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While the development of cities tends to focus on improving traffic mobility, it has gradually neglected people’s demand for safety and comfort walking on the streets. To address this problem, shared streets that can integrate traditional street life and traffic mobility are getting more attention as pedestrian-friendly development. In order to measure the performance of shared streets, it is essential to identify how people feel when driving and walking around. However, investigating the various factors that influence the real world is not straightforward because of cost, time-consuming, and safety problems. Virtual reality and the Human-in-the-loop (HITL) have become valuable tools for conducting experiments without compromising them. The experiments are performed on both pedestrians’ and drivers’ sides. The three shared street layouts in a virtual environment are designed according to Europe’s real shared street cases. To evaluate shared street effects, questions in five aspects: amenity, walking or driving experience, safety, economy or priority, and environmental perception are asked to participants, respectively. MPR, EWM, and Fuzzy Comprehension Evaluation methods are used to assess the performance. The result revealed that different groups of people have different sensitivity and preferences for each evaluation criteria. However, the results of the comprehensive evalutation showed that scenario C with the largest isolation measurement is preferable in both pedestrian and driver’s groups based on shared street design elements. The city planners can get help from this shared street analysis, where the new design and layout could be tested in advance.

While the development of cities tends to focus on improving traffic mobility, it has gradually neglected people's demand for safety and comfort walking on the streets. To address this problem, shared streets that can integrate traditional street life and traffic mobility are getting more attention as pedestrian-friendly development. In order to measure the performance of shared streets, it is essential to identify how people feel when driving and walking around. However, investigating the various factors that influence the real world is not straightforward because of cost, time-consuming, and safety problems. Virtual reality and the Human-in-the-loop (HITL) have become valuable tools for conducting experiments without compromising them. The experiments are performed on both pedestrians' and drivers' sides. The three shared street layouts in a virtual environment are designed according to Europe's real shared street cases. To evaluate shared street effects, questions in five aspects: amenity, walking or driving experience, safety, economy or priority, and environmental perception are asked to participants, respectively. MPR, EWM, and Fuzzy Comprehension Evaluation methods are used to assess the performance. The result revealed that different groups of people have different sensitivity and preferences for each evaluation criteria. However, the results of the comprehensive evalutation showed that scenario C with the largest isolation measurement is preferable in both pedestrian and driver's groups based on shared street design elements. The city planners can get help from this shared street analysis, where the new design and layout could be tested in advance.

Although dark tea is a unique microbial-fermented tea with a high reputation for having an antiobesity effect, little is known about the effect of microbial fermentation on tea leaves’ antiobesity properties. This study compared the antiobesity effects of microbial-fermented Qingzhuan tea (QZT) and unfermented Qingmao tea (QMT), providing insight into their underlying mechanisms associated with gut microbiota. Our results indicated that the supplementation of QMT extract (QMTe) and QZT extract (QZTe) displayed similar antiobesity effects in high-fat diet (HFD)-fed mice, but the hypolipidemic effect of QZTe was significantly stronger than that of QMTe. The microbiomic analysis indicated that QZTe was more effective than QMTe at regulating HFD-caused gut microbiota dysbiosis. Akkermansiaceae and Bifidobacteriaceae, which have negative correlations with obesity, were enhanced notably by QZTe, whereas Faecalibaculum and Erysipelotrichaceae, which are positively correlated with obesity, were decreased dramatically by QMTe and QZTe. A Tax4Fun analysis of QMTe/QZTe-mediated gut microbiota revealed that QMTe supplementation drastically reversed the HFD-induced upregulation of glycolysis and energy metabolism, whereas QZTe supplementation significantly restored the HFD-caused downregulation of pyruvate metabolism. Our findings suggested that microbial fermentation showed a limited effect on tea leaves’ antiobesity, but enhanced their hypolipidemic activity, and QZT could attenuate obesity and associated metabolic disorders by favorably modulating gut microbiota.

Many nitrogen-fixing bacteria can produce siderophores for iron acquisition in soil, but the impact of their siderophore-producing capabilities on the rhizosphere soil microecology is not well understood. To explore the effects of root inoculation with NFB strains with different siderophore-producing capabilities on the rhizosphere soil microecology and deeply evaluate the application value of a high-yielding siderophore strain in promoting crop growth, the wild-type nitrogen-fixing bacterial strain Kosakonia radicincitans GXGL-4A and its Tn5 mutants M107 (high siderophore-producing ability) and M246-2 (deficient in siderophore production) were used as biofertilizers in cucumber rhizosphere soil. Iron is important for the growth of bacterial cells, and the mutant M246-2 showed the slowest growth rate compared to the other strains when incubated in an A15 nitrogen-free medium supplied with different levels of iron. The mutant M107 had the strongest chelating ability for iron, with the largest yellow halo on the CAS detection plate. There were statistically significant differences in the halo diameters among the three NFB groups. Compared with the control group, the application of NFB significantly increased the activities of soil peroxidase and dehydrogenase and altered the soil nitrogen contents. Fertilization with the mutant M107 significantly improved the cucumber biomass and reduced the abundance and diversity of bacterial communities in the rhizosphere soil compared to the other groups. The contents of soil ammonium nitrogen and total nitrogen and soil dehydrogenase showed significant correlations with the abundance of the top 50 dominant genera in the soil. The soil TN content was the essential factor affecting the abundance of Kosakonia bacteria in the cucumber rhizosphere.

The application of nitrogen-fixing bacteria (NFB) as a biofertilizer can greatly reduce or even avoid environmental pollution caused by the excessive use of chemical nitrogen fertilizers. To explore the effect of short-term fertilization of GXGL-4A on the expression of functional genes in the roots of the cucumber (Cucumis sativus L.) cultivar “Xintaimici”, this study used transcriptome sequencing technology combined with fluorescent quantitative RT-PCR (qRT-PCR) verification to compare the gene transcription profiles of GXGL-4A-treated and control (sterile-water-treated) groups. A total of 418 differentially expressed genes (DEGs) were detected. The transcription levels of genes Csa5G161290 and Csa3G027720, which encode nitrate transporters, showed significant up-regulation (3.04- and 2.27-fold, respectively) in roots inoculated with GXGL-4A. The genes CsaV3_5G006200, encoding cytokinin dehydrogenase involved in the biosynthesis of zeatin, CsaV3_1G011730, encoding a wound-responsive protein, and CsaV3_6G015610, encoding a heat stress transcription factor, were significantly up-regulated at the transcriptional level (p < 0.05). However, the transcription of nitrogen cycling functional genes CsaV3_3G036500, CsaV3_1g008910, and CsaV3_3G018610, which encode nitrate reductase, high-affinity nitrate transporter (NRT), and ferredoxin-nitrite reductase, respectively, showed significant down-regulation (p < 0.05). Only the KEGG pathway of phenylpropanoid biosynthesis reached a significant level (p < 0.05). This study contributes to a deeper understanding of the interaction between NFB and plants and provides theoretical guidance for the development of GXGL-4A as a mature biological agent for sustainable agricultural production under drought stress.

Tomato leaf mold caused by Cladosporium fulvum ( C. fulvum ) is a serious fungal disease which results in huge yield losses in tomato cultivation worldwide. In our study, we discovered that ROS (reactive oxygen species) burst was triggered by C. fulvum treatment in tomato leaves. RNA-sequencing was used to identify differentially expressed genes (DEGs) induced by C. fulvum inoculation at the early stage of invasion in susceptible tomato plants. Gene ontology (GO) terms and Kyoto Encyclopedia of Genes and Genomes (KEGG) databases were used to annotate functions of DEGs in tomato plants. Based on our comparative analysis, DEGs related to plant-pathogen interaction pathway, plant hormone signal transduction pathway and the plant phenylpropanoid pathway were further analyzed. Our results discovered that a number of core defense genes against fungal invasion were induced and plant hormone signal transduction pathways were impacted by C. fulvum inoculation. Further, our results showed that SA (salicylic acid) and ABA (abscisic acid) contents were accumulated while JA (jasmonic acid) content decreased after C. fulvum inoculation in comparison with control, and quantitative real-time PCR to detect the relative expression of genes involved in SA, ABA and JA signaling pathway further confirmed our results. Together, results will contribute to understanding the mechanisms of C. fulvum and tomato interaction in future.

Monoterpenes are a class of volatile organic compounds that play crucial roles in imparting floral and fruity aromas to Muscat-type grapes. However, our understanding of the regulatory mechanisms underpinning monoterpene biosynthesis in grapes, particularly following abscisic acid (ABA) treatment, remains elusive. This study aimed to explore the impact of exogenous ABA on monoterpene biosynthesis in Ruiduhongyu grape berries by employing Headspace Solid-Phase Micro-Extraction Gas Chromatography–Mass Spectrometry (HS-SPME/GC–MS) analysis and transcriptome sequencing. The results suggested significant differences in total soluble solids (TSS), pH, and total acid content. ABA treatment resulted in a remarkable increase in endogenous ABA levels, with concentrations declining from veraison to ripening stages. ABA treatment notably enhanced monoterpene concentrations, particularly at the E_L37 and E_L38 stages, elevating the overall floral aroma of grape berries. According to the variable gene expression patterns across four developmental stages in response to ABA treatment, the E_L37 stage had the largest number of differential expressed genes (DEGs), which was correlated with a considerable change in free monoterpenes. Furthermore, functional annotation indicated that the DEGs were significantly enriched in primary and secondary metabolic pathways, underlining the relationship between ABA, sugar accumulation, and monoterpene biosynthesis. ABA treatment upregulated key genes involved in the methylerythritol phosphate (MEP) pathway, enhancing carbon allocation and subsequently impacting terpene synthesis. This study also identified transcription factors, including MYB and AP2/ERF families, potentially modulating monoterpene and aroma-related genes. Weighted gene co-expression network analysis (WGCNA) linked ABA-induced gene expression to monoterpene accumulation, highlighting specific modules enriched with genes associated with monoterpene biosynthesis; one of these modules (darkgreen) contained genes highly correlated with most monoterpenes, emphasizing the role of ABA in enhancing grape quality during berry maturation. Together, these findings provide valuable insights into the multifaceted effects of exogenous ABA on monoterpene compounds and grape berry flavor development, offering potential applications in viticulture and enology.

A total of 1467 mutants of the biocontrol bacterium Bacillus pumilus DX01 were obtained by Tn5 insertional mutagenesis and subjected to the determination of antagonistic capabilities. Compared with the wild-type strain DX01, the mutant M25 was identified to have the most significant reduction in antagonistic capability against the phytopathogen Bipolaris maydis and extracellular proteinase activity. The integration site of the exogenous T-DNA in the genome of mutant M25 was revealed in the coding region of malony CoA-ACP transacylase (MCAT) gene (mcat), which belongs to a polyketide synthase (PKS) gene cluster, DX01pks of B. pumilus DX01. Furthermore, the whole DX01pks gene cluster was cloned using Illumina Solexa sequencing technology, and it has a modular framework different from the other two gene clusters involved in polyketide synthesis in B. amyloliquefaciens FZB42 (pks1) and B. subtilis 168 (pksX). Finally, in order to gain more insights into the molecular mechanisms of biocontrol of B. pumilus DX01, the changes in the relative level of expression of total proteins between the original strain DX01 and the mutant M25 were detected by 2-DE-based proteomic analysis. A total of twenty differentially expressed proteins were identified upon the mcat gene transposition mutagenesis. Of these proteins, seven proteins were up-regulated in expression level and the other proteins were down-regulated. Taken together, the results in this study showed that Tn5 transposon mutagenesis of B. pumilus DX01 can lead to a significant change of antiphytopathogen ability, and the DX01pks gene cluster possibly play a potential role in the biocontrol processes of this bacterium.

The application of nitrogen-fixing bacteria (NFB) as a biofertilizer can greatly reduce or even avoid environmental pollution caused by the excessive use of chemical nitrogen fertilizers. To explore the effect of short-term fertilization of GXGL-4A on the expression of functional genes in the roots of the cucumber (Cucumis sativus L.) cultivar “Xintaimici”, this study used transcriptome sequencing technology combined with fluorescent quantitative RT-PCR (qRT-PCR) verification to compare the gene transcription profiles of GXGL-4A-treated and control (sterile-water-treated) groups. A total of 418 differentially expressed genes (DEGs) were detected. The transcription levels of genes Csa5G161290 and Csa3G027720, which encode nitrate transporters, showed significant up-regulation (3.04- and 2.27-fold, respectively) in roots inoculated with GXGL-4A. The genes CsaV3_5G006200, encoding cytokinin dehydrogenase involved in the biosynthesis of zeatin, CsaV3_1G011730, encoding a wound-responsive protein, and CsaV3_6G015610, encoding a heat stress transcription factor, were significantly up-regulated at the transcriptional level (p < 0.05). However, the transcription of nitrogen cycling functional genes CsaV3_3G036500, CsaV3_1g008910, and CsaV3_3G018610, which encode nitrate reductase, high-affinity nitrate transporter (NRT), and ferredoxin-nitrite reductase, respectively, showed significant down-regulation (p < 0.05). Only the KEGG pathway of phenylpropanoid biosynthesis reached a significant level (p < 0.05). This study contributes to a deeper understanding of the interaction between NFB and plants and provides theoretical guidance for the development of GXGL-4A as a mature biological agent for sustainable agricultural production under drought stress.