From Plants to Plants: Plant-Derived Biological Polymers as Sustainable and Safe Nanocarriers for Direct Delivery of DNA to Plant Cells
Kari Vinzant, Mohammad Mamunur Rashid, Delaney E. Clouse, Pratyusha Ghosh, Mohiuddin Quadir, Virginia A. Davis, Mariya V. Khodakovskaya
Abstract
Sustainable plant-derived biopolymers cellulose nanocrystals (CNC) and Zein protein were used to deliver plasmid DNA with a reporter GFP gene (pDNA) to plant cells. CNC and Zein were modified with the cationic agent 2,3-epoxypropyltrimethylammonium chloride (EPTMAC) to electrostatically bind the biopolymers to negatively charged pDNA. Established pDNA-CNC and pDNA-Zeins conjugates were delivered to tobacco cells by leaf injection and vacuum infiltration of tobacco leaves and seedlings. Both methods effectively provided transient GFP expression in exposed plant cells that was visualized by confocal microscopy and confirmed by qRT-PCR (GFP gene expression) and Western blot (GFP protein expression). Our findings support the idea that nanopolymers derived from agricultural waste residues can successfully be used to advance plant transformation and gene editing. Delivering genetic material using biocompatible, plant-based nanopolymers in large-scale vacuum infiltration of plant tissues reduces existing limitations of plant transformation and increases the speed of the transformation process.