Biosynthesis of iron oxide superparamagnetic nanoparticles and optimization of transient gene transfer to purslane (Portulaca oleracea L.) and safflower (Carthamus tinctorius L.)

Document Type : Research Paper

Authors

1 MSc Student, Department of Agricultural Biotechnology, Faculty of Agriculture, Shahid Bahonar University of Kerman, Kerman, Iran.

2 Associate Professor, Department of Agricultural Biotechnology, Faculty of Agriculture, Shahid Bahonar University of Kerman, Kerman, Iran.

3 PhD, Department of Agronomy and Plant Breeding, Faculty of Agriculture, Shahid Bahonar University of Kerman, Kerman, Iran.

Abstract

Objective
Despite significant advancements in biotechnology and genetic engineering over several decades, the genetic modification of many plant species remains challenging due to the presence of cell walls. Conventional gene transfer methods face various limitations. Recently, the unique properties of nanoparticles have attracted attention for their potential use in biotechnology as nanocarriers of biomolecules, including DNA, RNA, and proteins, due to their ability to penetrate living cells.
Materials and Methods
In this study, SPION nanoparticles were synthesized using a green synthesis method with turmeric rhizome extract. The nanoparticles were functionalized with the cationic polymer polyethyleneimine (PEI), and the plasmid pCAMBIA1304, containing GFP and GUS genes, was loaded onto the nanocarrier surface. Gene transfer was conducted using two methods: syringe infiltration and vacuum infiltration on safflower and purslane plant leaves. The presence of the mGFP protein fluorescent signal and PCR analysis were employed to evaluate gene transfer and expression.
Results
SPION nanoparticles were successfully synthesized using turmeric rhizome extract as a reducing agent for iron ions. A color change in the solution and subsequent analyses confirmed the successful synthesis of SPION nanoparticles. Additionally, the zeta potential shift from negative to positive indicated successful PEI functionalization on the nanoparticle surface. The presence of the mGFP protein fluorescent signal confirmed the nanocarrier's ability to deliver genes to plant cells.
Conclusions
The findings demonstrated that nanoparticle-mediated gene transfer to plant cells can be achieved efficiently without the use of bacteria and independent of plant species limitations. Given these advantages, optimizing and improving transfer efficiency could establish nanoparticles as a widely utilized carrier in plant gene transfer applications.

Keywords

References

Abedini S, Pourseyedi SH, Jafar J, Abdolshahi, R (2023) Optimization of transient gene delivery to catharanthus roseus using introduction of green synthesized superparamagnetic iron oxide nanoparticles and carbon nanotubes nanocarrier. Agric Biotechnol J 15(1), 61-80 (In Persian)
Adabavazeh F, Pourseyedi SH, Nadernejad N, et al. (2022) Evaluation of synthesized magnetic nanoparticles and salicylic acid effects on improvement of antioxidant properties and essential oils of Calotropis procera hairy roots and seedlings. Plant Cell Tiss Organ Cult 151, 133-148.
Bassim S, Mageed AK, AbdulRazak AA, Majdi HS (2022) Green synthesis of Fe3O4 nanoparticles and its applications in wastewater treatment. Inorganics 10(12), e260.
Bentahar S, Abada R, Ykhlef N (2023) Biotechnology: definitions, types and main applications. Ymer 22(4), 563-575.
Cardoso VF, Francesko A, Ribeiro C, et al. (2018) Advances in magnetic nanoparticles for biomedical applications. Adv Healthc Mater 7, e1700845.
Chang FP, Kuang LY, Huang CA, et al. (2013) A simple plant gene delivery system using mesoporous silica nanoparticles as carriers. J Mater Chem B 1(39), 5279-5287.
Chen F, Wang C, Yue L, et al. (2021) Cell walls are remodeled to alleviate nY2O3 cytotoxicity by elaborate regulation of de novo synthesis and vesicular transport. ACS Nano 15, 13166-13177.
Chen Z, Lv Z, Sun Y, et al. (2020) Recent advancements in polyethyleneimine-based materials and their biomedical, biotechnology, and biomaterial applications. J Mater Chem B 8, 2951-2973.
Chopra H, Bibi S, Singh I, et al. (2022) Green metallic nanoparticles: Biosynthesis to applications. Front Bioeng Biotechnol 10, e874742.
Dehghani J, Movafeghi A, Barzegari A, Barar J (2017) Efficient and stable transformation of Dunaliella pseudosalina by 3 strains of Agrobacterium tumefaciens. Bioimpacts 7(4), 247-254.
Demirer GS, Zhang H, Matos JL, et al. (2019) Carbon nanotube–mediated DNA delivery without transgene integration in intact plants. Nat Protoc 14, 456-464.
Dobson J (2006) Gene therapy progress and prospects: magnetic nanoparticle-based gene delivery. Gene Ther 13(4), 283-287.
Farooq N, Ather L, Shafiq M, et al. (2022) Magnetofection approach for the transformation of okra using green iron nanoparticles. Sci Rep 12, e16568
Hajiahmadi Z, Shirzadian-Khorramabad R, Kazemzad M, Sohani MM (2019) Enhancement of tomato resistance to Tuta absoluta using a new efficient mesoporous silica nanoparticle-mediated plant transient gene expression approach. Scientia Horticulturae 243, 367-375
Izuegbunam CL, Wijewantha N, Wone B, et al. (2021) A nano-biomimetic transformation system enables in planta expression of a reporter gene in mature plants and seeds. Nanoscale Adv 3(11), 3240-3250.
Joudeh J, Linke D (2022) Nanoparticle classification, physicochemical properties, characterization, and applications. J Nanobiotechnol 20, e262.
Karpagavinayagam P, Vedhi C (2019) Green synthesis of iron oxide nanoparticles using Avicennia marina flower extract. Vacuum 160, 286–292.
Liu Q, Chen B, Wang Q, Shi X, et al. (2009) Carbon nanotubes as molecular transporters for walled plant cells. Nano Lett 9(3), 1007-1010.
Lv Z, Jiang R, Chen J, Chen W (2020) Nanoparticle-mediated gene transformation strategies for plant genetic engineering. Plant J 104, 880-891.
Nitnaware KM, Naikawadi VB, Chavan SS, et al. (2021) Genetic Engineering in Safflower (Carthamus tinctorius L.): Retrospect and Prospect. In: Kavi Kishor, P.B., Rajam, M.V., Pullaiah, T. (eds) Genetically Modified Crops. Springer, Singapore. pp. 201-226.
Rana G, Dhiman P, Kumar A, et al. (2024) Phytomediated synthesis of Fe3O4 nanoparticles using Cannabis sativa root extract: photocatalytic activity and antibacterial efficacy. Biomass Conv Bioref 5785, 1-18.
Rohiwal SS, Dvorakova N, Klima J, et al. (2020) Polyethylenimine based magnetic nanoparticles mediated non-viral CRISPR/Cas9 system for genome editing. Sci Rep 10(1), e4619.
Rozita NASD (2020) Papain grafted into the silica coated iron-based magnetic nanoparticles “IONPs@SiO2–PPN” as a new delivery vehicle to the HeLa cells. J Nanotechnol 31, e195603.
Sosa‑Acosta JR, Iriarte‑Mesa C, Ortega GA, Díaz‑García AM (2020) DNA-iron oxide nanoparticles conjugates: Functional magnetic nanoplatforms in biomedical applications. Top Curr Chem 378, e13.
Su W, Xu M, Radani Y, Yang L (2023) Technological Development and Application of Plant Genetic Transformation. Int J Mol Sci 24, 10646. ‏
Tyurin AA, Suhorukova AV, Kabardaeva KV, Goldenkova IV (2020) Transient Gene Expression is an Effective Experimental Tool for the Research into the Fine Mechanisms of Plant Gene Function: Advantages, Limitations, and Solutions. Plants 9, e1187.
Zhao X, Meng Z, Wang Y, et al. (2017) Pollen magnetofection for genetic modification with magnetic nanoparticles as gene carriers. Nat Plants 3, 956–964.
Agricultural Biotechnology Journal
Volume 17, Issue 1
March 2025
Pages 161-176

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