A robust high-throughput functional screening assay for plant pathogen effectors using the TMV-GFP vector.

Uncovering the function of phytopathogen effectors is crucial for understanding mechanisms of pathogen pathogenicity and for improving our ability to protect plants from diseases. An increasing number of effectors have been predicted in various plant pathogens. Functional characterization of these effectors has become a major focus in the study of plant-pathogen interactions. In this study, we designed a novel screening system that combines the TMV (tobacco mosaic virus)-GFP vector and Agrobacterium-mediated transient expression in the model plant Nicotiana benthamiana. This system enables the rapid identification of effectors that interfere with plant immunity. The biological function of these effectors can be easily evaluated by observing the GFP fluorescence signal using a UV lamp within just a few days. To evaluate the TMV-GFP system, we initially tested it with well-described virulence and avirulence type III effectors from the bacterial pathogen Ralstonia solanacearum. After proving the accuracy and efficiency of the TMV-GFP system, we successfully screened a novel virulence effector, RipS1, using this approach. Furthermore, using the TMV-GFP system, we reproduced consistent results with previously known cytoplasmic effectors from a diverse array of pathogens. Additionally, we demonstrated the effectiveness of the TMV-GFP system in identifying apoplastic effectors. The easy operation, time-saving nature, broad effectiveness, and low technical requirements of the TMV-GFP system make it a promising approach for high-throughput screening of effectors with immune interference activity from various pathogens.

Ultra-Stable Ti Vacancies-Pt Atomic Clusters Structure on Titanium Oxycarbide Supports for High Current Density Hydrogen Evolution Reaction.

Electrocatalysts with low Pt loading mass to achieve high current density (≥1 A cm-2) for hydrogen evolution reaction (HER) are still extremely challenging due to the limited intrinsic activity and weak stability of catalytic sites. The modulation of the electronic microenvironment of the support-Pt structure is crucial to enhance the intrinsic activity and stability of catalytic sites. Herein, an innovative titanium oxycarbide (TiVCO) solid solution with Ti vacancies (TiV) is proposed as support to anchor sub-nanoscale Pt atomic clusters (Pt ACs) and a stable "TiV-Pt ACs" structure is carefully designed. The electronic microenvironment of "TiV-Pt ACs" is indirectly optimized by an unsaturated C/O site near TiV. Thanks to this, novel "TiV-Pt ACs" structure (Pt@TiVCO) with low Pt loading mass (2.44 wt.%) exhibits excellent HER activity in acidic solution and the mass activity is more than ten times that of commercial 20% Pt/C at the overpotentials of 50 and 100 mV. Particularly, Pt@TiVCO shows amazing stability at high and fluctuating current density of 1-2 A cm-2 for 120 h. This work provides a novel and promising method to develop stable and low-loading Pt-based catalysts adapting to high current density.

Au-allenylidene promoted decarboxylative annulation to access unsaturated γ-lactams/lactones.

A novel Au-allenylidene promoted decarboxylative annulation by intramolecular α-nucleophilic addition has been disclosed. The unsaturated cyclic ethynylethylene carbamates/carbonates can be converted to unique nucleophiles attached with alkylidene ketenes by sequential decarboxylation and oxidation processes. Such alkylidene ketenes can be rapidly trapped by intramolecular α-attacking annulation to generate potential biological active unsaturated γ-lactams/lactones with broad scope, facile post-modification, high regioselectivity and efficiency.

Variable angle of insulated shotcrete under different loading rates and temperature and humidity cycles- shear test and analysis.

The new thermal insulating shotcrete is of great significance for the management of thermal damage in deep mines, and its own strength has a greater impact on the roadway insulation and safe production, so it is very necessary to study the shear strength of the new thermal insulating shotcrete under the influence of the deep hot and humid environment and the stress of mining. For the heat-insulating shotcrete, firstly, we carried out the concrete variable angle shear test under different loading rates, which concluded that the shear rate and peak shear stress showed a trend of increasing and then decreasing; as the angle increases, the different rates have a greater impact on the peak shear stress of the specimen. Secondly, the concrete variable angle shear test was carried out under the temperature and humidity cycle, which revealed that the shear strength of thermal insulated shotcrete increased firstly and then decreased with the increase of temperature at the same number of cycles. Finally, the empirical equations between the cohesive force c, the angle of internal friction ϕ and the number of warm and wet cycles n and the temperature of warm and wet cycles T are fitted with the MATLAB software respectively, and the research results provide technical references for the management of geothermal temperature in deep well projects.

Integrated driving risk surrogate model and car-following behavior for freeway risk assessment.

Drivers' risk perception plays a crucial role in understanding vehicle interactions and car-following behavior under complex conditions and physical appearances. Therefore, it is imperative to evaluate the variability of risks involved. With advancements in communication technology and computing power, real-time risk assessment has become feasible for enhancing traffic safety. In this study, a novel approach for evaluating driving interaction risk on freeways is presented. The approach involves the integration of an interaction risk perception model with car-following behavior. The proposed model, named the driving risk surrogate (DRS), is based on the potential field theory and incorporates a virtual energy attribute that considers vehicle size and velocity. Risk factors are quantified through sub-models, including an interactive vehicle risk surrogate, a restrictions risk surrogate, and a speed risk surrogate. The DRS model is applied to assess driving risk in a typical scenario on freeways, and car-following behavior. A sensitivity analysis is conducted on the effect of different parameters in the DRS on the stability of traffic dynamics in car-following behavior. This behavior is then calibrated using a naturalistic driving dataset, and then car-following predictions are made. It was found that the DRS-simulated car-following behavior has a more accurate trajectory prediction and velocity estimation than other car-following methods. The accuracy of the DRS risk assessments was verified by comparing its performance to that of traditional risk models, including TTC, DRAC, MTTC, and DRPFM, and the results show that the DRS model can more accurately estimate risk levels in free-flow and congested traffic states. Thus the proposed risk assessment model provides a better approach for describing vehicle interactions and behavior in the digital world for both researchers and practitioners.

Efficient Near-Infrared Fluorophores Based on Cyanostyrene Derivatives for Two-Photon Fluorescence Bioimaging.

Organic fluorescent materials with red/near-infrared (NIR) emission are highly promising for use in biotechnology due to their exceptional advantages. However, traditional red/NIR fluorophores often exhibit weak emission at high concentrations or in an aggregated state due to the aggregate-caused quenching effect, which severely limits their applicability in biological imaging. To address this challenge, we developed a series of cyanostyrene derivatives with aggregation-induced emission characteristics, including 2,3-Bis-(4-styryl-phenyl)-but-2-enedinitrile (DPB), 2,3-Bis-{4-[2-(4-methoxy- phenyl)-vinyl]-phenyl}-but-2-enedinitrile (DOB), 2,3-Bis-{4-[2-(4-diphenylamino- phenyl)-vinyl]-phenyl}-but-2-enedinitrile (DTB), and 2,3-Bis-[4-(2-{4-[phenyl- (4-triphenylvinyl-phenyl)-amino]-phenyl}-vinyl)- phenyl]-but-2-enedinitrile (DTTB). Notably, these compounds exhibited intense solid state fluorescence owing to AIE effect, especially DTTB shows NIR emission with high solid state quantum efficiency (712 nm, ΦF=14.2 %). Then we prepared DTTB@PS-PEG NPs nanoparticles by encapsulating DTTB with the amphiphilic polymer polystyrene-polyethylene glycol (PS-PEG). Importantly, DTTB@PS-PEG NPs exhibited highly efficient NIR luminescence (ΦF=28.7 %) and a large two-photon absorption cross-section (1900 GM) under 800 nm laser excitation. The bright two-photon fluorescence of DTTB@PS-PEG indicated that it can be a highly promising candidate for two-photon fluorescence probe. Therefore, this work provides valuable insights for the design of highly efficient and NIR-emitting two-photon fluorescent probes.