Genetic Transformation of Plant Cells Mediated by Mesoporous Silica Nanoparticles

doi:10.13560/j.cnki.biotech.bull.1985.2021-1040

Abstract

Abstract:

With the rapid development of nanobiotechnology,nanoparticles have been applied as vectors for gene transformation in plant cells. In this study,mesoporous silica nanoparticles（MSNs）with different particle sizes were synthesized and then amine-functionalized. The transformation efficiencies of amine-functionalized MSNs（Am-MSNs）for delivering plasmid DNA encoding green fluorescent protein（smGFP）were investigated in the protoplasts of Arabidopsis thaliana. As results,the Am-MSN-20 exhibited monodispersed spheroids with average particle size of 20 nm,while Am-MSN-50 showed a flower-like structure and had an average particle size of 50 nm. Both amine-functionalized MSNs were positively charged. Am-MSN-50 had higher binding capacity of pDNA than that of Am-MSN-20. The Am-MSNs/pDNA complex showed good stability,which suggested that both Am-MSNs protected bounded pDNA efficiently against degradation by cellular nucleases. The MSNs also exhibited no cytotoxic effects on the protoplasts of A. thaliana. The Am-MSN-50 enabled much higher transformation efficiency of the pDNA encoding smGFP compared to that of Am-MSN-20. The results suggest that Am-MSNs can be employed as promising carriers for safe and effective gene delivery.

Key words: mesoporous silica nanoparticles, amine-functionalized, gene transfer, carrier, plant cells

LU Xin-hua, SUN De-quan, ZHANG Xiu-mei. Genetic Transformation of Plant Cells Mediated by Mesoporous Silica Nanoparticles[J]. Biotechnology Bulletin, 2022, 38(7): 194-204.

Figures/Tables 8

Fig.1 TEM images of different nanoparticles A：MSN-20;B：Am-MSN-20;C：MSN-50;D：Am-MSN-50

Fig.2 FTIR spectra of of different nanoparticles A：MSN-20;B：Am-MSN-20;C：MSN-50;D：Am-MSN-50

Fig.3 Zeta potential of different nanoparticles

Fig.4 Agarose gel electrophoresis assay of Am-MSNs/DNA complexes A：Am-MSN-20/pDNA;B：Am-MSN-50/pDNA;1：free pDNA（positive CK）;2：Am-MSNs（negative CK）;3-8 presented that the mass ratios of Am-MSNs to pDNA are 5∶1,10∶1,20∶1,30∶1,40∶1 and 50∶1,respectively

Fig.5 Protection of Am-MSNs against pDNA digestion 1：free pDNA;2：free pDNA + DNase I;3：Am-MSN-20/pDNA;4：Am-MSN-20/pDNA + DNase I;5：Am-MSN-50/pDNA;6：Am-MSN-50/pDNA + DNase I

Fig.6 Assessment of Am-MSNs cytotoxicity to A. thaliana mesophyll protoplasts A：Am-MSN-20;B：Am-MSN-50

Fig.7 Am-MSN-mediated transient smGFP expression in the protoplasts of A. thaliana A：pDNA only;B：Am-MSNs;C：pDNA/PEG;D：pDNA/Am-MSN-20;E：pDNA/Am-MSN-50. Scale bars=5 µm

Fig.8 Assessment of smGFP gene expression in the proto-plasts of A. thaliana 48 h post-transformation The error line in the figure refers to the standard deviation. Different lowercase letters indicate a significant difference（P < 0.05）

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