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


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