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…

Estimation of chemical age of red wines with the use of Fourier transform infrared spectroscopy (FT-IR) and chemometrics

The color of a red wine is one of the most important parameters of its quality, giving much information on its status, such as the grape variety used or the winemaking style. As the result of a complex equilibrium between different forms of anthocyanins and polymerization reactions which occur over the course of time, color can also serve as an indication of a wines’ age. For this purpose the “chemical age” i and ii indexes have been introduced by Somers in 1977. The chemical age index i measures the color absorbance after the addition of acetaldehyde while chemical index ii provides an indication of how much of the total red pigments are resistant to SO2 bleaching.

High-throughput screening of physical-mechanical berry skin traits facilitates targeted selection of breeding material with resistance to Botrytis bunch rot and grape sunburn

The ongoing climate change implies an increasing mean air temperature, which is signified by weather extremes or sudden changes between drought and local heavy rainfalls. These changing conditions are especially challenging for the established grapevine varieties growing under cool climate conditions due to an increased risk for fungal diseases like downy mildew (DM) and Botrytis bunch rot (BBR) as well as for grape sunburn. To meet that demand, the scope of most grapevine breeding programs is the selection of mildew fungus-resistant and climatic adapted grapevines with balanced, healthy yield and outstanding wine quality.

´Vinho Verde´ wines production from differential fermentation: the role of musts sulphitation as a preservation strategy to keep the musts character

High-volume mass-market white wines production method by means of harvest-deferred fermentation from desulphited musts allows an efficient business management by avoiding the seasonality in wine sector.

Monitoring of mannoprotein cessions during wine aging on lees: development of a simple enzymatic method

Mannoproteins are polysaccharides released by Saccharomyces cerevisiae yeast during alcoholic fermentation or by enzymatic action during aging on yeast lees (autolysis). These molecules play a major role in wine characteristics processing, namely, in the tartaric stabilization and protein haze prevention; moreover, they improve color stability and reduce astringency.

Berry maturity effects on physic and chemical characteristics of traditional sparkling wines produced from Chardonnay and Sauvignon blanc grapes.

One of the consequences of global warming is the quick berry development giving rise to a disconnection between sugar accumulation and the formation of important quality minor compounds such as phenolics and volatile compounds being a huge challenge for the oenologist [1]. Thus, this phenomenon is forcing the search on strategies for maintaining the quality of wines despite this situation. One possibility is to make an early harvest with a low sugar concentration (18ºbrix) and advanced harvest for sparkling wine (20-21ºbrix) and afterwards to combine base wines properly and carry out the second fermentation trying to compensate the lack of secondary metabolites due to the quick berry development and higher alcohol degree of the second one, not adequate itself for sparkling wine. The aim of this study was to assess the chemical and physical characteristics, mainly volatile profile, and foaming properties of sparkling wines from grapes of Chardonnay and Sauvignon blanc.