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  1. Abstract

    Current approaches for nanomaterial delivery in plants are unable to target specific subcellular compartments with high precision, limiting our ability to engineer plant function. We demonstrate a nanoscale platform that targets and delivers nanomaterials with biochemicals to plant photosynthetic organelles (chloroplasts) using a guiding peptide recognition motif. Quantum dot (QD) fluorescence emission in a low background window allows confocal microscopy imaging and quantitative detection by elemental analysis in plant cells and organelles. QD functionalization with β-cyclodextrin molecular baskets enables loading and delivery of diverse chemicals, and nanoparticle coating with a rationally designed and conserved guiding peptide targets their delivery to chloroplasts. Peptide biorecognition provides high delivery efficiency and specificity of QD with chemical cargoes to chloroplasts in plant cells in vivo (74.6 ± 10.8%) and more specific tunable changes of chloroplast redox function than chemicals alone. Targeted delivery of nanomaterials with chemical cargoes guided by biorecognition motifs has a broad range of nanotechnology applications in plant biology and bioengineering, nanoparticle-plant interactions, and nano-enabled agriculture.

     
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    Abstract Imaging of complex (biological) samples in the near-infrared (NIR) is beneficial due to reduced light scattering, absorption, phototoxicity, and autofluorescence. However, there are few NIR fluorescent materials known and suitable for biomedical applications. Here we exfoliate the layered pigment CaCuSi 4 O 10 (Egyptian Blue, EB) via ball milling and facile tip sonication into NIR fluorescent nanosheets (EB-NS). The size of EB-NS can be tailored to diameters <20 nm and heights down to 1 nm. EB-NS fluoresce at 910 nm and the fluorescence intensity correlates with the number of Cu 2+ ions. Furthermore, EB-NS display no bleaching and high brightness compared with other NIR fluorophores. The versatility of EB-NS is demonstrated by in-vivo single-particle tracking and microrheology measurements in Drosophila melanogaster embryos. EB-NS can be uptaken by plants and remotely detected in a low-cost stand-off detection setup. In summary, EB-NS have the potential for a wide range of bioimaging applications. 
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    Salinity is a widespread environmental stress that severely limits crop yield worldwide. Cerium oxide nanoparticles (nanoceria) have the unique capability of catalytically reducing levels of stress-induced reactive oxygen species (ROS) including hydroxyl radicals (˙OH) that lack enzymatic scavenging pathways. The underlying mechanisms of how nanoceria ROS scavenging augments plant tolerance to environmental stress are not well understood. Herein, we demonstrate that catalytic ˙OH scavenging by nanoceria in Arabidopsis thaliana leaves significantly improves mesophyll K + retention, a key trait associated with salinity stress tolerance. Leaves with mesophyll cells interfaced with 50 mg L −1 poly(acrylic acid) coated nanoceria (PNC) have significantly higher ( P < 0.05) carbon assimilation rates (85%), quantum efficiency of photosystem II (9%), and chlorophyll content (14%) compared to controls after being exposed to 100 mM NaCl for 3 days. PNC infiltrated leaves (PNC-leaves) under salinity stress exhibit lower ROS levels – including hydroxyl radical (41%) and its precursor hydrogen peroxide (44%) – and one fold higher ( P < 0.05) cytosolic K + dye intensity in leaf mesophyll cells relative to controls. Non-invasive microelectrode ion flux electrophysiological (MIFE) measurements indicated that PNC-leaves have about three-fold lower NaCl-induced K + efflux from leaf mesophyll cells compared to controls upon exposure to salinity stress. The ROS-activated nonselective cation channels (ROS-NSCC) in the plasma membrane of leaf mesophyll cells were identified as the main ˙OH-inducible K + efflux channels. Long term catalytic scavenging of ˙OH in leaves by PNC enhances plant photosynthetic performance under salinity stress by enabling plasma membrane channels/transporters to coordinately retain higher levels of K + in the leaf mesophyll cell cytosol. PNC augmented plant ROS scavenging provides a key tool for understanding and improving plant tolerance against abiotic stresses such as salinity. 
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