Unraveling the mechanism of flower color variation in Brassica napus by integrated metabolome and transcriptome analyses.

Brassica napus is one of the most important oil crops in the world. Breeding oilseed rape with colorful flowers can greatly enhance the ornamental value of B. napus and thus improve the economic benefits of planting. As water-soluble flavonoid secondary metabolites, anthocyanins are very important for the synthesis and accumulation of pigments in the petals of plants, giving them a wide range of bright colors. Despite the documentation of over 60 distinct flower shades in B. napus, the intricacies underlying flower color variation remain elusive. Particularly, the mechanisms driving color development across varying flower color backgrounds necessitate further comprehensive investigation. This research undertook a comprehensive exploration through the integration of transcriptome and metabolome analyses to pinpoint pivotal genes and metabolites underpinning an array of flower colors, including beige, beige-red, yellow, orange-red, deep orange-red, white, light-purple, and purple. First, we used a two-way BLAST search to find 275 genes in the reference genome of B. napus Darmor v10 that were involved in making anthocyanins. The subsequent scrutiny of RNA-seq outcomes underscored notable upregulation in the structural genes F3H and UGT, alongside the MYB75, GL3, and TTG1 transcriptional regulators within petals, showing anthocyanin accumulation. By synergizing this data with a weighted gene co-expression network analysis, we identified CHS, F3H, MYB75, MYB12, and MYB111 as the key players driving anthocyanin synthesis in beige-red, orange-red, deep orange-red, light-purple, and purple petals. By integrating transcriptome and weighted gene co-expression network analysis findings with anthocyanin metabolism data, it is hypothesized that the upregulation of MYB75, which, in turn, enhances F3H expression, plays a pivotal role in the development of pigmented oilseed rape flowers. These findings help to understand the transcriptional regulation of anthocyanin biosynthesis in B. napus and provide valuable genetic resources for breeding B. napus varieties with novel flower colors.

Complement factor C1q mediates vascular endothelial dysfunction in STZ-induced diabetic mice.

Diabetes is a significant global public health issue with implications for vascular endothelial cells (ECs) dysfunction and the subsequent development and advancement of diabetic complications. This study aims to compare the cellular and molecular properties of the aorta in normal and streptozotocin (STZ)-induced diabetic mice, with a focus on elucidating potential mechanism underlying EC dysfunction. Here, we performed a single-cell RNA sequencing survey of 32,573 cells from the aorta of normal and STZ-induced diabetic mice. We found a compendium of 10 distinct cell types, mainly ECs, smooth muscle cells (SMCs), fibroblast, pericyte, immune cells and stromal cells. As the diabetes condition progressed, we observed a subpopulation of aortic ECs that exhibited significantly elevated expression of complement (C) molecule C1qa compared to their healthy counterparts. This increased expression of C1qa was found to induce reactive oxygen species (ROS) production, facilitate EC migration and increased permeability, and impair the vasodilation within the aortic segment of mice. Furthermore, AAV-Tie2-shRNA-C1qa was administered into diabetic mice by tail vein injection, showing that inhibition of C1qa in the endothelium led to a reduction in ROS production, decreased vascular permeability, and improved vasodilation. Collectively, these findings highlight the crucial involvement of C1qa in endothelial dysfunction associated with diabetes.

Ru-regulated electronic structure CoNi-MOF nanosheets advance water electrolysis kinetics in alkaline and seawater media.

Herein, an ion-exchange strategy is utilized to greatly improve the kinetics of hydrogen evolution reaction (HER) and oxygen evolution reaction (OER) by Ru-modified CoNi- 1,3,5-Benzenetricarboxylic acid (BTC)-metal organic framework nanosheets (Ru@CoNi-MOF). Due to the higher Ni active sites and lower electron transfer impedance, Ru@CoNi-MOF catalyst requires the overpotential as low as 47 and 279 mV, at a current density of 10 mA/cm2 toward HER and OER, respectively. Significantly, the mass activity of Ru@CoNi-MOF for HER and OER are 25.9 and 10.6 mA mg-1, nearly 15.2 and 8.8 times higher than that of Ni-MOF. In addition, the electrolyzer of Ru@CoNi-MOF demonstrates exceptional electrolytic performance in both KOH and seawater environment, surpasses the commercial Pt/C||IrO2 couple. Theoretical calculations prove that introducing Ru atoms in - CoNi-MOF modulates the electronic structure of Ni, optimizes adsorption energy for H* and reduces energy barrier of metal organic frameworks (MOFs). This modification significantly improves the kinetic rate of the Ru@CoNi-MOF during water splitting. Certainly, this study highlights the utilization of MOF nanosheets as advanced HER/OER electrocatalysts with immense potential, and will paves a way to develop more efficient MOFs for catalytic applications.

Aminopoly(carboxylic acid)-Functionalized PolyHIPE Beads toward Eliminating Trace Heavy Metal Ions from Water.

Many advanced materials are designed for the removal of heavy metal ions from water. However, materials for eliminating trace heavy metal ions from wastewater to meet drinking water standards remain a major challenge. Herein, epoxy group-functionalized open-cellular beads are synthesized by UV polymerization of a water-in-oil-in-water system. The epoxy groups are further transformed into diethylenetriaminepentaacetic acid (DTPA) with hexamethylene diamine as a bridging agent. The resulting material (DTPA@polyHIPE beads) can eliminate trace Cu(II), Cr(III), Pb(II), Fe(III), or Cd(II) from water. When 0.15 g of DTPA@polyHIPE beads are used to adsorb metal ions of 20 mg in 100 mL of water, the residue concentrations of Cu(II), Cr(III), Pb(II), Fe(III), and Cd(II) are reduced to 0.08, 0.06, 0.02, 0.09, and 0.07 mg/L, respectively. The adsorption efficiencies of the beads for these ions are all higher than 99.55%. The adsorbent is durable and exhibits good recyclability by retaining an adsorption capacity of ≥91% after 5 cycles. The negative values of ΔG in the adsorption process indicate that the adsorption is feasible and spontaneous. The chemical adsorption follows the Freundlich adsorption model, indicating a multilayer heterogeneous adsorption. The DTPA@polyHIPE beads have a great potential application in dealing with trace heavy metal ion polluted water.

Single-Cell RNA-Seq Reveals Coronary Heterogeneity and Identifies CD133+TRPV4high Endothelial Subpopulation in Regulating Flow-Induced Vascular Tone in Mice.

BACKGROUND: Single-cell RNA-Seq analysis can determine the heterogeneity of cells between different tissues at a single-cell level. Coronary artery endothelial cells (ECs) are important to coronary blood flow. However, little is known about the heterogeneity of coronary artery ECs, and cellular identity responses to flow. Identifying endothelial subpopulations will contribute to the precise localization of vascular endothelial subpopulations, thus enabling the precision of vascular injury treatment. METHODS: Here, we performed a single-cell RNA sequencing of 31 962 cells and functional assays of 3 branches of the coronary arteries (right coronary artery/circumflex left coronary artery/anterior descending left coronary artery) in wild-type mice. RESULTS: We found a compendium of 7 distinct cell types in mouse coronary arteries, mainly ECs, granulocytes, cardiac myocytes, smooth muscle cells, lymphocytes, myeloid cells, and fibroblast cells, and showed spatial heterogeneity between arterial branches. Furthermore, we revealed a subpopulation of coronary artery ECs, CD133+TRPV4high ECs. TRPV4 (transient receptor potential vanilloid 4) in CD133+TRPV4high ECs is important for regulating vasodilation and coronary blood flow. CONCLUSIONS: Our study elucidates the nature and range of coronary arterial cell diversity and highlights the importance of coronary CD133+TRPV4high ECs in regulating coronary vascular tone.

Peptonics: A new family of cell-protecting surfactants for the recombinant expression of therapeutic proteins in mammalian cell cultures.

Polymer surfactants are key components of cell culture media as they prevent mechanical damage during fermentation in stirred bioreactors. Among cell-protecting surfactants, Pluronics are widely utilized in biomanufacturing to ensure high cell viability and productivity. Monodispersity of monomer sequence and length is critical for the effectiveness of Pluronics-since minor deviations can damage the cells-but is challenging to achieve due to the stochastic nature of polymerization. Responding to this challenge, this study introduces Peptonics, a novel family of peptide and peptoid surfactants whose monomer composition and sequence are designed to achieve high cell viability and productivity at a fraction of chain length and cost of Pluronics. A designed ensemble of Peptonics was initially characterized via light scattering and tensiometry to select sequences whose phase behavior and tensioactivity align with those of Pluronics. Selected sequences were evaluated as cell-protecting surfactants using Chinese hamster ovary (CHO) cells expressing therapeutic monoclonal antibodies (mAb). Peptonics IH-T1010, ih-T1010, and ih-T1020 afforded high cell density (up to 3 × 107 cells mL-1 ) and viability (up to 95% within 10 days of culture), while reducing the accumulation of ammonia (a toxic metabolite) by ≈10% compared to Pluronic F-68. Improved cell viability afforded high mAb titer (up to 5.5 mg mL-1 ) and extended the production window beyond 14 days; notably, Peptonic IH-T1020 decreased mAb fragmentation and aggregation ≈5%, and lowered the titer of host cell proteins by 16% compared to Pluronic F-68. These features can improve significantly the purification of mAbs, thus increasing their availability at a lower cost to patients.

Genetics in Ischemic Stroke: Current Perspectives and Future Directions.

Ischemic stroke is a heterogeneous condition influenced by a combination of genetic and environmental factors. Recent advancements have explored genetics in relation to various aspects of ischemic stroke, including the alteration of individual stroke occurrence risk, modulation of treatment response, and effectiveness of post-stroke functional recovery. This article aims to review the recent findings from genetic studies related to various clinical and molecular aspects of ischemic stroke. The potential clinical applications of these genetic insights in stratifying stroke risk, guiding personalized therapy, and identifying new therapeutic targets are discussed herein.

Analysis of Altered Flowering Related Genes in a Multi-Silique Rapeseed (Brassica napus L.) Line zws-ms Based on Combination of Genome, Transcriptome and Proteome Data.

Based on previous researches, we further investigated the multi-silique trait in rapeseed (Brassica napus L.) line zws-ms. In this study, we used a relatively comprehensive list of flowering related genes in rapeseed and compared them between zws-ms and its near-isogenic line (NIL) zws-217. Genes were studied on genome, transcriptome and proteome levels and then we focused on genes with non-synonymous single nucleotide polymorphism (SNP) or frame-shift insertion-deletion (InDel), finding some genes on the list which changes their sequences. Then, combined with their annotation and the information of their orthologs, certain genes such as BnaA09g05900D, ortholog of AGAMOUS-LIKE 42 (AGL42), which encodes an MADS-box protein, were assumed as probably responsible for the multi-silique trait. Also, we analyzed the Differentially Accumulated Proteins (DAPs) between zws-ms and zws-217, revealing some genes involved in homologous recombination and mismatch repair pathways. Since the development of flowers/siliques is crucial to crops and it influences the yield of rapeseed, this study paved a way to deeply understand the mechanism of the multi-pistil flower formation, which may facilitate researches on rapeseed production in future.

Relationship Between Platelets and the Clinical Efficacy of Umbilical Cord Mesenchymal Stem Cells for HBV-Related Acute-on-Chronic Liver Failure and Liver Cirrhosis: A Preliminary Clinical Study.

BACKGROUND: Previous studies have found that the production of platelets could enhance the therapeutic effects of stem cells. Nevertheless, there are still no articles reporting on the relationship between platelets and the clinical efficacy of umbilical cord mesenchymal stem cells (UCMSCs) for HBV-related acute-on-chronic liver failure (ACLF) and liver cirrhosis (LC). METHODS: In this retrospective observational study, patients who met the criteria were included. Patients were divided into subgroups according to the aims of this study. In the first part, the platelet count changes of ACLF and patients with LC after UCMSC therapy were compared and analyzed. Subgroup analysis based on UCMSC infusion times and patient age was also performed. In the second part, patients in the ACLF group and LC group were further divided into subgroups according to their platelet levels. Their clinical characteristics, demographics, and biochemical factors were compared. RESULTS: This study enrolled 64 patients with ACLF and 59 patients with LC. In both groups, platelet levels declined similarly. Compared with the short-course UCMSC treatment group (≤4 times), patients with ACLF and patients with LC with long-course UCMSC treatment (>4 times) showed an overall increasing trend. Younger patients with LC (<45 years) had significantly higher platelet levels than older patients with LC (≥45 years). However, this age difference was not present in the ACLF group. The median TBIL decrease and cumulative TBIL decrease were not significantly different between patients with high PLT and patients with low PLT after UCMSC transfusions. For patients with ACLF, the cumulative TBIL decrease and the median TBIL decrease were significantly greater than those of patients with LC at the same platelet level after UCMSC treatment. However, this difference was not observed at all time points. CONCLUSION: Trend of the platelet levels for HBV-related patients with ACLF and LC after UCMSC treatment did not parallel and varied according to treatment times and patients' age. Platelet levels did not affect the efficacy of MSCs for patients with ACLF or LC.

Functional role of coupling the endothelial TRPV4 and KCa 3.1 channels in regulating coronary vascular tone.

BACKGROUND AND PURPOSE: High-fat diet (HFD) induces dysregulated pathways in coronary artery endothelial cells (CAECs), which leads to altered regulation of vascular tone, tissue perfusion and increases the risk of coronary artery diseases. Ca2+ -activated K+ (KCa ) channels are known to be associated with transient receptor potential (TRP) channels, which are important for regulating endothelial function. But how TRPV4 channels interacts with KCa channels in regulating coronary vascular tone in HFD mice requires further exploration. EXPERIMENTAL APPROACH: TRPV4 channel activity was assessed by fluorescent Ca2+ imaging. Interactions between TRPV4 and KCa 3.1 channels were verified by co-immunoprecipitation and immunofluorescence resonance energy transfer (FRET), and their binding site was found by site-directed mutagenesis. Endothelium-specific TRPV4 knockout (TRPV4EC -/- ) mice were used to study the effect of the interactions between TRPV4-KCa 3.1 channels on coronary vascular tone. Coronary blood flow was measured by Doppler ultrasound device. KEY RESULTS: TRPV4 channels were involved in regulating coronary vascular tone, through coupling with a Ca2+ -sensitive K+ channel (KCa 3.1) in CAECs, affecting vasodilation and coronary blood flow. In mice fed a HFD diet, the coupling was damaged by a high concentration of plasma 1-heptadecanoyl-2-hydroxy-sn-glycero-3-phosphocholine. Using a bridging approach, we then identified folic acid as an effective drug to repair the uncoupled TRPV4-KCa 3.1 channels and to improve coronary arterial function. CONCLUSION AND IMPLICATIONS: Our data highlight the importance of coupling between TRPV4 and KCa 3.1 channels in the regulation of coronary vascular tone and provide a novel strategy for developing new drugs to reduce the incidence of cardiovascular events.

Altered electronic structure of trimetallic FeNiCo-MOF nanosheets for efficient oxygen evolution.

Theoretical calculation results unveil that the reconstructed Co(Ni)OOH on FeNiCo-MOF during OER processes is beneficial to improve the OER activity. Experimentally, to achieve 2D trimetallic FeNiCo-MOF nanosheets, a facile room-temperature dispersion approach is employed. Such 2D nanosheets reveal an OER overpotential as low as 239 mV at 10 mA cm-2 and excellent long-term stability in 1M KOH. Undoubtedly, this work highlights the great potential of directly utilizing MOF nanosheets as OER electrocatalysts.

The Role of Hydrogen Sulfide in the Development and Progression of Lung Cancer.

Lung cancer is one of the 10 most common cancers in the world, which seriously affects the normal life and health of patients. According to the investigation report, the 3-year survival rate of patients with lung cancer is less than 20%. Heredity, the environment, and long-term smoking or secondhand smoke greatly promote the development and progress of the disease. The mechanisms of action of the occurrence and development of lung cancer have not been fully clarified. As a new type of gas signal molecule, hydrogen sulfide (H2S) has received great attention for its physiological and pathological roles in mammalian cells. It has been found that H2S is widely involved in the regulation of the respiratory system and digestive system, and plays an important role in the occurrence and development of lung cancer. H2S has the characteristics of dissolving in water and passing through the cell membrane, and is widely expressed in body tissues, which determines the possibility of its participation in the occurrence of lung cancer. Both endogenous and exogenous H2S may be involved in the inhibition of lung cancer cells by regulating mitochondrial energy metabolism, mitochondrial DNA integrity, and phosphoinositide 3-kinase/protein kinase B co-pathway hypoxia-inducible factor-1α (HIF-1α). This article reviews and discusses the molecular mechanism of H2S in the development of lung cancer, and provides novel insights for the prevention and targeted therapy of lung cancer.

Insulin resistance in school-aged girls with overweight and obesity is strongly associated with elevated white blood cell count and absolute neutrophil count.

Background: The primary objective of the study was to discuss the sex differences in insulin resistance-induced changes in metabolic and inflammatory markers in school-aged children with overweight and obesity. Methods: A cross-sectional study of 800 children aged seven and twelve years was performed. Questionnaires, anthropometric data and fasting blood samples were collected. Results: Children with overweight and obesity showed statistically significant differences in multiple metabolic and inflammatory markers compared with children with normal BMI. The correlation coefficient (r) between white blood cell count, absolute neutrophil count, fasting plasma insulin, HOMA-IR, HOMA-ß, triglyceride, HDL-C, triglyceride/HDL ratio, alanine transaminase, serum uric acid, systolic blood pressure and BMI were higher in all children, but the linear relationships between white blood cell count, absolute neutrophil count and BMI were stronger in girls with overweight and obesity than in boys with overweight and obesity. Subsequently, HOMA-IR was shown to be more strongly associated with increased white blood cell count and absolute neutrophil count in school-aged girls with overweight and obesity by partial correlation analysis and the multiple linear regression analysis. Conclusions: Elevated white blood cell count and absolute neutrophil count in children with overweight and obesity, especially girls, can serve as markers of insulin resistance.

Interferon add-on therapy increased clinical cure significantly for interferon-experienced chronic hepatitis B patients with low HBsAg.

Currently, interferon add-on therapy brings hope for clinical cure of chronic hepatitis B patients with low HBsAg. However, in clinical practice patients with poor responses to their first interferon therapy were often switched to nucleos(t)ide analog therapy and then labeled as unsuitable patients for interferon therapy. Even if their HBsAg levels dropped to a low level, they were reluctant or not recommended to take interferon again, which caused them to miss out on interferon add-on therapy and clinical cure. Therefore, it is urgent to elucidate the effectiveness of interferon add-on therapy to get clinical cure for these interferon-experienced patients with low HBsAg. The purpose of this study was to investigate whether interferon-experienced patients could achieve the same HBsAg clearance and HBsAg seroconversion rates as interferon-naive patients. Also, the associated factor of HBsAg clearance and seroconversion were aimed to be clarified. 292 patients, including 85 interferon-experienced patients, were enrolled with HBsAg< 1500 IU/ml, HBeAg negative and HBV-DNA negative. And then, peg-interferon α-2b add-on therapy was performed. The results showed that the week 48 HBsAg clearance and seroconversion rates of all patients were 29.8% and 22.0%. There was no statistically significant difference between interferon-experienced and interferon-naive patients in week 48 HBsAg clearance and seroconversion rates, suggesting satisfactory clinical cure of the interferon add-on therapy for interferon-experienced patients. The age, baseline HBsAg, and week 12 HBsAg were negative correlated factors for week 48 HBsAg clearance and seroconversion. Furthermore, the age, baseline HBsAg and week 12 HBsAg for predicting the week 48 HBsAg clearance were cut off at 40.5 years, at 152.0 IU/ml and at 34.99 IU/ml, and for predicting seroconversion were cut off at 40.5 years, at 181.9 IU/ml and at 34.99 IU/ml, correspondingly. Significantly, interferon-experienced patients with low HBsAg were suggested with interferon add-on therapy to achieve clinical cure as soon as possible. This research provided evidences and cut-offs for the interferon add-on therapy against chronic hepatitis B.

Co0.9Co0.1S Nanorods with an Internal Electric Field and Photothermal Effect Synergistically for Boosting Photocatalytic H2 Evolution.

The paper reports a strategy to synthesize Cd0.9Co0.1S nanorods (NRs) via a one-pot solvothermal method. Remarkably, the pencil-shaped Cd0.9Co0.1S NRs with a large aspect ratio and good polycrystalline plane structure significantly shorten the photogenerated carrier transfer path and achieve fast separation. An appropriate amount of Co addition enhances visible light-harvesting and generates a photothermal effect to improve the surface reaction kinetics and increases the charge transfer rate. Moreover, the internal electric field facilitates the separation and transfer of carriers and effectively impedes their recombination. As a result, the optimized Cd0.9Co0.1S NRs yield a remarkable H2 evolution rate of 8.009 mmol·g-1·h-1, which is approximately 7.2 times higher than that of pristine CdS. This work improves the photocatalytic hydrogen production rate by tuning and optimizing electronic structures through element addition and using the photothermal synergistic effect.

Physiological levels of fluid shear stress modulate vascular function through TRPV4 sparklets.

Endothelial calcium (Ca 2+) signaling plays a major role in regulating vasodilation in response to fluid shear stress (FSS) generated by blood flow. Local Ca 2+ influx through single transient receptor potential channel subfamily V member 4 (TRPV4) (termed "sparklets") activated by low concentrations of chemical and biological stimuli has been revealed to modulate vascular function. However, the range in which FSS can initiate TRPV4 sparklets to induce vasodilation is unknown. Here, we assess the activity of TPRV4 sparklets induced by various physiological levels of FSS and investigate the mechanisms involving these Ca 2+ signals in FSS-induced vasodilation. Intact small mesenteric arteries are used for Ca 2+ imaging with a GCaMP2(TRPV4-KO) mouse model and high-speed confocal systems. Markedly increased local Ca 2+ signals are observed in the endothelium under 4-8 dyne/cm 2 FSS, whereas FSS >8 dyne/cm 2 causes global Ca 2+ influx. Further analysis shows that TRPV4 channels form a four-channel group to mediate Ca 2+ sparklets under certain levels of FSS. The large Ca 2+ influx hyperpolarizes endothelial cells by stimulating intermediate (IK)- and small (SK)-conductance Ca 2+-sensitive potassium channels, leading to hyperpolarization of the surrounding smooth muscle cells and ultimately causing endothelium-dependent vasodilation. In conclusion, Ca 2+ influx transits through a small number of endothelial TRPV4 channels opened by certain levels of FSS, which activates the Ca 2+-sensitive IK and SK channels to cause vasodilation.

Transcription factor WRKY28 curbs WRKY33-mediated resistance to Sclerotinia sclerotiorum in Brassica napus.

Sclerotinia sclerotiorum causes substantial damage and loss of yield in oilseed rape (Brassica napus). The molecular mechanisms of oilseed rape defense against Sclerotinia remain elusive. In this study, we found that in the early stages of B. napus infection a conserved mitogen-activated protein kinase (MAPK) cascade mediated by BnaA03.MKK5-BnaA06.MPK3/BnaC03.MPK3 module phosphorylates the substrate BnWRKY33, enhancing its transcriptional activity. The activated BnWRKY33 binds to its own promoter and triggers a transcriptional burst of BnWRKY33, thus helping plants effectively resist the pathogenic fungi by enhancing the expression of phytoalexin synthesis-related genes. The expression of BnWRKY33 is fine-tuned during defense. Ongoing Sclerotinia infection induces BnaA03.WRKY28 and BnaA09.VQ12 expression. BnaA09.VQ12 interacts physically with BnaA03.WRKY28 to form a protein complex, causing BnaA03.WRKY28 to outcompete BnWRKY33 and bind to the BnWRKY33 promoter. BnaA03.WRKY28 induction suppresses BnWRKY33 expression in the later stages of infection but promotes branch formation in the leaf axils by regulating the expression of branching-related genes such as BnBRC1. BnaA03.WRKY28 participates in the trade-off between defense and growth. These findings suggest that oilseed rape plants may modulate defense-response strength and develop alternative reproduction and survival strategies in the face of lethal pathogens.

TRPC5 mediates endothelium-dependent contraction in the carotid artery of diet-induced obese mice.

Little is known about the contribution of the transient receptor potential canonical channel isoform 5 (TRPC5), a Ca2+-sensitive channel, to vasoconstriction in obesity. In this study, we found that the TRPC5 expression and carotid artery contraction of diet-induced obese (DIO) mice were significantly higher than those of wild-type mice. Endothelium-dependent vasocontraction was inhibited by the TRPC5 inhibitor clemizole and the knockout of TRPC5 in DIO mouse carotid arteries, while activation of TRPC5 enhanced contraction in wild-type mice. TRPC5-regulated vasocontraction can be inhibited by the ROS scavenger NAC and the COX-2 inhibitor NS-398. Our study suggested that upregulation of TRPC5 contributes to endothelium-dependent contraction, which is involved in ROS production and COX-2 expression in DIO mouse carotid arteries. From these results, we speculated that TRPC5 mediated endothelium-dependent contraction in the carotid artery of DIO mice, which was achieved by increasing the levels of ROS and COX-2 expression.

BnaA03.MKK5-BnaA06.MPK3/BnaC03.MPK3 Module Positively Contributes to Sclerotinia sclerotiorum Resistance in Brassica napus.

Brassica napus (oilseed rape) is one of the most important oil crops worldwide, but its growth is seriously threatened by Sclerotinia sclerotiorum. The mechanism of oilseed rape response to this pathogen has rarely been studied. Here, it was identified that BnaA03.MKK5 whose expression was induced by S. sclerotiorum infection was involved in plant immunity. BnaA03.MKK5 overexpression lines exhibited decreased disease symptoms compared to wild-type plants, accompanied by the increased expression of camalexin-biosynthesis-related genes, including BnPAD3 and BnCYP71A13. In addition, two copies of BnMPK3 (BnA06.MPK3 and BnC03.MPK3) were induced by Sclerotinia incubation, and BnaA03.MKK5 interacted with BnaA06.MPK3/BnaC03.MPK3 in yeast. These interactions were confirmed using in vivo co-immunoprecipitation assays. In vitro phosphorylation assays showed that BnaA06.MPK3 and BnaC03.MPK3 were the direct phosphorylation substrates of BnaA03.MKK5. The transgenic oilseed rape plants including BnaA06.MPK3 and BnaC03.MPK3 overexpression lines and BnMPK3 gene editing lines mediated by CRISPR/Cas9 were generated; the results of the genetic transformation of BnaA06.MPK3/BnaC03.MPK3 indicate that BnMPK3 also has a positive role in Sclerotinia resistance. This study provides information about the potential mechanism of B. napus defense against S. Sclerotiorum mediated by a detailed BnaA03.MKK5-BnaA06.MPK3/BnaC03.MPK3 module.