Yilin Zhang

Postharvest management of leafy vegetables requires refrigeration to control their rapid deterioration and loss, which accounts for ∼30% of total food waste. Here, silk microneedles with a length of 700 μm were used to deliver physiological doses of melatonin (approximately 22 μg) in the leafy vegetable Pak choy (Brassica rapa subsp. chinensis) and successfully extended the shelf life of the harvested crop by 4 and 10 days at room temperature (25 °C) and under refrigeration (4 °C), respectively, compared to nontreated control plants. The exogenous dose of melatonin did not alter the natural concentration of the hormone in the harvested plants. Transcriptome analysis showed that melatonin regulates senescence by modulating auxin synthesis, the antioxidant system, and chlorophyll degradation. Overall, silk microneedles can be used to precisely deliver in harvested products compounds that extend their shelf life outside the cold chain.

Biomaterials bridging the biotic–abiotic interface in plants offer the opportunity to precisely deliver agrochemicals and continuously monitor plant health, with the goals of increasing resilience to climate change, enhancing crop production and mitigating environmental impact. In this study we report the manipulation of silk fibroin assembly with inorganics nucleation at their phase front to nanomanufacture porous and hollow microneedles that can be interfaced with plants. Plant growth analysis and quantification of wounding gene expression show a non-significant systemic wounding response to the injection of silk microneedles in tomato plants. Microneedles with a hollow structure enable the systemic delivery of plant micronutrients to treat chlorosis in tomato plants and crop biofortification through transport of human micronutrients injected in the petiole and loaded into tomato fruits. Hollow microneedles also provide access to plant vasculature for sap sampling, enabling continuous monitoring and early detection of phytoaccumulation of environmental contaminants such as cadmium.

Plant genetic engineering methods are critical for food security and biofuel production and to enable molecular farming. Here, we elucidated how polymeric high aspect ratio nanocarriers can enable DNA delivery to Nicotiana benthamiana plants and transient expression. We demonstrated that a nanocarrier with 20 nm width, 80 nm length, and a polymer-to-DNA ratio of N/P = 3.0 afforded the most efficient DNA delivery and expression among the parameter space investigated. Additionally, we showed that polymer-DNA complexes with a moderate positive charge of ∼14 mV favored penetration through the cell wall and membranes with the assistance of cell wall degrading enzymes. Together, these results establish a narrow window of aspect ratios and charges of the nanocarrier-DNA complex that enables DNA delivery to plants using polymeric nanocarriers. This fundamental nanocarrier structure-function relationship informs the design of soft-material nanocarriers for nucleic acid delivery in plant cells to facilitate a wide range of plant biotechnology applications.

Emerging contaminants, including per- and polyfluoroalkyl substances and heavy metals, are threatening the health of humans and ecosystems. Their removal from the environment remains challenging. Here, we designed silk fibroin-cellulose nanocrystal (silk-CNC) nanofibrillar and nanoporous membranes for emerging contaminant remediation. The protein-polysaccharide nanofibrils were fabricated by templating the assembly of silk fibroin using CNCs. Silk fibroin polymorphic nature combined with surface charge modulation of CNCs produced cationic silk-CNC(+) and anionic silk-CNC(-) nanofibrils that can target a broad spectrum of contaminants. Silk-CNC(+) nanofibrils and membranes exhibited antimicrobial properties and captured both short-chain heptafluorobutyric acid, perfluorobutanesulfonic acid, and long-chain perfluorooctanoic acid by virtue of hydrophobic attraction from β-sheeted silk fibroin and electrostatic interactions with CNC(+). Silk-CNC(-) provided the opportunity to target cations such as heavy metal cocontaminants. The nanofabrication of biopolymer-based membranes combines high performance with environmentally benign and cost-effective removal of emerging contaminants for water purification, wastewater treatment, and remediation.

Delivery of proteins in plant cells can facilitate the design of desired functions by modulation of biological processes and plant traits but is currently limited by narrow host range, tissue damage, and poor scalability. Physical barriers in plants, including cell walls and membranes, limit protein delivery to desired plant tissues. Herein, a cationic high aspect ratio polymeric nanocarriers (PNCs) platform is developed to enable efficient protein delivery to plants. The cationic nature of PNCs binds proteins through electrostatic. The ability to precisely design PNCs’ size and aspect ratio allowed us to find a cutoff of ≈14 nm in the cell wall, below which cationic PNCs can autonomously overcome the barrier and carry their cargo into plant cells. To exploit these findings, a reduction‐oxidation sensitive green fluorescent protein (roGFP) is deployed as a stress sensor protein cargo in a model plant Nicotiana benthamiana and common crop plants, including tomato and maize. In vivo imaging of PNC‐roGFP enabled optical monitoring of plant response to wounding, biotic, and heat stressors. These results show that PNCs can be precisely designed below the size exclusion limit of cell walls to overcome current limitations in protein delivery to plants and facilitate species‐independent plant engineering.

An incomplete understanding of how agrochemical nanocarrier properties affect their uptake and translocation in plants limits their application for promoting sustainable agriculture. Herein, we investigated how the nanocarrier aspect ratio and charge affect uptake and translocation in monocot wheat (Triticum aestivum) and dicot tomato (Solanum lycopersicum) after foliar application. Leaf uptake and distribution to plant organs were quantified for polymer nanocarriers with the same diameter (∼10 nm) but different aspect ratios (low (L), medium (M), and high (H), 10-300 nm long) and charges (-50 to +15 mV). In tomato, anionic nanocarrier translocation (20.7 ± 6.7 wt %) was higher than for cationic nanocarriers (13.3 ± 4.1 wt %). In wheat, only anionic nanocarriers were transported (8.7 ± 3.8 wt %). Both low and high aspect ratio polymers translocated in tomato, but the longest nanocarrier did not translocate in wheat, suggesting a phloem transport size cutoff. Differences in translocation correlated with leaf uptake and interactions with mesophyll cells. The positive charge decreases nanocarrier penetration through the leaf epidermis and promotes uptake into mesophyll cells, decreasing apoplastic transport and phloem loading. These results suggest design parameters to provide agrochemical nanocarriers with rapid and complete leaf uptake and an ability to target agrochemicals to specific plant organs, with the potential to lower agrochemical use and the associated environmental impacts.

Anticipated increases in the frequency and intensity of extreme temperatures will damage crops. Methods that efficiently deliver stress-regulating agents to crops can mitigate these effects. Here, we describe high aspect ratio polymer bottlebrushes for temperature-controlled agent delivery in plants. The foliar-applied bottlebrush polymers had near complete uptake into the leaf and resided in both the apoplastic regions of the leaf mesophyll and in cells surrounding the vasculature. Elevated temperature enhanced the in vivo release of spermidine (a stress-regulating agent) from the bottlebrushes, promoting tomato plant (Solanum lycopersicum) photosynthesis under heat and light stress. The bottlebrushes continued to provide protection against heat stress for at least 15 days after foliar application, whereas free spermidine did not. About 30% of the ∼80 nm short and ∼300 nm long bottlebrushes entered the phloem and moved to other plant organs, enabling heat-activated release of plant protection agents in phloem. These results indicate the ability of the polymer bottlebrushes to release encapsulated stress relief agents when triggered by heat to provide long-term protection to plants and the potential to manage plant phloem pathogens. Overall, this temperature-responsive delivery platform provides a new tool for protecting plants against climate-induced damage and yield loss.

Plant abiotic stress induces reactive oxygen species (ROS) accumulation in leaves that can decrease photosynthetic performance and crop yield. Materials that scavenge ROS and simultaneously provide nutrients in vivo are needed to manage this stress. Here, we incorporated both ROS scavenging and ROS triggered agent release functionality into an ∼20 nm ROS responsive star polymer (RSP) poly(acrylic acid)-block-poly((2-(methylsulfinyl)ethyl acrylate)-co-(2-(methylthio)ethyl acrylate)) (PAA-b-P(MSEA-co-MTEA)) that alleviated plant stress by simultaneous ROS scavenging and nutrient agent release. Hyperspectral imaging indicates that all of the RSP penetrates through the tomato leaf epidermis, and 32.7% of the applied RSP associates with chloroplasts in mesophyll. RSP scavenged up to 10 μmol mg^–1 ROS in vitro and suppressed ROS in vivo in stressed tomato (Solanum lycopersicum) leaves. Reaction of the RSP with H₂O₂in vitro enhanced the release of nutrient agent (Mg²⁺) from star polymers. Foliar applied RSP increased photosynthesis in plants under heat and light stress compared to untreated controls, enhancing the carbon assimilation, quantum yield of CO₂ assimilation, Rubisco carboxylation rate, and photosystem II quantum yield. Mg loaded RSP improved photosynthesis in Mg deficient plants, mainly by promoting Rubisco activity. These results indicate the potential of ROS scavenging nanocarriers like RSP to alleviate abiotic stress in crop plants, allowing crop plants to be more resilient to heat stress, and potentially other climate change induced abiotic stressors.

Demand for rare earth elements (REEs) is increasing, and REE production from ores is energy-intensive. Recovering REEs from waste streams can provide a more sustainable approach to help meet REE demand but requires materials with high selectivity and capacity for REEs due to the low concentration of REEs and high competing ion concentrations. Here, we developed a phosphate polymer nanogel (PPN) to selectively recover REEs from low REE content waste streams, including leached fly ash. A high phosphorus content (16.2 wt % P as phosphate groups) in the PPN provides an abundance of coordination sites for REE binding. In model solutions, the distribution coefficient (K_d) for all REEs ranged from 1.3 × 10⁵ to 3.1 × 10⁵ mL g^–1 at pH = 7, and the sorption capacity (q_m) for Nd, Gd, and Ho were ∼300 mg g^–1. The PPN was selective toward REEs, outcompeting cations (Ca, Mg, Fe, Al) at up to 1000-fold excess concentration. The PPN had a K_d of ∼10⁵–10⁶ mL g^–1 for lanthanides in coal fly ash leachate (pH = 5), orders of magnitude higher than the K_d of major competing ions (∼10³–10⁴ mL g^–1). REEs were recovered from the PPN using 3.5% HNO₃, and the material remained effective over three sorption–elution cycles. The high REE capacity and selectivity and good durability in a real waste stream matrix suggest its potential to recover REEs from a broad range of secondary REE stocks.

Determination of how the properties of nanocarriers of agrochemicals affect their uptake and translocation in plants would enable more efficient agent delivery. Here, we synthesized star polymer nanocarriers poly(acrylic acid)-block-poly(2-(methylsulfinyl)ethyl acrylate) (PAA-b-PMSEA) and poly(acrylic acid)-block-poly((2-(methylsulfinyl)ethyl acrylate)-co-(2-(methylthio)ethyl acrylate)) (PAA-b-P(MSEA-co-MTEA)) with well-controlled sizes (from 6 to 35 nm), negative charge content (from 17% to 83% PAA), and hydrophobicity and quantified their leaf uptake, phloem loading, and distribution in tomato (Solanum lycopersicum) plants 3 days after foliar application of 20 μL of a 1g L^–1 star polymer solution. In spite of their property differences, ∼30% of the applied star polymers translocated to other plant organs, higher than uptake of conventional foliar applied agrochemicals (<5%). The property differences affected their distribution in the plant. The ∼6 nm star polymers exhibited 3 times higher transport to younger leaves than larger ones, while the ∼35 nm star polymer had over 2 times higher transport to roots than smaller ones, suggesting small star polymers favor symplastic unloading in young leaves, while larger polymers favor apoplastic unloading in roots. For the same sized star polymer, a smaller negative charge content (yielding ζ ∼ −12 mV) enhanced translocation to young leaves and roots, whereas a larger negative charge (ζ < −26 mV) had lower mobility. Hydrophobicity only affected leaf uptake pathways, but not translocation. This study can help design agrochemical nanocarriers for efficient foliar uptake and targeting to desired plant organs, which may decrease agrochemical use and environmental impacts of agriculture.