terclim by ICS banner
IVES 9 IVES Conference Series 9 Defining gene regulation and co-regulation at single cell resolution in grapevine

Defining gene regulation and co-regulation at single cell resolution in grapevine

Abstract

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. By bioinformatics approaches we will identify cell and gene clusters, interpreting the heterogeneity from single-cell transcriptomes; subsequently, we will perform in situ hybridizations to corroborate already predicted cell-type annotations and to identify new cell-type marker genes, required for the cell identity definition, and for the experimental validations of scRNA-seq data. The realization of a single cell resolution spatiotemporal transcriptomic and chromatin accessibility map of grapevine berry will allow to link gene expression profiles to cellular and developmental processes, uncovering part of the molecular mechanisms of ripening and slowly providing the key in maintaining high quality grapes and wine. Building organ-scale gene expression maps is essential to drive technological innovation such as reprogramming cell identity and inducing phenotypic changes via cell-type-specific gene editing.

DOI:

Publication date: June 14, 2024

Issue: Open GPB 2024

Type: Poster

Authors

Chiara Foresti1*, Michele Morgante2, Paola Paci3, Sara Zenoni1

1Department of Biotechnology, University of Verona, Verona, Italy
2IGA and Department of Agri-food, Environmental and Animal Sciences, University of Udine, Italy
3Institute for System Analysis and Computer Science “Antonio Ruberti”, National Research Council, Rome, Italy

Contact the author*

Keywords

Single-cell RNA-seq, single nucleus ATAC-seq, gene expression regulation, gene network, developmental trajectories

Tags

IVES Conference Series | Open GPB | Open GPB 2024

Citation

Related articles…

Comparison of aroma-related compounds of carbonic maceration and traditional young red winemaking in case of Merlot by means of targeted metabolomic approach

Winemaking decisions and techniques are known to affect the final aromatic composition of red wines. Winemakers put a constant effort into the improved controlling of vinification procedures to achieve better quality. Anyway an increased customer’s demand for uniqueness is often forcing them to adjust and offer new and new interesting products. To support the producers, an improved knowledge on aromatic potential as affected by classical and alternative strategies is needed.

Mobile device to induce heat-stress on grapevine berries

Studying heat stress response of grapevine berries in the field often relies on weather conditions during the growing season. We constructed a mobile heating device, able to induce controlled heat stress on grapes in vineyards. The heater consisted of six 150 W infrared lamps mounted in a profile frame. Heating power of the lamps could be controlled individually by a control unit consisting of a single board computer and six temperature sensors to reach a pre-set temperature. The heat energy applied to individual berries within a cluster decreases by the squared distance to the heat source, enabling the establishment of temperature profiles within individual clusters. These profiles can be measured by infrared thermography once a steady state has been reached. Radiant flux density received by a berry depending on the distance was calculated based on a view factor and measured lamp surface temperature and resulted to 665 Wm-2 at 7cm. Infrared thermography of the fruit surface was in good agreement with measurements conducted with a thermocouple inserted at epidermis level. In combination with infrared thermography, the presented device offers possibilities for a wide range of applications like phenotyping for heat tolerance in the field to proceed in the understanding of the complex response of plants to heat stress. Sunburn necrosis symptoms were artificially induced with the aid of the device for cv. Bacchus and cv. Sylvaner in the 2020 and 2021 growing season. Threshold temperatures for sunburn induction (LT5030min) were derived from temperature data of single berries and visual sunburn assessment, applying logistic regression. A comparison of threshold temperatures for the occurrence of sunburn necrosis confirmed the higher susceptibility of cv. Bacchus. The lower susceptibility of cv. Sylvaner did not seem to be related to its phenolic composition, rendering a thermoprotective role of berry phenolic compounds unlikely.

Influence of agronomic practices in soil water content in mid-mountain vineyards

In the context of LIFE project MIDMACC (LIFE18 CCA/ES/001099), several pilots have been installed in vineyards in mid mountain areas of Catalonia (NE Spain) to test well stablished agronomic practices to increase the adaptation of Mediterranean mid mountain to climate change. Soil water content (SWC) at three different depths (15, 30 and 45cm) was measured in continuum from August 2020. One pilot (WC) included a well-established green cover (GC), a new GC (NC) and a conventional soil management (CM, tilling+herbicides). NC presented an intermediate state between WC and CM, responding similarly to CM in autumn but quickly reaching similar SWC to WC, then following the same evolution till next spring, with CM presenting lower values along autumn and winter. Then vegetation activation decreased SWC in all plots, (much slower in CM, lacking GC). Sensibility to spring rains is again intermediate for NC, which joins SWC evolution of CM by the end of spring till next autumn. It is expected that NC will resemble WC more and more as its GC develops. In the pilot combining vine training (VSP vs Gobelet) and hillside management (slope vs terrace), no clear pattern could be related with these conditions. However, both terraces seem to be more sensitive to spring rains. A third pilot included new vineyards (7 and 1 year old). In the new vineyard (N), higher canopy development, a spontaneous green cover and row straw resulted in a slower SWC dynamic, not so sensitive to rains but conserving more soil water in spring and most of summer, even with presumably a higher water extraction by vines. In the newest vineyard (VN) the deepest sensor is still sensitive to rain events all over the year and SWC is always highest at this depth, revealing small water capture by vines.

Exploring the impact of yeast derivatives on aromatic and sensory profiles of white and red wines: a multifactorial analysis

Specific inactivated yeast derivatives (SYDS) are obtained from s. cerevisiae yeasts by various processes (thermal, mechanical, and enzymatic) and have diverse oenological applications to improve wine quality. However, different impacts on wine sensory and aromas were reported, depending on syds types and fractions, wine matrices, and experimental settings. Few works have examined the impact of SYDS on aromas considering also those on wine macromolecules influencing organoleptic properties.

Oxidability of wines made from Spanish minority grape varieties

The phenolic profile of a wine plays an essential role in its oxidative capacity and in both white and red wines it defines its shelf life[1]. The study of minority varieties to produce wines with peculiar characteristics necessarily includes the phenolic and oxidative characterization of the wines produced. This paper presents the study of wines made from 24 minority and majority white and red grape varieties, focusing on phenolic characteristics (total phenols, slightly polymerized phenols, highly polymerized phenols, anthocyanins…), color, as well as parameters related to the oxidability of the wines and their capacity to consume oxygen [2].