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Cotton production is hampered by a variety of abiotic stresses that wreak havoc on the growth and development of plants, resulting in significant financial losses. According to reports, cotton production areas have declined around the world as a result of the ongoing stress. Therefore, plant breeding programs are concentrating on abiotic stress-tolerant cotton varieties. Mitogen-activated protein kinase (MAPK) cascades are involved in plant growth, stress responses, and the hormonal signaling pathway. In this research, three abiotic stresses (cold, drought, and salt) were analyzed on GhMPK3 transformed Arabidopsis plants. The transgenic plant’s gene expression and morphologic analysis were studied under cold, drought, and salt stress. Physiological parameters such as relative leaf water content, excised leaf water loss, chlorophyll content, and ion leakage showed that overexpressed plants possess more stable content under stress conditions compared with the WT plants. Furthermore, GhMPK3 overexpressed plants had greater antioxidant activities and weaker oxidant activities. Silencing GhMPK3 in cotton inhibited its tolerance to drought stress. Our research findings strongly suggest that GhMPK3 can be regarded as an essential gene for abiotic stress tolerance in cotton plants.

Main conclusion: Transcriptional regulation of stress-responsive genes is a crucial step in establishing the mechanisms behind plant abiotic stress tolerance. A sensitive method of regulating transcription factors activity, stability, protein interaction, and subcellular localization is through phosphorylation. This review highlights a widespread regulation mechanism that involves phosphorylation of plant TFs in response to abiotic stress. Abiotic stress is one of the main components limiting crop yield and sustainability on a global scale. It greatly reduces the land area that is planted and lowers crop production globally. In all living organisms, transcription factors (TFs) play a crucial role in regulating gene expression. They participate in cell signaling, cell cycle, development, and plant stress response. Plant resilience to diverse abiotic stressors is largely influenced by TFs. Transcription factors modulate gene expression by binding to their target gene's cis-elements, which are impacted by genomic characteristics, DNA structure, and TF interconnections. In this review, we focus on the six major TFs implicated in abiotic stress tolerance, namely, DREB, bZIP, WRKY, ABF, MYB, and NAC, and the cruciality of phosphorylation of these transcription factors in abiotic stress signaling, as protein phosphorylation has emerged as one of the key post-translational modifications, playing a critical role in cell signaling, DNA amplification, gene expression and differentiation, and modification of other biological configurations. These TFs have been discovered after extensive study as stress-responsive transcription factors which may be major targets for crop development and important contributors to stress tolerance and crop production. [ABSTRACT FROM AUTHOR]

The gastric tissue is often damaged by ingesting large amounts of alcohol. Scutellarin (SCU) is a flavonoid with anti-inflammatory and antioxidant properties. However, its potential effect on alcohol-induced gastric injury has not been investigated. Therefore, the present study investigated its protective effects on gastric injury. Twenty-five male BALB/c mice were divided into five groups, including the control group, alcohol group, SCU (10 mg kg-1, 25 mg kg-1, 50 mg kg-1) combined with alcohol gavage groups. Haematoxylin and eosin (H&E) were used for histopathological observation in gastric tissues. The levels of superoxide dismutase (SOD), catalase (CAT), malondialdehyde (MDA), and inducible nitric oxide synthase (iNOS) in gastric tissues were measured by respective biochemical kits. We further investigated the gene expression of interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) using real-time quantitative PCR (RT-qPCR). The results demonstrated that alcohol stimulation leads to the production of inflammatory cells. In addition, alcohol stimulation resulted in a significant increase in MDA levels and an obvious decrease in SOD and CAT levels. SCU improves oxidative stress in the gastric by reducing MDA and increasing SOD and CAT levels. It also showed a mitigating effect on the inflammatory response by reducing the transcript levels of Il-1β, Il-6, and Tnf-α, as well as the activity of iNOS. SCU enhances the gastric response to oxidative stress and effectively reduces the inflammatory response. Overall, our data suggest that SCU may be an attractive therapeutic strategy to mitigate the development of gastric tissue injury. [ABSTRACT FROM AUTHOR]

Introduction: Neospora caninum, an obligate intracellular parasite of the phylum Apicomplexa, typically causes an illness known as neosporosis. Dense granule proteins (GRAs) are secreted by apicomplexan and constitute the parasitophorous vacuoles (PVs) structure where tachyzoites proliferate after invasion into host cells. In Toxoplasma gondii, TgGRA3 proteins are strongly associated with PVs membrane and enhance its virulence in vivo, however, research on NcGRA3 has not been reported. Methods: Here, a novel NcGRA3 protein in N. caninum was discovered using bioinformatics analysis; the location of NcGRA3 was determined in the extracellular Nc-1 tachyzoites and intracellular PVs after invasion using immunofluorescence assays; the NcGRA3 protein existing form in the PVs membranes was analyzed using western blotting. Results: NcGRA3 shared 41.67% nucleotide homology and 22.17% amino acid homology with TgGRA3. Amino acid sequences ranging from 1 to 25 were signal peptide regions and 135–157 were transmembrane domains. The immunofluorescence assays showed that NcGRA3 was an apical organ secreted dense granule protein and expressed at the posterior end of tachyzoites; the partial co-localization with NcGRA6 in PVs demonstrated that NcGRA3 were expressed in the intravacuolar network structure and PVs membrane. The western blotting assays showed that NcGRA3 protein behaved as an integrated transmembrane protein in PVs. Overall, characterization of the newly discovered NcGRA3 protein will lay a foundation for its function research in the further. [ABSTRACT FROM AUTHOR]

Vibrio alginolyticus is a food-borne marine Vibrio that causes gastroenteritis, otitis media, otitis externa, and septicemia in humans. The pathogenic mechanisms of V. alginolyticus have previously been studied in aquaculture animals; however, the underlying mechanisms in mammals remain unknown. In this study, an in vitro model of mouse peritoneal macrophages infected with V. alginolyticus was established. qPCR results revealed that V. alginolyticus induced the transcription levels of various cytokines, including IL-1β, IL-12, IL-18, TNF-α, IL-17, IL-6, IFN-γ, and IL-10, and the secretion level of IL-1β is the most significant. Inhibition assays with Ac-YVAD-CHO (a caspase-1 inhibitor) and Z-VAD-FMK (a pan-caspase inhibitor) were conducted to determine whether caspase-1 or caspase - 11 is involved in V. alginolyticus- triggered IL-1β secretion. Results showed that IL-1β secretion was partly inhibited by Ac-YVAD-CHO and absolutely blocked by Z-VAD-FMK. To explore the sensed pattern recognition receptors, several NLR family members and the AIM2 receptor were detected and many receptors were upregulated especially NLRP3. Moreover, the NLRP3 protein displayed a puncta-like surrounding cell nucleus, which signified that the NLRP3 inflammasome was activated in response to V. alginolyticus infection. Inhibition assays with glyburide and CA-074 methyl ester (K+ outflow inhibitor and cathepsin B inhibitor) blocked IL-1β secretion, which demonstrated the essential role of the NLRP3 inflammasome in inflammatory response. To better understand how V. alginolyticus affects IL-1β release, the NLRP3 inflammasome was detected with doses ranging from 0.1 to 10 MOIs and time periods ranging from 3 to 12 h. Results showed that V. alginolyticus -mediated NLRP3 inflammasome activation was in a time- and dose-dependent manner and IL-1β release peaked at MOI of 1 for 12 h. Most importantly, blocking the NLRP3 inflammasome with inhibitors and the use of NLRP3-/- and caspase-1/11-/- mice could attenuate pro-inflammatory cytokine secretion, such as IL-1β, IL-6, IL-12, and TNF-α. Taken together, our study first found that the NLRP3 inflammasome plays vital roles in V. alginolyticus triggered inflammatory response in mouse peritoneal macrophages. This may provide reference information for the development of potential anti-inflammatory treatments against V. alginolyticus infection. [ABSTRACT FROM AUTHOR]

Aeromonas sobria , a common conditional pathogenic bacteria, is widely distributed in the environment and causes gastroenteritis in humans or septicemia in fish. Of all Aeromonas species, A. sobria is the most frequently isolated from human infections especially in immunocompromised subjects. Innate immunity is the first protection system of organism to resist non-specific pathogens invasion; however, the immune response process of hosts against A. sobria infection re\mains unexplored. The present study established an A. sobria infection model using primary mouse peritoneal macrophages (PMφs). The adherence and cytotoxicity of A. sobria on PMφs were determined by May-Grünwald Giemsa staining and LDH release measurement. Pro-inflammatory cytokine expression levels were measured using qPCR, western blotting, and ELISA methods. We also investigated the levels of ASC oligomerization and determined the roles of active caspase-1 in IL-1β secretion through inhibition assays and explored the activated pattern recognition receptors through immunofluorescence. We further elucidated the roles of activated inflammasome in regulating the host's inflammatory response through inhibition combined with ELISA assays. Our results showed that A. sobria induced lytic cell death and LDH release, whereas it had no adhesive properties on PMφs. A. sobria triggered various proinflammatory cytokine transcription level upregulation, and IL-1β occupied the highest levels. The pro-IL-1β protein expression levels increased in a dose-dependent manner with MOI ranging from 1 to 100. This process was regulated by ASC-dependent inflammasome, which cleavage pro-IL-1β into active IL-1β p17 with activated caspase-1 p20. Meanwhile, the expression levels of NLRP3 receptor significantly increased, location analysis revealed puncta-like surrounding nuclear, and inhibition of NLRP3 inflammasome downregulated caspase-1 activation and IL-1β secretion. Blocking of NLRP3 inflammasome activation through K+ efflux and cathepsin B or caspase approaches downregulated A. sobria –induced proinflammatory cytokine production. Overall, these data indicated that A. sobria induced proinflammatory cytokine production in PMφs through activating NLRP3 inflammasome signaling pathways. [ABSTRACT FROM AUTHOR]

Summary: Identification of seed development regulatory genes is the key for the genetic improvement in rice grain quality. NF‐Ys are the important transcription factors, but their roles in rice grain quality control and the underlying molecular mechanism remain largely unknown. Here, we report the functional characterization a rice NF‐Y heterotrimer complex NF‐YB1‐YC12‐bHLH144, which is formed by the binding of NF‐YB1 to NF‐YC12 and then bHLH144 in a sequential order. Knock‐out of each of the complex genes resulted in alteration of grain qualities in all the mutants as well as reduced grain size in crnf‐yb1 and crnf‐yc12. RNA‐seq analysis identified 1496 genes that were commonly regulated by NF‐YB1 and NF‐YC12, including the key granule‐bound starch synthase gene Wx. NF‐YC12 and bHLH144 maintain NF‐YB1 stability from the degradation mediated by ubiquitin/26S proteasome, while NF‐YB1 directly binds to the 'G‐box' domain of Wx promoter and activates Wx transcription, hence to regulate rice grain quality. Finally, we revealed a novel grain quality regulatory pathway controlled by NF‐YB1‐YC12‐bHLH144 complex, which has great potential for rice genetic improvement. [ABSTRACT FROM AUTHOR]

Excessive use of hard-to-degrade pesticides threatens the ecological health of aquatic systems. This study aimed to investigate difenoconazole (DFZ) residues in the environment induced neurotoxicity in carp and the underlying mechanisms. A total of thirty-six carps were divided into three groups and exposed to 0, 0.5, and 2.0 mg/L DFZ for 96 h, respectively. The alterations in behavior and blood-brain barrier (BBB) were examined, and potential mechanisms were explored using immunological assays and biochemical methods. The results showed that DFZ exposure caused behavioral freezing, reduced feeding, and neuronal necrosis in carp. Mechanistically, DFZ triggered ROS accumulation and destroyed the balance between oxidation and antioxidation with increased lipid peroxidation product MDA contents and reduced antioxidant enzymes SOD and CAT activities in the carp brain by inhibiting the NF-E2-related factor 2 (Nrf2) pathway. The activation of oxidative stress further reduced tight junction proteins and MMP levels, thereby destroying BBB and leading to DFZ leakage into the brain. Increased BBB permeability additionally led to DFZ activation of nuclear factor kappa-B signaling-mediated inflammatory cytokine storm, exacerbating neuroinflammation. Meanwhile, DFZ exposure activated mitochondria-associated apoptosis in the carp's brain by up-regulating Bcl-2 associated X protein, cleaved-caspase3, and cytochrome C and decreasing B-cell lymphoma-2 levels. Interestingly, the carp's brain initiated a protective autophagic response via the PI3K/AKT/TOR pathway intending to counteract the neurotoxicity of DFZ. Overall, we concluded that accumulation of DFZ at high concentrations in the aquatic systems disrupted the BBB and resulted in neurotoxicity in carp through inhibition of Nrf2 pathway-mediated ROS accumulation. This study provides a reference for monitoring DFZ residues in the environment and a new target for the treatment of DFZ-induced neurotoxicity in carp. [Display omitted] • Difenoconazole (DFZ) resulted in altered behavior and brain tissue damage in carp. • Acute DFZ exposure inhibited Nrf2/Keap1 pathway and caused oxidative stress. • DFZ exposure caused ROS accumulation, BBB disruption, and DFZ leakage into the brain. • Disruption of BBB led to DFZ activation of inflammation via the NF-κB pathway. • Disruption of BBB led to DFZ activation of apoptosis via the mitochondrial pathway. [ABSTRACT FROM AUTHOR]

Notched belly grain (NBG) is a type of deformed grain shape that has been associated with inferior appearance and tastes in rice. NBG is coordinated by both environments and genetics. In this study, we report on the first map-based cloning of an NBG gene on chromosome 4, denoted NBG4, which is a novel allele of Dwarf 11 encoding a cytochrome P450 (CYP724B1) involved in brassinosteroid (BR) biosynthesis. A 10-bp deletion in the 7th exon knocked down the level of the NBG4 transcript and shifted the reading frame of the resulting protein. In addition to the dwarf and clustered panicle as previously reported in the allelic mutants, nbg4 grains also displayed retarded germination and NBG due to the physical constraint of deformed hulls caused by abnormal hull elongation. NBG4 is constitutively expressed with the highest level of expression in immature inflorescences. In all, 2294 genes were differentially expressed in nbg4 and wild-type (WT), and evidence is presented that NBG4 regulates OsPPS-2, OsPRA2, OsYUCCA1, sped1-D, and Dwarf that play critical roles in determining plant architecture, panicle development, and seed germination. This study demonstrated that NBG4 is a key node in the brassinosteroid-mediated regulation of rice grain shape. [ABSTRACT FROM AUTHOR]

PKA (protein lysine acetylation) is a key post-translational modification involved in the regulation of various biological processes in rice. So far, rice acetylome data is very limited due to the highly-dynamic pattern of protein expression and PKA modification. In this study, we performed a comprehensive quantitative acetylome profile on four typical rice tissues, i.e., the callus, root, leaf, and panicle, by using a mass spectrometry (MS)-based, label-free approach. The identification of 1536 acetylsites on 1454 acetylpeptides from 890 acetylproteins represented one of the largest acetylome datasets on rice. A total of 1445 peptides on 887 proteins were differentially acetylated, and are extensively involved in protein translation, chloroplast development, and photosynthesis, flowering and pollen fertility, and root meristem activity, indicating the important roles of PKA in rice tissue development and functions. The current study provides an overall view of the acetylation events in rice tissues, as well as clues to reveal the function of PKA proteins in physiologically-relevant tissues. [ABSTRACT FROM AUTHOR]