An upstream signaling gene calmodulin regulates the synthesis of insect wax via activating fatty acid biosynthesis pathway.

Insect wax accumulates on the surface of insect cuticle, which acts as an important protective barrier against rain, ultraviolet light radiation, pathogens, etc. The waxing behavior, wax composition and molecular mechanism underling wax biosynthesis are unclear in dustywings. Herein, the current study determined the vital developmental stage for waxing behavior in dustywings, examined the components of waxy secretions, and identified key regulatory genes for wax biosynthesis. The wax glands were mainly located on the thorax and abdomen of dustywing adults. The adults spread the waxy secretions over their entire body surface. The metabolomics analysis identified 32 lipids and lipid-like molecules, 15 organic acids and derivatives, 7 benzenoids, etc. as the main components of waxy secretions. The fatty acids represented the largest proportion of the category of lipid and lipid-like molecules. The conjoint analysis of metabolomics and transcriptomics identified two crucial genes fatty acyl-CoA reductase (CsFAR) and calmodulin (CsCaM) for wax biosynthesis. The down-regulation of these genes via nanocarrier-mediated RNA interference technology significantly reduced the amount of wax particles. Notably, the RNAi of CsCaM apparently suppressed the expression of most genes in fatty acid biosynthesis pathway, indicating the CsCaM might act as a main upstream regulator of fatty acid biosynthesis pathway.

Perfect cooperative pest control via nano-pesticide and natural predator: High predation selectivity and negligible toxicity toward predatory stinkbug.

The improper use of synthetic pesticides has caused adverse effects on global ecosystems and human health. As a part of sustainable pest management strategy, natural predators, along with nano-pesticides, have made significant contributions to ecological agriculture. The cooperative application of both approaches may overcome their limitations, substantially reducing pesticide application while controlling insect pests efficiently. Herein, the current study introduced a cationic star polymer (SPc) to prepare two types of nano-pesticides, which were co-applied with predatory stinkbugs Picromerus lewisi to achieve perfect cooperative pest control. The SPc exhibited nearly no toxicity against predatory stinkbugs at the working concentration, but it led to the death of predatory stinkbugs at extremely high concentration with the lethal concentration 50 (LC50) value of 13.57 mg/mL through oral feeding method. RNA-seq analysis revealed that the oral feeding of SPc could induce obvious stress responses, leading to stronger phagocytosis, exocytosis, and energy synthesis to ultimately result in the death of predatory stinkbugs. Then, the broflanilide and chlorobenzuron were employed to prepare the self-assembled nano-pesticides via hydrogen bond and Van der Waals force, and the complexation with SPc broke the self-aggregated structures of pesticides and reduced their particle sizes down to nanoscale. The bioactivities of prepared nano-pesticides were significantly improved toward common cutworm Spodoptera litura with the corrected mortality increase by approximately 30%. Importantly, predatory stinkbugs exhibited a strong predation selectivity for alive common cutworms to reduce the exposure risk of nano-pesticides, and the nano-pesticides showed negligible toxicity against predators. Thus, the nano-pesticides and predatory stinkbugs could be applied simultaneously for efficient and sustainable pest management. The current study provides an excellent precedent for perfect cooperative pest control via nano-pesticide and natural predator.

Self-assembled co-delivery nanoplatform for increasing the broad-spectrum susceptibility of fall armyworm toward insecticides.

INTRODUCTION: Unscientific application of insecticides has led to severe resistance of pests to almost all classes of insecticides. Enhanced detoxification is the most common mechanism for this kind of resistance. OBJECT: Fall armyworm (FAW) has developed insecticide resistance, which is often linked to the overexpression of detoxification genes. Herein, a multicomponent nano-pesticide is designed to increase its broad-spectrum susceptibility toward insecticides. METHOD: Regulatory function of nuclear factor erythroid 2-related factor 2 (Nrf2) in detoxification was confirmed using transcriptome sequencing, quantitative real-time PCR and enzyme activity measurement. A star polycation (SPc) was adopted to construct the pesticide/SPc/complex, whose self-assembly mechanism and characterization were examined using isothermal titration calorimetry, dynamic light scattering and transmission electron microscope. The delivery efficiency of SPc-loaded dsRNA was examined in vitro and in vivo using fluorescent tracer technique. A multicomponent nano-pesticide was created through the integration of bacterial expression system and nano-delivery system, and its bioactivity was tested in laboratory and field. RESULTS: We confirmed the crucial role of Nrf2 in regulating the detoxification in FAW, and silencing Nrf2 could decrease detoxification gene expression and increase insecticide susceptibility. We then applied the SPc to self-assemble a nanoplatform for delivering Nrf2 double-stranded RNA (dsRNA) and pesticide simultaneously. Nano-sized pesticide/SPc/dsRNA complex exhibited high delivery efficiency in vitro and in vivo. Excitingly, the insecticidal activities of pesticide/SPc/dsNrf2 complexes were remarkably improved with the normalized synergistic ratios of 5.43-6.25 for chlorantraniliprole, 4.45-15.00 for emamectin benzoate, and 6.75-15.00 for spinetoram. Finally, we developed a multicomponent nano-pesticide (pesticide/SPc/dsNrf2 complex) using a bacterial expression system and nano-delivery system. This approach exhibited excellent leaf protection and pest control efficacy. CONCLUSION: The integration between the pesticide nanometerization and insecticide susceptibility improvement offers a promising strategy to increase insecticidal activity. Our study provides a revolutionary and universal strategy to increase insecticidal activity and decease application doses.

Shedding light on cellular dynamics: the progress in developing photoactivated fluorophores.

Photoactivated fluorophores (PAFs) are highly effective imaging tools that exhibit a removal of caging groups upon light excitation, resulting in the restoration of their bright fluorescence. This unique property allows for precise control over the spatiotemporal aspects of small molecule substances, making them indispensable for studying protein labeling and small molecule signaling within live cells. In this comprehensive review, we explore the historical background of this field and emphasize recent advancements based on various reaction mechanisms. Additionally, we discuss the structures and applications of the PAFs. We firmly believe that the development of more novel PAFs will provide powerful tools to dynamically investigate cells and expand the applications of these techniques into new domains.

Self-Assembled Nanonematicide Induces Adverse Effects on Oxidative Stress, Succinate Dehydrogenase Activity, and ATP Generation.

Long-term overuse of chemical nematicides has resulted in low control efficacy toward destructive root-knot nematodes, and continuous development in nanotechnology is supposed to enhance the utilization efficiency of nematicides to meet practical needs. Herein, a cationic star polymer (SPc) was constructed to load fluopyram (flu) and prepare a flu nanoagent. Hydrogen bonding and van der Waals forces facilitated the self-assembly of the flu nanoagent, leading to the breakdown of self-aggregated flu and reducing its particle size to 60 nm. The bioactivity of flu was remarkably improved, with the half lethal concentration 50 from 8.63 to 5.70 mg/L due to the help of SPc. Transcriptome analysis found that a large number of transport-related genes were upregulated in flu nanoagent-exposed nematodes, while the expression of many energy-related genes was disturbed, suggesting that the enhanced uptake of flu nanoagents by nematodes might lead to the disturbance of energy synthesis and metabolism. Subsequent experiments confirmed that exposure to flu nanoagents markedly increased the reactive oxygen species (ROS) level of nematodes. Compared to flu treatment alone, succinate dehydrogenase (SDH) activity was inhibited in flu nanoagent-exposed nematodes with an increase in the pIC50 from 8.81 to 11.04, which further interfered with adenosine triphosphate (ATP) biosynthesis. Furthermore, the persistence of SPc-loaded flu in soil was prolonged by 2.33 times at 50 days after application. The protective effects of flu nanoagents on eggplant seedlings were significantly improved in both greenhouse and field trials, and the root-knot number was consistently smaller in roots treated with flu nanoagents than in those treated with flu alone. Overall, this study successfully constructed a self-assembled flu nanoagent with amplified effects on oxidative stress, SDH activity, and ATP generation, leading to highly effective control of root-knot nematodes in the field.

Self-assembled thiophanate-methyl/star polycation complex prevents plant cell-wall penetration and fungal carbon utilization during cotton infection by Verticillium dahliae.

No effective fungicides are available for the management of Verticillium dahliae, which causes vascular wilt disease. In this study, a star polycation (SPc)-based nanodelivery system was used for the first time to develop a thiophanate-methyl (TM) nanoagent for the management of V. dahliae. SPc spontaneously assembled with TM through hydrogen bonding and Van der Waals forces to decrease the particle size of TM from 834 to 86 nm. Compared to TM alone, the SPc-loaded TM further reduced the colony diameter of V. dahliae to 1.12 and 0.64 cm, and the spore number to 1.13 × 108 and 0.72 × 108 cfu/mL at the concentrations of 3.77 and 4.71 mg/L, respectively. The TM nanoagents disturbed the expression of various crucial genes in V. dahliae, and contributed to preventing plant cell-wall degradation and carbon utilization by V. dahliae, which mainly impaired the infective interaction between pathogens and plants. TM nanoagents remarkably decreased the plant disease index and the fungal biomass in the root compared to TM alone, and its control efficacy was the best (61.20 %) among the various formulations tested in the field. Furthermore, SPc showed negligible acute toxicity toward cotton seeds. To the best of our knowledge, this study is the first to design a self-assembled nanofungicide that efficiently inhibits V. dahliae growth and protects cotton from the destructive Verticillium wilt.

A nano-delivery system expands the insecticidal target of thiamethoxam to include a devastating pest, the fall armyworm.

Nano-delivery systems have been applied to deliver various synthetic/botanical pesticides to increase the efficiency of pesticide use and reduce the volumes of pesticides applied. Previous studies have supported the hypothesis that the nanocarriers can help expand the insecticidal target of pesticides to include non-target pests. However, the potential mechanism underlying this interesting phenomenon remains unclear. Herein, a widely applied star polycation (SPc) nanocarrier was synthesized to construct a thiamethoxam (TMX) nano-delivery system. The SPc-based delivery system could promote the translocation of exogenous substances across the membrane of Sf9 cells, increase the cytotoxicity of TMX against Sf9 cells by nearly 20%, and expand the insecticidal target of TMX to include Spodoptera frugiperda (the fall armyworm), with a 27.5% mortality increase at a concentration of 0.25 mg/mL. Moreover, the RNA-seq analysis demonstrated that the SPc could upregulate various transport-related genes, such as Rab, SORT1, CYTH, and PIKfyve, for the enhanced cellular uptake of TMX. Furthermore, enhanced cell death in larvae treated with the TMX-SPc complex was observed through changes in the expression levels of death-related genes, such as Casp7, BIRC5, MSK1, and PGAM5. The SPc-based nano-delivery system improved the cellular uptake of TMX and expanded its insecticidal target by adjusting the expression levels of death-related genes. The current study mainly identified the transport and cell death genes related to nanocarrier-based insecticidal target expansion, which is beneficial for understanding the bioactivity enhancement of the nano-delivery system.

Efficient polymer-mediated delivery system for thiocyclam: Nanometerization remarkably improves the bioactivity toward green peach aphids.

The unscientific application of synthetic pesticides has brought various negative effects on the environment, hindering the sustainable development of agriculture. Nanoparticles can be applied as carriers to improve pesticide delivery, showing great potential in the development of pesticide formulation in recent years. Herein, a star polymer (SPc) was constructed as an efficient pesticide nanocarrier/adjuvant that could spontaneously assemble with thiocyclam or monosultap into a complex, through hydrophobic association and hydrogen bonding, respectively, with the pesticide-loading contents of 42.54% and 19.3%. This complexation reduced the particle sizes of thiocyclam from 543.54 to 52.74 nm for pure thiocyclam, and 3 814.16 to 1 185.89 nm for commercial preparation (cp) of thiocyclam. Interestingly, the introduction of SPc decreased the contact angles of both pure and cp thiocyclam on plant leaves, and increased the plant uptake of cp thiocyclam to 2.4-1.9 times of that without SPc. Meanwhile, the SPc could promote the bioactivity of pure/cp thiocyclam against green peach aphids through leaf dipping method and root application. For leaf dipping method, the 50% lethal concentration decreased from 0.532 to 0.221 g/L after the complexation of pure thiocyclam with SPc, and that decreased from 0.390 to 0.251 g/L for cp thiocyclam. SPc seems a promising adjuvant for nanometerization of both pure and cp insecticides, which is beneficial for improving the delivery efficiency and utilization rate of pesticides.

Calcium nutrition nanoagent rescues tomatoes from mosaic virus disease by accelerating calcium transport and activating antiviral immunity.

As an essential structural, metabolic and signaling element, calcium shows low remobilization from old to young tissues in plants, restricting the nutrient-use efficiency and control efficacy against mosaic virus disease. Nanotechnology has been applied to prevent/minimize nutrient losses and improve the accessibility of poorly-available nutrients. Herein, the current study applied a star polycation (SPc) to prepare a calcium nutrition nanoagent. The SPc could assemble with calcium glycinate through hydrogen bond and Van der Waals force, forming stable spherical particles with nanoscale size (17.72 nm). Transcriptomic results revealed that the calcium glycinate/SPc complex could activate the expression of many transport-related genes and disease resistance genes in tomatoes, suggesting the enhanced transport and antiviral immunity of SPc-loaded calcium glycinate. Reasonably, the calcium transport was accelerated by 3.17 times into tomato leaves with the help of SPc, and the protective effect of calcium glycinate was remarkably improved to 77.40% and 67.31% toward tomato mosaic virus with the help of SPc after the third and fifth applications. Furthermore, SPc-loaded calcium glycinate could be applied to increase the leaf photosynthetic rate and control the unusual fast growth of tomatoes. The current study is the first success to apply nano-delivery system for enhanced calcium transport and antiviral immunity, which is beneficial for increasing nutrient-use efficiency and shows good prospects for field application.

A Preparation Method of Nano-Pesticide Improves the Selective Toxicity toward Natural Enemies.

Various nano-delivery systems have been designed to deliver synthetic/botanical pesticides for improved bioactivity. However, the enhanced toxicity of nanocarrier-loaded pesticides may injure the natural enemies, and their selective toxicity should be evaluated before the large-scale application. In this context, a star polymer (SPc)-based cyantraniliprole (CNAP) nano-delivery system was constructed, and its selective toxicity was evaluated using pest Frankliniella occidentalis (WFT) and predator Orius sauteri. The amide NH of CNAP could assemble with carbonyl groups or tertiary amines of SPc through hydrogen bonds to form CNAP/SPc complex spontaneously. The above self-assembly decreased the particle size of CNAP from 808 to 299 nm. With the help of SPc, the lethal concentration 50 (LC50) values of CNAP decreased from 99 to 54 mg/L and 230 to 173 mg/L toward WFTs and O. sauteri due to the enhancement of broad-spectrum bioactivity. Interestingly, the toxicity selective ratio (TSR) of CNAP increased from 2.33 to 3.23 with the help of SPc, revealing the higher selectivity of SPc-loaded CNAP. To our knowledge, it was the first successful exploration of the selective toxicity of nanocarrier-loaded pesticides, and the higher selective toxicity of SPc-loaded CNAP was beneficial for alleviating the negative impacts on predators.

Nanocarrier-Loaded Imidaclothiz Promotes Plant Uptake and Decreases Pesticide Residue.

There is a great demand for improving the effective utilization of pesticides and reducing their application for sustainable agriculture, and polymeric nanoparticles have provided strong technical support for the efficient delivery of pesticides. To this context, we tried to construct a relatively safe imidaclothiz nano-delivery system for enhanced plant uptake, reduced pesticide residue and improved bioactivity toward green peach aphids. The imidaclothiz could be assembled with the hydrophobic core of SPc through hydrophobic association, which led to the self-assembly of nanoscale imidaclothiz/SPc complex consisting of nearly spherical particles. The SPc decreased the contact angle of imidaclothiz drops and remarkably increased the plant uptake. Furthermore, the bioactivity and control efficacy of imidaclothiz were significantly improved with the help of SPc in both laboratory and field. Excitingly, the residue of imidaclothiz decreased with the help of SPc 7 d after the treatment due to the faster degradation of nanoscale imidaclothiz/SPc complex, which exhibited no negative effects on agronomic traits of tobacco plants. The current study successfully constructed a nano-delivery system for imidaclothiz, which can not only increase the effective utilization of pesticides, but also decrease the pesticide residue.

A Star Polyamine-Based Nanocarrier Delivery System for Enhanced Avermectin Contact and Stomach Toxicity against Green Peach Aphids.

The unscientific application of synthesized/botanical pesticides has not only brought the resistance of plant pathogens and pests, but also led to serious environmental pollution. In recent years, various nano-delivery systems have been used for the development of environmental-friendly pesticides with improved efficacy. Herein, the current study constructed an efficient avermectin B1a (AVM) nano-delivery system based on a star polyamine (SPc) and focused on the characterization and bioactivity of SPc-loaded AVM at various mass ratios. The hydroxyl groups of AVM could assemble with carbonyl groups of SPc through hydrogen bond and van der Waals forces, and the self-assembly of AVM and SPc formed nearly spherical particles of AVM/SPc complex with nanoscale size. The contact angle of SPc-loaded AVM decreased with the increasing mass ratio of SPc, revealing the easier distribution and spreading of the AVM/SPc complex. Furthermore, the stomach and contact toxicity of AVM/SPc complex also increased along with the increasing SPc mass ratio, which could be attributed to the enhanced systemic transportation in plants, enlarged contact area to insect pests and stronger permeability across the insect cuticle. The current study provides an efficient nano-delivery system for increasing stomach and contact toxicity of pesticides with wide applications in the agricultural field.

Shape Prior Guided Instance Disparity Estimation for 3D Object Detection.

In this paper, we propose a novel system named Disp R-CNN for 3D object detection from stereo images. Many recent works solve this problem by first recovering point clouds with disparity estimation and then apply a 3D detector. The disparity map is computed for the entire image, which is costly and fails to leverage category-specific prior. In contrast, we design an instance disparity estimation network (iDispNet) that predicts disparity only for pixels on objects of interest and learns a category-specific shape prior for more accurate disparity estimation. To address the challenge from scarcity of disparity annotation in training, we propose to use a statistical shape model to generate dense disparity pseudo-ground-truth without the need of LiDAR point clouds, which makes our system more widely applicable. Experiments on the KITTI dataset show that, when LiDAR ground-truth is not used at training time, Disp R-CNN outperforms previous state-of-the-art methods based on stereo input by 20 percent in terms of average precision for all categories. The code and pseudo-ground-truth data are available at the project page: https://github.com/zju3dv/disprcnn.

MonoEF: Extrinsic Parameter Free Monocular 3D Object Detection.

Monocular 3D object detection is an important task in autonomous driving. It can be easily intractable where there exists ego-car pose change w.r.t. ground plane. This is common due to the slight fluctuation of road smoothness and slope. Due to the lack of insight in industrial application, existing methods on open datasets neglect the camera pose information, which inevitably results in the detector being susceptible to camera extrinsic parameters. The perturbation of objects is very popular in most autonomous driving cases for industrial products. To this end, we propose a novel method to capture camera pose to formulate the detector free from extrinsic perturbation. Specifically, the proposed framework predicts camera extrinsic parameters by detecting vanishing point and horizon change. A converter is designed to rectify perturbative features in the latent space. By doing so, our 3D detector works independent of the extrinsic parameter variations and produces accurate results in realistic cases, e.g., potholed and uneven roads, where almost all existing monocular detectors fail to handle. Experiments demonstrate our method yields the best performance compared with the other state-of-the-arts by a large margin on both KITTI 3D and nuScenes datasets.

Simple Osthole/Nanocarrier Pesticide Efficiently Controls Both Pests and Diseases Fulfilling the Need of Green Production of Strawberry.

The application of botanical pesticides is a good choice in organic agriculture. However, most botanical pesticides have limitations of slow action and short persistence for pest and disease management, which constrain their further application. With the objective of exploring a green pesticide for controlling strawberry pests and diseases simultaneously, a star polymer (SPc) with a low production cost was synthesized as a pesticide nanocarrier through simple reactions. The SPc complexed with osthole quickly through electrostatic interaction and hydrophobic association, which decreased the particle size of osthole down to the nanoscale (17.66 nm). With the help of SPc, more nano-sized osthole was delivered into cytoplasm through endocytosis, leading to the enhanced cytotoxicity against insect cells. As a green botanical pesticide, the control efficacy of the osthole/SPc complex was improved against main strawberry pests (green peach aphid and two-spotted spider mite) and disease (powdery mildew), which fulfilled the need of both pest and disease management in sustainable production of strawberry. Meanwhile, the introduction of SPc not only improved plant-uptake but also decreased the residue of osthole due to the higher degradation rate. Furthermore, the application of the osthole/SPc complex exhibited no influence on the strawberry fruit quality and nontarget predators. To our knowledge, it is the first success to control plant pests and diseases simultaneously for sustainable agriculture by only one pesticidal formulation based on nanoparticle-delivered botanical pesticides.