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…

Effect of Quercus Alba oak barrels from different forests on the polyphenolic composition of Tempranillo red wines

The species and origin used for red wine oak aging determines the physiological composition of the wood and thus the finished wines. In America, oak is grown primarily in the states of Virginia, Missouri, Kentucky, Oregon, Ohio, Minnesota, Wisconsin and California. The aim of this study was to analyze how the choice of barrels made with Quercus Alba oak from different geographic areas of the United States (Missouri, Kentucky, Ohio and Pennsylvania) influences the polyphenolic composition of Tempranillo red wines.

IMPACT OF NEW BIO STIMULANTS ON GRAPE SECONDARY METABOLITES UNDER CLIMATE CHANGE CONDITIONS

In a context of climate change and excessive use of agrochemical products, sustainable approaches for environmental and human health such as the use of bio stimulants in viticulture represent a potential option, against abiotic and biotic threats. Bio stimulants are organic compounds, microbes, or a combination of both, that stimulate plant’s vital processes, allowing high yields and good quality products. In vines, may trigger an innate immune response leading to the synthesis of secondary metabolites, key compounds for the organoleptic properties of grapes and wines.

Organic volatile compounds as suitable markers of grapevine response to defense elicitors in the vineyard

In greenhouse, emission of volatile organic compounds (VOC) by grapevine leaves has already been reported in response to the defence elicitor sulfated laminarin (PS3) [1]. In order to check that this response was not specific to PS3, experiments were conducted on Vitis cv Marselan plantlets with several other elicitors of different chemical structures: i.e. Bastid® (COS-OGA),

Grape byproducts as source of resveratrol oligomers for the development of antifungal extracts

Grape canes are a non-recycled byproduct of wine industry (1-5 tons per hectare per year) containing valuable phytochemicals of medicine and agronomical interest. Resveratrol and wine polyphenols are known to exert a plethora of health-promoting effects including antioxidant capacity, cardioprotection, anticancer activity, anti-inflammatory effects, and estrogenic/antiestrogenic properties (Guerrero et al. 2009). Additionally, resveratrol is a major phytoalexin produced by plants in response to various stresses and promotes disease resistance (Chang et al. 2011). Our project aims to develop polyphenol-rich grape cane extracts to fight phytopathogenic or clinically relevant fungi. We initiate the project with the development of analytical methods to analyze resveratrol mono- and oligomers (dimers, trimers and tetramers) from grape canes and we evaluate their potential activity against clinically relevant opportunistic fungal pathogens (Houillé et al. 2014).

Berry carbon (δ13C) and nitrogen (δ15N) isotopic ratio reflects within farm terroir diffferences

ÂThe natural abundance of carbon stable isotopes has been reported to be related to water availability in grapevines quite widely. In the case of nitrogen, the natural abundance of its stable isotopes is mainly affected by the nature of the source of nitrogen (organic vs. inorganic) used by the plant, though the bibliography available for grapevine is very scarce.