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…

Influence of cell-cell contact on yeast interactions and exo-metabolome

Alcoholic fermentation is the main step for winemaking, mainly performed by the yeast Saccharomyces cerevisiae. But other wine yeasts called non-Saccharomyces may contribute to alcoholic fermentation and modulate the wine aroma complexity. The recurrent problem with the use of these non-Saccharomyces yeasts is their trend to die off prematurely during alcoholic fermentation, leading to a lack of their interesting aromatic properties searched in the desired wine. This phenomenon appears to be mainly due to interactions with S. cerevisiae. These interactions are often negatives but remain unclear because of the species and strain specific response. Among the non-Saccharomyces yeasts, Lachancea thermotolerans is a wine yeast naturally found in grape must and well known as a great L-lactic acid producer and an aromatic molecules enhancer, but its behavior during alcoholic fermentation can be completely different in co-fermentation with S. cerevisiae in function of strain used.

LC-MS based metabolomics and target analysis to study the chemical evolution of wines stored under different redox conditions

Oxygen is a key player in oenology, since its effects can be a blessing, benefiting wine quality, or a curse causing irreversible damage.

The generation of suspended cell wall material may limit the effect of ultrasound in some varieties

The disruptive effect exerted by high-power ultrasound (US) on plant cell walls, natural barriers to the diffusion of compounds of interest during the maceration of red wines, is established as the reason behind the chromatic improvement that its treatment causes. However, sometimes this improvement is not observed, especially with short maceration times. The presence of a high quantity of suspended cell wall material, which formation is favored by the sonication, could be the cause of this lack of positive results since this cell wall material has a high affinity for phenolic compounds.

Searching for the sweet spot: a focus on wine dealcoholization

It is well known that the vinification of grapes at full maturation can produce rich, full-bodied wines,
with intense and complex flavour profiles. However, the juice obtained from such grapes may have very
high sugar concentration, resulting in wines with an excessive concentration of ethanol. In addition, the decoupling between technological maturity and phenolic/aromatic one due to global warming, exacerbates this problem in some wine-growing regions. In parallel with the increase of the mean alcohol content of wines on the market, also the demand for reduced alcohol beverages has increased in recent years, mainly as a result of health and social concerns about the risks related to the consumption of alcohol.

Mathematical modeling of fermentation kinetics: a tool to better understand interactions between Torulaspora delbrueckii and Saccharomyces cerevisiae in mixed cultures

Nowadays the use of Torulaspora delbrueckii is more and more common in winemaking. However, its behavior in presence of Saccharomyces cerevisiae is not always predictable.