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…

Is complex nutrition more advantageous than mineral nitrogen for the fermentative capacities of S. cerevisiae?

During alcoholic fermentation, nitrogen is an essential nutrient for yeast as it plays a key role in sugar transport and biosynthesis of wine aromatic compounds (thiols, esters, higher alcohols). The main issue of a lack in yeast assimilable nitrogen (yan) in winemaking is sluggish or stuck fermentations promoting the growth of alteration species which may lead to economic losses. However, correcting this nitrogen deficiency is sometimes not enough to restore proper fermentation performance. This suggests the existence of other nutritional shortages.

Long-term flooding effects on the physiological and productive performance of Montepulciano and Sangiovese cultivars

Extreme climatic events, such as prolonged drought followed by intense flooding, increasingly impact viticulture, affecting vine physiology, productivity, and grape composition.

Bioprotection en phase pré-fermentaire, synthèse de 3 ans d’expérimentations dans différentes régions viticoles

With growing consumer interest in products without chemical additives, limiting the use of sulfites is a priority for the wine industry. Bioprotection is a biological alternative that avoids or reduces the risks of alterations that have a negative impact on the organoleptic quality of wines and, ultimately, on their acceptability to consumers. bioprotection can also provide a response to the risks of microbiological deviations, which are increased both by climate change and by the organization of harvesting operations, which increasingly include the use of multi-bins filled at the vine, exposing the harvest to sometimes high temperatures for longer periods of time.

AOC Saint-Romain, Hautes-Côtes-de-Beaune, Burgundy: analysis of a “terroir”

The aim of this study is to provide an overview of the terroir of Saint-Romain, Burgundy, based on three main information sources: official data relating to vines (CVI), soil cartography and a survey of winegrowers’ practices.

Model-assisted analysis of the root traits underlying RSA genotypic diversity in Vitis: a promising approach for rootstock selection?

By dissecting the root system architecture (RSA) into its underpinning components (e.g. root emission, axial growth, radial growth, branching, root direction or tropism) and identifying the relationships between them, functional-structural 3D root models are promising tools for analyzing the diversity and complexity of root system phenotypes with Genotype × Environment interactions. The model parameters are assumed to be synthetic traits, less influenced by the environment, and consequently with less polygenic architectures than the integrative RSA traits they drive. Root models can serve as a basis for in silico development of root system ideotypes by highlighting the developmental processes and parameters that most likely influence RSA fitness.