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.
Open GPB
How sensor technologies combined with artificial intelligence increase the efficiency in grapevine breeding (research): current developments and future perspectives
Viticulture and grapevine breeding programs have to face and adapt to the rapidly changing growing conditions due to the ongoing climate change, the scarcity of resources and the demand for sustainability within the whole value chain of wine production. In times of highly effective and cost-efficient genotyping technologies routinely applied in plant research and breeding, the need for comparable high-speed and high-resolution phenotyping tools has increased substantially. The disciplines of grapevine research, breeding and precision viticulture picked up this demand – mostly independent from each other – by the development, validation and establishment of different sensor technologies in order to extend management strategies or to transform labor-intensive and expensive phenotyping.
The grapevine single-berry clock, practical tools and outcomes
The dynamic sequence of physiological events along the three-months of berry development from anthesis to ripe stage has been thoroughly investigated. Most studies were performed on average samples, taking care to crush enough fruits to fairly represent the overall trend of the future harvest. However, phenological stages like 30% caps off (EL25) highlights the asynchronous nature of this population. Consequently, softening, onset of sugar accumulation and coloration were melted by asynchrony in a developmental mumbo jumbo, until their respective timing could be clarified by single berries approaches.
Unveiling the Grapevine Red Blotch Virus (GRBV) host-pathogen arms-race via multi-omics for enhanced viral defense
The Grapevine Red Blotch Virus (GRBV) poses a critical challenge to the wine sector, lacking a uniquely identified vector. Current control methods involve costly and labor-intensive vine removal, emphasizing the urgency for targeted alternatives. The limited understanding of intricate host-virus interactions underscores the need for foundational knowledge to develop innovative disease control strategies. These include efforts to boost the plant’s RNA interference (RNAi) response, including RNA-based topical applications.
Activation of retrotransposition in grapevine
Retrotransposons, particularly of the Ty-Copia and Ty-Gypsy superfamilies, represent the most abundant and widespread transposons in many plant genomes. Grapevine is no exception and it is clear that these mobile elements have played a major role in the evolution of Vitaceae genomes. While speculation abounds around the possible role of transposons in plant genomes, outside of the rather obvious involvement of retrotransposition in fueling genome expansion, there is little clarity of the actual role these elements have in both developing new genetic variation and in modulating epigenetic responses within genomes to changing climate. To this end we have been exploring de-novo assembled Sauvignon blanc and Pinot noir genomes with a view to catalogue retrotransposon loci to determine the structural intactness and thus age of insertion variation across a small number of clonal linages of these 2 varietals in an attempt to identify ‘live’ TE loci.
Reduced bunch compactness in a clone of Tempranillo associates with a complex reciprocal translocation detected by long-read sequencing genomics
Grapevine cultivars are vegetatively propagated to maintain their varietal attributes. However, spontaneous somatic variation emerges during prolonged periods of vegetative growth, providing an opportunity for the natural improvement of traditional grapevine cultivars. Notably, reduction in bunch compactness is a favorable trait in viticulture, offering advantages such as decreased susceptibility to bunch fungal diseases, and a more uniform ripening of berries. To unravel the genetic and developmental mechanisms behind bunch compactness variation, we examined a somatic variant of Tempranillo Tinto cultivar with loose bunches. We found that the mutant clone exhibits a ~50% reduction in pollen viability compared to typical Tempranillo clones.
The grapevine QTLome is ripe: QTL survey, databasing, and first applications
Overarching surveys of QTL (Quantitative Trait Loci) studies in both model plants and staple crops have facilitated the access to information and boosted the impact of existing data on plant improvement activities. Today, the grapevine community is ready to take up the challenge of making the wealth of QTL information F.A.I.R.. To ensure that all valuable published data can be used more effectively, the myriad of identified QTLs have to be captured, standardised and stored in a dedicated public database.
As an outcome of the GRAPEDIA initiative, QTL-dedicated experts from around the world have gathered to compile the grapevine QTLome: the complete information (e.g., map positions, associated phenotypes) describing all experimentally supported QTLs for a specific trait.
Genomic characterization of terpene biosynthetic genes in seven Vitis vinifera L. varieties
Grapes (Vitis vinifera L.) are a fruit crop of high economic significance globally. Each grapevine cultivar is characterized by its distinctive grape aroma, affecting the wine quality. In several cultivars, the aroma is shaped by terpenoid (mono- and sesqui-terpenoids). Their profile is controlled by terpene synthases (TPS), which are part of a largely expanded gene family. How the variation in TPS copy number and sequence among cultivars determines terpenoid profiles of grapes remains largely unexplored. We annotated TPS in the haplotypes of seven genomes (Riesling, Albariño, Fiano, Gewürztraminer, Pinot Noir, Cabernet Sauvignon, and Viognier) using BLAST, GMAP, PFAM, and phylogenetic analyses. Further, TPS expression patterns and terpenoid accumulation during berry development and ripening were characterized using RNA-Seq and SPME/GC-MS platforms, respectively. Variation in TPS copy number exists among cultivars. Specifically, the TPS counts span a range of 251 to 150 for Riesling and Fiano, respectively, when considering combined haplotypes within each cultivar. Total terpenoid accumulation patterns throughout development were consistent among the five aromatic cultivars, marked by high concentrations in flowers, followed by a decline and subsequent rise during berry development and ripening, respectively. Conversely, non-aromatic cultivars exhibited no substantial increase in terpenoid concentration during ripening. Transcriptome and network analyses are currently employed to determine which TPS are expressed in the berry and determine the terpenoid profile of the specific cultivar. These findings shed light on the genomic determinants of grape aroma in major cultivars, and allow future studies focused on cultivar-specific responses of terpenoid biosynthesis to environmental stresses.
Exploring the gene regulatory networks of WRKY family in grapevine (Vitis vinifera L.) using DAP-Seq
The recent development of regulatory genomics has raised increasing interest in plant research since transcriptional regulation of genes plays a pivotal role in many biological processes. By shedding light on the target genes of the various transcription factors (TFs), it is therefore possible to infer the influence they exert on the different molecular mechanisms. In this regard, the attention was focused on WRKYs, a family of TFs almost exclusively found in plant species. In grapevine, WRKYs are involved in several biological processes, playing a key role in berry development, hormonal balance and signalling, biotic and abiotic stresses responses, and secondary metabolites biosynthesis.
Somatic embryogenesis and polyploidy in grapevine: morphological shoot and leaf traits variations
Somatic embryogenesis (SE) has been used in a variety of biotechnology applications such as virus elimination, cryopreservation, induced mutagenesis and genetic transformation. The SE induction process may cause DNA alterations and ploidy changes, which may provide a source of genetic variability useful for the improvement of agronomic characteristics of plants. This research aims at investigating the spontaneous alterations of the genome in grapevine plants regenerated through SE. Regenerants obtained from different embryogenic events from three different grapevine genotypes (Catarratto, Frappato and Nero d’Avola) were analysed.
Optimized protocol for high-quality RNA extraction from grape tissues using sorbitol pre-wash
Obtaining high-quality RNA from grape tissues, including berry pulp, berry skins, stems, rachis, or roots, is challenging due to their composition, which includes polysaccharides, phenolic compounds, sugars, and organic acids that can negatively affect RNA extraction. For instance, polyphenols and other secondary metabolites can bind to RNA, making it difficult to extract a pure sample. Additionally, RNA can co-precipitate with polysaccharides, leading to lower extraction yield. Also, sugars and organic acids can interfere with the pH and ionic properties of the extraction buffer. To address these challenges, we optimized a protocol for RNA isolation from grape tissues.
Preliminary steps of a protocol to isolate transcription factors bound to a specific DNA locus in grapevine using CRISPR-dCas9 system
Cis-acting regulatory elements are DNA sequences that can be bound by transcription factors to regulate the expression of genes in a condition-dependent and tissue-specific way. It is nowadays possible to search for DNA motives and sequences that a given transcription factor is binding or at least can, but it is still hard to have a glance at all the transcription factors that are contemporaneously located at the same locus. Inspired by an existing technique that uses the CRISPR-Cas system in mammal cells, we are trying to develop a protocol to study such regulation in Vitis vinifera. Using the highly sequence-specific binding capacity of a catalytically inactive Cas9 protein (dCas9), our idea is to set up a system to target a desired sequence and precipitate all the crosslinked proteins and distantly interacting chromatin at this locus and analyze them.
Enhancing plant defense: carbon dots for efficient spray-induced gene silencing
Ectopic RNA application for plant defense faces challenges in tree crops, including size, diffusion, and stability of active compounds such as ribonucleoproteins and nucleic acids. While existing strategies involve expressing dsRNA in transgenic plants targeting pathogens, our research strives to develop a transient RNAi system based on Spray-Induced Gene Silencing (SIGS). This approach aims to circumvent legal barriers and public concerns associated with genetically modified organisms (GMOs). Our strategy integrates SIGS with branched polyethyleneimine-functionalized Carbon Dots (bPEI-CDs) as nanocarriers, effectively addressing unique delivery challenges in plant defense as RNA stability and uptake enhancement
Plant regeneration via somatic embryogenesis and preliminary trials for the application of the DNA-free genome editing in grapevine cv. Corvina veronese
Grapevine (Vitis spp.) is a globally significant fruit crop, and enhancing its agronomic and oenological traits is crucial to meet changing agricultural conditions and consumer demands. Conventional breeding has played a key role in domesticating grapevine varieties, but it is a time-consuming process to develop new cultivars with desirable traits for cultivation.
New plant breeding techniques (NpBTs) offer a potential revolution in grapevine cultivation, and genome editing has shown promise for targeted mutagenesis. The success of these biotechnological approaches relies on efficient in vitro regeneration protocols, particularly through somatic embryogenesis (SE).
Using nanopore skim-sequencing to characterise regional epigenetic variability in New Zealand Sauvignon Blanc
Recent advancements in genomic sequencing technologies have enabled more detailed and direct studies of DNA methylation, which can help characterise epigenetic variations in plants. The Grapevine Improvement team at the Bragato Research Institute is studying the use of Oxford Nanopore sequencing to identify epigenetic changes associated with environmental differences among clonally-propagated grapevines.
This study involved sequencing DNA from the same Sauvignon Blanc clone, sourced from diverse New Zealand viticultural regions, using the PromethION platform.
Exogenous dsRNA applications to identify novel candidate susceptibility genes to downy mildew
One of the major threats to viticulture is represented by fungal pathogens. Plasmopara viticola, an oomycete causing grapevine downy mildew, is one of the principal causes of grape production losses. The most efficient management strategies are represented by a combination of agronomical practices, fungicides’ applications, and use of resistant varieties. Plant resistance is conferred by the presence of resistance (R) genes. Opposed to them, susceptibility (S) genes are encoded by plants and exploited by pathogens to promote infection. Loss or mutation of S genes can limit the ability of pathogens to infect the host. By exploiting post-transcriptional gene silencing, known as RNA intereference (RNAi), it is possible to knock-down the expression of S genes, promoting plant resistance.
Novel approaches and promising perspectives for enhancing grapevine editing and regeneration
Grapevine (Vitis vinifera L.) is a challenging plant species to transform and regenerate due to its complex genome and biological characteristics. This limits the development of cisgenic and gene-edited varieties. One hurdle is selecting the best starting tissue for the transformation process, much like isolating suitable tissue for protoplasts. One promising method involves delivering CRISPR/Cas components to protoplasts isolated from embryogenic calli, which are then induced to regenerate. However, this process is inefficient, time-consuming, and only applicable to a few genotypes. To enhance grapevine regeneration efficiency, the expression of developmental and plant growth regulators shows promise in escaping the recalcitrance encountered in traditional tissue culture methods.
A versatile genome editing platform for grapevine: improving biotic and abiotic stress resilience
New Plant Breeding Techniques (NPBTs) have arisen with the objective of surmounting the constraints inherent in conventional breeding methodologies, thereby enhancing plant resilience against both biotic and abiotic stresses. To date the application of genome editing in grapevine is still limited by the necessity to overcome recalcitrance to produce embryogenic calli and to regenerate plants. In our studies, we developed a smart and versatile genetic transformation system carrying all the most promising features of different genome editing approaches. In specific, we joined the GRF-GIF expression to improve regeneration, the systemic movement of the editing transcripts through tRNA-like sequences (TLS) and the cisgenic-like approach to remove transgenes.