terclim by ICS banner
IVES 9 IVES Conference Series 9 Enhancing plant defense: carbon dots for efficient spray-induced gene silencing 

Enhancing plant defense: carbon dots for efficient spray-induced gene silencing 

Abstract

Ectopic RNA application for plant defense faces challenges in tree crops, including size, diffusion, and stability of active compounds such as ribonucleoproteins and nucleic acids. While existing strategies involve expressing dsRNA in transgenic plants targeting pathogens, our research strives to develop a transient RNAi system based on Spray-Induced Gene Silencing (SIGS). This approach aims to circumvent legal barriers and public concerns associated with genetically modified organisms (GMOs). Our strategy integrates SIGS with branched polyethyleneimine-functionalized Carbon Dots (bPEI-CDs) as nanocarriers, effectively addressing unique delivery challenges in plant defense as RNA stability and uptake enhancement. We developed a cost-effective microwave-assisted protocol for the synthesis of 10 nm monodispersed bPEI-CDs. These molecules showed over 50% dsRNA protection against RNase III degradation at a 200:1 (w/w) ratio. We further demonstrated their capability to enhance cellular uptake by successfully delivering fluorolabeled-dsRNAs (Cy3-RNA) complexed with CDs into intact plant tissue-cultured cells. Notably, Cy3-RNA sprayed with CD on the surface of greenhouse grapevine leaves through a low-pressure spray application penetrated stomata cells and neighboring cells. Ultimately, the ability of dsRNA complexed with CD to trigger RNA interference was confirmed using a 21 nucleotide-dsRNA targeting eGFP . Ongoing work quantitatively compares, through qPCR, the reduced expression of GFP on eGFP-expressing transgenic microvine leaves sprayed with CD-complexed and naked siRNA. This innovative approach, leveraging CDs, aims to tackle critical barriers in dsRNA delivery, particularly by addressing cell wall-related limitations in plant. This research marks a pivotal step in optimizing RNA-based grapevine defense strategies, and propelling sustainable viticulture practices forward.

DOI:

Publication date: June 13, 2024

Issue: Open GPB 2024

Type: Poster

Authors

Christian Mandelli1*, Laurent G. Deluc1,2

1 Department of Horticulture, Oregon State University, Corvallis, OR, United States
2 Oregon Wine Research Institute, Oregon State University, Corvallis, OR, United States

Contact the author*

Keywords

RNA interference, Spray-Induced Gene Silencing (SIGS), Carbon dots, dsRNA delivery, Sustainable agriculture

Tags

IVES Conference Series | Open GPB | Open GPB 2024

Citation

Related articles…

INTEGRAPE guidelines and tools: an effort of COST Action CA17111

INTEGRAPE was a European interdisciplinary network for “data integration to maximize the power of omics for grapevine improvement” (CA17111, https://integrape.eu/), funded by the European COST Association from September 2018 to 2022. This Action successfully developed guidelines and tools for data management and promoted the best practices in grapevine omics studies with a holistic future vision of: “Imagine having all data on grapevine accessible in a single place”.

Physiological means to curb the enthusiasm of viruses from infecting grapevines  

The two most deadly viruses infecting and threatening the productivity of grapevines worldwide are leafroll and red blotch viruses. There is no cure for viral diseases other than roguing the symptomatic vines and replacing them with certified vines derived from clean, virus-tested stocks.
Given that phloem plays a central role in virus infection, this study aimed to purge the virus by girdling the phloem of leafroll-infected vines at different phenological stages of infected grapevines. Phloem-girdling was performed on canes at veraison to varying regions between the proximal and distal clusters.

Correlation between skin cell wall composition and phenolic extractability in Cabernet sauvignon wines

The phenolic component of red wine is responsible for important elements of flavor and mouthfeel, and thus quality of the finished wine. Additionally, many of these phenolics have been associated with health benefits such as reduction of the risk of developing cardiovascular disease, cancer, osteoporosis and preventing Alzheimer’s disease. While the origins, concentrations, and chemistries of the phenolics in a finished red wine are well known, the fundamental mechanisms and kinetics of extraction of these phenolics from grape skins and seeds during red wine fermentation are poorly understood. This lack of knowledge regarding the extraction mechanisms of phenolics during red wine fermentation makes informed manipulations of the finished wine’s phenolic composition difficult.

Pure wine vs natural wine

S’il n’existe pas de réglementation officielle, la démarche des vins naturels prône un retour aux pratiques dites ancestrales préconisant notamment un mode d’élaboration des vins utilisant le moins d’intrants possible. Le seul autorisé reste l’anhydride sulfureux (SO2) à des doses quatre à cinq fois moins importantes que pour les vins dits conventionnels. Ce désir de renouer avec

Impact of the fumaric acid/glutathione pair addition before bottling on Cabernet Sauvignon wine quality

Over the last decades, climate change and rising temperatures have impacted the wine industry. Wines from warm regions tend to have a higher pH and lower total acidity.