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…

Effects of soil water content and environmental conditions on vine water status and gas exchange of Vitis vinifera L. cv. chardonnay

Vine water status has a significant influence on vineyard yield and berry composition (Williams and Matthews, 1990; Williams et al., 1994). It has been hypothesized that the response of plants to soil water deficits may be due to some sort of “root signal” (Davies and Zhang, 1991). This signal probably arises due to the roots sensing a reduction in soil water content or an increase in the mecanical impedance as the soil dries out.

The wine country, between landscape and promoting tool. The example of Chinon and Saint-Nicolas-de-Bourgueil vineyards (France)

When talking about wine, terroirs are never too far. The National Institute of Apellation d’Origine (INAO) defines it as a system inside of which interact a group of human factors, an agricultural production and a physical environment.

NADES extraction of anthocyanins derivatives from grape pomace

Grape pomace is one of the main by-products generated after pressing in wine-making. It’s valorization through the extraction of bioactive compounds is the answer for the development of sustainable processes. Nevertheless, in the recovery of anthocyanins derivatives, the extraction stage continues to be a limiting step. The nature of the sample and the type of solvent determine the efficiency of the process

New methods and technologies to describe the environment in terroir studies

The concept of terroir in viticulture deals with the influence of environmental factors on vine behaviour and grape ripening. Recent advances in technology, in particular computer technology, allow a more in-depth study of the environment. Geomorphology can be studied with digital Elevation Models (DEM). Soils can be surveyed with geophysics.

Differences in the chemical composition and “fruity” aromas of Auxerrois sparkling wines from the use of cane and beet sugar during wine production.

The main objective of this study was to establish if beet sugar produces a different concentration of “fruity” volatile aroma compounds (VOCs), compared to cane sugar when used for second alcoholic fermentation of Auxerrois sparkling wines. Auxerrois base wine from the 2020 vintage was separated into two lots; half was fermented with cane sugar and half with beet sugar (both sucrose products and tested for sugar purity). These sugars were used in yeast acclimation (IOC 2007), and base wines for the second fermentation (12 bottles each). Base wines were manually bottled at the Cool Climate Oenology and Viticulture Institute (CCOVI) research winery. The standard chemical analysis took place at intervals of 0, 4 weeks, and 8 weeks post-bottling. Acidity and pH measurements were carried out by an auto-titrator. Residual Sugar (g/L) (glucose (g/L), fructose (g/L)), YAN (mg N/L), malic acid, and acetic acid (g/L) were analyzed by Megazyme assay kits. parameters were analyzed by Megazyme assay kits. Alcohol (% v/v) was assessed by GC-FID. VOC analysis of base wines, finished sparkling wines, as well as the two sugars in model sparkling wine solutions, was carried out by GC-MS. VOCs included ethyl octanoate, ethyl hexanoate, ethyl butanoate, ethyl decanoate, ethyl-2-methylbutyrate, ethyl-3-methylbutyrate, ethyl 2-methyl propanoate, ethyl 2- hydroxy propanoate, 1-hexanol, 2-phenylethan-1-ol, ethyl acetate, hexyl acetate, isoamyl acetate and 2-phenylethyl acetate.