Abstract
Key message
We report the size dependent uptake of dsRNA loaded MSNPs into the leaves and roots of Nicotiana benthamiana plants and accessed for their relative reduction in Tomato leaf curl New Delhi viral load.
Abstract
A non-GMO method of RNA interference (RNAi) has been recently in practice through direct delivery of double stranded RNA into the plant cells. Tomato leaf curl New Delhi virus (ToLCNDV), a bipartitie begomovirus, is a significant viral pathogen of many crops in the Indian subcontinent. Conventional RNAi cargo delivery strategies for instance uses viral vectors and Agrobacterium-facilitated delivery, exhibiting specific host responses from the plant system. In the present study, we synthesized three different sizes of amine-functionalized mesoporous silica nanoparticles (amino-MSNPs) to mediate the delivery of dsRNA derived from the AC2 (dsAC2) gene of ToLCNDV and showed that these dsRNA loaded nanoparticles enabled effective reduction in viral load. Furthermore, we demonstrate that amino-MSNPs protected the dsRNA molecules from nuclease degradation, while the complex was efficiently taken up by the leaves and roots of Nicotiana benthamiana. The real time gene expression evaluation showed that plants treated with nanoparticles of different sizes ~ 10 nm (MSNPDEA), ~ 32 nm (MSNPTEA) and ~ 66 nm (MSNPNH3) showed five-, eleven- and threefold reduction of ToLCNDV in N. benthamiana, respectively compared to the plants treated with naked dsRNA. This work clearly demonstrates the size dependent internalization of amino-MSNPs and relative efficacy in transporting dsRNA into the plant system, which will be useful in convenient topical treatment to protect plants against their pathogens including viruses.
Graphical abstract
Mesoporous silica nanoparticles loaded with FITC, checked for its uptake into Nicotiana benthamiana.
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References
Abdo GG, Zagho MM, Khalil A (2020) Recent advances in stimuli-responsive drug release and targeting concepts using mesoporous silica nanoparticles. Emerg Mater 3:407–425
Avellan A, Yun J, Zhang Y et al (2019) Nanoparticle size and coating chemistry control foliar uptake pathways, translocation, and leaf-to-rhizosphere transport in wheat. ACS Nano 13:5291–5305. https://doi.org/10.1021/acsnano.8b09781
Bailey-Serres J, Parker JE, Ainsworth EA et al (2019) Genetic strategies for improving crop yields. Nature 575:109–118. https://doi.org/10.1038/s41586-019-1679-0
Bocos-Asenjo IT, Niño-Sánchez J, Ginésy M, Diez JJ (2022) New insights on the integrated management of plant diseases by RNA strategies: mycoviruses and RNA interference. Int J Mol Sci 23:9236. https://doi.org/10.3390/ijms23169236
Boualem A, Dogimont C, Bendahmane A (2016) The battle for survival between viruses and their host plants. Curr Opin Virol 17:32–38. https://doi.org/10.1016/j.coviro.2015.12.001
Calil IP, Fontes EPB (2017) Plant immunity against viruses: antiviral immune receptors in focus. Ann Bot 119:711–723. https://doi.org/10.1093/aob/mcw200
Castellanos NL, Smagghe G, Sharma R et al (2019) Liposome encapsulation and EDTA formulation of dsRNA targeting essential genes increase oral RNAi-caused mortality in the Neotropical stink bug Euschistus heros. Pest Manag Sci 75:537–548. https://doi.org/10.1002/ps.5167
Corredor E, Testillano PS, Coronado M, et al Nanoparticle penetration and transport in living pumpkin plants : in situ subcellular identification. BMC Plant Biol 11:1–11. https://doi.org/10.1186/1471-2229-9-45
Dangl JL, Horvath DM, Staskawich BJ (2013) Pivoting the plant immune system. Science (80-) 341:745–751
Darsan Singh JK, Mat Jalaluddin NS, Sanan-Mishra N, Harikrishna JA (2019) Genetic modification in Malaysia and India: current regulatory framework and the special case of non-transformative RNAi in agriculture. Plant Cell Rep 38:1449–1463. https://doi.org/10.1007/s00299-019-02446-6
Das S, Debnath N, Cui Y et al (2015) Chitosan, carbon quantum dot, and silica nanoparticle mediated dsRNA delivery for gene silencing in Aedes aegypti: a comparative analysis. ACS Appl Mater Interfaces 7:19530–19535. https://doi.org/10.1021/acsami.5b05232
Demirer GS, Zhang H, Goh NS et al (2020) Carbon nanocarriers deliver siRNA to intact plant cells for efficient gene knockdown. Sci Adv. https://doi.org/10.1126/sciadv.aaz0495
Douglas AE (2018) Strategies for enhanced crop resistance to insect pests. Annu Rev Plant Biol 69:637–660. https://doi.org/10.1146/annurev-arplant-042817-040248
Edwards CH, Christie CR, Masotti A et al (2020) Dendrimer-coated carbon nanotubes deliver dsRNA and increase the efficacy of gene knockdown in the red flour beetle Tribolium castaneum. Sci Rep 10:1–11. https://doi.org/10.1038/s41598-020-69068-x
Farooq T, Adeel M, He Z et al (2021) Nanotechnology and plant viruses: an emerging disease management approach for resistant pathogens. ACS Nano 15:6030–6037. https://doi.org/10.1021/acsnano.0c10910
Fiallo-Olivé E, Pan LL, Liu SS, Navas-Castillo J (2020) Transmission of begomoviruses and other whitefly-borne viruses: dependence on the vector species. Phytopathology 110:10–17. https://doi.org/10.1094/PHYTO-07-19-0273-FI
Ghassemi-Golezani K, Abdoli S (2021) Improving ATPase and PPase activities, nutrient uptake and growth of salt stressed ajowan plants by salicylic acid and iron-oxide nanoparticles. Plant Cell Rep 40:559–573. https://doi.org/10.1007/s00299-020-02652-7
Guerrero J, Regedanz E, Lu L et al (2020) Manipulation of the plant host by the Geminivirus AC2/C2 protein, a central player in the infection cycle. Front Plant Sci 11:1–18. https://doi.org/10.3389/fpls.2020.00591
He B, Chu Y, Yin M et al (2013) Fluorescent nanoparticle delivered dsRNA toward genetic control of insect pests. Adv Mater 25:4580–4584. https://doi.org/10.1002/adma.201301201
Jones RAC (2021) Global plant virus disease pandemics and epidemics
Kah M, Tufenkji N, White JC (2019) Nano-enabled strategies to enhance crop nutrition and protection. Nat Nanotechnol 14:532–540. https://doi.org/10.1038/s41565-019-0439-5
Khan MR, Siddiqui ZA, Fang X (2022) Potential of metal and metal oxide nanoparticles in plant disease diagnostics and management: recent advances and challenges. Chemosphere 297:134114. https://doi.org/10.1016/j.chemosphere.2022.134114
Kwak SY, Lew TTS, Sweeney CJ et al (2019) Chloroplast-selective gene delivery and expression in planta using chitosan-complexed single-walled carbon nanotube carriers. Nat Nanotechnol 14:447–455. https://doi.org/10.1038/s41565-019-0375-4
Lei WX, An ZS, Zhang BH et al (2020) Construction of gold-siRNANPR1 nanoparticles for effective and quick silencing ofNPR1 in Arabidopsis thaliana. RSC Adv 10:19300–19308. https://doi.org/10.1039/d0ra02156c
Li K, Wu G, Li M et al (2018) Transcriptome analysis of Nicotiana benthamiana infected by tobacco curly shoot virus. Virol J 15:1–15. https://doi.org/10.1186/s12985-018-1044-1
Liu C, Tian S, Lv X et al (2022) Nicotiana benthamiana asparagine synthetase associates with IP-L and confers resistance against tobacco mosaic virus via the asparagine-induced salicylic acid signalling pathway. Mol Plant Pathol 23:60–77. https://doi.org/10.1111/mpp.13143
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method. Methods 25:402–408. https://doi.org/10.1006/meth.2001.1262
Ma X, Zhao Y, Ng KW, Zhao Y (2013) Integrated hollow mesoporous silica nanoparticles for target drug/siRNA co-delivery. Chem A Eur J 19:15593–15603. https://doi.org/10.1002/chem.201302736
Meroni D, Lo Presti L, Di Liberto G et al (2017) A close look at the structure of the TiO2–APTES interface in hybrid nanomaterials and its degradation pathway: an experimental and theoretical study. J Phys Chem C 121:430–440. https://doi.org/10.1021/acs.jpcc.6b10720
Miller RNG, Alves GSC, Van Sluys MA (2017) Plant immunity: unravelling the complexity of plant responses to biotic stresses. Ann Bot 119:681–687. https://doi.org/10.1093/aob/mcw284
Mitter N, Worrall EA, Robinson KE et al (2017) Clay nanosheets for topical delivery of RNAi for sustained protection against plant viruses. Nat Plants. https://doi.org/10.1038/nplants.2016.207
Mohan C, Shibao PYT, de Paula FFP et al (2021) hRNAi-mediated knock-down of Sphenophorus levis V-ATPase E in transgenic sugarcane (Saccharum spp interspecific hybrid) affects the insect growth and survival. Plant Cell Rep 40:507–516. https://doi.org/10.1007/s00299-020-02646-5
Moriones E, Praveen S, Chakraborty S (2017) Tomato leaf curl New Delhi virus: an emerging virus complex threatening vegetable and fiber crops. Viruses. https://doi.org/10.3390/v9100264
Narayan R, Nayak UY (2018) Mesoporous silica nanoparticles : a comprehensive review on synthesis and recent advances. Pharmaceutics. https://doi.org/10.3390/pharmaceutics10030118
Niazian M, Molaahmad Nalousi A, Azadi P et al (2021) Perspectives on new opportunities for nano-enabled strategies for gene delivery to plants using nanoporous materials. Planta 254:1–20. https://doi.org/10.1007/s00425-021-03734-w
Palocci C, Valletta A, Chronopoulou L et al (2017) Endocytic pathways involved in PLGA nanoparticle uptake by grapevine cells and role of cell wall and membrane in size selection. Plant Cell Rep 36:1917–1928. https://doi.org/10.1007/s00299-017-2206-0
Rahman A, Sinha KV, Sopory SK, Sanan-Mishra N (2021) Influence of virus–host interactions on plant response to abiotic stress. Plant Cell Rep 40:2225–2245. https://doi.org/10.1007/s00299-021-02718-0
Sapino S, Ugazio E, Gastaldi L et al (2015) Mesoporous silica as topical nanocarriers for quercetin: characterization and in vitro studies. Eur J Pharm Biopharm 89:116–125. https://doi.org/10.1016/j.ejpb.2014.11.022
Schmittgen TD, Livak KJ (2008) Analyzing real-time PCR data by the comparative CT method. Nat Protoc 3:1101–1108. https://doi.org/10.1038/nprot.2008.73
Sharma N, Prasad M (2020) Silencing AC1 of Tomato leaf curl virus using artificial microRNA confers resistance to leaf curl disease in transgenic tomato. Plant Cell Rep 39:1565–1579. https://doi.org/10.1007/s00299-020-02584-2
Sharma VK, Basu S, Chakraborty S (2015) RNAi mediated broad-spectrum transgenic resistance in Nicotiana benthamiana to chilli-infecting begomoviruses. Plant Cell Rep 34:1389–1399. https://doi.org/10.1007/s00299-015-1795-8
Sharma A, Kumar V, Shahzad B et al (2019) Worldwide pesticide usage and its impacts on ecosystem. SN Appl Sci 1:1–16. https://doi.org/10.1007/s42452-019-1485-1
Singh OW, Gupta D, Joshi B et al (2022) Spray application of a cocktail of dsRNAs reduces infection of chilli leaf curl virus in Nicotiana benthamiana. J Plant Dis Prot 129:433–438. https://doi.org/10.1007/s41348-021-00549-5
Stoeckel D, Wallacher D, Zickler GA et al (2014) Coherent analysis of disordered mesoporous adsorbents using small angle X-ray scattering and physisorption experiments. Phys Chem Chem Phys 16:6583–6592. https://doi.org/10.1039/c3cp55072a
Strange RN, Scott PR (2005) Plant disease: a threat to global food security. Annu Rev Phytopathol 43:83–116. https://doi.org/10.1146/annurev.phyto.43.113004.133839
Sun D, Hussain HI, Yi Z et al (2014) Uptake and cellular distribution, in four plant species, of fluorescently labeled mesoporous silica nanoparticles. Plant Cell Rep 33:1389–1402. https://doi.org/10.1007/s00299-014-1624-5
Tabashnik BE, Carrière Y (2017) Surge in insect resistance to transgenic crops and prospects for sustainability. Nat Biotechnol 35:926–935. https://doi.org/10.1038/nbt.3974
Tarn D, Ashley CE, Xue M et al (2013) Mesoporous silica nanoparticle nanocarriers. Acc Chem Res 46:792
Wang JW, Grandio EG, Newkirk GM et al (2019) Nanoparticle-mediated genetic engineering of plants. Mol Plant 12:1037–1040. https://doi.org/10.1016/j.molp.2019.06.010
Wang K, Peng Y, Chen J et al (2020) Comparison of efficacy of RNAi mediated by various nanoparticles in the rice striped stem borer (Chilo suppressalis). Pestic Biochem Physiol. https://doi.org/10.1016/j.pestbp.2019.10.005
Worrall EA, Bravo-Cazar A, Nilon AT et al (2019) Exogenous application of RNAi-inducing double-stranded RNA inhibits aphid-mediated transmission of a plant virus. Front Plant Sci. https://doi.org/10.3389/fpls.2019.00265
Wu SH, Lin HP (2013) Synthesis of mesoporous silica nanoparticles. Chem Soc Rev 42:3862–3875. https://doi.org/10.1039/c3cs35405a
Xue G, Yurun F, Li M et al (2017) Phosphoryl functionalized mesoporous silica for uranium adsorption. Appl Surf Sci 402:53–60. https://doi.org/10.1016/j.apsusc.2017.01.050
Zeng W, Bai H (2014) Swelling-agent-free synthesis of rice husk derived silica materials with large mesopores for efficient CO2 capture. Chem Eng J 251:1–9. https://doi.org/10.1016/j.cej.2014.04.041
Zhang X, Zheng X, Zhang S et al (2012) AM-TEPA impregnated disordered mesoporous silica as CO2 capture adsorbent for balanced adsorption–desorption properties. Ind Eng Chem Res 51:15163–15169. https://doi.org/10.1021/ie300180u
Zhang H, Cao Y, Xu D et al (2021) Gold-nanocluster-mediated delivery of siRNA to intact plant cells for efficient gene knockdown. Nano Lett 21:5859–5866. https://doi.org/10.1021/acs.nanolett.1c01792
Zotti M, dos Santos EA, Cagliari D et al (2018) RNA interference technology in crop protection against arthropod pests, pathogens and nematodes. Pest Manag Sci 74:1239–1250. https://doi.org/10.1002/ps.4813
Acknowledgements
A.S. acknowledges University Grants Commission (UGC), for financial support as research fellowship. N.S. and B.M acknowledge research funding from National Agricultural Science Fund (NASF), ICAR, India (NASF/ABP-7021/2018-19/256). We thank CRF (IITD) for HR-TEM, FE-SEM, FTIR, XRD and SAXS facilities. We specially thank Gurdeep Kaur (TERI Gram) and Dr P.M. Reddy (TERI Gram) for their help in the final revision of the manuscript.
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This study was supported by the ICAR-National Agricultural Science Fund (Grant No. NASF/ABP-7021/2018-19/256).
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AS: data curation, methodology, visualization, original draft; DG: methodology, visualization, review and editing, OSW: review and editing; AR: review and editing; SKM: review and editing; BM: conceptualization, visualization, supervision, review and editing; NS: conceptualization, visualization, supervision, review and editing.
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Communicated by Om Parkash Parkash Dhankher.
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Sangwan, A., Gupta, D., Singh, O.W. 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). https://doi.org/10.1007/s00299-023-03048-z
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DOI: https://doi.org/10.1007/s00299-023-03048-z