Expression Of Synthesis-related Genes Research Articles

Giant Triton Snail Charonia tritonis Macrophage-Expressed Gene 1 Protein Ct-Mpeg1: Molecular Identification, Expression Analysis, and Antimicrobial Activity

Macrophage-expressed gene 1 proteins (Mpeg1/Perforin-2 (PRF2)) are a family of pore-forming proteins (PFPs) which can form pores and destroy the cell membrane of invading pathogens. However, little information is available regarding the function of Mpeg1 in the giant triton snail Charonia tritonis. In this study, a homolog of Mpeg1 (Ct-Mpeg1) was identified in C. tritonis. The predicted protein of Ct-Mpeg1 contains several structural features known in Mpegs, including a membrane attack complex/perforin (MACPF) domain and single transmembrane region. The Ct-Mpeg1 gene was constitutively expressed in almost all tissues examined except in the proboscis, with the highest expression level observed in the mantle. As a typical pore-forming protein, Ct-Mpeg1 has antibacterial activities against Vibrio (including Vibrio alginolyticus and Vibrio parahaemolyticus). In addition, rCt-Mpeg1 challenge to V. alginolyticus represses the expression of most outer membrane protein synthesis-related genes and genes involved in the TCA cycle pathway, which will lead to reduced outer membrane protein synthesis and less energy capacity. This is the first report to characterize the macrophage-expressed gene 1 protein in C. tritonis, and these results suggest that macrophage-expressed gene 1 protein Ct-Mpeg1 is an important immune molecule of C. tritonis that is involved in the bacterial infection resistance of Vibrio, and this study may provide crucial basic data for the understanding of the innate immunity system of C. tritonis.

SIRT4 Expression Ameliorates the Detrimental Effect of Heat Stress via AMPK/mTOR Signaling Pathway in BMECs.

Sirtuin 4 (SIRT4), a member of the SIRT family, has been reported to be a key factor involved in antioxidant defense in mitochondria. This study aimed to explore the potential molecular mechanism via which SIRT4 regulates heat stress-induced oxidative stress and lactoprotein synthesis in bovine mammary epithelial cells (BMECs). Our results showed that SIRT4 was significantly decreased in heat stressed mammary tissue. Depletion of SIRT4 in BMECs induced the generation of ROS, which, as exhibited by the decreased activity of antioxidant enzymes, changed mitochondrial morphology through mediating protein and mRNA levels related to mitochondrial fission and fusion. Moreover, we found that depletion of SIRT4 or stress conditions inhibited the expression of milk proteins, as well as lipid and glucose synthesis-related genes, and activated the AMPK/mTOR signaling pathway. Increased SIRT4 expression was found to have the opposite effect. However, blocking the AMPK/mTOR signaling pathway could inhibit the regulatory function of SIRT4 in milk synthesis-related gene expression. In summary, our results suggest that SIRT4 may play critical roles in maintaining mammary gland function by regulating the AMPK/mTOR signaling pathway in dairy cows, indicating that SIRT4 may be a potential molecular target for curing heat stress-induced BMEC injury and low milk production in dairy cows.

Comparative analysis of amino acid content and protein synthesis-related genes expression levels in breast muscle among different duck breeds/strains

Evidences have found important effects of breeds/strains on the content of amino acids (AAs) which is an important substrate for protein synthesis and contributes greatly to meat quality. Therefore, the objective of the present study was to compare the AAs content and protein synthesis-related genes expression levels in breast muscle of native breed (Jianchang duck (J)), hybrid strains (BH1, BH2, and MC♂×(BGF2♂×GF2♀)♀ (MC)), and commercial breed (Cherry Verry duck). Results showed that a total of 17 AAs (TAAs) was detected from breast muscle among 5 duck breeds/strains including 11 essential AAs (EAAs). Among these AAs, the contents of Proline, Threonine, Glutamine, Serine, Methionine, Phenylalanine, Histidine, and Cysteine were significant difference among 5 duck breeds/strains. The contents of EAAs, TAAs, and flavor AAs were higher in breast muscle of J and BH2 than those in other duck breeds/strains, and the ratio of EAAs/TAAs was higher in breast muscle of BH2. Furthermore, the expression levels of eukaryotic translation initiation factor 4E-binding protein 1, mammalian target of rapamycin, and proton-coupled amino acid transporter 1 were the highest in breast muscle of BH2, and that of solute carrier family 38 member 2 was the highest in breast muscle of J. Meanwhile, principal component analysis results showed that principal component 1 of BH1, principal component 3 of BH2, and principal component 2 of MC were positively corelated with EAAs/TAAs, and principal component 1 was positively correlated with flavor AAs and EAAs. In conclusion, compared to BH1, MC, and Cherry Verry duck, AA content was higher in breast muscle of BH2 and J, which might be associated with the higher expression levels of mammalian target of rapamycin, eukaryotic translation initiation factor 4E-binding protein 1, and proton-coupled amino acid transporter 1 in breast muscle of BH2 and solute carrier family 38 member 2 in breast muscle of J. The comparative analysis of AA content in breast muscle among different duck breeds/strains could provide an important basis for improving the nutritional value of duck meat in the breeding process.

Hydrogen sulfide, regulated by VvWRKY30, promotes berry color changes in grapevine cabernet sauvignon

Since hydrogen sulfide (H2S) is a newly discovered gas signal molecule in plants, minimal studies are available regarding its role in grape berries color changes. This study found that 0.1 mM NaHS (H2S donor) could significantly promote color changes in grape berries, mainly by affecting the upregulated expression of anthocyanin synthesis-related genes. Further studies showed that the endogenous H2S content gradually accumulated as the grape berries ripened and mainly depended on the upregulation of the H2S synthetase gene VvSiR and its enhanced enzyme activity. More results showed that VvWRKY30 gene expression was also significantly upregulated during veraison. The overexpression of VvWRKY30 could induce VvSiR expression and H2S accumulation. Experiments involving the yeast one-hybrid and dual-luciferase reporter assay system showed that VvWRKY30 could directly interact with the VvSiR promoter. The overexpression of VvWRKY30 and VvSiR during the grape berry development stage, both promoted the anthocyanin accumulation and berry color changes. These findings indicated that H2S encouraged anthocyanin accumulation and color changes in grape berries, while the H2S accumulation in vivo depended on VvWRKY30 promoting the expression of its synthetase gene, VvSiR.

OPAQUE3, encoding a transmembrane bZIP transcription factor, regulates endosperm storage protein and starch biosynthesis in rice

Starch and storage proteins are the main components of rice (Oryza sativa L.) grains. Despite their importance, the molecular regulatory mechanisms of storage protein and starch biosynthesis remain largely elusive. Here, we identified a rice opaque endosperm mutant, opaque3 (o3), that overaccumulates 57-kDa proglutelins and has significantly lower protein and starch contents than the wild type. The o3 mutant also has abnormal protein body structures and compound starch grains in its endosperm cells. OPAQUE3 (O3) encodes a transmembrane basic leucine zipper (bZIP) transcription factor (OsbZIP60) and is localized in the endoplasmic reticulum (ER) and the nucleus, but it is localized mostly in the nucleus under ER stress. We demonstrated that O3 could activate the expression of several starch synthesis-related genes (GBSSI, AGPL2, SBEI, and ISA2) and storage protein synthesis-related genes (OsGluA2, Prol14, and Glb1). O3 also plays an important role in protein processing and export in the ER by directly binding to the promoters and activating the expression of OsBIP1 and PDIL1-1, two major chaperones that assist with folding of immature secretory proteins in the ER of rice endosperm cells. High-temperature conditions aggravate ER stress and result in more abnormal grain development in o3 mutants. We also revealed that OsbZIP50 can assist O3 in response to ER stress, especially under high-temperature conditions. We thus demonstrate that O3 plays a central role in rice grain development by participating simultaneously in the regulation of storage protein and starch biosynthesis and the maintenance of ER homeostasis in endosperm cells.

CEF3 is involved in membrane trafficking and essential for secondary cell wall biosynthesis and its mutation enhanced biomass enzymatic saccharification in rice

BackgroundAs one of the most important staple food crops, rice produces large of agronomic biomass residues that contain lots of secondary cell walls (SCWs). Membrane trafficking plays key roles in SCWs biosynthesis, but information association membrane trafficking and SCWs formation in plants is limited.ResultsIn this study, we report the function characterization of a rice mutant, culm easily fragile 3 (cef3), that exhibits growth retardation and fragile culm phenotype with significantly altered cell wall composition and reduced secondary wall thickness. Map-based cloning revealed that CEF3 encodes a homologous protein of Arabidopsis STOMATAL CYTOKINESIS DEFECTIVE2 (SCD2). The saccharification assays revealed that CEF3 mutation can improve biomass enzymatic saccharification. Expression pattern analysis indicated that CEF3 is ubiquitously expressed in many organs at different developmental stages. Subcellular localization revealed that CEF3 is a Golgi-localized protein. The FM4-64 uptake assay revealed CEF3 is involved in endocytosis. Furthermore, mutation of CEF3 not only affected cellulose synthesis-related genes expression, but also altered the abundance of cellulose synthase catalytic subunit 9 (OsCESA9) in the PM and in the endomembrane systems.ConclusionsThis study has demonstrated that CEF3 participates in the membrane trafficking that is essential for normal cellulose and other polysaccharides biosynthesis of the secondary cell wall, thereby manipulation of CEF3 could alter cellulose content and enhance biomass enzymatic saccharification in rice plants. Therefore, the study of the function of CEF3 can provide a strategy for genetic modification of SCWs in bioenergy crops.

Dietary phosphatidylcholine improved the survival, growth performance, antioxidant, and osmoregulation ability of early juvenile mud crab Scylla paramamosain

A 60-day culture experiment was designed to investigate the effect of dietary phosphatidylcholine/soybean lecithin (PC/SL) levels on survival, growth, body composition, antioxidant, and osmoregulation ability of early juvenile mud crab Scylla paramamosain. Six isonitrogenous and isolipidic experimental diets were formulated with five graded PC levels (0.1% (PC0.1), 0.5% (PC0.5), 1.0% (PC1.0), 2.0% (PC2.0), and 4.0% (PC4.0)) and one SL level (2.0%, SL2.0) to fed early juvenile S. paramamosain (8.70 ± 0.19 mg/crab). The results showed a significantly low survival in the PC0.1 group. Mud crabs obtained the highest growth performance in the PC2.0 and SL2.0 groups, but there was no significant difference with crabs in the PC0.5, PC1.0, and PC4.0 groups. The expression of the target of rapamycin (tor) gene was positively correlated with growth performance. The crab's whole-body crude lipid, triglyceride (TG), and total cholesterol (T-CHO) content rose significantly with increasing dietary PC levels. At the same time, the crab whole-body long-chain polyunsaturated fatty acids (lc-PUFA) proportion increased significantly, accompanied by a decrease in the proportion of C18:2n6. The expression of fatty acid synthesis-related genes decreased significantly with the increase in dietary PC levels. Dietary PC levels ≥1% significantly increased the antioxidant capacity of the mud crabs. Crabs obtained the highest total antioxidant capacity (T-AOC) and superoxide dismutase (SOD) activities in the PC1.0 group; the catalase (CAT) activity was positively correlated with dietary PC levels; significantly lower malondialdehyde (MDA) content was found when dietary PC ≥ 1%. Na+/K+-ATPase (NKA) activity increased with increasing dietary PC levels, and the expression of Na+/K+/2Cl−-cotransporter (nkcc) and nka genes were also significantly elevated. The results of the salinity challenge showed that the crabs obtained the highest survival in the PC4.0 group. In summary, the second-order polynomial regression analysis of weight gain and specific growth rate showed that early juvenile S. paramamosain obtained the highest growth at a dietary PC level of 2.60–2.62%.

Transcription factor AacmrA mediated melanin synthesis regulates the growth, appressorium formation, stress response and pathogenicity of pear fungal Alternaria alternata.

CmrA, as transcription factor for regulating DHN-melanin synthesis, controls melanin synthesis gene expression, and also regulate growth, development, stress response and virulence of plant fungi. However, little is known about the roles of CmrA on infection structure formation, penetration and pathogenicity of pear fungal Alternaria alternata. Here, we identified cmrA gene in A.alternata and assigned as AacmrA, sequence alignment and phylogenetic analysis revealed that AacmrA is highly conserved among fungi and encoded protein contain two Cys2His2 zinc finger motifs and one Zn(II)2Cys6 zinc cluster protein motif. ΔAacmrA severely decreased melanin production and DHN melanin synthesis related genes expression. Deletion of AacmrA impaired the morphology of spore and hyphae. Spore germination and appressorium formation induced by hydrophobicity surfaces and fruit wax significantly decreased in ΔAacmrA mutant. ΔAacmrA mutants were more sensitive than the wild type to osmotic stress and cell wall inhibitors, especially more sensitive to oxidative stress. In addition, lesion diameter of pear fruit wound inoculated with the ΔAacmrA mutant was reduced by 40.8% compared with the wild type 12d after inoculation. All findings of this study suggested that AacmrA is required for melanin biosynthesis, infection structure formation, and pathogenicity in A.alternata.

Fibroblast growth factor 21 (FGF21) promotes porcine granulosa cell estradiol production and proliferation via PI3K/AKT/mTOR signaling

The proliferation and steroidogenesis of mammalian ovarian granulosa cells (GCs) are related to follicular development. Previous studies found that fibroblast growth factor 21 (FGF21) regulated female fertility through the hypothalamic–pituitary–gonad axis. However, FGF21 receptors are expressed on GCs, so we speculate that it might affect female reproduction by regulating their physiological activities. Here, we showed that FGF21, fibroblast growth factor receptor-1(FGFR1), and beta-klotho (KLB) were expressed in porcine GCs. ELISA assays showed that estradiol (E2) production was increased significantly when treating GCs with recombinant FGF21 (rFGF21). In addition, rFGF21 upregulated the mRNA and protein levels of E2 synthesis-related genes including StAR, CYP11A1, and CYP19A1 in porcine GCs. Correspondingly, FGF21 siRNA inhibited E2 levels and its synthesis-related gene expression. After rFGF21 treatment, CCK8 showed increased cell viability, and flow cytometry showed that the number of S phase increased, and cycle-related genes also increased. However, treatment with FGF21 siRNA to porcine GCs suppressed the cell cycle, viability, and EdU positive cell number. Consequently, FGF21/FGFR1/KLB forms a complex to activate the phosphatidylinositol-3-kinase (PI3K)/AKT/mTOR signaling pathway and further promote the proliferation and E2 synthesis in porcine GCs. Collectively, these findings suggests that FGF21 regulates porcine ovarian folliculogenesis.

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.

Pasteurized Akkermansia muciniphila Reduces Fat Accumulation via nhr-49-Mediated Nuclear Hormone Signaling Pathway in Caenorhabditis elegans.

Pasteurized Akkermansia muciniphila (p-AKK) is related to lipid metabolism and helps control obesity. The main goal of this study was to investigate the role and mechanism of p-AKK in lipid metabolism using Caenorhabditis elegans. The results showed that p-AKK increased the healthy lifespan of nematodes and helped maintain exercise ability in aging, suggesting a potential increase in energy expenditure. The overall fat deposition and triglyceride level were significantly decreased and the p-AKK anti-oxidative stress helped to regulate fatty acid composition. Additionally, the transcriptome results showed that p-AKK increased the expression of lipo-hydrolase and fatty acid β-oxidation-related genes, including lipl-4, nhr-49, acs-2 and acdh-8, while it decreased the expression of fat synthesis-related genes, including fat-7, elo-2 and men-1. These results partially explain the mechanisms underlying the fact that p-AKK decreases fat accumulation of C. elegans via nhr-49/acs-2-mediated signaling involved in fatty acid β-oxidation and synthesis.

Yinchenhao decoction alleviates high‐carbohydrate diet‐induced hepatic lipids deposition by strengthening lipids metabolism and transport in largemouth bass

Yinchenhao decoction (YCHD) is a famous traditional Chinese medicine for improving hepatic metabolism disorder. The aim of this study was to investigate the effects of YCHD on lipid metabolism of juvenile largemouth bass fed on high-carbohydrate diet. Triplicate groups of fish (5.6 ± 0.2 g) were fed six experimental diets (one low-carbohydrate diet and five high-carbohydrate diets with gradients YCHD) for 10 weeks. The results showed that appropriate YCHD administration reversed the adverse effects of high-carbohydrate diet on the growth performance. Meanwhile, YCHD decreased the depositions of hepatic T-CHO, TG and NEFA, and alleviated lipids deposition in a dose-dependent manner. Importantly, as the key energy regulating element, AMPK was significantly increased by YCHD, coupled with high expression of β-oxidation (PPARα and CPT1) and lipids transport (APoB100, MTP, PNPLA7 CD36, CETP and FABP1) genes. Furthermore, PI3K-AKT pathway was prominently suppressed by YCHD, and the expression of lipids synthesis-related genes (ACCA, FAS and SCD1) was notably down-regulated. Additionally, YCHD up-regulated the expression of glycogen metabolism-related genes (GSK3β, PGY and GSY). In summary, our results demonstrated that YCHD promoted lipids β-oxidation and transport, enhanced glucose metabolism and reduced lipids deposition, indicating its beneficial effects on juvenile largemouth bass fed on high-carbohydrate diet.

Effect of light intensity on growth, accumulation of ginsenosides, and expression of related enzyme genes of Panax quinquefolius

Appropriate light intensity is favorable for the photosynthesis, biomass accumulation, key enzyme activity, and secondary metabolite synthesis of medicinal plants. This study aims to explore the influence of light intensity on growth and quality of Panax quinquefolius. To be specific, sand culture experiment was carried out in a greenhouse under the light intensity of 40, 80, 120, and 160 μmol·m~(-2)·s~(-1), respectively. The growth indexes, photosynthetic characteristics, content of 6 ginsenosides of the 3-year-old P. quinquefolius were determined, and the expression of ginsenoside synthesis-related enzyme genes in leaves, main roots, and fibrous roots was determined. The results showed that the P. quinquefolius growing at 80 μmol·m~(-2)·s~(-1) light intensity had the most biomass and the highest net photosynthetic rate. The total biomass of P. quinquefolius treated with 120 μmol·m~(-2)·s~(-1) light intensity was slightly lower than that with 80 μmol·m~(-2)·s~(-1). The root-to-shoot ratio in the treatment with 120 μmol·m~(-2)·s~(-1) light intensity was up to 6.86, higher than those in other treatments(P<0.05),and the ginsenoside content in both aboveground and underground parts of P. quinquefolius in this treatment was the highest, which was possibly associated with the high expression of farnesylpyrophosphate synthase(FPS), squalene synthase(SQS), squalene epoxidase(SQE), oxidosqualene cyclase(OSC), dammarenediol-Ⅱ synthase(DS), and P450 genes in leaves and SQE and DS genes in main roots. In addition, light intensities of 120 and 160 μmol·m~(-2)·s~(-1) could promote PPD-type ginsenoside synthesis in leaves by triggering up-regulation of the expression of upstream ginsenoside synthesis genes. The decrease in underground biomass accumulation of the P. quinquefolius grown under weak light(40 μmol·m~(-2)·s~(-1)) and strong light(160 μmol·m~(-2)·s~(-1)) was possibly attributed to the low net photosynthetic rate, stomatal conductance, and transpiration rate in leaves. In the meantime, the low expression of SQS, SQE, OSC, and DS genes in the main roots might led to the decrease in ginsenoside content. However, there was no significant correlation between the ginsenoside content and the expression of synthesis-related genes in the fibrous roots of P. quinquefolius. Therefore, the light intensity of 80 and 120 μmol·m~(-2)·s~(-1) is beneficial to improving yield and quality of P. quinquefolius. The above findings contributed to a theoretical basis for reasonable shading in P. quinquefolius cultivation, which is of great significance for improving the yield and quality of P. quinquefolius through light regulation.

Effect of dietary supplementation with mulberry and moringa leaves on the chicken reproductive performance

The effects of dietary supplementation with moringa (Moringa oleifera Lam.) and mulberry (Morus nigra L.) leaf powder on the clutch trait, reproductive organs, follicles, steroid hormones, and gene expressions of laying chickens were investigated during a 42-day experiment. Two hundred and ten Chinese local strain hens aged 37 weeks were randomly allocated to three groups, each group included five replicates, with 14 hens per replicate. The hens in the control group were fed with the basal diet, whereas those in the test groups were fed with a basal diet supplemented with 2.5% moringa leaf powder and 2.5% mulberry leaf powder (MOLP2.5+MLP2.5), 5% MOLP and 2.5% MLP (MOLP5+MLP2.5). The clutch traits (clutches, clutch length, delay days) were recorded during the trial. At the end of the experiment, the effect of the dietary supplementation with MOLP and MLP on the reproductive organs and tissues was estimated, and the gene expressions of ESR1, ESR2, CYP19A1 and STAR were analysed. Compared with the control group, the clutch performed worse in the treatment groups, however, the laying rate, reproductive organs and tissues in the MOLP2.5+MLP2.5 group showed no significant difference. Though the oestrogen levels did not significantly increase, they were elevated in the MOLP2.5+MLP2.5 group. Furthermore, it was identified that the expression levels of the ESR1 and CYP19A1 mRNA were significantly upregulated in the MOLP2.5+MLP2.5 group compared with those in the control group. In conclusion, a low addition level of MOLP and MLP can be used in laying hens to improve the steroid hormones synthesis-related gene expression which might improve the reproductive performance over the long term.

CIDEA Regulates De Novo Fatty Acid Synthesis in Bovine Mammary Epithelial Cells by Targeting the AMPK/PPARγ Axis and Regulating SREBP1.

Cell-death-inducing DNA fragmentation factor-α-like effector A (CIDEA) is a lipid-droplet-associated protein that helps to promote lipid metabolism in adipocytes of mice and humans. However, studies on the regulatory mechanism of CIDEA on lipid metabolism in the mammary glands of dairy cows are rare. Therefore, the role of CIDEA in bovine mammary epithelial cells (bMECs) was investigated in this study. The CIDEA expression levels in the mammary glands of high-fat-milk-producing cows were significantly higher compared to those in low-fat-milk-producing cows. Results of in vitro studies in bMECs showed that the inhibition of CIDEA inhibited the expression of fatty acid synthesis-related genes and triglyceride (TAG) synthesis-related genes. Conversely, the overexpression of CIDEA leads to an increase in the content of TAG and fatty acid. The results of mechanistic studies indicated that the overexpression of CIDEA inhibits AMP-activated protein kinase (AMPK) activity, which enhances the expression of peroxisome proliferator-activated receptor-γ (PPARγ) and consequently increases the TAG content. Furthermore, the overexpression of CIDEA promoted the nuclear translocation of sterol regulatory element-binding protein 1 (SREBP1). Therefore, a theoretical framework is provided by this study for the regulation of lipid metabolism in dairy cows by means of nutrition and the hormone targeting of CIDEA.

Functional characterization of transcription factor WIN1 genes associated with lipid biosynthesis and stress tolerance in soybean (Glycine max)

Transcription factor WAX INDUCER1 (WIN1) plays an important role in wax and cutin biosynthesis in plants. In this study, six GmWIN1 genes were identified from soybean ( Glycine max ), with GmWIN1–5 having high expression in flowers and seeds, particularly under cold stress. GmWIN1–5′s functions in lipid/oil biosynthesis and stress tolerance were characterized by its overexpression in soybean hairy roots and tobacco ( Nicotiana benthamiana ) plants. Under cold stress, GmWIN1–5 overexpression ( GmWIN1–5- OE) resulted in large oil enhancement and high level of phospholipids involved in cell membranes formation. Malondialdehyde content and elcetronlyte leakage in GmWIN1–5-OE plants was smaller than that in the wild-type control under cold stress, indicating that GmWIN1–5 promote plant stress tolerance. Moreover, GmWIN1–5-OE also led to upregulated expression of numerous genes involved in de novo fatty acid biosynthesis and triacylglycerol (TAG) production, including genes encoding phospholipase D ( PLD ), biotin carboxyl carrier protein 1 ( BCCP1 ), lysophosphatidic acid acyltransferase 5 ( LPAT5 ), and diacylglycerol acyltransferase 2 ( DGAT2 ). GmWIN1–5 directly bound to the promoter region of PLD gene. Overall, the results revealed that GmWIN1–5 can upregulate the biosynthesis and accumulation of lipids/oils, especially unsaturated fatty acids and phospholipids benefiting membrane integrity, and ultimately confer plants with high stress tolerance. • Six GmWIN1 members with structural integrity are identified in soybean ( Glycine max ). • Of the six GmWIN1s, GmWIN1-5 expression level was the highest in flower and seed tissues. • Overexpression of GmWiN1-5 in soybean hair roots increased total lipids and cold stress tolerance. • Ectopic expression of GmWIN1-5 in tobacco plant boosted oil content and stress tolerance. • GmWIN1-5 promoted expression of several lipid synthesis-related genes, particularly PLD.

Antibiofilm and Antiquorum Sensing Potential of Lactiplantibacillus plantarum Z057 against Vibrio parahaemolyticus.

Vibrio parahaemolyticus is a widespread foodborne pathogen that causes serious seafood-borne gastrointestinal infections. Biofilm and quorum sensing (QS) are critical in regulating these infections. In this study, first, the ability of Lactiplantibacillus plantarum Z057 to compete, exclude, and displace V. parahaemolyticus biofilm was evaluated. Then, the inhibitory effects of L. plantarum Z057 extract (Z057-E) on V. parahaemolyticus biofilm and QS were explored from the aspects of biofilm biomass, metabolic activity, physicochemical properties, extracellular polymer matrix content, QS signal AI-2 activity, biofilm microstructure, and the expression levels of biofilm and QS-related genes. Results showed that L. plantarum Z057 effectively inhibited biofilm formation of V. parahaemolyticus and interfered with the adhesion of V. parahaemolyticus on the carrier surface. In addition, the Z057-E could significantly reduce the biofilm biomass, metabolic activity, hydrophobicity, auto-aggregation ability, swimming and swarming migration diameter, AI-2 activity, extracellular polysaccharide (EPS), and extracellular protein content of V. parahaemolyticus. Fluorescence microscope and scanning electron microscope (SEM) images demonstrated that the Z057-E could efficiently inactivate the living cells, destroy the dense and complete biofilm architectures, and reduce the essential component of the extracellular polymer matrix. Real-time fluorescence quantitative PCR revealed that the Z057-E treatment down-regulated the expression of flagellum synthesis-related genes (flaA, flgM), EPS, and extracellular protein synthesis-related genes (cpsA, cpsQ, cpsR, ompW), QS-related genes (luxS, aphA, opaR), and hemolysin secretion-related genes (toxS, toxR) of V. parahaemolyticus. Thus, our results suggested that L. plantarum Z057 could represent an alternative biocontrol strategy against foodborne pathogens with anti-adhesive, antibiofilm, and antiquorum sensing activities.

Akkermansia muciniphila Enhances Egg Quality and the Lipid Profile of Egg Yolk by Improving Lipid Metabolism.

Akkermansia muciniphila (A. muciniphila) has shown potential as a probiotic for the prevention and treatment of non-alcoholic fatty liver disease in both humans and mice. However, relatively little is known about the effects of A. muciniphila on lipid metabolism, productivity, and product quality in laying hens. In this study, we explored whether A. muciniphila supplementation could improve lipid metabolism and egg quality in laying hens and sought to identify the underlying mechanism. In the first experiment, 80 Hy-Line Brown laying hens were divided into four groups, one of which was fed a normal diet (control group), while the other three groups were administered a high-energy, low-protein diet to induce fatty liver hemorrhagic syndrome (FLHS). Among the three FLHS groups, one was treated with phosphate-buffered saline, one with live A. muciniphila, and one with pasteurized A. muciniphila. In the second experiment, 140 Hy-Line Brown laying hens were divided into two groups and respectively fed a basal diet supplemented or not with A. muciniphila lyophilized powder. The results showed that, in laying hens with FLHS, treatment with either live or pasteurized A. muciniphila efficiently decreased body weight, abdominal fat deposition, and lipid content in both serum and the liver; downregulated the mRNA expression of lipid synthesis-related genes and upregulated that of lipid transport-related genes in the liver; promoted the growth of short-chain fatty acids (SCFAs)-producing microorganisms and increased the cecal SCFAs content; and improved the yolk lipid profile. Additionally, the supplementation of lyophilized powder of A. muciniphila to aged laying hens reduced abdominal fat deposition and total cholesterol (TC) levels in both serum and the liver, suppressed the mRNA expression of cholesterol synthesis-related genes in the liver, reduced TC content in the yolk, increased eggshell thickness, and reshaped the composition of the gut microbiota. Collectively, our findings demonstrated that A. muciniphila can modulate lipid metabolism, thereby, promoting laying hen health as well as egg quality and nutritive value. Live, pasteurized, and lyophilized A. muciniphila preparations all have the potential for use as additives for improving laying hen production.

Dietary resistant starch ameliorating lipopolysaccharide-induced inflammation in meat ducks associated with the alteration in gut microbiome and glucagon-like peptide 1 signaling

BackgroundConsumption of resistant starch (RS) has been associated with various intestinal and systemic health benefits, but knowledge of its effects on intestinal health and inflammatory response in stressed birds is limited. Here, we examined how dietary RS supplementation from 12% raw potato starch (RPS) modulated inflammatory severity induced by lipopolysaccharide (LPS) in meat ducks.ResultsLPS administration at 14, 16, and 18 d (chronic challenge) decreased body weight (BW) and glucagon-like peptide 1 receptor (GLP-1R) level with higher intestinal permeability and inflammation, evident by higher pro-inflammatory cytokine levels. Dietary 12% RPS supplementation enhanced Claudin-1 and GLP-1R expression, along with lower levels of inflammatory factors in both ileum and serum. Microbiome analysis showed that RS treatment shifted microbial structure reflected by enriched the proportion of Firmicutes, Bifidobacterium, Ruminococcus, etc. Dietary RS addition also significantly increased the concentrations of propionate and butyrate during chronic LPS challenge. Furthermore, response to acute challenge, the ducks received 2 mg/kg BW LPS at 14 d had higher concentrations of serum endotoxins and inflammatory cytokines, as well as upregulated transcription of toll like receptor 4 (TLR4) in ileum when compared to control birds. Analogous to GLP-1 agonist liraglutide, dietary RS addition decreased endotoxins and inflammation cytokines, whereas it upregulated the GLP-1 synthesis related genes expression. Meanwhile, dietary RS supplementation suppressed the acute LPS challenge-induced TLR4 transcription.ConclusionsThese data suggest that dietary 12% RPS supplementation could attenuate the LPS-induced inflammation as well as intestinal injury of meat ducks, which might involve in the alteration in gut microbiota, SCFAs production and the signaling pathways of TLR4 and GLP-1/GLP-1R.

Study on the Influencing Mechanism of Human Chorionic Gonadotropin (hCG) on Oocyte Maturation in Patients with Polycystic Ovary Syndrome.

The study was aimed at investigating the influence of human chorionic gonadotropin (hCG) hormone on oocyte maturation in the patients with polycystic ovary syndrome (PCOS). A total of 54 patients with PCOS who received in vitro maturation (IVM) treatment in the Cheeloo College of Medicine, Shandong University, were divided into two groups: one group who underwent hCG injections was the observation group (OG; n = 27) and other was the control group (CG; n = 27) with no hCG injection. The oocyte development and the expression of steroid hormone synthesis-related genes including gonadotropin-releasing hormone receptor (GnRHR), Conexin43, epidermal growth factor-related genes, luteinizing hormone/choriogonadotropin receptor (LHCGR), epiregulin (EREG), and vascular endothelial growth factor (VEGF) were examined. The human ovarian granulosa cell line (SVOG cells) and ovarian epithelial cell line (HOSEpiC cells) were employed to analyze the effect of hCG on the biological behaviour of cells. As a result, OG showed higher normal fertilization, cleavage, and high-qualified embryo rate than CG. Expression levels of GnRHR, Cx43, LHCGR, EREG, and VEGF were significantly elevated in granulosa cells in the OG group. Western blot revealed that phosphoinositide 3-kinase (PI3K), protein kinase B (AKT), and rapamycin (mTOR) proteins were decreased in granulosa cells under hCG intervention. A biological behaviour test indicated that the multiplication capacity of hCG-intervened SVOG and HOSEpiC was increased, while the apoptosis was decreased. In conclusion, hCG could accelerate follicular development and oocyte maturation by activating oocyte maturation genes in PCOS patients, which could significantly improve and popularize the application of IVM technology.