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…

Screening of soil yeasts with fermentative capacity from the antarctic continent for their application in the wine industry

AIM: In the last years, many wineries are increasing experimentation to produce more distinguishable beverages. In this sense, the reduction of the fermentation temperature could be a useful tool because it preserves volatile compounds and prevents wines from browning, particularly in the case of white wines.

Chinese localization of wine aroma descriptors

Wine aroma descriptors are important tools for wine evaluation. The present well-known wine aroma descriptor system was created and based on Western culture, which makes difficult for Chinese consumers to recognize and learn wine. AIM: The aim of this study was to update the wine aroma descriptor system for Chinese.

Methods: Fifty-four wine aroma descriptors of ‘Le nez du vin’ was used as substitution candidates. Firstly, a survey on unfamiliar aromas was distributed to 150 untrained Chinese wine consumers. Twenty attributors, such as blackcurrent buds, quince, linden, were selected as the most 17 unfamiliar. Then, a descriptive analysis was performed by trained tasting panel to substitute the targeted twenty aromas perfume. Furthermore, reference standards were looked and new le nez du vin were made. Finally, a substitution analysis was performed to replace the unknown wine aroma to the Chinese local aromas.

Preliminary steps of a protocol to isolate transcription factors bound to a specific DNA locus in grapevine using CRISPR-dCas9 system

Cis-acting regulatory elements are DNA sequences that can be bound by transcription factors to regulate the expression of genes in a condition-dependent and tissue-specific way. It is nowadays possible to search for DNA motives and sequences that a given transcription factor is binding or at least can, but it is still hard to have a glance at all the transcription factors that are contemporaneously located at the same locus. Inspired by an existing technique that uses the CRISPR-Cas system in mammal cells, we are trying to develop a protocol to study such regulation in Vitis vinifera. Using the highly sequence-specific binding capacity of a catalytically inactive Cas9 protein (dCas9), our idea is to set up a system to target a desired sequence and precipitate all the crosslinked proteins and distantly interacting chromatin at this locus and analyze them.

Combined use of Lachancea thermotolerans and Schizosaccharomyces pombe in winemaking

Commercial red wines use the malolactic fermentation process to ensure stability from a microbiological point of view. In this second fermentation, malic acid is converted into L-lactic acid under controlled steps.

Brettanomyces bruxellensis and off-odours: genetic and proteomic approaches to unravel the molecular mechanism of ethyl-phenols production

Brettanomyces/Dekkera yeasts in wine are able to produce various spoilage compounds that are, at high concentration, detrimental to wine quality. The principal spoiler compounds associated with Brettanomyces spp. are vinyl and ethyl-phenols that are responsible for off- odours described as “animal”, “medicinal”, “sweaty leather”, “barnyard”, “spicy” and “clove-like”.