Targeted Carbon Nanostructures for Chemical and Gene Delivery to Plant Chloroplasts

Nanotechnology approaches for improving the delivery efficiency of chemicals and molecular cargoes in plants through plant biorecognition mechanisms remain relatively unexplored. We developed targeted carbon-based nanomaterials as tools for precise chemical delivery (carbon dots, CDs) and gene delivery platforms (single-walled carbon nanotubes, SWCNTs) to chloroplasts, key organelles involved in efforts to improve plant photosynthesis, assimilation of nutrients, and delivery of agrochemicals. A biorecognition approach of coating the nanomaterials with a rationally designed chloroplast targeting peptide improved the delivery of CDs with molecular baskets (TP-β-CD) for delivery of agrochemicals and of plasmid DNA coated SWCNT (TP-pATV1-SWCNT) from 47% to 70% and from 39% to 57% of chloroplasts in leaves, respectively. Plants treated with TP-β-CD (20 mg/L) and TP-pATV1-SWCNT (2 mg/L) had a low percentage of dead cells, 6% and 8%, respectively, similar to controls without nanoparticles, and no permanent cell and chloroplast membrane damage after 5 days of exposure. However, targeted nanomaterials transiently increased leaf H 2 O 2 (0.3225 μmol gFW –1 ) above control plant levels (0.03441 μmol gFW –1 ) but within the normal range reported in land plants. The increase in leaf H 2 O 2 levels was associated with oxidative damage in whole plant cell DNA, a transient effect on chloroplast DNA, and a decrease in leaf chlorophyll content (−17%) and carbon assimilation rates at saturation light levels (−32%) with no impact on photosystem II quantum yield. This work provides targeted delivery approaches for carbon-based nanomaterials mediated by biorecognition and a comprehensive understanding of their impact on plant cell and molecular biology for engineering safer and efficient agrochemical and biomolecule delivery tools.