Poster – Advances in omics and big data

Addressing the opportunities and challenges of genomics methods in grapevine (Vitis vinifera L.) requires the development of a comprehensive and accurate reference genome and annotation. We aimed to create a new gene annotation for the PN40024 grapevine reference genome by integrating the highly accurate and complete T2T assembly and the manually curated PN40024.v4 annotation. Here, we present a novel workflow to enhance the annotation of the T2T genome by incorporating past community input found in PN40024.v4. The pipeline's containerization will improve the workflow's reproducibility and flexibility, facilitating its inclusion as a shared workflow on the Grapedia portal, the grapevine genomics encyclopedia.

Genetic variants known to produce single residue missense mutations have been associated with phenotypic traits of commercial interest in grapevine. This is the case of the K284N substitution in VviDXS1 associated with muscat aroma, or the R197L in VviAGL11 causing stenospermocarpic seedless grapes. The impact of such mutations on protein structure, stability, dynamics, interactions, or functional mechanism can be studied by computational methods, including our pyDock scoring, previously developed. For this, knowledge on the 3D structure of the protein and its complexes with other proteins and biomolecules is required, but such knowledge is not available for virtually none of the proteins and complexes in grapevine.

Uruguay is known for the production of Tannat wines, which is a neutral variety from an aroma point of view, but capable of providing aromatic precursors that are of interest in the production of wines for ageing. The main aromatic precursors present are glycosidic compounds and carotenoids. The contribution of carotenoid degradation by-products such as norisoprenoids to wine aroma is fundamental, as they are associated with pleasant aroma descriptors and very low olfactory perception thresholds. Several factors have been shown to influence carotenoid concentrations in grapes, such as cultivar, climatic conditions, viticultural region, plant water status, exposure to sunlight and ripening stage.

Conventional molecular analyses provide bulk genomic/transcriptomic data that are unable to reveal the cellular heterogeneity and to precisely define how gene networks orchestrate organ development. We will profile gene expression and identify open chromatin regions at the individual cells level, allowing to define cell-type specific regulatory elements, developmental trajectories and transcriptional networks orchestrating organ development and function. We will perform scRNA-seq and snATAC-seq on leaf/berry protoplasts and nuclei and combine them with the leaf/berry bulk tissues obtained results, where the analysis of transcripts, chromatin accessibility, histone modification and transcription factor binding sites showed that a large fraction of phenotypic variation appears to be determined by regulatory rather than coding variation and that many variants have an organ-specific effect.

The maintenance and valorization of genetic diversity is an undoubtable resource for the viticulture of the future, since the climate crisis is forcing us to think of new, more resilient varieties. For this reason, the grapevine germplasm of the Fondazione Edmund Mach has been continuously expanded in the last decade to a total of 3,120 accessions, whose trueness-to-type has been verified by means of the universal set of nine microsatellites. About two thirds are V. vinifera subsp. vinifera accessions, while the rest consists of naturalized and selected hybrids, V. vinifera subsp. sylvestris, and pure species. The genetic material has also been characterized over three consecutive years for ampelographic, vine development, and biotic stress response traits to be exploited for experimental purposes.

One of the most important effects of climate change in wine-growing areas is the advance of phenological stages, especially concerning early berry ripening. In the hottest seasons, this results in a lack of synchrony between sugar and phenolic ripeness. In order to cope with this fact, a general effort is being made by researchers and growers aiming at delaying ripening through different strategies. One of the proposed approaches is the application of elicitors. This study aims to assess the effect at the transcriptomic level of the application of three hormone- and natural-based elicitors in Tempranillo.

Haplotype-resolved genome assemblies were produced for Chasselas and Ugni Blanc, two heterozygous real-field genetic pool Vitis vinifera cultivars by combining high-fidelity long-read sequencing (HiFi) and high‐throughput chromosome conformation capture (Hi-C). The telomere-to-telomere full coverage of the chromosomes allowed us to assemble separately the two haplo-genomes of both cultivars and revealed structural variations between the two haplotypes of a given cultivar.

Developing a tractable genetic engineering and gene editing system is an essential tool for grapevine. We initiated a plant transformation and biotechnology program at Oregon State University using the grape microvine system (V. vinifera) in 2018 to interrogate gene-to-trait relationships using traditional genetic engineering and gene editing. The microvine model is also used for nanomaterial-assisted RNP, DNA, and RNA delivery. Most reference genomes and annotations for grapevine are collapsed assemblies of homologous chromosomes and do not represent the specific microvine cultivar ‘043023V004’ under study at our institution.

Grapevine (Vitis vinifera) is amongst the world’s most cultivated fruit crops, and of global and economic significance, producing a wide variety of grape-derived products, including wine, and table grapes. The genus Vitis, encompassing approximately 70 naturally occurring inter-fertile species, exhibits extensive genetic and phenotypic diversity, highlighted by the global cultivation of thousands of predominantly Vitis vinifera cultivars. Despite the importance of harnessing its naturally occurring genetic diversity to pursue traits of interest, especially considering the continued and growing demand for sustainable high-quality grape production, the systematic characterization of available functional genetic variants remains limited.

From fruit set to ripening, the grape berry mesocarp experiences a wide range of dynamic physical, physiological, and biochemical changes, such as mesocarp cell death (MCD) and hydraulic isolation. The premature occurrence of such events is a characteristic of the Niagara Rosada (NR) variety, utilised as table grapes and winemaking. In our opinion, the onset of ripening would not cause MCD, but a down-regulation of respiratory enzymes during the early loss of cell viability, while maintaining membrane integrity. For this, we investigated three distinct developmental stages (green (E-L33), veraison (E-L35), and ripe (E-L39)) of NR berries by label-free proteomics, enzymatic respiratory activity and outer mesocarp imaging. Cell wall-modifying proteins were found to accumulate differently throughout ripening, while cytoplasmic membranes continue intact.

Grapevine cultivars are vegetatively propagated to maintain their varietal characteristics. This process of multiplication leads to spontaneous somatic mutations that can eventually generate a variant phenotype, of potential interest for cultivar improvement and innovation. However, regardless their phenotypic effect, somatic mutations stack in the genome, and they can be used to reveal the origin and dissemination history of ancient cultivars. Here, a stringent somatic variant calling over whole genome resequencing data from 35 ‘Tempranillo Tinto’ clones or old vines from seven Iberian winemaking regions revealed 135 single nucleotide variations (SNVs) shared by some of the clonal lines.

The quality of wine is assessed, among other things, by its color, which is mainly due to its anthocyanin content. These pigments are polyphenols that give red, purple and blue hues depending on the relative proportion of anthocyanins produced by the action of flavonoid 3'5' hydroxylase (delphinidin-3-glucoside, petunidin-3-glucoside, malvidin-3-glucoside) or flavonoid 3' hydroxylase (cyanidin-3-glucoside, peonidin-3-glucoside). To study the genes involved in this biosynthetic pathway, we focused on Vitis vinifera cv. Tannat, known for producing wines with higher anthocyanin content and darker purple color compared to most red grape varieties. In this work, we have performed RNA-Seq analysis of skins during berry development, taking green and red berries at 50% veraison as separate samples, as an experimental strategy to focus on the differential expression of genes of interest.

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