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…

Effects of post-veraison irrigation dose on Cabernet-Sauvignon vines in a dry and warm season in Valencia, Spain

In the old-world viticulture, there is a common but most often not scientifically proved consideration that supplemental irrigation should detrimentally affect berry and wine composition. In the semi-arid

Application of uv-led in wine as an alternative to sulphur dioxide

Sulfites (SO2) are commonly used in the wine industry to preserve products during storage for antiseptic and antioxidant purposes (Oliveira et al., 2011).

Moscatel vine-shoot extracts as grapevine biostimulant to increase the varietal aroma of Airén wines

There is a growing interest in the exploitation of vine-shoots waste, since they are often left or burned. Sánchez-Gómez et al. [1] have shown that vines-shoots aqueous extracts have significant contents of bioactive compounds, among which several polyphenols and volatiles are highlighted. Recent studied had demonstrated that the chemical composition of vine-shoots is enhanced when vine-shoots are toasted
[2,3]. The application of vegetable products in the vineyards has led to significant changes towards a more “Sustainable Viticulture”. An innovative foliar application for Airén vine-shoot extracts have been carried out to the vineyard. It has been shown that they act as grape biostimulants, improving certain wine quality characteristics [4].

Low and zero alcohol “wines”: impact of different dealcoholization processes on phenol profile and health benefits

Consumers’ demand for non-alcoholic wine has notably increased in the last years: this trend is a consequence of a growing interest in more healthy habits, and as a response to higher alcohol levels in wine due to climate change. In addition, drinking limitations due to physiological/pathological conditions (e.g., pregnancy, diabetes, hepatic disorders), driving regulations, ethical/religious considerations, and high import taxes on alcoholic beverages have positively influenced this marked (us$ 1.6 billion in 2021). International organisation of vine and wine (OIV) established that alcohol content defining wines must not be less than 8.5% vol, (OIV, 2017).

Fractal analysis as a tool for delimiting guarantee of quality areas

The pioneering work of Mandelbrot in the 70’s for building the fractal theory lead rapidly to many interesting applications in different fields such as earth sciences and economy.