Date of Award

6-2-2025

Document Type

Dissertation

Degree Name

Doctor of Philosophy (PhD)

Department

Biology

First Advisor

Mariya Khodakovskaya

Abstract

Safe and efficient agricultural practices require innovative technologies. Nanotechnology offers promising solutions for advancing plant agriculture by enabling the targeted delivery of biomolecules such as agrochemicals and nucleic acids. This study focuses on developing a safe nanotechnological approach for delivering molecules of interest into plant cells. We investigated the ability of four types of polymeric nanoparticles such as Polyethylene glycol N, Nʹ-dibutylethylenediamine (PEG-DB), Polyethylene glycol 2-pyrrolidin-1-yl-ethyl-amine (PEG-PY), Polyethyleneimine (PEI), Arabinoxylan (AX)+, and Zein+ to form conjugates with plasmid DNA (pDNA). Our findings confirm that plant-derived cationic nanopolymers, AX+ and Zein+, effectively bind negatively charged pDNA (pGFPGUSPlus) carrying a GFP reporter gene. Using confocal microscopy, we demonstrated the successful intracellular delivery of pDNA by AX+ and Zein+ into plant cells. Additionally, we noted that both leaf injection and vacuum infiltration efficiently introduced these conjugates into plant tissues. The pDNA delivery was further validated through qPCR (GFP gene expression) and Western blot analysis (GFP protein expression). Furthermore, a comprehensive analysis of plant phenotype and transcriptomic profiling (RNA-seq) in tomato model plants exposed to a broad range of AX+ concentrations confirmed the absence of phytotoxic and genotoxic effects. These findings establish AX+ and Zein+ as effective carriers for the delivery of agrochemicals and biomolecules in agricultural applications. Overall, our study highlights the potential of soft, plant-based polymeric nanoparticles as promising vehicles for nucleic acid delivery, paving the way for advanced nanotechnological solutions in sustainable agriculture.

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