RuSe2-CoTe Heterogeneous Surfaces Coated with NC Layer for Excellent HER Performance under Alkaline Condition.

Electrocatalytic hydrogen production has been a promising high-purity hydrogen production technology, attracting a large number of researchers' research interest. Ru has a hydrogen binding capacity similar to Pt, but its price is far lower than Pt, making it a promising alternative to Pt. However, a single Se electronic structure modulation is not sufficient to enable RuSe2 to be used for practical applications on a large scale due to the lack of electrons. Therefore, choosing a suitable way to electronically modulate the Ru atoms in RuSe2 can effectively improve the activity of the catalyst. Cobalt telluride (CoTe) can significantly enhance electrocatalytic performance due to tellurium's low electronegativity and excellent metal properties. In this work, the NC layer possesses excellent electrical conductivity and CoTe acts as an electron donor to optimize the electronic structure locally and trigger electron transfer efficiently. The RuSe2-CoTe/NC electrode requires an overpotential of only 25.4 mV (10 mA cm-2), which is superior to that of RuSe2/NF (65 mV) and CoTe/NC (115 mV). Meanwhile, the Tafel slope of RuSe2-CoTe/NC (67.8 mV dec-1) was better than that of RuSe2/NF (113.6 mV dec-1) and CoTe/NC (209.5 mV dec-1), showing that the build-up of the superior heterojunction makes the RuSe2-CoTe/NC with better hydrogen evolution reaction (HER) reaction kinetics. In addition, after 30 h of long-term stability testing, no significant decrease in catalytic activity was observed, proving the good stability of the RuSe2-CoTe/NC catalyst.

Spindle Nanorods of CeO2 and NiS Heterointerface Coated by the NC Layer: A High-Performance Bifunctional Electrocatalyst for Water Splitting.

Currently, nickel sulfides are widely employed in the oxygen evolution reaction (OER) and hydrogen evolution reaction (HER), thanks to the narrow electronegativity difference of only 0.67 eV between nickel and sulfur. Among them, NiS stands out in terms of the OER performance; however, its HER performance and stability remain somewhat inadequate. The construction of heterogeneous interfaces can efficiently improve the HER performance and regulate the electronic structure of the NiS catalyst. CeO2 has been discovered to possess exceptional electronic modulation capabilities, which may lead to the effective enhancement of both HER and OER of the NiS catalyst. As a result, a nitrogen-doped carbon-coated CeO2-NiS heterogeneous interface catalyst (NC/NiS-CeO2) is designed as a bifunctional electrocatalyst for HER and OER with high performance. The NC/NiS-CeO2 catalyst demonstrates excellent HER (47 mV at 10 mA cm-2) and OER (92 mV at 10 mA cm-2) performances in a 1 M KOH alkaline solution. Characterization analysis reveals that the coupling of the heterostructure interface, which consists of CeO2 and NiS, significantly enhances electron conduction, the synergistic effect, and the electrocatalytic activity of the electrode. This study demonstrates that the HER and OER activity can be effectively improved by constructing a rational heterogeneous interface.

Z-Scheme CdIn2S4/Bi2WO6 Heterojunction for High Piezo-Photo Synergetic Performance.

The piezoelectric effect triggered by mechanical energy could establish an internal electric field to effectively modulate the separation behavior of carriers. Herein, a novel CdIn2S4/Bi2WO6 (CIS/BWO) piezo-photocatalyst for removing diclofenac (DCF) from water was constructed for the first time. Encouragingly, the photocatalytic degradation activity of CIS/BWO was effectively promoted through the piezoelectric effect. Specifically, 10% CIS/BWO exhibited promising DCF degradation performance under co-excitation of light irradiation and ultrasonic vibration, with a degradation efficiency of 99.9% within 40 min, much higher than that of pure photocatalysts (72.3%) and piezocatalysts (60.3%). Meanwhile, an in-depth study of the charge carrier separation mechanism of the CIS/BWO composite under the piezo-photo synergy condition was proposed. Both the built-in electric field induced by the piezoelectric effect in BWO and the Z-scheme transfer path of the CIS/BWO heterojunction are beneficial to interfacial charge transfer. Moreover, the Z-scheme mechanism was further demonstrated by trapping experiments and the electron spin resonance (ESR) technique. Finally, the corresponding intermediates of DCF over CIS/BWO composites and possible degradation pathways were also investigated by DFT calculations and liquid chromatography-mass spectrometry.

Efficient Charge Separation via MoSe2 Nanosheets with Tunable 1T Phase Contents: Piezoreduction of Cr(VI) to Cr(III) and Piezodegradation of RhB.

Piezocatalysis is a promising technology to address environmental pollution by converting mechanical energy into chemical energy. Herein, MoSe2 nanosheets with different 1T phase percentages (ranging from 30 to 80%) were constructed by adjusting hydrothermal temperature. Moreover, the roles of phase engineering in the piezocatalysis were thoroughly investigated by degrading rhodamine B and reducing Cr(VI) in ultrasonic vibration conditions. In particular, MoSe2 prepared at 220 °C (MoSe2-220) exhibits ultrahigh observed constant kobs and degradation rate k, which is superior to most reported catalysts to date. The experimental results indicate that the introduction of the 1T phase increases the active sites of the material, improves the conductivity, and inhibits the recombination of electrons and holes. Moreover, an internal electric field in the 2H phase induced by piezoelectric polarization is facilitated to separate electron-hole pairs, enabling the degradation and reduction to proceed. The capture experiments and EPR tests further confirm that •O2- and •OH are main reactive species, and a rational mechanism is finally put forward. This study offers a clear understanding of phase engineering in piezocatalysis and provides an efficiency strategy to construct highly efficient piezocatalysts.

Identification of genes related to tipburn resistance in Chinese cabbage and preliminary exploration of its molecular mechanism.

BACKGROUND: Tipburn, also known as leaf tip necrosis, is a severe issue in Chinese cabbage production. One known cause is that plants are unable to provide adequate Ca2+ to rapidly expanding leaves. Bacterial infection is also a contributing factor. Different cultivars have varying degrees of tolerance to tipburn. Two inbred lines of Chinese cabbage were employed as resources in this research. RESULTS: We determined that the inbred line 'J39290' was the tipburn resistant material and the inbred line 'J95822' was the tipburn sensitive material based on the severity of tipburn, and the integrity of cell membrane structure. Ca2+ concentration measurements revealed no significant difference in Ca2+ concentration between the two materials inner leaves. Transcriptome sequencing technology was also used to find the differentially expressed genes (DEGs) of 'J95822' and 'J39290', and there was no significant difference in the previously reported Ca2+ uptake and transport related genes in the two materials. However, it is evident through DEG screening and classification that 23 genes are highly linked to plant-pathogen interactions, and they encode three different types of proteins: CaM/CML, Rboh, and CDPK. These 23 genes mainly function through Ca2+-CaM/CML-CDPK signal pathway based on KEGG pathway analysis, protein interaction prediction, and quantitative real-time PCR (qRT-PCR) of key genes. CONCLUSIONS: By analyzing the Ca2+ concentration in the above two materials, the transcription of previously reported genes related to Ca2+ uptake and transport, the functional annotation and KEGG pathway of DEGs, it was found that Ca2+ deficiency was not the main cause of tipburn in 'J95822', but was probably caused by bacterial infection. This study lays a theoretical foundation for exploring the molecular mechanism of resistance to tipburn in Chinese cabbage, and has important guiding significance for genetics and breeding.

The role of transcription factor Ap1 in the activation of the Nrf2/ARE pathway through TET1 in diabetic nephropathy.

TET1 mediates demethylation in tumors, but its role in diabetic nephropathy (DN), a prevalent diabetic complication, is unclear. We attempted to probe the possible mechanism of TET1 in DN. A DN rat model was established and verified by marker detection and histopathological observation. The in vitro model was established on human mesangial cells (HMCs) induced by high glucose (HG), and verified by evaluation of fibrosis and inflammation. The differentially expressed mRNA was screened out by microarray analysis. The most differentially expressed mRNA (TET1) was reduced in DN rats and HG-HMCs. The upstream and downstream factors of TET1 were verified, and their roles in DN were analyzed by gain- and loss-function assays. TET1 was decreased in DN rats and HG-HMCs. High expression of TET1 decreased biochemical indexes and renal injury of DN rats and hampered the activity, fibrosis, and inflammation of HG-HMCs. Ap1 lowered TET1 expression, and enhanced inflammation in HG-HMCs, and accentuated renal injury in DN rats. TET1 overexpression inhibited the effect of Ap1 on DN. TET1 promoted the transcription of Nrf2. The Ap1/TET1 axis mediated the Nrf2/ARE pathway activity. Overall, TET1 overexpression weakened the inhibitory effect of Ap1 on the Nrf2/ARE pathway, thus alleviating inflammation and renal injury in DN.

Characterization of antiviral immune response induced by poly(I:C) in macrophages of farmed large yellow croaker (Larimichthys crocea).

Fish tend to rely more on their innate immunity to executing defense against viral infection by inducing antiviral gene production. However, the expression pattern and underlying mechanism of fish antiviral responses have yet to be fully defined. In the present study, an in vitro viral infection model was established by exposing head kidney-derived macrophages of large yellow croaker to virus analog, poly(I:C). Transcriptome analysis indicated that poly(I:C) appeared to induce potent antiviral activity featuring dominant interferon a3 (IFNa3) expression through activation of toll-like receptors (TLRs)/TIR-domain-containing adapter-inducing interferon-ß (TRIF) and retinoic acid-inducible gene I-like receptors (RLRs)/mitochondrial antiviral signaling protein (MAVS) pathways. Inhibition of nuclear factor κB (NF-κB) and stimulator of interferon genes (STING)/interferon regulatory factor 3 (IRF3) pathways diminished the expression of IFNa3. Mechanistically, transcription factors including p65 and IRF3 could promote expression of IRF3, and activated IRF3 alone further increased the transcriptional activity of IFNa3. We also characterized the promoter of IFNa3 with direct IRF3 binding site which was sufficient to render the transcription of IFNa3. This effect was attenuated after deletion or mutation of the IRF3 binding sites. Taken together, our findings illustrate the distinct transcriptional profiling of fish macrophages triggered by poly(I:C). Also, this work provides new insights into the molecular mechanism underpinning coordinated activation of pathogen recognition and signaling transduction in the antiviral responses of non-model fish species.

Effect of replacement of dietary fish oil with four vegetable oils on prostaglandin E2 synthetic pathway and expression of inflammatory genes in marine fish Larimichthys crocea.

As a lipid mediator with important immune function, prostaglandin E2 (PGE2) has been widely studied in mammals, whereas its synthetic pathway and immune function in fish have yet to be fully studied. To investigate the regulation of PGE2 synthetic pathway and inflammatory genes expression by dietary different oils and the underlying relationship, a 10-week feeding experiment and an immune challenge were carried out in marine fish Larimichthys crocea. Replacement of dietary fish oil (FO) with four vegetable oils (VO), including soybean oil, linseed oil, palm oil, and olive oil, all reduced PGE2 levels, and the decrease of arachidonic acid (ARA, substrate for PGE2) could account for this decline. Meanwhile, the expression of PGE2 synthesis related genes was basically upregulated, which seemed to be a feedback regulation, but it cannot compensate the deficiency of ARA. In addition, mRNA expression of inflammatory genes, including interleukin (IL)-1ß, IL-6, tumor necrosis factor (TNF)α and interferon (IFN)γ was all upregulated in four VO groups compared with FO group, which was the opposite of PGE2 levels. To verify the inflammatory regulation of PGE2, an immune challenge was conducted, and PGE2 alleviated LPS-induced expression of inflammatory genes, including IL-6, TNFα and IFNγ, and the similar downregulation of toll-like receptor (TLR) genes expression revealed that TLR signaling pathway participated in the anti-inflammatory regulation of PGE2. In conclusion, replacement of dietary FO with four VO (lack of ARA) reduced the levels of PGE2 that could alleviate LPS-induced inflammatory genes expression via TLR signaling pathway, which could be one of the reasons that VO induced inflammation in marine fish.

Allelism analysis of BrRfp locus in different restorer lines and map-based cloning of a fertility restorer gene, BrRfp1, for pol CMS in Chinese cabbage (Brassica rapa L.).

KEY MESSAGE: In Chinese cabbage, there are two Rf loci for pol CMS and one of them was mapped to a 12.6-kb region containing a potential candidate gene encoding PPR protein. In Chinese cabbage (Brassica rapa), polima cytoplasmic male sterility (pol CMS) is an important CMS type and is widely used for hybrid breeding. By extensive test crossing in Chinese cabbage, four restorer lines (92s105, 01s325, 00s109, and 88s148) for pol CMS were screened. By analyzing the allelism of the four restorer lines, it was found that 92s105, 01s325, and 00s109 had the same "restorers of fertility" (Rf) locus (designated as BrRfp1), but 88s148 had a different Rf locus (designated as BrRfp2). For fine mapping the BrRfp1 locus of 92s105, a BC1F1 population with 487 individuals and a BC1F2 population with 2485 individuals were successively constructed. Using simple sequence repeat (SSR) markers developed from Brassica rapa reference genome and InDel markers derived from whole-genome resequencing data of 94c9 and 92s105, BrRfp1 was mapped to a 12.6-kb region containing a potential candidate gene encoding pentatricopeptide repeat-containing protein. Based on the nucleotide polymorphisms of the candidate gene sequence between the restoring and nonrestoring alleles, a co-segregating marker SC718 was developed, which would be helpful for hybrid breeding by marker-assisted screening and for detecting new restorer lines.

Preparation of Yeast Extract from Brewer's Yeast Waste and Its Potential Application as a Medium Constituent.

Yeast extract serves as a source of nutritional components essential for human dietary requirements, feed formulations, and the vital growth factors and nutrients necessary for microorganisms. However, the production cost of yeast extract using cultivated active dry yeast is relatively high. This study aims to utilize the autolysis of discarded yeast post beer brewing to produce yeast extract. The concentration, temperature, pH, and time conditions are systematically optimized. It reveals that the yield of amino nitrogen and solids in the extract was increased by 3.3% and 20.9% under the optimized conditions (1.2% wall-breaking enzyme, 1% yeast extract enzyme, and a hydrolysis time of 24 h) than that of the documented 4.03% and 69.05%. Additionally, a comparative analysis with commercially available yeast powder demonstrates that the yeast extract derived from this study adequately fulfills the nutritional requirements for microbial growth. Hence, the utilization of discarded beer yeast presents an opportunity for the valuable reclamation of waste yeast, showcasing promising potential applications.

A comparative study: In vitro effects of EPA and DHA on immune functions of head-kidney macrophages isolated from large yellow croaker (Larmichthys crocea).

Comparative effects of different concentrations of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) on immune responses of head-kidney macrophages isolated from large yellow croaker were studied in vitro. After exposing to serum-free medium for 1 day, cultured cells were incubated in medium supplemented with graded levels of EPA or DHA (0, 5, 25, 100, 200 and 1000 µM, respectively) in the form of fatty acid bovine serum albumin (FA-BSA) complex for 12 h, 24 h and 36 h, respectively. Control samples were incubated in the absence of EPA or DHA (2% bovine serum albumin, BSA). Following stimulation, cell viability, lipid peroxidation, secretary phopholipase A2 (sPLA2) and prostaglandin E2 (PGE2) production as well as some immune parameters including phagocytosis, respiratory burst activity and interleukin 1ß (IL-1ß) production were determined. Results showed that EPA and DHA affected cell viability in dose-dependent and time-dependent manners. In particular, cell viability was significantly decreased after 24 h and 36 h incubation with 1000 µM EPA or DHA (P < 0.05). Higher levels of EPA (200 and 1000 µM) caused a significant increase in the production of malondialdehyde (MDA) (P < 0.05), while DHA did not significantly affect the MDA production. EPA significantly increased the intracellular superoxide anion synthesis which, on the contrary, was significantly reduced by DHA. Phagocytosis percentage (PP) values were significantly higher in treatments with 5 µM DHA (P < 0.05), but significantly decreased by 200 and 1000 µM EPA and DHA compared to the control group (P < 0.05). Decreased PGE2 production was produced by cells treated with relatively low doses of EPA or DHA. When high levels of stimulants (1000 µM EPA or DHA) were used, PGE2 levels were elevated and reached a significant level (P < 0.05). Both EPA and DHA significantly inhibited the production of sPLA2, where DHA exerted the more potent inhibitory effects than EPA. No pronounced effect was observed on IL-1ß production among all the treatments, and IL-1ß level in cell culture supernatant was fairly low (only approximately 6 pg/ml). Those findings suggested that EPA and DHA could influence the immunity and physiological conditions of macrophages from head kidney of large yellow croaker in vitro.

A vacuolar processing enzyme RsVPE1 gene of radish is involved in floral bud abortion under heat stress.

Radish floral bud abortion (FBA) is an adverse biological phenomenon that occurs during reproduction. Although FBA is a frequent occurrence, its molecular mechanism remains unknown. A transcript-derived fragment (TDF72), which was obtained by cDNA amplified fragment length polymorphism (cDNA-AFLP), was up-regulated in the aborted buds and exhibited 89% sequence homology with the AtγVPE gene. In this study, TDF72 was used to clarify the role of VPE in FBA by isolation of the VPE gene RsVPE1 from radish flower buds. The full-length genomic DNA was 2346 bp including nine exons and eight introns. The full-length cDNA was 1825 bp, containing a complete open reading frame (ORF) of 1470 bp, which encoded a predicted protein containing 489 amino acid residues, with a calculated molecular mass of 53.735 kDa. Expression analysis demonstrated that RsVPE1 was expressed in all tested organs of radish at different levels. Highest expression was detected in aborted flower buds, suggesting that RsVPE1 has a role in FBA. In order to analyze the role of RsVPE1 in FBA, RsVPE1 was overexpressed in transgenic Arabidopsis thaliana plants. Aborted flower buds appeared in transgenic plants subjected to heat stress. In addition, RsVPE1 expression in the transgenic plants reached a maximum when subjected to heat stress for 24 h and increased by 2.1-fold to 2.8-fold in three homozygous transgenic lines. These results indicated that RsVPE1 led to FBA when its expression levels exceeded a particular threshold, and provided evidence for the involvement of RsVPE1 in promoting FBA under heat stress.

Removal of algae from raw water by ultrasonic irradiation and flocculation: a pilot scale experiment.

Using Chl alpha removal rate as index, a 28 kHz/900w ultrasonic cleaning machine was applied to testify algal removal by ultrasonic irradiation from raw water of a pool, where Microcystis aeruginosa colonies is absolutely dominated with temperature being over 20 degrees C, and the irradiation lasted for 5 min. PAC was used as flocculant at the dose of 60 mgl(-1), jar tests were done to investigate the Chl alpha removal by flocculation. The results showed that ultrasound raised the water temperature instantly but did not lead to a regular pH change pattern in all the treated samples. Ultrasound could remove more than 90% of Chl alpha from raw water with temperature over 31 degrees C, but less than 20% of Chl alpha or even increased Chl alpha concentration in some samples with temperature lower than 31 degrees C. Compared with the algal removal effects by direct raw water flocculation with PAC, ultrasound did not enhance markedly the flocculation effects on algae removal, which was not in agreement with the findings reported. The reason might be due to morphology and characteristics of natural algae differed greatly from that of algae cultured in laboratory.

Tentacle-like core-shell CoNi2S4/C3N4 bifunctional electrocatalysts for efficient overall alkaline water splitting.

Stable and efficient bifunctional electrocatalysts are of great significance for sustainable energy conversion and human society sustainability. However, conventional electrocatalytic materials tend to exhibit high overpotentials and unsatisfactory chemical activities. Herein, we construct novel CoNi2S4/C3N4 nanowires on a nickel foam (NF) electrode as a bifunctional electrocatalyst for alkaline water splitting by a two-step hydrothermal and thermal annealing process. The prepared CoNi2S4/C3N4 electrocatalyst exhibits superior HER (e.g. 40 mV (ηH210)) and OER (e.g. 110 mV (ηO210)) activities in a 1 M KOH electrolyte, which are much smaller than those of bare NF, Co@NF, NiCoO@NF and most reported materials. Furthermore, the stability test at 10 mA cm-2 for 20 h for the CoNi2S4/C3N4 electrocatalyst shows no obvious decay and proves the excellent stability of CoNi2S4/C3N4. In this work, the unique tentacle-like CoNi2S4/C3N4 nanowire nanostructure leads to minimized interfacial resistance and abundant channels during electrocatalysis. Moreover, comprehensive analysis results show that Ni(Co)OOH active sites, which are beneficial for excellent OER activity, partially form on the surface of CoNi2S4/C3N4 during electrocatalysis. Finally, the CoNi2S4/C3N4∥CoNi2S4/C3N4 two-electrode system is constructed and it exhibits a low-voltage water splitting capability of 1.40 V.

Fabrication of multiphase MoSe2 modified BiOCl nanosheets for efficient piezo-photoelectric hydrogen evolution and antibiotic degradation.

Efficient spatial charge separation plays a crucial role in improving the photocatalytic performance. Therefore, 1T/2H MoSe2/BiOCl (1T/2H MS/BOC) and 2H MoSe2/BiOCl (2H MS/BOC) piezo-photocatalysts are synthesized. By combining piezoelectric catalysis and photocatalysis, a highly active piezo-photocatalytic process is realized. The optimal 1T/2H MS/BOC piezo-photocatalyst displays superior diclofenac (DCF) degradation and hydrogen (H2) evolution activity under the combined action of ultrasound and light. In particular, the DCF degradation kinetic constant (k) of optimal 0.5% 1T/2H MS/BOC under the synergistic effect of ultrasound and light is 0.057 min-1, which is 8.1 and 6.3 times higher than those of BiOCl (0.007 min-1) and 0.5% 2H MS/BOC (0.009 min-1). Moreover, the H2 evolution rate of 0.5% 1T/2H MS/BOC is 122.5 µmol g-1 h-1, which is also higher than those of BiOCl (45.8 µmol g-1 h-1) and 2H MS/BOC (49.5 µmol g-1 h-1). The dramatic improvement in the DCF degradation and H2 evolution piezo-photocatalytic performance of 1T/2H MS/BOC catalysts is ascribed to the built-in polarization electric field and abundance of active sites of 1T/2H MS/BOC as well as the advantageous band structure between BiOCl and 1T/2H MoSe2. Additionally, three probable degradation pathways of DCF were put forward from the results of liquid chromatography-mass spectrometry (LCMS) and density functional theory (DFT) calculations. This study provides the design strategy of high efficiency piezo-photocatalysts in environmental purification and energy-generation fields based on phase and band structure engineering.

Expression and Characterization of a GH38 α-Mannosidase from the Hyperthermophile Pseudothermotoga thermarum.

This study focused on the bio-characterization of a GH38 α-mannosidase from the hyperthermophile Pseudothermotoga thermarum DSM 5069. We aimed to successfully express and characterize this thermophilic α-mannosidase and to assess its functional properties. Subsequently, recombinant α-mannosidase PtαMan was expressed in Escherichia coli BL21(DE3) and purified via affinity chromatography, and native protein was verified as a tetramer by size exclusion chromatography. In addition, the activity of α-mannosidase PtαMan was relatively stable at pH 5.0-6.5 and temperatures up to 75 ℃. α-Mannosidase PtαMan was active toward Co2+ and had a good catalytic efficiency deduced from the kinetic parameters. However, its activity was strongly inhibited by Cu2+, Zn2+, SDS, and swainsonine. In summary, this cobalt-required α-mannosidase is putatively involved in the direct modification of glycoproteins.

Cloning, expression and characterization of a glycoside hydrolase family 51 α-l-arabinofuranosidase from Thermoanaerobacterium thermosaccharolyticum DSM 571.

The present study focused on the characterization of a glycoside hydrolase 51 family α-l-arabinofuranosidase named TtAbf51 from thermophile Thermoanaerobacterium thermosaccharolyticum DSM 571. The recombinant TtAbf51 with 497 amino acids was successfully expressed in Escherichia coli BL21(DE3) and purified via nickel affinity chromatography, and native protein was a dimer verified by size exclusion chromatography. The TtAbf51 showed an optimum pH and temperature of 5.5 and 55 °C, and was relatively stable at pH 5.0-8.0 and up to 60 °C for 2 h of incubation. In addition, TtAbf51 was significantly inhibited by Cu2+, Zn2+ and 1 mM or 10 mM SDS. In the presence of 800 mM arabinose, the residual activity remained over 40% of the initial activity. In addition, the recombinant enzyme possessed a good catalytic effect for both synthesized and natural substrates, and the specific enzyme activity toward CM-linear arabinan reached 426.5 µmol min-1 mg-1. In summary, this study provides an α-l-arabinofuranosidase with potential in the synergistic hydrolysis of hemicellulose to fermentable sugars in applications such as liquid biofuels, food and beverages, and related industries.

Integrated transcriptome and hormone analyses provide insights into silver thiosulfate-induced "maleness" responses in the floral sex differentiation of pumpkin (Cucurbita moschata D.).

The perfect mating of male and female flowers is the key to successful pollination. The regulation of ethylene with chemicals is a good option for inducing staminate or female flowers. Silver thiosulfate is often used to induce the formation of male flowers in subgynoecious and gynoecious crops, which is important to maintain their progenies. However, its effects on flower sex differentiation in pumpkin (Cucurbita moschata Duch.) and the underlying mechanism remain unclear. In this study, the application of silver thiosulfate to pumpkin seedlings significantly delayed the occurrence of the first female flower and increased the number of male flowers. We next investigated the underlying mechanism by employing transcriptome and endogenous hormone analyses of the treated plants. In total, 1,304 annotated differentially expressed genes (DEGs)were identified by comparing silver thiosulfate-treated and control plants. Among these genes, 835 were upregulated and 469 were downregulated. The DEGs were mainly enriched in the phenylpropanoid biosynthesis and metabolism pathways (phenylalanine ammonia-lyase, peroxidase) and plant hormone signal transduction pathways (auxin signaling, indole-3-acetic acid-amido synthetase, ethylene response factor). Silver thiosulfate significantly reduced the levels of 2-oxindole-3-acetic acid, para-topolin riboside, dihydrozeatin-O-glucoside riboside, and jasmonoyl-l-isoleucine but increased the levels of trans-zeatin-O-glucoside, cis-zeatin riboside, and salicylic acid 2-O-ß-glucoside. The levels of auxin and jasmonic acid were decreased, whereas those of salicylic acid were increased. Different trends were observed for different types of cytokinins. We concluded that silver thiosulfate treatment not only affects the expression of auxin synthesis and signaling genes but also that of ethylene response factor genes and regulates the levels of auxin, salicylic acid, jasmonic acid, and cytokinins, which together might contribute to the maleness of pumpkin. This study provides useful information for understanding the mechanism underlying the effect of silver thiosulfate on floral sex differentiation in pumpkin, a widely cultivated vegetable crop worldwide, and gives a production guidance for the induction of maleness using STS for the reproduction of gynoecious lines of Cucurbitaceae crops.

Genome-Wide Identification of APETALA2/ETHYLENE RESPONSIVE FACTOR Transcription Factors in Cucurbita moschata and Their Involvement in Ethylene Response.

APETALA2/ETHYLENE RESPONSIVE FACTOR (AP2/ERF), a plant-specific transcription factor (TF) family, plays an essential role in the growth and development of plants, and in their response to biotic and abiotic stresses. However, information on AP2/ERF in Cucurbita moschata (pumpkin), an edible and medicinal vegetable used worldwide, is scarce. A total of 212 AP2/ERF genes were identified in the C. moschata genome (CmoAP2/ERFs). Based on phylogenetic analysis, they were divided into four groups-28 AP2s, 92 ERFs, 86 dehydration-responsive element-binding (DREB) factors, and 6 ABI3/VPs (RAV). The 212 AP2/ERF genes were unevenly distributed on the 20 chromosomes of C. moschata. The results of structural analysis showed the absence of introns on 132 CmoAP2/ERFs. Four pairs of tandem duplication and 155 pairs of segmental duplication events were identified, which indicated that segmental duplications might be the main reason for the expansion of the CmoAP2/ERF family. The analysis of cis-regulatory elements (CREs) showed that most of the CmoAP2/ERFs contained hormone response elements (ABREs, EREs) in their promoters, suggesting that AP2/ERFs could contribute to the processes regulated by ethylene and abscisic acid. By comparing the transcriptome of ethephon-treated and control plants, we found that 16 CmoAP2/ERFs were significantly upregulated after ethephon treatment. Furthermore, we determined the expression patterns of these genes at different developmental stages of female and male flowers. This study provides insights into the identification, classification, physicochemical property, phylogenetic analysis, chromosomal location, gene structure, motif identification, and CRE prediction of the AP2/ERF superfamily in C. moschata. Sixteen CmoAP2/ERF genes were identified as ethylene-inducible genes. The results of this study will be valuable for understanding the roles of CmoAP2/ERFs in ethylene response and should provide a foundation for elucidating the function of AP2/ERF TFs in C. moschata.

Functional analysis and regulation mechanism of interferon gamma in macrophages of large yellow croaker (Larimichthys crocea).

Interferon gamma (IFN-γ) is a widely expressed cytokine that has potent antiviral and immunomodulatory effects. The expression and bioactivity of IFN-γ have been reported in several fish species. However, the molecular mechanism mediated by IFN-γ in fish macrophages has not been completely elucidated. This study used the macrophage cell line to investigate the functional activities and regulation mechanism of large yellow croaker IFN-γ (LcIFN-γ). Herein, the mRNA expression of Lcifn-γ was significantly upregulated in macrophages after LPS and poly(I:C) treatment. Recombinant LcIFN-γ protein (rLcIFN-γ) significantly enhanced the phagocytic ability and respiratory burst activity of macrophages. Meanwhile, rLcIFN-γ induced M1 phenotype polarization of macrophages with the upregulated expressions of pro-inflammatory gene. Moreover, rLcIFN-γ upregulated the IFN-stimulated genes (ISGs) expression and activated JAK (Janus tyrosine kinases)-STAT (signal transducer and activator of transcription) signaling pathway by causing the phosphorylation of JAK1 and STAT1Tyr701. Furthermore, the promoter activity of IFN-regulatory factor 1 (IRF1) was significantly upregulated by the phosphorylated transcription factor STAT1 through binding to its promoter region. In addition to the classical JAK-STAT pathway, rLcIFN-γ also activated multiple distinct signaling cascades such as mitogen-activated protein kinase (MAPK) and protein kinase B (AKT) pathways. Overall, this study indicated the powerful effects of LcIFN-γ on macrophage activation of large yellow croaker and its molecular mechanism.