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…

Litchi tomato as a fumigation alternative in Washington state wine grape vineyards

The northern root-knot nematode (Meloidogyne hapla) is one of the most prevalent plant-parasitic nematodes affecting Washington State Vitis vinifera vineyards. This nematode induces small galls on roots, restricting water and nutrient uptake. In new vineyards this can impede establishment. In existing vineyards, it can exacerbate decline in chronically stressed vines. While preplant fumigation is a common strategy for M. hapla management, its efficacy is temporary and relies on broad-spectrum chemicals that undergo frequent regulatory scrutiny. The trap crop litchi tomato (Solanum sisymbriifolium) showed promise in reducing plant-parasitic nematode densities in potato. This prompted field greenhouse experiments to evaluate its potential to reduce M. hapla in V. vinifera.

The effects of cane girdling on berry texture properties and the concentration of some aroma compounds in three table grape cultivars

The marketability of the table grapes is highly influenced by the consumer demand; therefore the market value of the table grapes is mainly characterized by its berry size, colour, taste and texture. Girdling could cause accumulation of several components in plants above the ringing of the phloem including clusters and resulting improved maturity. The aim of the experiments was to examine the effect of girdling on berry texture characteristics and aroma concentration.

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.

Data integration via modeling for adaptation to climate change and efficiency breeding in grapevine

Climate can greatly affect grape yield and quality (van Leeuwen et al., 2024). Growing suitable cultivars in a given region and or breed environmental resilient cultivars are essential for maintaining viticulture sustainability, particularly in the face of climate change (Wolkovich et al., 2018).

Non-destructive his based analysis for shelf-life evaluation of table grape 

Fast, accurate, and non-destructive analytical techniques based on hyperspectral imaging (hsi) represent effective tools for food quality evaluation. A visible change in the appearance of a fresh product often negatively impacts the perceived quality from a consumer’s point of view.