(Invited) Carbon Nanostructures for Targeted Delivery and Health Monitoring in Plants

Abstract

To meet the demand for food and bioenergy of a rapidly growing global population requires novel and convergent approaches to enhance photosynthetic organism function. Nanotechnology is enabling the development of targeted and species independent tools for advancing plant bioengineering and precision agriculture.My research group has developed carbon nanotube-based sensors capable of real-time sensing of plant signaling molecules. These nanosensors enabled the creation of plant sentinels that communicate the health status of plants to electronic devices. Near-infrared (nIR) fluorescent single-walled carbon nanotubes (SWCNTs) were designed and interfaced with plant leaves to report hydrogen peroxide (H2O2), a key signaling molecule associated with the onset of plant stress. The sensor nIR fluorescence response (>900 nm) is quenched by H2O2 with selectivity against other stress-associated signaling molecules and within the plant physiological range (10–100 H2O2 μM). In vivo remote nIR imaging of H2O2 sensors enabled optical monitoring of plant health in response to stresses including UV-B light (−11%), high light (−6%), and a pathogen-related peptide (flg22) (−10%), but not mechanical leaf wounding (<3%). The sensor’s high biocompatibility was reflected on similar leaf cell death (<5%) and photosynthetic rates to controls without SWCNT. These optical nanosensors report early signs of stress and will improve our understanding of plant stress communication, provide novel tools for precision agriculture, and optimize the use of agrochemicals in the environment.We also developed targeted carbon-based nanomaterials as tools for precise chemical delivery (carbon dots, CDs) and gene delivery platforms (SWCNTs) to plant organelles (chloroplasts) and vasculature (phloem). 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. Orthogonally, the surfaces of carbon dot nanocarriers were functionalized with sucrose, enabling rapid and efficient foliar delivery into the plant phloem, a vascular tissue that transports sugars, signaling molecules, and agrochemicals through the whole plant. The chemical affinity of sucrose molecules to sugar membrane transporters on the phloem cells enhances the uptake of biocompatible carbon dots with β-cyclodextrin molecular baskets (suc-β-CD) that can carry a wide range of agrochemicals. The nanoparticle fluorescence emission properties allowed detection and monitoring of rapid translocation (<40 min) in the vasculature of wheat leaves by confocal and epifluorescence microscopy. The suc-β-CDs more than doubled the delivery of chemical cargoes into the leaf vascular tissue. The sucrose coating of nanoparticles approach enables unprecedented targeted delivery to roots with ≈70% of phloem-loaded nanoparticles delivered to roots.The use of plant biorecognition molecules mediated delivery provides an approach for guiding nanocarriers with their cargoes to plant organelles, cells and tissues and whole plants and engineering safer and efficient agrochemical and biomolecule delivery tools. Communication and actuation of plants mediated by nanosensors and targeted nanostructures can turn plants into technology and lead to a more precise and sustainable agriculture with reduced environmental impact.