Fostering sustainable agriculture: genotype-independent plant cell transfection mediated by piezoelectric nanomaterials via sonoporation

Abstract

In the pursuit of advancing sustainable agriculture through innovative biotechnological approaches, our research focuses on the transformative potential of nanoparticle-mediated cargo delivery via sonoporation that can potentially create reversible pores in plasma membranes and rigid cell walls, which in turn enhances the gene transfection efficiency. In this research study, we fabricated core–shell piezoelectric nanoparticles – specifically, barium titanate nanoparticles (BTNPs) encased in a silica shell, exhibiting an approximate diameter of 60 nm. These engineered nanoparticles were specifically crafted to proficiently transport nucleic acids into suspension cells of both monocots and dicots, employing the technique of sonoporation. In this study, we conducted comparative analyses between BTNPs and SNPs, employing the latter as a control, with the objective of establishing a highly efficient system for delivering diverse cargoes into plant suspension cells. The DNA adsorption capacity was around 40.1 μg per mg of BTNP@SiO₂. By keeping the intensity and frequency constant, time variations have been optimized for both type of cells by taking cell viability into consideration. At 160 W intensity and frequency of 40 kHz, the best ultrasonic treatment for transfection is obtained as 5 and 8 min for dicot and monocot cells, respectively. GUS quantitative analysis shows that in the presence of ultrasound, piezoelectric nanoparticles have enhanced the transfection efficiency in comparison to baseline naked pDNA delivery by 56.9 ± 2.29 and 72.6 ± 3.01% in dicot and monocot suspension cells, respectively. Our findings not only demonstrate the enhanced transfection efficiency of piezoelectric nanoparticles, but also underscore the significance of this approach for genotype-independent transfection. By offering a universal and streamlined method that is effective across various plant genotypes, this research presents a significant step forward in the field of plant genetic engineering. The genotype-independent transfection potential of our methodology has the power to transform the way genetic material is delivered into plant cells, with profound implications for addressing global challenges in agriculture and plant biology.