terclim by ICS banner
IVES 9 IVES Conference Series 9 Open-GPB 9 Open-GPB-2024 9 Flash - New biotechnological tools 9 Exploring the impact of NPR3 gene silencing on the interaction between grapevine and mycorrhizal fungi through genome editing

Exploring the impact of NPR3 gene silencing on the interaction between grapevine and mycorrhizal fungi through genome editing

Abstract

One of the main plant defence mechanisms is the Systemic Acquired Resistance (SAR) mediated by Salicylic Acid (SA). This is a heightened and broad-spectrum immune response initiated by the exposure to pathogens, inducing resistance not only in the infected site, but also throughout the entire plant. It was demonstrated that plant immune system can be regulated by two classes of SA receptors: NONEXPRESSOR OF PR GENES 1 (NPR1) and NPR1-LIKE PROTEIN 3 and 4 (NPR3/NPR4). While NPR1 is required for SA-induction followed by the expression of pathogenesis-related (PR) protein and resistance against pathogens, NPR3/NPR4 serve as transcriptional co-repressors of SA-responsive genes. The aim of this work was to trigger SAR by suppressing NPR3, and to investigate how plant response affects its ability to recruit beneficial microorganisms, specifically arbuscular mycorrhizal fungi (AMF). To this aim, embryogenic calli were obtained from anther and ovaries of grapevine (cultivar Chardonnay) and NPR3 knock out lines were achieved using CRISPR/Cas9 technique. Three regenerated lines, along with a backbone and a wild-type lines, were inoculated in axenic condition with the AMF Rhizophagus irregularis to test their recruitment ability. After the acclimatation, plants were transferred in the greenhouse and forty-five days later both roots and leaves were collected. Root colonization was evaluated using the Trouvelot method and significant differences in colonization level were observed among plants. Consequently, root DNA and RNA were extracted for metabarcoding and RNAseq analysis. Additionally, metabolomic analysis targeting metabolites involved in plant-microorganism interactions are ongoing.

DOI:

Publication date: June 13, 2024

Issue: Open GPB 2024

Type: Article

Authors

Ivan Bevilacqua1,2, Loredana Moffa1, Serena Varotto2, Raffaella Balestrini3, Walter Chitarra1,3, Luca Nerva1,3

1Council for Agricultural Research and Economics – Research Centre for Viticulture and Enology (CREA-VE). Via XXVIII Aprile, 26, 31015 Conegliano (TV), Italy
2University of Padua, Department of Agronomy, Food, Natural Resources, Animals and Environment, Agripolis, Viale dell’Università 16 – 35020 Legnaro (Pd)
3National Research Council of Italy – Institute for Sustainable Plant Protection (IPSP-CNR). Strada delle Cacce, 73, 10135 Torino (TO), Italy 

Contact the author*

Keywords

CRISPR/Cas9, grapevine, arbuscular mycorrhizal fungi (AMF), Systemic Acquired Resistance (SAR), multi-omics

Tags

IVES Conference Series | Open GPB | Open GPB 2024

Citation

Related articles…

Biotype diversity within the autochthonous ‘Bobal’ grapevine variety

Bobal is the second most widely grown Spanish red grape variety (54,165 has), mainly cultivated in the Valencian Community and especially, in Utiel-Requena region (about 67% of 34,000 has). In this study, agronomic and enological parameters were determined in 98 biotypes selected during 2018 and 2019 in more than 50 vineyards over 50 years-old in the Utiel-Requena region. Moreover, a multi-criteria approach considering temperature and rainfall (Fig. 1A), among other parameters, was made to establish three different zones within the region (Fig. 1B), where in the future the selected biotypes will evaluated. In fact, in 2020, 4 replicates and 12 vines per biotype were planted in an experimental vineyard to preserve this important intra-cultivar diversity.

Proteomic profiling of grape berry presenting early loss of mesocarp cell vitality

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.

A comparative analysis of regions worldwide with Pinot noir

This study examines the growing season climates of selected wine regions worldwide that have significant areas under Pinot noir.

Highlighting the several chemical situations of Dimethyl sulfide in wine

Dimethyl sulfide (DMS) is a compound that accumulate in wine for the early years of ageing 1. During this stage, which is often carried out in the bottle, the environmental conditions are conducive to the release of DMS from its precursors, already present in grapes2

Parcours de découverte des terroirs viticoles

A partir des recherches conduites sur la caractérisation des terroirs viticoles par des chercheurs de l’Unité de Recherches Vigne et Vin (1, 2, 3, 4, 5) du Centre INRA d’Angers, Terre des Sciences, le Centre de Culture Scientifique et Technique d’Angers (CCSTA) a mis au point un parcours de découverte d’une journée dans le vignoble angevin avec une approche pluridisciplinaire.