Abstract
The management of emerging plant viruses presents significant challenges for global agriculture, requiring innovative approaches beyond conventional control strategies. Traditional methods rely on cultural practices, vector management, and breeding for genetic resistance, and these approaches are often time-consuming and may have limited effectiveness against emerging viral strains. Spray-Induced Gene Silencing (SIGS), involving topical application of virus-derived double-stranded RNA (dsRNA) to trigger plant defense mechanisms, offers a promising alternative strategy. However, the application of SIGS faces challenges due to inefficient dsRNA uptake by the plant, among other issues. In this study, we developed and characterized nanocomposite formulations using carbon dots (CDs) and polyethylenimine-functionalized mesoporous silica nanoparticles (PMSNs) to enhance dsRNA delivery and stability for the control of turnip mosaic virus (TuMV) and beet curly top virus (BCTV), an RNA and a DNA virus, respectively, in Nicotiana benthamiana. Our results demonstrated that dsRNA delivery was significantly enhanced (up to 5-fold) when formulated with nanoparticles compared to naked dsRNA. For TuMV-infected plants, both nanocomposite formulations significantly reduced viral titers (by 13.5-fold for PMSNs and 17.3-fold for CDs) and maintained photosynthetic capacity similar to uninfected controls even at 66 days post-inoculation. Regarding BCTV, the nanocomposite treatments significantly delayed disease symptom appearance and reduced viral DNA accumulation by 8-28-fold compared to control plants. The enhanced antiviral efficacy observed with nanoparticle formulations correlates with improved dsRNA delivery and persistence in plant tissues, making the nanoparticle-based dsRNA delivery systems represent a viable approach for developing sustainable, environmentally friendly strategies to protect crops against economically important viral diseases.
Data availability
Data from RT-qPCR and qPCR analyses, which evaluated the NPs-mediated dsRNA delivery and efficacy of topical dsRNA nanocomposites against beet curly top virus and turnip mosaic virus, have been deposited in Zenodo at https://doi.org/10.5281/zenodo.17543127.
References
Savary, S. et al. The global burden of pathogens and pests on major food crops. Nat. Ecol. Evol. 3, 430–439 (2019).
Velasco, L., Ruiz, L., Galipienso, L., Rubio, L. & Janssen, D. A. Historical account of viruses in intensive horticultural crops in the Spanish mediterranean arc: new challenges for a sustainable agriculture. Agronomy 10, 860 (2020).
Niu, D. et al. RNAs - a new frontier in crop protection. Curr. Opin. Biotechnol. 70, 204–212 (2021).
Mat Jalaluddin, N. S., Asem, M. & Harikrishna, J. A. Ahmad Fuaad, A. A. H. Recent progress on nanocarriers for Topical-Mediated RNAi strategies for crop Protection—A review. Molecules 28, 2700 (2023).
Taliansky, M. et al. Rna-based technologies for engineering plant virus resistance. Plants 10, 1–19 (2021).
Mitter, N., Worrall, E. A., Robinson, K. E., Xu, Z. P. & Carroll, B. J. Induction of virus resistance by exogenous application of double-stranded RNA. Curr. Opin. Virol. 26, 49–55 (2017).
Mitter, N. et al. Clay nanosheets for topical delivery of RNAi for sustained protection against plant viruses. Nat. Plants. 3, 1–10 (2017).
Demirer, G. S. et al. High aspect ratio nanomaterials enable delivery of functional genetic material without DNA integration in mature plants. Nat. Nanotechnol. 14, 456–464 (2019).
Das, R., Bandyopadhyay, R. & Pramanik, P. Carbon quantum Dots from natural resource: A review. Mater. Today Chem. 8, 96–109 (2018).
Maholiya, A. et al. An insight into the role of carbon Dots in the agriculture system: a review. Environ. Sci. Nano. 10, 959–995 (2023).
Wang, Z. et al. Functionalized carbon Dot-Delivered RNA nano fungicides as superior tools to control Phytophthora pathogens through plant RdRP1 mediated Spray‐Induced gene Silencing. Adv. Funct. Mater. 33, 2213143 (2023).
Liu, Z. et al. Non-viral nanoparticles for RNA interference: principles of design and practical guidelines. Adv. Drug Deliv Rev. 174, 576–612 (2021).
Sangwan, A. et al. Size variations of mesoporous silica nanoparticle control uptake efficiency and delivery of AC2-derived DsRNA for protection against tomato leaf curl new Delhi virus. Plant. Cell. Rep. 42, 1571–1587 (2023).
Xu, Y. et al. Size effect of mesoporous silica nanoparticles on pesticide Loading, Release, and delivery in cucumber plants. Appl. Sci. 11, 575 (2021).
Cai, Y., Liu, Z., Wang, H., Meng, H. & Cao, Y. Mesoporous silica nanoparticles mediate SiRNA delivery for Long-Term Multi‐Gene Silencing in intact plants. Adv. Sci. 11, e2301358 (2024).
Chen, L. F., Brannigan, K., Clark, R. & Gilbertson, R. L. Characterization of curtoviruses associated with Curly top disease of tomato in California and monitoring for these viruses in beet leafhoppers. Plant. Dis. 94, 99–108 (2010).
Ohshima, K. et al. Patterns of recombination in turnip mosaic virus genomic sequences indicate hotspots of recombination. J. Gen. Virol. 88, 298–315 (2007).
Strausbaugh, C. A., Wintermantel, W. M. & Gillen, A. M. Eujayl, I. A. Curly top survey in the Western united States. Phytopathology 98, 1212–1217 (2008).
Soto, M. J. & Gilbertson, R. L. Distribution and rate of movement of the curtovirus beet mild Curly top virus (Family Geminiviridae) in the beet leafhopper. Phytopathology 93, 478–484 (2003).
Majidi, A., Hamzehzarghani, H., Izadpanah, I. & Behjatnia, S. A. A. Interaction between beet Curly top Iran virus and the severe isolate of beet Curly top virus in three selective sugar beet cultivars. 99, 381–389 (2017).
Lecoq, H. & Desbiez, C. Viruses of cucurbit crops in the mediterranean region: an ever-changing picture. Adv. Virus Res. 84, 67–126 (2012).
Nomura, K., Ohshima, K., Anai, T. & Uekusa, H. Kita, N. RNA Silencing of the introduced coat protein gene of turnip mosaic virus confers Broad-Spectrum resistance in Transgenic Arabidopsis. Phytopathology 94, 730–736 (2004).
Jan, F. J., Pang, S. Z., Fagoaga, C. & Gonsalves, D. Turnip mosaic potyvirus resistance in Nicotiana benthamiana derived by post-transcriptional gene Silencing. Transgenic Res. 8, 203–213 (1999).
Frischmuth, T. & Stanley, J. Beet Curly top virus symptom amelioration in Nicotiana benthamiana transformed with a naturally occurring viral subgenomic DNA. Virology 200, 826–830 (1994).
Ali, Z. et al. CRISPR/Cas9-mediated viral interference in plants. Genome Biol. 16, 1–11 (2015).
Delgado-Martín, J., Ruiz, L., Janssen, D. & Velasco, L. Exogenous application of DsRNA for the control of viruses in cucurbits. Front. Plant. Sci. 13, 895953 (2022).
Frascati, F. et al. Exogenous application of DsRNA for protection against tomato leaf curl new Delhi virus. Viruses 16, 436 (2024).
Delgado-Martín, J., Delgado-olidén, A. & Velasco, L. Carbon Dots boost DsRNA delivery in plants and increase local and systemic SiRNA production. Int. J. Mol. Sci. 23, 5338 (2022).
Weinberger, C. et al. The structure of water in silica Mesopores – Influence of the pore wall Polarity. Adv. Mater. Interfaces. 9, 2200245 (2022).
Keil, W. et al. Thermostable water reservoirs in the interlayer space of a sodium Hectorite clay through the intercalation of γ-aminopropyl(dimethyl)ethoxysilane in toluene. Phys. Chem. Chem. Phys. 24, 477–487 (2021).
Behzadi, F. et al. Stability and antimicrobial activity of Nisin-Loaded mesoporous silica nanoparticles: A Game-Changer in the war against maleficent microbes. J. Agric. Food Chem. 66, 4233–4243 (2018).
Heidari, R., Khosravian, P., Mirzaei, S. A. & Elahian, F. SiRNA delivery using intelligent chitosan-capped mesoporous silica nanoparticles for overcoming multidrug resistance in malignant carcinoma cells. Sci. Rep. 11, 20531 (2021).
Choi, K. et al. Chromium removal from aqueous solution by a PEI-silica nanocomposite. Sci. Rep. 8, 1–10 (2018).
Gisbert-Garzarán, M. & Vallet-Regí, M. Influence of the surface functionalization on the fate and performance of mesoporous silica nanoparticles. Nanomaterials 2020. 10, Page 916 (10), 916 (2020).
Jeppu, G. P. & Clement, T. P. A modified Langmuir-Freundlich isotherm model for simulating pH-dependent adsorption effects. J. Contam. Hydrol. 129–130, 46–53 (2012).
Zolghadrnasab, M., Mousavi, A., Farmany, A. & Arpanaei, A. Ultrasound-mediated gene delivery into suspended plant cells using polyethyleneimine-coated mesoporous silica nanoparticles. Ultrason. Sonochem. 73, 105507 (2021).
Schwartz, S. H., Hendrix, B., Hoffer, P., Sanders, R. A. & Zheng, W. Carbon Dots for efficient small interfering RNA delivery and gene Silencing in plants. Plant. Physiol. 184, 647–657 (2020).
Xu, X. et al. Evaluation of the anti-viral efficacy of three different DsRNA nanoparticles against potato virus Y using various delivery methods. Ecotoxicol. Environ. Saf. 255, 114775 (2023).
Melita, O. et al. Topical application of double-stranded RNA molecules deriving from tomato yellow leaf curl virus reduces cognate virus infection in tomato. Biol. Plant. 65, 100–110 (2021).
Namgial, T., Kaldis, A., Chakraborty, S. & Voloudakis, A. Topical application of double-stranded RNA molecules containing sequences of tomato leaf curl virus and cucumber mosaic virus confers protection against the cognate viruses. Physiol. Mol. Plant. Pathol. 108, 101432 (2019).
Rego-Machado, C. M. et al. SiRNA biogenesis and advances in topically applied DsRNA for controlling virus infections in tomato plants. Sci. Rep. 10, 22277 (2020).
Montazeri, R. & Malboobi, M. A. Shams-bakhsh, M. Resistance induced by viral sense, anti-sense, and hairpin constructs against beet Curly top virus and beet Curly top Iran virus. Iran. J. Biotechnol. 22, 50–61 (2024).
Yang, X., Li, Y. & Wang, A. Research advances in potyviruses: from the laboratory bench to the field. Annu. Rev. Phytopathol. 59, 1–29 (2021).
Sun, Z. N., Song, Y. Z., Yin, G. H., Zhu, C. X. & Wen, F. J. Hprnas derived from different regions of the Nib gene have different abilities to protect tobacco from infection with potato virus y. J. Phytopathol. 158, 566–568 (2010).
Worrall, E. A. et al. Exogenous application of RNAi-inducing double-stranded RNA inhibits aphid-mediated transmission of a plant virus. Front. Plant. Sci. 10, 265 (2019).
Rêgo-Machado, C. M., Inoue-Nagata, A. K. & Nakasu, E. Y. T. Topical application of DsRNA for plant virus control: a review. Trop. Plant. Pathol. 48, 11–22 (2022).
Tenllado, F. & Dı́az-Ruı́z, J. R. Double-Stranded RNA-Mediated interference with plant virus infection. J. Virol. 75, 12288–12297 (2001).
Yoon, J. et al. Double-stranded RNA confers resistance to pepper mottle virus in Nicotiana benthamiana. Appl. Biol. Chem. 64, 1–8 (2021).
Hu, J., Xiong, J., Zhao, Z. & Wang, X. Carbon Dots promote plant growth via coordinated regulation of nutrient uptake and photosynthesis: evidence from multivariate modeling. Bioresour Technol. 440, 133515 (2026).
Cheng, W. et al. Closed-loop enhancement of plant photosynthesis via biomass-derived carbon Dots in biohybrids. Commun Mater 6, (2025).
Shivashakarappa, K. et al. DNA delivery into plant tissues using carbon Dots made from citric acid and β-alanine. Front Chem 13 (2025).
Bachman, P., Fischer, J., Song, Z., Urbanczyk-Wochniak, E. & Watson, G. Environmental fate and dissipation of applied DsRNA in Soil, aquatic Systems, and plants. Front. Plant. Sci. 11, 21 (2020).
Ahn, S. J., Donahue, K., Koh, Y., Martin, R. R. & Choi, M. Y. Microbial-Based Double-Stranded RNA production to develop Cost-Effective RNA interference application for insect pest management. Int. J. Insect Sci. 11, 117954331984032 (2019).
Sánchez, F., Martínez-Herrera, D., Aguilar, I. & Ponz, F. Infectivity of turnip mosaic potyvirus cDNA clones and transcripts on the systemic host Arabidopsis Thaliana and local lesion hosts. Virus Res. 55, 207–219 (1998).
Walsh, J. A. Genetic control of immunity to turnip mosaic virus in winter oilseed rape (Brassica Napus ssp. oleifera) and the effect of foreign isolates of the virus. Ann. Appl. Biol. 115, 89–99 (1989).
Briddon, R. W. et al. Comparison of a beet Curly top virus isolate originating from the old world with those from the new world. Eur. J. Plant. Pathol. 104, 77–84 (1998).
Taebnia, N. et al. The effect of mesoporous silica nanoparticle surface chemistry and concentration on the α-synuclein fibrillation. RSC Adv. 5, 60966–60974 (2015).
Abramoff, M. D., Magalhães, P. J. & Ram, S. J. Image processing with ImageJ. Biophotonics Int. 11, 36–42 (2004).
Campbell, A., Cardine, V., Hines, D. & Kerns, S. Fitting ODE parameters to data using excel- using regression to fit complex models in excel. (2007). https://eng.libretexts.org/@go/page/22371
Livak, K. J. & Schmittgen, T. D. Analysis of relative gene expression data using real-time quantitative PCR and the 2-∆∆CT method. Methods 25, 402–408 (2001).
Delgado-Martín, J. & Velasco, L. An efficient DsRNA constitutive expression system in Escherichia coli. Appl. Microbiol. Biotechnol. 105, 6381–6393 (2021).
Accotto, G. P., Navas-Castillo, J., Noris, E., Moriones, E. & Louro, D. Typing of tomato yellow leaf curl viruses in Europe. Eur. J. Plant. Pathol. 106 (2000).
Funding
This work was supported by grant PID2021-125787OR-C32 financed by MICIU/AEI/10.13039/501100011033 and FEDER, EU, and by the Center for International Scientific Studies & Collaboration (CISSC), Ministry of Science, Research and Technology of Iran.
Author information
Authors and Affiliations
Contributions
SZ, AA, MS-B and LV designed the research. SZ, CR, EM-C, JD-M, AA and LV performed the experiments and analyzed the data and results. SZ and LV wrote the original draft. All the authors contributed to the final manuscript and approved the submitted version. LV, MS-B, and AA provided funds for the project.
Corresponding authors
Ethics declarations
Competing interests
The authors declare no competing interests.
Additional information
Publisher’s note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Below is the link to the electronic supplementary material.
Rights and permissions
Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if you modified the licensed material. You do not have permission under this licence to share adapted material derived from this article or parts of it. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by-nc-nd/4.0/.
About this article
Cite this article
Zarrabi, S., Rangel, C., Martínez-Campos, E. et al. Carbon Dots and mesoporous silica nanocomposites improve spray-induced gene silencing to suppress plant RNA and DNA viruses. Sci Rep (2026). https://doi.org/10.1038/s41598-026-36331-6
Received:
Accepted:
Published:
DOI: https://doi.org/10.1038/s41598-026-36331-6