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…

How do different oak treatment affect the sensory composition of Chenin blanc wines over time?

Wooden barrels have been the preferred method for oak maturation for wines, but the use of alternative oak products, such as staves and oak chips have increased in South Africa due to lower production costs. This study investigated the effect of different oak products used during fermentation and ageing on the sensory profile, degree of liking and perceived quality of a South African Chenin blanc wine. The different wine treatments included an unoaked tank control wine, wines matured in 5th fill barrels, wines matured in new barrels from three different cooperages, and wines matured in 5th fill barrels with stave inserts from two different cooperages.

MONITOR SOME KEY PARAMETERS THROUGH THE IMPLEMENTATION OFCONTINUOUS CONTROL SYSTEMS OF THE MUST-WINE DURING MACERATION-FERMENTATION IN RED WINEMAKING TO MANAGE OPERATIONS IN “AUTOMATION”

This study is aimed to develop a complete tool for the winemaker with, complete and targeted “winemaking recipes” that can be adapted to criteria set by the winemaker, such as: grape variety, grape health status, degree of ripening, desired wine, redox status throughout the alcoholic fermentation.
To get such aim, specific sets of experiments using red grape juices from different varieties (Nebbiolo, Barbera, Pinot noir, etc.) collected at different technological and phenolic maturity points, will be held with “automatized 4.0 tanks” equipped with sensors for measuring: redox potential, dissolved oxygen, relative density, temperature, and color in order to collect a sufficient amount of data preparatory to the creation of operating models in the most widely winemaking situations in which the automatized 4.0 tanks “will be able to independently respond” with the right corrective actions (opening/closing aeration valve, execution/block pumping overs , etc.) if the key parameters exceed the limits of the recommended ranges set in the selected recipe.

Vineyard’s ozone application to induce secondary metabolites accumulation in grapes and wine

In viticulture sector to find new tools for pest management has become an urgent necessity. Hence, grapevines cultivation has high production rate demand and to meet the intensive market request, a massive use of pesticides is often required. In addition to the environmental problems associated with large use of chemicals, there is an increasing number of consumers which are asking for

For a phenomenology of terroir. A consumers’ perspective

This study investigates the notion of terroir by applying a phenomenological approach, focusing on the subjective experience of consumers. We will consider how terroir is described by consumers in order to gauge their subjective viewpoint and understand their way of describing and defining this spatiality.

Study of the effect of native vineyard bacteria on the expression of Plasmopara viticolaeffectors

Downy mildew, caused by the oomycete Plasmopara viticola (Berk. & M.A. Curtis) Berl. & De Toni, is one of the most destructive grapevine diseases mostly affecting Vitis vinifera L. and impacting on viticulture. The pathogen invasion can induce in grapevine multiple defense reactions, first PAMP-Triggered Immunity and secondly Effector-Triggered Immunity. Plasmopara viticola can overcome these defense mechanisms through the secretion of effectors, such as RxLR, into the plant cells, making it easier for the oomycete to infect grapevines. Currently, the use of chemical pesticides remains the most effective way to control the pathogen with severe negative side effects on the environment and animal health.