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.

Download the abstract

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…

Significance of factors making Riesling an iconic grape variety

Riesling is the iconic grape variety of Germany and accounts for 23% of the German viticulture acreage, which comprises 45% of the worldwide Riesling plantings. Riesling wines offer a wide array of styles from crisp sparkling wines to highly concentrated and sweet Trockenbeerenauslese or Icewines. However, its thin berry skin makes Riesling more vulnerable to detrimental environmental threats than other white wine varieties.  

Read More

Response of red grape varieties irrigated during the summer to water availability at the end of winter in four Spanish wine-growing regions: berry phenolic composition

Water availability is the most limiting factor for vineyard productivity under Mediterranean conditions. Due to the effects caused by the current climate change, wine-growing regions may face serious soil moisture conservation problems, due to the lower water retention capacity of the soil and higher soil irradiation. The aim of this work was to evaluate the effects of soil recharge irrigation in pre-sprouting and summer irrigation every week (30 % ETo) from the pea size state until the end of ripening (RP) compared to exclusively summer irrigation every week (R) in the same way that RP, on berry phenolic composition at harvest.

Read More

From average to individual fruit, a paradigm shift for accurate analysis of water accumulation and primary metabolism in developing berries

Presentknowledge about grape development is mainly driven by the premise that a typical berry would follow the same kinetics as the population average

Read More

Enological evaluation of the attitude of the grapevine fumin to give varietal wines

Initiatives have been ongoing in recent years to safeguard biodiversity in the oenological sector via a process of enhancement of ancient varieties, under a pressure of a market strongly oriented towards production deriving from native vines of specific geographical zones. In that sense, Aosta Valley
(Italy) has raised the need to preserve and characterize its minority vine varieties which have the potentiality to give varietal wines. Fumin represents the 7% of the production of the region with 16 hectares of vineyards and 753 hectolitres of derived wine. Due to its large phenolic potential, strong astringency and deep colour, it has long been, and is still today, assembled or blended with other varieties as occurs, for example, for the Torrette.

Read More

Investigation of the biostimulant activity of naringenin on anthocyanins biosynthesis: from an explanatory transcriptomic approach on Gamay callus towards a future vineyard application

Context and purpose of the study. Anthocyanins are essential phenolic compounds in red wine, contributing significantly to colour intensity, stability, and sensory quality.

Read More