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…

Anthropogenic factors in modulations of fungal populations from grapes to wines and their repercussions on wine characteristics

The effects of anthropogenic activities on vineyard (different plant protections) and in winery
(pressing/clarification step, addition of sulfur dioxide) on fungal populations from grape to wine were studied. The studied anthropogenic activities modify the fungal diversity. Thus, lower biodiversity of grapes from organic modality was measured for the three vintages considered compared to biodiversity from ecophyto modality and conventional modality. The pressing / clarification steps strongly modify fungal populations and the influence of the winery flora is highlighted.

Evaluation of the effect of regulated deficit irrigation on Vitis vinifera Cabernet-Sauvignon physiological traits and final fruit composition

Climate change establishes challenges, as well as opportunities for many sectors, and markedly the wine sector.

Colloids in red wines: new insights from recent research

Despite their significant impact on wine quality and stability, colloids in red wine remain relatively under-researched. A series of studies, developed in the context of the d-wines project, aimed to provide a comprehensive understanding of the structure, composition, and formation mechanisms of red wine colloids by studying monovarietal wines from 10 of the most significant Italian red grape varieties. Starting from the idea that proteins, polysaccharides, and tannins should be involved in colloid formation, 110 monovarietal red wines were analysed for these components, revealing high inter- and intra-varietal diversity [1].

Proanthocyanin composition in new varieties from monastrell

AIM: Proanthocyanidins are responsible in an important way for positive aspects in wines, such as body and color stability in red wines, but they are also responsible for sensory characteristics that can be negative for their quality when found in excessive concentrations.

Juvenile-to-adult vegetative phase transition in grapevine 

The sequential activity of miR156 and miR172 controls the juvenile to adult phase transition in many plant species, where miR156 abundance decreases while miR172 increases along plant development. Very little is known about phase transition in horticultural woody species, which show substantially long vegetative phases. In grapevine, phase transition seems to be dissociated, displaying a first transition from juvenile to adult vegetative state in the first year, coincident with tendril differentiation and a subsequent induction of inflorescences in place of some of tendrils in later years under flowering inductive environmental conditions. Since grapevine is a highly heterozygous species, the generation of genetically homogeneous material for replicated transcriptomic analyses from seed-derived plants was a main challenge.