The Vitis yeshanensis U-box E3 ubiquitin ligase VyPUB21 enhances resistance to powdery mildew by targeting degradation of NIM1-interacting (NIMIN) protein.

KEY MESSAGE: VyPUB21 plays a key role during the defense against powdery mildew in grapes. Ubiquitin-ligating enzyme (E3), a type of protein widely found in plants, plays a key role in their resistance to disease. Yet how E3 participates in the disease-resistant response of Chinese wild grapevine (Vitis yeshanensis) remains unclear. Here we isolated and identified a U-box type E3 ubiquitin ligase, VyPUB21, from V. yeshanensis. This gene's expression level rose rapidly after induction by exogenous salicylic acid (SA), jasmonic acid (JA), and ethylene (ETH) and powdery mildew. In vitro ubiquitination assay results revealed VyPUB21 could produce ubiquitination bands after co-incubation with ubiquitin, ubiquitin-activating enzyme (E1), and ubiquitin-conjugating enzyme (E2); further, mutation of the conserved amino acid site in the U-box can inhibit the ubiquitination. Transgenic VyPUB21 Arabidopsis had low susceptibility to powdery mildew, and significantly fewer conidiophores and spores on its leaves. Expression levels of disease resistance-related genes were also augmented in transgenic Arabidopsis, and its SA concentration also significantly increased. VyPUB21 interacts with VyNIMIN and targets VyNIMIN protein hydrolysis through the 26S proteasome system. Thus, the repressive effect of the NIMIN-NPR complex on the late systemic acquired resistance (SAR) gene was attenuated, resulting in enhanced resistance to powdery mildew. These results indicate that VyPUB21 encoding ubiquitin ligase U-box E3 activates the SA signaling pathway, and VyPUB21 promotes the expression of late SAR gene by degrading the important protein VyNIMIN of SA signaling pathway, thus enhancing grape resistance to powdery mildew.

Transcription factor VviWOX13C regulates fruit set by directly activating VviEXPA37/38/39 in grape (Vitis vinifera L).

KEY MESSAGE: VviWOX13C plays a key regulatory role in the expansin during fruit set. Expansins as a type of non-enzymatic cell wall proteins, are responsible for the loosening and extension in cell walls leading to the enlargement of the plant cells. However, the current studies are still lacking in expansin genes associated with promoting fruit set. Here, 29 members of the expansin gene family were identified in the whole genome of grapes (Vitis vinifera L.), and the functional prediction of expansins was based on the gene annotated information. Results showed that the 29 members of grape expansin gene family could be mainly divided into four subfamilies (EXPA, EXPB, LIKE A, and LIKE B), distributed on 16 chromosomes. Replication analysis showed that there were four segmental duplications and two tandem duplications. Each expansins sequence contained two conserved domain features of grape EXPs (DPBB_1 and Expansin_C) through protein sequence analysis. The transcriptome sequencing results revealed that VviEXPA37, VviEXPA38, and VviEXPA39 were induced and upregulated by CPPU. Furthermore, transcriptional regulatory prediction network indicated that VviWOX13C targeted regulates VviEXPA37, VviEXPA38, and VviEXPA39 simultaneously. EMSA and dual luciferase assays demonstrated that VviWOX13C directly activated the expression of VviEXPA37, VviEXPA38, and VviEXPA39 by directly binding to its promoter. These results provide a basis for further studies on the function and regulatory mechanisms of expansin genes in fruit set.

Inter-Frame-Relationship Protected Signal: A New Design for Radio Frequency Fingerprint Authentication.

Utilizing a multi-frame signal (MFS) rather than a single-frame signal (SFS) for radio frequency fingerprint authentication (RFFA) shows the advantage of higher accuracy. However, previous studies have often overlooked the associated security threats in MFS-based RFFA. In this paper, we focus on the carrier-sense multiple access with collision avoidance channel and identify a potential security threat, in that an attacker may inject a forged frame into valid traffic, making it more likely to be accepted alongside legitimate frames. To counter such a security threat, we propose an innovative design called the inter-frame-relationship protected signal (IfrPS), which enables the receiver to determine whether two consecutively received frames originate from the same transmitter to safeguard the MFS-based RFFA. To demonstrate the applicability of our proposition, we analyze and numerically evaluate two important properties: its impact on message demodulation and the accuracy gain in IfrPS-aided, MFS-based RFFA compared with the SFS-based RFFA. Our results show that the proposed scheme has a minimal impact of only -0.5 dB on message demodulation, while achieving up to 5 dB gain for RFFA accuracy.

Fe3O4 quantum dots mediated P-g-C3N4/BiOI as an efficient and recyclable Z-scheme photo-Fenton catalyst for tetracycline degradation and bacterial inactivation.

Photo-Fenton technology integrated by photocatalysis and Fenton reaction is a favorable strategy for water remediation. Nevertheless, the development of visible-light-assisted efficient and recyclable photo-Fenton catalysts still faces challenges. This study successfully constructed a novel separable Z-scheme P-g-C3N4/Fe3O4QDs/BiOI (PCN/FOQDs/BOI) heterojunction via in-situ deposition method. The results showed that the photo-Fenton degradation efficiency for tetracycline over optimal ternary catalyst reached 96.5% within 40 min at visible illumination, which was 7.1 and 9.6 times higher than its single photocatalysis and Fenton system, respectively. Moreover, PCN/FOQDs/BOI possessed excellent photo-Fenton antibacterial activity, which could completely inactivate 108 CFU·mL-1 of E. coli and S. aureus within 20 and 40 min, respectively. Theoretical calculation and in-situ characterization revealed that the enhanced catalysis behavior resulted from the FOQDs mediated Z-scheme electronic system, which not only facilitated photocreated carrier separation of PCN and BOI while maintaining maximum redox capacity, but also accelerated H2O2 activation and Fe3+/Fe2+ cycle, thus synergistically forming more active species in system. Additionally, PCN/FOQDs/BOI/Vis/H2O2 system displayed extensive adaptability at pH range of 3-11, removal universality for various organic pollutants and attractive magnetic separation property. This work would provide an inspiration for design of efficient and multifunctional Z-scheme photo-Fenton catalyst in water purification.

Genome-wide identification and expression analysis of JmjC domain-containing genes in grape under MTA treatment.

Histone demethylases containing the JmjC domain play an extremely important role in maintaining the homeostasis of histone methylation and are closely related to plant growth and development. Currently, the JmjC domain-containing proteins have been reported in many species; however, they have not been systematically studied in grapes. In this paper, 21 VviJMJ gene family members were identified from the whole grape genome, and the VviJMJ genes were classified into five subfamilies: KDM3, KDM4, KDM5, JMJD6, and JMJ-only based on the phylogenetic relationship and structural features of Arabidopsis and grape. After that, the conserved sites of VviJMJ genes were revealed by protein sequence analysis. In addition, chromosomal localization and gene structure analysis revealed the heterogeneous distribution of VviJMJ genes on grape chromosomes and the structural features of VviJMJ genes, respectively. Analysis of promoter cis-acting elements demonstrated numerous hormone, light, and stress response elements in the promoter region of the VviJMJ genes. Subsequently, the grape fruit was treated with MTA (an H3K4 methylation inhibitor), which significantly resulted in the early ripening of grape fruits. The qRT-PCR analysis showed that VviJMJ genes (except VviJMJ13c) had different expression patterns during grape fruit development. The expression of VviJMJ genes in the treatment group was significantly higher than that in the control group. The results indicate that VviJMJ genes are closely related to grape fruit ripening.

Overexpression of VvPPR1, a DYW-type PPR protein in grape, affects the phenotype of Arabidopsis thaliana leaves.

Pentatricopeptide repeat (PPR) proteins play important roles in plant growth and development. However, little is known about their functions in the leaf morphogenesis of Jingxiu grape (Vitis vinifera L.). Here, we explored the function of VvPPR1, which encodes a DYW-type PPR protein in grape. We showed that VvPPR1 is involved in the regulation of leaf rolling, anthocyanin accumulation, and trichome formation in Arabidopsis thaliana. Analysis of structural characteristics showed that VvPPR1 is a DYW-type PPR gene in the PLS subfamily consisting of 15 PPR motifs. The N-terminal had a targeted chloroplast site, and the C-terminal had a DYW domain. Quantitative PCR analysis revealed that the expression level of VvPPR1 was highest in grape leaves. Subcellular localization revealed that VvPPR1 is localized in the cytoplasm and chloroplast. VvPPR1-overexpressing plants had rolled leaves, high degrees of anthocyanin accumulation, and longer trichomes. The expression levels of genes related to these phenotypes were either significantly up-regulated or down-regulated. These results demonstrate that VvPPR1 is involved in leaf rolling, anthocyanin accumulation, and trichome formation in Arabidopsis; more generally, our findings indicate that VvPPR1 could be a target for improving the cultivation of horticultural crops.

Resistance risk induced by quorum sensing inhibitors and their combined use with antibiotics: Mechanism and its relationship with toxicity.

The abuse of antibiotics has brought out serious bacterial resistance, which threatens the ecological environment and human health. Quorum sensing inhibitors (QSIs), as a new kind of potential antibiotic substitutes that are theoretically difficult to trigger bacterial resistance, are recommended to individually use or jointly use with traditional antibiotics. However, there are few studies on the resistance risk in the use of QSIs. In this study, the influence of QSIs alone or in combination with sulfonamides (SAs) on conjugative transfer and mutation of Escherichia coli (E. coli) was investigated to explore whether QSIs have the potential to induce bacterial resistance. The results show that QSIs may facilitate plasmid RP4 conjugative transfer by binding with SdiA protein to regulate pilus expression, and interact with LsrR protein to increase SOS gene expression, inducing gene mutation. The QSIs-SAs mixtures could promote plasmid RP4 conjugative transfer and mutation in E. coli, and the main joint effects are synergism and antagonism. Furthermore, there is a good correlation among conjugative transfer, mutation, and growth inhibition of QSIs-SAs to E. coli. It could be speculated that bacteria may delay cell division to provide sufficient energy and time for regulating conjugative transfer and mutation under the stress of QSIs and their combined exposure with antibiotics, which is essentially a balance between bacterial resistance and toxicity. This study provides a reference for the resistance risk assessment of QSIs and benefits the clinical application of QSIs.

Hormetic dose-dependent response about typical antibiotics and their mixtures on plasmid conjugative transfer of Escherichia coli and its relationship with toxic effects on growth.

Bacterial resistance caused by the abuse of antibiotics has attracted worldwide attention. However, there are few studies exploring bacterial resistance under the environmental exposure condition of antibiotics that is featured by low-dose and mixture. In this study, sulfonamides (SAs), sulfonamide potentiators (SAPs) and tetracyclines (TCs) were used to determine the effects of antibiotics on plasmid RP4 conjugative transfer of Escherichia coli (E. coli) under single or combined exposure, and the relationship between the effects of antibiotics on conjugative transfer and growth was investigated. The results show that the effects of single or binary antibiotics on plasmid RP4 conjugative transfer all exhibit a hormetic phenomenon. The linear regression reveals that the concentrations of the three antibiotics promoting conjugative transfer are correlated with the concentrations promoting growth and the physicochemical properties of the compounds. The combined effects of SAs-SAPs and SAs-TCs on plasmid conjugative transfer are mainly synergistic and antagonistic. While SAPs provide more effective concentrations for the promotion of conjugative transfer in SAs-SAPs mixtures, SAs play a more important role in promoting conjugative transfer in SAs-TCs mixtures. Mechanism explanation shows that SAs, SAPs and TCs inhibit bacterial growth by acting on their target proteins DHPS, DHFR and 30S ribosomal subunit, respectively. This study indicates that toxic stress stimulates the occurrence of conjugative transfer and promotes the development of bacterial resistance, which will provide a reference for resistance risk assessment of antibiotic exposure.

Antibacterial and anticorrosive properties of CuZnO@RGO waterborne polyurethane coating in circulating cooling water.

In order to control bacterial adhesion and metal corrosion in the circulating cooling water system, it is necessary to prepare a nanocomposite-modified coating with antibacterial and anticorrosive functions. Copper and zinc composite oxide (CuZnO) was synthesized to prepare CuZnO@RGO nanocomposites. The antibacterial mechanism of CuZnO@RGO nanocomposites was investigated using gram-negative bacteria E. coli and gram-positive bacteria S. aureus as the two model microorganisms. The antibacterial properties of CuZnO@RGO nanocomposites on mixed bacteria were researched in the cooling water system. In addition, the CuZnO@RGO waterborne polyurethane (WPU) composite coating (CuZnO@RGO/WPU) was synthesized. The antibacterial performance, hardness, and corrosion inhibition performance of CuZnO@RGO/WPU composite coating in the cooling water system were also investigated. The results showed that after adding CuZnO@RGO nanocomposites to E. coli or S. aureus suspension, the protein leakage after 20 h was 9.3 times or 7.2 times higher than that in the blank experiment. The antibacterial rate of CuZnO@RGO nanocomposites in circulating cooling water reached 99.70% when the mass fraction of RGO was 15%. When the mass fraction of CuZnO@RGO accounting for CuZnO@RGO/WPU composite coating was 2%, the antibacterial rate, hardness, and corrosion inhibition efficiency were 94.35%, 5H, and 93.30%, respectively.

Antibacterial properties of chitosan chloride-graphene oxide composites modified quartz sand filter media in water treatment.

In order to control bacterial pollution in water treatment, it is necessary to prepare efficient antibacterial materials. Novel chitosan chloride-graphene oxide (CSCl@GO) composites were prepared via a one-step solution blending method. The composites were characterized by FT-IR, XRD and TEM techniques. The antibacterial mechanism of CSCl@GO composites was investigated using gram-negative bacteria E. coli and gram-positive bacteria S. aureus as two model microorganisms. The antibacterial properties of CSCl@GO composites for complex bacteria in the cooling water system were researched. In addition, the CSCl@GO composites modified quartz sand filter media (CSCl@GO/QS) were synthesized. The antibacterial performance of CSCl@GO/QS filter media in secondary effluent of domestic sewage was also investigated. The results showed that E. coli and S. aureus were completely inactivated after treatment with 100 mg·L-1 of CSCl@GO composites for 15 min. When adding CSCl@GO composites to E. coli or S. aureus suspension, the protein leakage was 12.7 or 9.9 times higher than that in the blank experiment after 12 h. The antibacterial rate of CSCl@GO composites in circulating cooling water reached 95.74% when the mass fraction of GO was 0.6%. After the CSCl@GO/QS filter media were backwashed for 3 times, the antibacterial rate could still be above 90%.

Three-dimensional graphene networks and RGO-based counter electrode for DSSCs.

Graphene is considered to be a potential replacement for the traditional Pt counter electrode (CE) in dye-sensitized solar cells (DSSCs). Besides a high electron transport ability, a close contact between the CE and electrolyte is crucial to its outstanding catalytic activity for the I3 -/I redox reaction. In this study, reduced graphene oxide (RGO) and three-dimensional graphene networks (3DGNs) were used to fabricate the CE, and the graphene-based CE endowed the resulting DSSC with excellent photovoltaic performance features. The high quality and continuous structure of the 3DGNs provided a channel amenable to fast transport of electrons, while the RGO afforded a close contact at the interface between the graphene basal plane and electrolyte. The obtained energy conversion efficiency (η) was closely related to the mass fraction and reduction degree of the RGO that was used. Corresponding optimization yielded, for the DSSCs based on the 3DGN-RGO CE, a value of η as high as 9.79%, comparable to that of the device using a Pt CE and hence implying promising prospects for the as-prepared CE.

Graphene-Assisted Thermal Interface Materials with a Satisfied Interface Contact Level Between the Matrix and Fillers.

Reduced graphene oxide (RGO) and three-dimensional graphene networks (3DGNs) are adopted to improve the performance of thermal interface materials (TIMs). Therein, the 3DGNs provide a fast transport network for phonons, while the RGO plays as a bridge to enhance the phonon transport ability at the interface between the filler and matrix. The types of surface functional groups of the RGO are found to exert a remarkable influence on the resulting thermal performance; the carboxyl groups are found in the optimal selection to promote the transport process at the interface area because a strong chemical bond will form between the graphene basal plane and epoxy resin (ER) through this kind of group. The resulting thermal conductivity reaches 6.7 Wm-1 K-1 after optimizing the mass fraction and morphology of the filler, which is 3250% higher than that of the pristine ER. Moreover, the mechanical properties of these as-prepared TIMs are also detected, and the specimens by using the RGO(OOH) filler display the better performances.

[Design, synthesis and biological evaluation of novel [1,3] dioxolo [4,5-f]isoindolone derivatives].

A series of [1,3]dioxolo[4,5-f]isoindolone derivatives were designed, synthesized and evaluated as inhibitors of acetylcholinesterases (AChE). Furthermore, their effects on memory impairment of mice induced by scopolamine were investigated with step-through test. The results suggested that most of the target compounds exhibited potential inhibition on AChE with IC50 values at micromolar range. Compounds I1 (IC50 value of 0.086 µmol · L(-1)) and I2 (IC50 value of 0.080 µmol · L(-1)) showed the strongest AChE inhibitory activity, which are equipotent to donepezil (IC50 value of 0.094 µmol · L(-1)). Moreover, compounds I1-I4 could improve the memory impairment induced by scopolamine in mice.

[Design, synthesis and biological evaluation of novel 4-substituted-3-nitrobenzamide derivatives].

A series of novel 4-substituted-3-nitrobenzamide derivatives were designed and synthesized. The structures of the target compounds were confirmed with 1H NMR, 13C NMR, MS and element analysis. Anti-tumor activities against HCT-116, MDA-MB435 and HL-60 cell lines in vitro were evaluated by SRB assay. The results indicated most of the target compounds exhibited potent anti-tumor activity. Compound 4a showed the most potent inhibitory activities against three cancer cell lines with the GI50 values of 1.904-2.111 micromol x L(-1). Compounds 4g, 41-4n exhibited more potent inhibitory activities against MDA-MB435 and HL-60 cell lines with the GI50 values of 1.008-3.586 micromol x L(-1) and 1.993-3.778 micromol x L(-1), respectively. The structure-activity relationship of these compounds is discussed preliminarily.

[Simultaneous determination of chlorpromazine and promethazine and their main metabolites by capillary electrophoresis with electrochemiluminescence].

Based on the phenomenon that each of chlorpromazine (CPZ), promethazine (PMZ), chlorpromazine sulfoxide (CPZSO) and promethazine sulfoxide (PMZSO) could enhance the electrochemiluminescence (ECL) intensity of tris(2,2'-bipyridyl) ruthenium, a novel and sensitive method was proposed for the simultaneous determination of CPZ, PMZ and their main metabolites using capillary electrophoresis (CE) coupled with ECL detection. The influences of several experimental parameters were explored. The optimum experimental conditions were as follows: detection potential of 1. 20 V (Ag/AgCl), 40 mmol/L of phosphate buffer solution (pH 6.5) containing 5 mmol/L tris(2,2'-bipyridyl) ruthenium in ECL detection cell, running buffer solution of 18 mmol/L (pH 4.8), sample injection of 8 s at 11 kV, and separation voltage of 13.5 kV. The detection limits (3sigma) of this method were 8.3 x 10(-7) g/L for CPZ, 7.2 x 10(-6) g/L for PMZ, 1.9 x 10(-5) g/L for CPZSO and 3.7 x 10(-6) g/L for PMZSO. The linear ranges of ECL intensity versus mass concentration of medicaments were 7. 1 x 10(-6) - 6. 3 x 10(-3) g/L for CPZ, 7.5 x 10(-5) - 4.6 x 10(-3) g/L for PMZ, 9.7 x 10(-5) - 3.6 x 10(-3) g/L for CPZSO and 8.1 x 10(-5) - 7.7 x 10(-3) g/L for PMZSO. The relative standard deviations (RSDs) of the four target compounds were not more than 3% for ECL intensity and 1% for migration time. This method has the merits of simplicity, speediness, sensitivity, small sample injection, and free from interference. This method was successfully utilized to directly and simultaneously detect CPZ, PMZ, CPZSO and PMZSO in urine samples of pet dogs.