Analysis of the response regulatory network of pepper genes under hydrogen peroxide stress

Hydrogen peroxide (H2O2) is a regulatory component related to plant signal transduction. To better understand the genome-wide gene expression response to H2O2 stress in pepper plants, a regulatory network of H2O2 stress-gene expression in pepper leaves and roots was constructed in the present study. We collected the normal tissues of leaves and roots of pepper plants after 40 days of H2O2 treatment and obtained the RNA-seq data of leaves and roots exposed to H2O2 for 0.5–24 h. By comparing the gene responses of pepper leaves and roots exposed to H2O2 stress for different time periods, we found that the response in roots reached the peak at 3 h, whereas the response in leaves reached the peak at 24 h after treatment, and the response degree in the roots was higher than that in the leaves. We used all datasets for K-means analysis and network analysis identified the clusters related to stress response and related genes. In addition, CaEBS1, CaRAP2, and CabHLH029 were identified through a co-expression analysis and were found to be strongly related to several reactive oxygen species-scavenging enzyme genes; their homologous genes in Arabidopsis showed important functions in response to hypoxia or iron uptake. This study provides a theoretical basis for determining the dynamic response process of pepper plants to H2O2 stress in leaves and roots, as well as for determining the critical time and the molecular mechanism of H2O2 stress response in leaves and roots. The candidate transcription factors identified in this study can be used as a reference for further experimental verification

Genome-Wide Identification and Capsaicinoid Biosynthesis-Related Expression Analysis of the R2R3-MYB Gene Family in Capsicum annuum L.

Capsaicinoids are naturally specialized metabolites in pepper and are the main reason that Capsicum fruits have a pungent smell. During the synthesis of capsaicin, MYB transcription factors play key regulatory roles. In particular, R2R3-MYB subfamily genes are the most important members of the MYB family and are critical candidate factors in capsaicinoid biosynthesis. The 108 R2R3-MYB genes in pepper were identified in this study and all are shown to have two highly conserved MYB binding domains. Phylogenetic and structural analyses clustered CaR2R3-MYB genes into seven groups. Interspecies collinearity analysis found that the R2R3-MYB family contains 16 duplicated gene pairs and the highest gene density is on chromosome 00 and 03. The expression levels of CaR2R3-MYB differentially expressed genes (DEGs) and capsaicinoid-biosynthetic genes (CBGs) in fruit development stages were obtained via RNA-seq and quantitative polymerase chain reaction (qRT-PCR). Co-expression analyses reveal that highly expressed CaR2R3-MYB genes are co-expressed with CBGs during early stages of pericarp and placenta development processes. It is speculated that six candidate CaR2R3-MYB genes are involved in regulating the synthesis of capsaicin and dihydrocapsaicin. This study is the first systematic analysis of the CaR2R3-MYB gene family and provided references for studying their molecular functions. At the same time, these results also laid the foundation for further research on the capsaicin characteristics of CaR2R3-MYB genes in pepper

Self-Powered Photoelectrochemical Assay for Hg²⁺ Detection Based on g-C₃N₄-CdS-CuO Composites and Redox Cycle Signal Amplification Strategy

A highly sensitive self-powered photoelectrochemical (spPEC) sensing platform was constructed for Hg2+ determination based on the g-C3N4-CdS-CuO co-sensitized photoelectrode and a visible light-induced redox cycle for signal amplification. Through successively coating the single-layer g-C3N4, CdS, and CuO onto the surface of an electrode, the modified electrode exhibited significantly enhanced PEC activity. The microstructure of the material was characterized by scanning electron microscopy (SEM), X-ray diffraction (XRD) and Fourier transform infrared spectroscopy (FT-IR). However, the boost in photocurrent could be noticeably suppressed due to the consumption of hole-scavenging agents (reduced glutathione) by the added Hg2+. Under optimal conditions, we discovered that the photocurrent was linearly related to the Hg2+ concentration in the range of 5 pM–100 nM. The detection limit for Hg2+ was 0.84 pM. Moreover, the spPEC sensor demonstrated good performance for the detection of mercury ions in human urine and artificial saliva

Self-Powered Photoelectrochemical Assay for Hg2+ Detection Based on g-C3N4-CdS-CuO Composites and Redox Cycle Signal Amplification Strategy

A highly sensitive self-powered photoelectrochemical (spPEC) sensing platform was constructed for Hg2+ determination based on the g-C3N4-CdS-CuO co-sensitized photoelectrode and a visible light-induced redox cycle for signal amplification. Through successively coating the single-layer g-C3N4, CdS, and CuO onto the surface of an electrode, the modified electrode exhibited significantly enhanced PEC activity. The microstructure of the material was characterized by scanning electron microscopy (SEM), X-ray diffraction (XRD) and Fourier transform infrared spectroscopy (FT-IR). However, the boost in photocurrent could be noticeably suppressed due to the consumption of hole-scavenging agents (reduced glutathione) by the added Hg2+. Under optimal conditions, we discovered that the photocurrent was linearly related to the Hg2+ concentration in the range of 5 pM–100 nM. The detection limit for Hg2+ was 0.84 pM. Moreover, the spPEC sensor demonstrated good performance for the detection of mercury ions in human urine and artificial saliva

Cleavage of Metal-Ion-Induced DNAzymes Released from Nanolabels for Highly Sensitive and Specific Immunoassay

This work reports a novel electrochemical immunoassay protocol with signal amplification for determination of low-abundance protein (free prostate-specific antigen, PSA, used as a model) with high sensitivity and high selectivity by coupling metal sulfide (PbS)-based nanolabels with cleavage of the corresponding lead ion-induced DNAzymes. The assay mainly consists of an antigen–antibody immunoreaction with metal nanolabel in a transparent 96-well polystyrene microplate, the release of metal ions from the nanolabel, and cleavage of metal ion-induced DNAzyme. The signal is amplified by the labeled redox tag (ferrocene) on the DNAzyme-based sensor. In the presence of target analyte, the sandwiched immunocomplex can be formed between the primary antibody on the microplate and the corresponding metal sulfide nanolabel. The carried nanolabel can release numerous metal ions by acid, and induce the cleavage of the corresponding DNAzyme, thus resulting in the change of electrochemical signal. Under optimal conditions, the DNAzyme-based immunoassay presents an obvious electrochemical response for the detection of PSA, and allows detection of PSA at a concentration as low as 0.1 pg mL^–1. Intra-assay and interassay coefficients of variation (CV) were less than 9.5% and 10%, respectively. No significant differences at the 0.05 significance level were encountered in the analysis of 12 clinical serum specimens between the developed immunoassay and a commercially available enzyme-linked immunosorbent assay (ELISA)

Preparation and evaluation of goose reovirus inactivated vaccine

Abstract Background Infection with Goose Reovirus (GRV) can cause serious economic losses in the goose breeding industry. In this study, the GRV allantoic fluid was concentrated and used as an antigen in a formalin-inactivated oil-emulsion vaccine. Results When 6 day-old geese were inoculated, antibodies against GRV became detectable at 6 days post-vaccination, their concentration peaked at 3 weeks. These antibodies were maintained for longer than 2 weeks. As the most susceptible age for GRV infection is birds under 2 weeks of age this vaccine should provide adequate cover for the most at risk birds. When geese were exposed to reovirus at different time intervals after immunization, the data revealed that the vaccine can provide a protection rate of 80%. The developed vaccine has good stability and could be stored at 4 °C for at least 12 months. Conclusion These results indicate that the developed GRV vaccine is safe, effectively absorbed, efficacious in inducing a rapid immune response, and effective in controlling GRV infection. Our results should be useful for the application of vaccines for controlling GRV in different goose flocks

Genome-Wide Identification and Characterization of the Mitochondrial Transcription Termination Factors (mTERFs) in Capsicum annuum L.

Mitochondrial transcription termination factors (mTERFs) regulate the expression of mitochondrial genes and are closely related to the function of the mitochondrion and chloroplast. In this study, the mTERF gene family in capsicum (Capsicum annuum L.) was identified and characterized through genomic and bioinformatic analyses. Capsicum was found to possess at least 35 mTERF genes (CamTERFs), which were divided into eight major groups following phylogenetic analysis. Analysis of CamTERF promoters revealed the presence of many cis-elements related to the regulation of cellular respiration and photosynthesis. In addition, CamTERF promoters contained cis-elements related to phytohormone regulation and stress responses. Differentially expressed genes in different tissues and developmental phases were identified using RNA-seq data, which revealed that CamTERFs exhibit various expression and co-expression patterns. Gene ontology (GO) annotations associated CamTERFs primarily with mitochondrion and chloroplast function and composition. These results contribute towards understanding the role of mTERFs in capsicum growth, development, and stress responses. Moreover, our data assist in the identification of CamTERFs with important functions, which opens avenues for future studies

Reduced Ectopic Pregnancy Rate on Day 5 Embryo Transfer Compared with Day 3: A Meta-Analysis

<div><p>Objective</p><p>To compare the risk of ectopic pregnancy (EP) after embryo transfer on day 3(D3-ET) and day 5(D5-ET).</p><p>Design</p><p>Meta-analysis</p><p>Patients</p><p>Women with pregnancy resulting from in vitro undergoing in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI)</p><p>Result(s)</p><p>Twenty-two studies were identified through research conducted using the PubMed, Embase, and Cochrane databases and ClinicalTrials.gov. All studies were conducted prior to October 2016. Adding the reproductive data from our center, a total of 143 643 pregnancies were reviewed(D3-ET: n = 62027,D5-ET:n = 81616). A lower EP rate was found in women undergoing D5-ET than in those undergoing D3-ET [relative risk (RR), 0.67;95% confidence interval (CI), 0.54–0.85;143643 pregnancies in 23 studies; I² = 67%]. These results were validated in subgroups of fresh embryo-transfer (Fre-ET) cycles [RR, 0.78; 95%CI, 0.69–0.88; 91 871 pregnancies in 21 studies; I² = 29%] and frozen-thawed embryo-transfer (Fro-ET) cycles [RR, 0.43; 95%CI, 0.36–0.51; 51 772 pregnancies in 10 studies; I² = 33%]. After separating out the randomized controlled trials (RCTs), a significant difference was found in the retrospective studies in both subgroups [both Fre-ET (RR,0.78;95% CI 0.69–0.88);91182 pregnancies in 14 studies; I² = 45%] and Fro-ET(RR,0.43;95% CI 0.36–0.51; 51751pregnancies in 9 studies;I² = 33%)], while the RCTs showed no statistical significance for Fre-ET cycles[RR,0.86;95% CI 0.32–2.26); 689 pregnancies in 7 studies; I² = 0%].</p><p>Conclusion(s)</p><p>The present study indicates that D5-ET reduces the risk for EP in cycles that use IVF or ICSI, compared with D3-ET. It suggests that D5-ET may be a better choice for decreasing the EP rate in assisted reproductive technology. Further high-quality randomized controlled trials are anticipated.</p></div