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…

Stable or dynamic? How phenotypic plasticity could be key to select for grapevine adaptation?

Climate change will require the adaptation of agricultural systems and among the different means of adaptation, changing plant material is a promising strategy. In viticulture, different levels of diversity are currently exploited: clonal and varietal diversity for rootstocks and scions. A huge quantity of research aims to evaluate different genotypes in different environmental conditions to identify which ones are the best adapted and the most tolerant to future environmental conditions.

Sensory profile: a tool to characterize originality of wines produced without sulfites

A trend to reduce chemical inputs in wines exists, especially sulfur dioxide (SO2). This additive is widely used due to its antioxidant, antiseptic and antioxidasic properties. During without sulfites vinification, bioprotection by adding yeast on harvest could be a sulfites alternative. With extension of this wine market, sensory impact linked to sulfites absence and/or sulfites alternative should be evaluated. That’s what this approach proposes to do, focusing on sensory characteristics of wines produced with or without SO2 addition during the winemaking process. METHODS: Wines were elaborated from Merlot grapes of two maturity levels according to three modalities: SO2, without SO2 and bioprotection on harvest (mix of Torulaspora delbrueckii and Metschnikowia pulcherrima). SO2 modality was sulfited throughout the winemaking and aging processes whether other modalities received any addition. After two years of aging, sensory studies were carried out with a specific panel for one month. First, descriptors were generated to differentiate the wines, then panelists were trained on these specific descriptors for five sessions and finally wines sensory profiles were elaborated

Characterization and application of silicon carbide (SiC) membranes to oenology

After fermentations, the crude wine is a turbid medium not accepted by the consumer therefore, it needs to be filtered

Arbuscular mycorrhizal fungi as biomarkers of vineyard yield in Champagne

The vine is colonized by a multitude of micro-organisms (fungi, bacteria, oomycetes) mainly coming from the microbial reservoir constituted by the soil. These microorganisms have positive or negative effects on the vine (protection against pathogens, resistance to abiotic stress, nutrition, but also triggering of diseases) (Fournier, Pellan et al. 2022). In addition to these functional roles, they respond quickly to environmental changes (climate, cultural practices) which could make them good bioindicators of the functioning of the wine ecosystem.

The combined use of Lachancea thermotolerans and lactic bacteria in wine technology

The production of most red wines that are sold involves an alcoholic fermentation carried out by yeasts of the Saccharomyces genus, and a subsequent fermentation carried out by lactic bacteria of the Oenococus oeni species after the first one is fully completed. However, the traditional process can face complications, which can be more likely in grape juices with high levels of sugar and pH. Because of climate change, these situations are more frequent in the wine industry. The main hazards in those scenarios are halts or delays in the alcoholic fermentation or the growth of unwanted bacteria while the alcoholic fermentation is not done yet and the wine still has residual sugars.