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…

Landscape marketing and landscape reality: what is the relationship? The case of the Loire Valley vineyards

This issue poses two questions: the relationship between beauty and taste (is landscape quality an index of wine quality ?), and the gap or the conformity between our image of the “terroir” and the visible reality. The landscape is both an object and a representation.

Correspondence between physiological plant variables and carbon isotope composition in different climate winegrape regions

The climate is the environmental factor that contributes with greater weight in the variability of the yield and the composition of the grape, therefore, it is key in the determination of the typicity of the product. Of the environmental factors, the evolution of water availability conditions, among other things, the biochemical evolution of the compounds of the grape and the type of wine to be elaborated. An integrating parameter of the hydric state of the plant is the carbon isotopic composition (δ13C). This indicator is a useful parameter to characterize the water status during the maturation period and estimate the transpiration efficiency or water use efficiency (EUA) in the vine.

From varietal and terroir expression to off-odors: chemical background of wine aroma evolution during aging

Expression of sensory attributes that reflect the varietal and geographical origin of wines (aka terroir) is central to perceived wine quality and reputation of wine producing regions.

PERCEPTUAL INTERACTIONS PHENOMENA INVOLVING VARIOUS VOLATILE COMPOUND FAMILIES LINKED TO SOME FRUITY NOTES IN BORDEAUX RED WINES

Fruity notes play a key role in the consumer’s appreciation of Bordeaux red wines. If literature provides a lot of knowledge about the nature of volatile compounds involved in this fruity expression, the sensory phenomena involving these compounds in mixture still need to be explored. Considering previous sensory works about the impact of esters and some overripening compounds, the goal of this work was to study the implication of perceptual interactions involving red wine odorant compounds of diverse origins and described as potentially affecting fruity aromatic expression.

Determination of Aroma Compounds in Grape Mash under Conditions of Tasting by On-line Near-Infrared Spectroscopy

The production of high-quality wines requires the use of high-quality grapes. Some compounds originating from grapes may negatively influence the odour and flavour of the resulting wine in their original form or as precursors for off-odours and –flavours. Therefore, a rapid evaluation of the grapes directly upon receival at the winery is advantageous. Up to now, grape aroma is mainly evaluated by tasting, however, this leads to subjective results. The use of near-infrared (NIR) spectroscopy allows a rapid, objective and destruction-free analysis without previous sample preparation. Moreover, the measurement can be integrated into an existing process without additional sampling.