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…

Characterization of berry softening and sugar accumulation dynamics in a slow-ripening genotype and its response to abscisic acid treatments

In the current viticultural context, global warming leads to advanced and possibly accelerated ripening which can alter the balance among desirable grape quality traits sought for winemaking. Evaluation of genetic material that displays delayed and/or slower ripening could uncover a potential “slow ripening” trait for incorporation into commercial varieties through breeding. In this study, we evaluated a white-fruited selection discovered in the Grape Breeding and Genetics program at E. & J. Gallo Winery that displayed an unusual ripening pattern compared to standard varieties. Vines of the slow-ripening selection did not differ in their visual appearance, water status or gas exchange characteristics compared to vines of its normal-ripening sibling.

Prediction of astringency in red wine using tribology approach to study in-mouth perception

AIM Astringency is described as a ‘dry puckering‐like sensation’ following consumption of tannins1 that affect consumer preference of foods and beverages, including red wine2. To improve the understanding of astringency, which is a complex interaction due to multiple mechanisms occurring simultaneously, further studies are needed. In this view, oral tribology is considered a useful technique for beverage study to evaluate the thin-film lubrication properties of saliva resulting in oral friction‐related sensations3. The aim of this study was to examine the film behavior of selected protein-based fluids under controlled friction conditions, to understand polyphenol-protein interactions involved in the sensation of astringency.

Pedicel lenticel diversity of cultivars and their influence on cell death and berry shrivel

Shrinking berries are the common symptoms of Berry Shrivel (BS) and Late-Season-Dehydration (LSD) in grapevine cultivars.

Phenological stage dependency of Cabernet Sauvignon and Grenache response to water and nutrient limitation 

As the frequency and intensity of drought events increase, understanding the mechanisms of plant resilience to water deficit is crucial. To maintain an appropriate plant yield, a common practice is the application of high amounts of fertilizers with negative environmental impacts. The single and combined effect of water deficit and nutrient availability, namely nitrogen (N) and potassium (K), in Vitis Vinifera L. cv. Cabernet Sauvignon and Grenache was evaluated. Two-year-old grapevine plants grafted on SO4 rootstock were transferred in pots under semi-environmental conditions. During the growing season, plants were either maintained well-watered (100% ETc) or subjected to a controlled water deficit irrigation (33% ETc).

Yeast derivatives: an innovative approach to produce Oenococcus oeni under biofilm form?

The malolactic fermentation can occur naturally or be induced by inoculation of selected bacterial strains, most commonly of Oenococcus oeni.