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…

Simulating the impact of climate change on grapevine behaviour and viticultural activities

Global climate change affects regional climates and hold implications for wine growing regions worldwide

Climatic zoning of viticultural production periods over the year in the tropical zone: application of the methodology of the Geoviticulture MCC system

L’objectif de cette recherche est le zonage climatique des périodes viticoles de l’année dans la Vallée du São Francisco, région brésilienne productrice de vins située en climat tropical semi-aride. Dans cette région, la production peut être échelonnée sur tous les mois de l’année.

AOC Saint-Romain, Hautes-Côtes-de-Beaune, Burgundy: analysis of a “terroir”

The aim of this study is to provide an overview of the terroir of Saint-Romain, Burgundy, based on three main information sources: official data relating to vines (CVI), soil cartography and a survey of winegrowers’ practices.

Characterization of commercial enological tannins and its effect on human saliva diffusion

Commercial oenological tannins (TECs) are widely used in the wine industry. TECs are rich in condensed tannins, hydrolyzable tannins or a mixture of both. Wine grapes are a important source of proanthocyanidins or condensed tannins while oak wood possess a high concentration of hydrolyzable tannins (Obreque-Slier et al., 2009). TECs contribute with the antioxidant capacity of wine, catalyze oxide-reduction reactions and participate in the removal of sulfur compounds and metals.

How distinctive are single vineyard Gewürztraminer musts and wines from Alto Adige (Italy) based on untargeted analysis, sensory profiling, and chemometric elaboration?

Vitis vinifera L. ‘Gewürztraminer’ is a historical grape variety of Alto Adige (Südtirol), Italy, which is widely grown in the area of Tramin an der Weinstraße, but is also grown globally. It produces highly aromatic wines that are strongly influenced by the terroir of the vineyard sites where they are grown. This study looked at musts and young wines from ‘Gewürztraminer’ grapes harvested in seven distinct vineyards near Tramin and then processed at Cantina di Termeno, minimizing winemaking protocol variability. Samples were profiled using bidimensional gas chromatography–time-of-flight mass spectrometry, liquid chromatography coupled to electrochemical detection, and near-IR spectrometry. The data were subjected to Principle Component Analysis and Hierarchical Clustering Analysis. Sensory discriminant testing was undertaken using the sorting method with a semi-trained panel, and the data were processed using Multidimensional Scaling. Seven must/wine pairs could be distinguished based on their untargeted volatilome profiles and on sensory evaluation. As expected, there were greater differences in the volatile compounds between the wines than between the musts. The wines from vineyards 4 and 5 were nonetheless quite homogenous in terms of chemical and sensory analyses, as were the wines from vineyards 1 and 3. For the phenolic profile, differences were noted between the musts and wines of vineyards 2, 3, and 4, but the musts from vineyards 5 and 7 were similar. Sensory analysis showed the wines from vineyards 6 and 7 to be distinct from the rest. These results reinforce that the composition of ‘Gewürztraminer’ musts and wines is strongly determined by vineyard site, even in a small geographic area with high variability of the terroir (soil and microclimate), and that these differences are apparent in the flavours and aromas of the finished wines. Further confirmation would require a larger sample of wines, preferably from several vintages.