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…

Vineyard Landscape in Vale dos Vinhedos: Pressure and Protection

The region with Origin Indication Vale dos Vinhedos (IPVV) is located in the cities of Bento Gonçalves, Garibaldi and Monte Belo do Sul. It was established in November 2002, through an act of Instituto Nacional de Propriedade Industrial, according to Resolução INPI Nº. 75, of 2000, and Law Nº. 9.279 of 1996. The changes on the grape growing and wine making in the last 15 years, with the expansion of grapevines Vitis viníferas cultivation, the installation of several modern family wine companies, making wine with competitive quality, among other aspects, have enabled the implementation of concepts of Geographical Indications.

INSIGHT THE IMPACT OF GRAPE PRESSING ON MUST COMPOSITION

The pre-fermentative steps play a relevant role for the characteristics of white wine [1]. In particular, the grape pressing can affect the chemical composition and sensory profile and its optimized management leads to the desired extraction of aromas and their precursors, and phenols resulting in a balanced wine [2-4]. These aspects are important especially for must addressed to the sparkling wine as appropriate extraction of phenols is expected being dependent to grape composition, as well.

Recent advancements in understanding the terroir effect on aromas in grapes and wines

Terroir is about the link between wine and its origin. It has long been understood by sensory evaluation that the taste of wine from a given variety can be related to its origins. Specific organoleptic characteristics of wine are influenced by environmental factors such as soil and climate. By deconstructing the effect of measurable soil and climate parameters on grape and wine aroma compounds,

Winery by-products as potential bioresources for green valorization and sustainable biotechnological applications

The wine and distillery industries are among the most prominent sectors in EU agriculture, where 75% of grape production is dedicated to winemaking.

Modelling vine water stress during a critical period and potential yield reduction rate in European wine regions: a retrospective analysis

Most European vineyards are managed under rainfed conditions, where seasonal water deficit has become increasingly important. The flowering-veraison phenophase represents an important period for vine response to water stress, which is seldomly thoroughly evaluated. Therefore, we aim to quantify the flowering-veraison water stress levels using Crop Water Stress Indicator (CWSI) over 1986–2015 for important European wine regions, and to assess the respective potential Yield Lose Rate (YLR). Additionally, we also investigate whether an advanced flowering-veraison phase may help alleviating the water stress with improved yield. A process-based grapevine model STICS is employed, which has been extensively calibrated for flowering and veraison stages using observed data at 38 locations with 10 different grapevine varieties. Subsequently, the model is being implemented at the regional level, considering site-specific calibration results and gridded climate and soil datasets. The findings suggest wine regions with stronger flowering-veraison CWSI tend to have higher potential YLR. However, contrasting patterns are found between wine regions in France-Germany-Luxembourg and Italy-Portugal-Spain. The former tends to have slight-to-moderate drought conditions (CWSI<0.5) and a negligible-to-moderate YLR (<30%), whereas the latter possesses severe-to-extreme CWSI (>0.5) and substantial YLR (>40%). Wine regions prone to a high drought risk (CWSI>0.75) are also identified, which are concentrated in southern Mediterranean Europe. An advanced flowering-veraison phase may have benefited from cooler temperatures and a higher fraction of spring precipitation in wine regions of Italy-Portugal-Spain, resulting in alleviated CWSI and moderate reductions of YLR. For those of France-Germany-Luxembourg, this can have reduced flowering-veraison precipitation, but prevalent alleviations of YLR are also found, possibly because of shifted phase towards a cooler growing season with reduced evaporative demands. Overall, such a retrospective analysis might provide new insights towards better management of seasonal water deficit for conventionally vulnerable Mediterranean wine regions, but also for relatively cooler and wetter Central European regions.