterocolitica, Pseudomonas aeroginosa, Staphylococcus aureus, Listeria monocytogenes, Candida albicans, Candida krusei, and Candida tropicalis. The best antimicrobial activity was found against Enterococcus faecalis in variety Weisser Riesling.

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Thanks to current advances in sequencing technologies, novel bioinformatics tools, and efficient modeling solutions, association mapping has become a widely accepted approach to unravel the link between genotype and phenotype diversity in numerous crops. In grapevine, this strategy has been used in the last decades to understand the genetic basis of traits of agronomic interest (fruit quality, crop yield, biotic and abiotic resistance), of special relevance nowadays to improve crop resilience to cope with future climate scenarios. Genome-wide association studies have identified many putative causative loci for different traits, some of them overlapping well-known causal genes identified by conventional quantitative trait loci studies in biparental progenies, and/or validated by functional approaches. In addition, candidate-gene association studies have been useful to pinpoint the causal mutation underlying phenotypic variation for several traits of high interest in breeding programs (like berry color, seedlessness, and muscat flavor), information that has been used to develop highly informative and useful markers already in use in marker-assisted selection processes. Thus, association mapping has proved to represent a valuable step towards high quality and sustainable grape production. This review summarizes current applications of association mapping in grapevine research and discusses future prospects in view of current viticulture challenges.