Functional characterization

功能表征
  • 文章类型: Journal Article
    超长链脂肪酸(Elovls)的延伸酶是关键的限速酶,其通过催化预先存在的脂肪酰基链的两个碳延伸而参与LC-PUFA生物合成。到目前为止,在teleost中已经对几种Elovls进行了广泛的研究。然而,伊洛夫在软骨病中的功能和生理作用鲜有报道。在这项研究中,我们通过全基因组扫描从濒临灭绝的中华st(Acipensersinensis)中鉴定并表征了elovl2。结果表明,elovl2的编码序列长度为894bp,对于297个羊膜酸的推定蛋白质。比较基因组分析表明,中国st鱼elovl2在进化上是保守的。在酵母中的功能表征表明,中国stElovl2可以有效地延长C20(ARA和EPA)和C22(22:4n-6和22:5n-3)底物,证实其在LC-PUFA生物合成中的关键作用。时空表达分析显示,在肝脏和大脑中检测到高elovl2mRNA水平,并且在胚胎和孵化后阶段均显示出增加趋势。有趣的是,以植物油为脂类来源的日粮可以显著诱导中国st中elovl2的高表达,这意味着内源性LC-PUFA生物合成途径受到饮食中缺乏LC-PUFA的刺激。我们的发现将增强我们对elovl2的进化和功能作用的理解,并为脊椎动物中LC-PUFA生物合成机制提供新的见解。
    Elongases of very-long-chain fatty acids (Elovls) are critical rate-limiting enzymes that are involved in LC-PUFA biosynthesis through catalyzing the two-carbon elongation of a pre-existing fatty acyl chain. Thus far, several Elovls have been extensively studied in teleost. However, the functional and physiological roles of Elovls in chondrichthyans have rarely been reported. In this study, we identified and characterized elovl2 from the endangered Chinese sturgeon (Acipenser sinensis) by whole genome scanning. The results show that the coding sequence of elovl2 was 894 bp in length, for a putative protein of 297 amnio acids. Comparative genomic analyses indicated that Chinese sturgeon elovl2 was evolutionarily conserved. Functional characterization in yeast demonstrated that the Chinese sturgeon Elovl2 could efficiently elongate C20 (ARA and EPA) and C22 (22:4n-6 and 22:5n-3) substrates, confirming its critical roles in LC-PUFA biosynthesis. Spatial and temporal expression analyses showed high elovl2 mRNA levels were detected in the liver and brain and showed an increase trend both in embryonic and post-hatching stages. Interestingly, diets with vegetable oils as lipid sources could significantly induce the high expression of elovl2 in Chinese sturgeon, implying that the endogenous LC-PUFA biosynthesis pathway was stimulated by lack of LC-PUFA in their diets. Our findings will enhance our understanding about the evolutionary and functional roles of elovl2 and provide novel insights into the LC-PUFA biosynthesis mechanism in vertebrates.
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  • 文章类型: Journal Article
    肽/组氨酸转运蛋白PHT1(SLC15A4)在免疫细胞的溶酶体膜中表达,在代谢和炎症信号传导中起重要作用。PHT1是一个H+偶联/组氨酸转运体,可以转运多种寡肽,包括各种细菌衍生的肽。此外,它使各种代谢信号分子的支架,并与免疫反应的关键调节元件相互作用。毫不奇怪,PHT1与自身免疫性疾病如系统性红斑狼疮(SLE)的发病机理有关。不幸的是,PHT1调节剂的药理学开发由于缺乏合适的转运试验而受到阻碍。为了解决这个缺点,提出了一种基于固体支持膜的电生理学(SSME)的新型转运测定法。目前SSME研究的主要发现包括电生理特性的首次记录,pH依赖性分析,对PHT1底物选择性的评估,以及所识别底物的传输动力学。与以前的工作相比,PHT1在其天然溶酶体环境中进行研究。此外,通过分子对接验证了观察到的底物选择性。总的来说,这种新的基于SSME的检测方法有望有助于释放PHT1的药理学潜力,并加深对其功能特性的理解。
    The peptide/histidine transporter PHT1 (SLC15A4) is expressed in the lysosomal membranes of immune cells where it plays an important role in metabolic and inflammatory signaling. PHT1 is an H+-coupled/histidine symporter that can transport a wide range of oligopeptides, including a variety of bacterial-derived peptides. Moreover, it enables the scaffolding of various metabolic signaling molecules and interacts with key regulatory elements of the immune response. Not surprisingly, PHT1 has been implicated in the pathogenesis of autoimmune diseases such as systemic lupus erythematosus (SLE). Unfortunately, the pharmacological development of PHT1 modulators has been hampered by the lack of suitable transport assays. To address this shortcoming, a novel transport assay based on solid-supported membrane-based electrophysiology (SSME) is presented. Key findings of the present SSME studies include the first recordings of electrophysiological properties, a pH dependence analysis, an assessment of PHT1 substrate selectivity, as well as the transport kinetics of the identified substrates. In contrast to previous work, PHT1 is studied in its native lysosomal environment. Moreover, observed substrate selectivity is validated by molecular docking. Overall, this new SSME-based assay is expected to contribute to unlocking the pharmacological potential of PHT1 and to deepen the understanding of its functional properties.
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  • 文章类型: Journal Article
    糖最终将外溢的转运蛋白(SWEET)已被证实在植物生长中起着不同的生理作用,发育和应激反应。然而,Hemerocalliscitrina中SWEET基因的特征和功能仍不清楚,也不清楚。在这项研究中,利用全基因组进行生物信息学分析,成功鉴定出19个HcSWEET基因。物理化学性质的分析表明这些HcSWEET之间存在主要差异。系统发育分析表明,HcSWEET蛋白可以分为4个进化枝,从进化枝I到IV,其中同一进化枝内的蛋白质表现出共享的保守基序和基因结构。大多数HcSWEET基因中含有5到6个外显子,它们在11条染色体上分布不均。基因重复分析显示存在4个基因对。比较性图谱显示,与双子叶植物相比,HcSWEET基因家族在单子叶植物中可能表现出更多的封闭同源性。HcSWEET基因的顺式作用元件分析表明对各种激素的关键反应,光,和压力。此外,转录组测序分析表明,大多数HcSWEET基因在根中的表达相对较高,HcSWEET4a在盐胁迫下显著上调。过表达进一步证实了HcSWEET4a参与盐胁迫反应的可能性,这提供了新颖的见解,并促进了对HcSWEET在抵抗非生物胁迫方面的功能分析的深入研究。
    Sugars will be eventually effluxed transporters (SWEETs) have been confirmed to play diverse physiological roles in plant growth, development and stress response. However, the characteristics and functions of the SWEET genes in Hemerocallis citrina remain unclear and poorly elucidated. In this study, the whole genome of Hemerocallis citrina was utilized to conduct bioinformatics analysis and a total of 19 HcSWEET genes were successfully identified. Analysis of the physicochemical properties indicated dominant differences among these HcSWEETs. A phylogenetic analysis revealed that HcSWEET proteins can be divided into 4 clades ranging from Clade I to IV, where proteins within the same clade exhibited shared conserved motifs and gene structures. Five to six exons were contained in the majority of HcSWEET genes, which were unevenly distributed across 11 chromosomes. The gene duplication analysis showed the presence of 4 gene pairs. Comparative syntenic maps revealed that the HcSWEET gene family might present more closed homology in monocotyledons than dicotyledons. Cis-acting element analysis of HcSWEET genes indicated key responsiveness to various hormones, light, and stresses. Additionally, transcriptome sequencing analysis suggested that most HcSWEET genes had a relatively higher expression in roots, and HcSWEET4a was significantly up-regulated under salt stress. Overexpression further verified the possibility that HcSWEET4a was involved in response to salt stress, which provides novel insights and facilitates in-depth studies of the functional analysis of HcSWEETs in resistance to abiotic stress.
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  • 文章类型: Journal Article
    背景:炎症性肠病(IBD)是一个主要的公共卫生问题,病因尚不清楚。肠道微生物群的组成和功能的变化与这些病理有关,包括耗竭严格的厌氧菌,如普氏双杆菌。在其他厌氧菌中观察到的消耗证据较少,其中双歧杆菌。这项研究通过文化方法表征了从活性和非活性IBD患者的人类肠道微生物群分离的双歧杆菌的分类学和功能多样性,并评估了这些双歧杆菌是否可用作肠道健康的益生菌。
    结果:从IBD患者(52例克罗恩病和26例溃疡性结肠炎患者)的肠道菌群中分离出341株双歧杆菌,与高比例的双歧杆菌菌株(分离的双歧杆菌28%)。在溃疡性结肠炎中,鉴定出的主要物种是牙本质芽孢杆菌(占分离的双歧杆菌的39%),在活动性和非活动性溃疡性结肠炎中。在克罗恩病,青少年芽孢杆菌是从非活跃患者中分离出来的主要物种(40%),而在活跃的克罗恩病患者中发现了相似量的牙本质芽孢杆菌和青春期芽孢杆菌。牙菌根的相对丰度随着年龄的增长而增加,克罗恩病和溃疡性结肠炎以及活动性和非活动性IBD患者。与从活性溃疡性结肠炎中分离的菌株相比,从非活性溃疡性结肠炎中分离的双歧杆菌对大肠杆菌LF82和肠道沙门氏菌ATCC14028的抗菌能力更为常见。最后,长芽孢杆菌被保留为具有最高益生菌潜力的菌株,因为它们是呈现胞外多糖合成的主要菌株。抗菌活性,和抗炎能力。通过计算机模拟分析,抗菌活性和EPS合成进一步与抗菌和EPS基因簇的存在相关。
    结论:在IBD患者的微生物群中鉴定出不同的双歧杆菌分类谱。最丰富的物种是牙本质芽孢杆菌,主要与溃疡性结肠炎患者和青少年芽孢杆菌的微生物群有关,在克罗恩病患者的肠道微生物群中。此外,牙菌根的相对丰度随着年龄的增长而显著增加。此外,这项研究证明,双歧杆菌具有益生菌潜力(抗致病活性,胞外多糖的产生和抗炎活性),尤其是B.longum菌株,可以从活动性和非活动性克罗恩病和溃疡性结肠炎患者的肠道菌群中分离。
    BACKGROUND: Inflammatory Bowel Diseases (IBD) are a major public health issue with unclear aetiology. Changes in the composition and functionality of the intestinal microbiota are associated with these pathologies, including the depletion of strict anaerobes such as Feacalibacterium prausnitzii. Less evidence is observed for depletion in other anaerobes, among which bifidobacteria. This study characterized the taxonomic and functional diversity of bifidobacteria isolated from the human intestinal microbiota in active and non-active IBD patients by a culturomics approach and evaluated if these bifidobacteria might be used as probiotics for gut health.
    RESULTS: A total of 341 bifidobacteria were isolated from the intestinal microbiota of IBD patients (52 Crohn\'s disease and 26 ulcerative colitis patients), with a high proportion of Bifidobacterium dentium strains (28% of isolated bifidobacteria). In ulcerative colitis, the major species identified was B. dentium (39% of isolated bifidobacteria), in active and non-active ulcerative colitis. In Crohn\'s disease, B. adolescentis was the major species isolated from non-active patients (40%), while similar amounts of B. dentium and B. adolescentis were found in active Crohn\'s disease patients. The relative abundance of B. dentium was increased with age, both in Crohn\'s disease and ulcerative colitis and active and non-active IBD patients. Antibacterial capacities of bifidobacteria isolated from non-active ulcerative colitis against Escherichia coli LF82 and Salmonella enterica ATCC 14028 were observed more often compared to strains isolated from active ulcerative colitis. Finally, B. longum were retained as strains with the highest probiotic potential as they were the major strains presenting exopolysaccharide synthesis, antibacterial activity, and anti-inflammatory capacities. Antimicrobial activity and EPS synthesis were further correlated to the presence of antimicrobial and EPS gene clusters by in silico analysis.
    CONCLUSIONS: Different bifidobacterial taxonomic profiles were identified in the microbiota of IBD patients. The most abundant species were B. dentium, mainly associated to the microbiota of ulcerative colitis patients and B. adolescentis, in the intestinal microbiota of Crohn\'s disease patients. Additionally, the relative abundance of B. dentium significantly increased with age. Furthermore, this study evidenced that bifidobacteria with probiotic potential (antipathogenic activity, exopolysaccharide production and anti-inflammatory activity), especially B. longum strains, can be isolated from the intestinal microbiota of both active and non-active Crohn\'s disease and ulcerative colitis patients.
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  • 文章类型: Journal Article
    丝氨酸蛋白酶抑制剂是调节各种生理过程的蛋白质超家族,包括纤维蛋白溶解,炎症和免疫反应。在寄生虫系统中,serpins被认为在寄生虫定植中起重要作用,抑制宿主免疫丝氨酸蛋白酶和穿透防御屏障。然而,serpin在血吸虫中的特征较差。在这项研究中,包含1360个碱基对开放阅读框的曼氏血吸虫serpin(Smserpin-p46),被克隆,表达和功能特征。生物信息学分析显示Smserpin-p46含有关键残基,抑制性serpin的结构域和基序特征。基因表达谱证明了Smserpin-p46的阶段特异性表达,在成年雄性蠕虫中表达最高。重组Smserpin-p46(rSmserpin-p46)抑制人中性粒细胞组织蛋白酶G和弹性蛋白酶,参与NETosis的关键丝氨酸蛋白酶,形成中性粒细胞胞外陷阱的程序。使用特定的兔抗血清,在可溶性蠕虫抗原制剂中检测到Smserpin-p46,并定位在成虫被膜上。累计,Smserpin-p46在寄生虫被膜上的表达及其抑制NETosis相关蛋白酶的能力突出了该serpin在寄生虫-宿主相互作用中的重要性,并鼓励其作为控制血吸虫病的候选疫苗抗原进行进一步研究.
    Serine protease inhibitors are a superfamily of proteins that regulate various physiological processes including fibrinolysis, inflammation and immune responses. In parasite systems, serpins are believed to play important roles in parasite colonization, inhibition of host immune serine proteases and penetration of defensive barriers. However, serpins are less well characterized in schistosomes. In this study, a Schistosoma mansoni serpin (Smserpin-p46) containing a 1360 base pair open reading frame, was cloned, expressed and functionally characterized. Bioinformatics analysis revealed that Smserpin-p46 contains the key residues, structural domains and motifs characteristic of inhibitory serpins. Gene expression profiling demonstrated stage-specific expression of Smserpin-p46 with the highest expression in adult male worms. Recombinant Smserpin-p46 (rSmserpin-p46) inhibited both human neutrophil cathepsin G and elastase, key serine proteases involved in NETosis, a program for the formation of neutrophil extracellular traps. Using specific rabbit antiserum, Smserpin-p46 was detected in soluble worm antigen preparation and was localized to the adult worm tegument. Cumulatively, the expression of Smserpin-p46 on the parasite tegument and its ability to inhibit proteases involved in NETosis highlights the importance of this serpin in parasite-host interactions and encourages its further investigation as a candidate vaccine antigen for the control of schistosomiasis.
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  • 文章类型: Journal Article
    通过由植物烯合酶(PSY)催化的两个香叶基香叶基香叶基二磷酸酯分子缩合形成植物烯是类胡萝卜素生物合成中的第一个承诺和限速步骤,已经在细菌中进行了广泛的研究,陆地植物和微藻。然而,大型藻类的这一步仍然未知。在本研究中,从经济红藻Pyropiayezoensis克隆了一种编码推定的植物烯合酶的基因,该物种长期以来一直用于食品和药品。使用生物信息学工具预测的保守基序/结构域和三级结构表明,克隆的PyPSY应编码八烯合酶;这在经验上通过大肠杆菌中的色素互补得到了证实。这种八烯合成酶由单拷贝基因编码,其表达可能受多种因素调控。来自不同生物的PSY的系统发育关系表明,红藻可能是初级内共生的后代和次级内共生的质体供体。
    The formation of phytoene by condensing two geranylgeranyl diphosphate molecules catalyzed by phytoene synthase (PSY) is the first committed and rate-limiting step in carotenoid biosynthesis, which has been extensively investigated in bacteria, land plants and microalgae. However, this step in macroalgae remains unknown. In the present study, a gene encoding putative phytoene synthase was cloned from the economic red alga Pyropia yezoensis-a species that has long been used in food and pharmaceuticals. The conservative motifs/domains and the tertiary structure predicted using bioinformatic tools suggested that the cloned PyPSY should encode a phytoene synthase; this was empirically confirmed by pigment complementation in E. coli. This phytoene synthase was encoded by a single copy gene, whose expression was presumably regulated by many factors. The phylogenetic relationship of PSYs from different organisms suggested that red algae are probably the progeny of primary endosymbiosis and plastid donors of secondary endosymbiosis.
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  • 文章类型: Journal Article
    透明质酸(HA)水凝胶通常用于面部真皮填充和用于替代医学美学目的。商业配方中存在高度多样性,特别是对于成品稳定性的优化,功能,和性能。多价成分如羟基磷灰石钙(CaHA)或维生素B3(烟酰胺)值得注意地用作生物刺激剂以改善施用部位的皮肤质量属性。本研究的目的是对两种新型交联真皮填充剂配方(HAR-1“InstantRefine”和HAR-3“MaxiLift”)进行多参数表征,以阐明维生素B3掺入的各种功能影响。因此,首先在体外流变学方面对HAR产品进行了比较表征,粘结性,可注射性,和对化学或酶降解的抗性(暴露于H2O2、AAPH、透明质酸酶,或黄嘌呤氧化酶)。然后,在原代真皮成纤维细胞模型中评估HAR产品的细胞相容性和体外生物刺激属性.结果显示,在受控降解测定面板中,与JUV_DERM®VOLBELLA®和VOLUMA®参考产品相比,粘性HAR水凝胶的弹性增强。此外,对于HAR-1和HAR-3,记录了初级真皮成纤维细胞培养物中总胶原合成的显著诱导,表明固有生物刺激效应与Radiesse®和Sculptra™参考产品相当或优于这些。本文使用稳健和正交实验方法(水凝胶降解,功能基准)和研究设计。总的来说,报告的结果证实了维生素B3在交联HA真皮填充剂中的双重功能化作用,具有水凝胶系统稳定性属性的显着增强和有效的生物刺激能力的部署。
    Hyaluronic acid (HA) hydrogels are commonly used for facial dermal filling and for alternative medical aesthetic purposes. High diversity exists in commercial formulations, notably for the optimization of finished product stability, functionality, and performance. Polyvalent ingredients such as calcium hydroxylapatite (CaHA) or vitamin B3 (niacinamide) are notably used as bio-stimulants to improve skin quality attributes at the administration site. The aim of the present study was to perform multi-parametric characterization of two novel cross-linked dermal filler formulas (HAR-1 \"Instant Refine\" and HAR-3 \"Maxi Lift\") for elucidation of the various functional impacts of vitamin B3 incorporation. Therefore, the HAR products were firstly comparatively characterized in terms of in vitro rheology, cohesivity, injectability, and resistance to chemical or enzymatic degradation (exposition to H2O2, AAPH, hyaluronidases, or xanthine oxidase). Then, the HAR products were assessed for cytocompatibility and in vitro bio-stimulation attributes in a primary dermal fibroblast model. The results showed enhanced resilience of the cohesive HAR hydrogels as compared to JUVÉDERM® VOLBELLA® and VOLUMA® reference products in a controlled degradation assay panel. Furthermore, significant induction of total collagen synthesis in primary dermal fibroblast cultures was recorded for HAR-1 and HAR-3, denoting intrinsic bio-stimulatory effects comparable or superior to those of the Radiesse® and Sculptra™ reference products. Original results of high translational relevance were generated herein using robust and orthogonal experimental methodologies (hydrogel degradation, functional benchmarking) and study designs. Overall, the reported results confirmed the dual functionalization role of vitamin B3 in cross-linked HA dermal fillers, with a significant enhancement of hydrogel system stability attributes and the deployment of potent bio-stimulatory capacities.
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  • 文章类型: Journal Article
    糖转运蛋白(STPs)是高亲和力H+偶联的己糖转运体。最近,STP13对多种植物物种的细菌和真菌病原体抗性的贡献已经引起了极大的兴趣。来源叶的定量PCR分析,发育中的胚胎,菜豆(普通豆)的种皮表明,在整个种子发育过程中,PvSTP13.1在源叶和种皮中表达。相比之下,在发育中的胚胎中检测到极低水平的PvSTP13.1转录本。为了表征传输机制,PvSTP13.1在非洲爪狼卵母细胞中表达,使用双电极电压钳制分析了向内电流。PvSTP13.1显示出作为H偶联的单糖转运体的功能,在去极化的膜电位下对己糖和醛固酮具有独特的高亲和力。具体来说,在评估的31种基质中,其中包括己醛糖,脱氧己糖,果糖,3-O-甲基-D-葡萄糖,醛固酮,多元醇糖苷,二糖,三糖,和葡萄糖醛酸,PvSTP13.1对葡萄糖(43μM)表现出最高的亲和力(K0.5),甘露糖(92μM),半乳糖(145μM),果糖(224μM),木糖(1.0mM),和岩藻糖(3.7mM)在pH5.6,去极化膜电位为-40mV。此处显示的结果表明,PvSTP13.1有助于从源叶片和发育中种子的外皮中的外质空间中回收己糖。
    Sugar transport proteins (STPs) are high-affinity H+-coupled hexose symporters. Recently, the contribution of STP13 to bacterial and fungal pathogen resistance across multiple plant species has garnered significant interest. Quantitative PCR analysis of source leaves, developing embryos, and seed coats of Phaseolus vulgaris L. (common bean) revealed that PvSTP13.1 was expressed in source leaves and seed coats throughout seed development. In contrast, PvSTP13.1 transcripts were detected at exceedingly low levels in developing embryos. To characterize the transport mechanism, PvSTP13.1 was expressed in Xenopus laevis oocytes, and inward-directed currents were analyzed using two-electrode voltage clamping. PvSTP13.1 was shown to function as an H+-coupled monosaccharide symporter exhibiting a unique high affinity for hexoses and aldopentoses at depolarized membrane potentials. Specifically, of the 31 assessed substrates, which included aldohexoses, deoxyhexoses, fructose, 3-O-methyl-D-glucose, aldopentoses, polyols, glycosides, disaccharides, trisaccharides, and glucuronic acid, PvSTP13.1 displayed the highest affinity (K 0.5) for glucose (43 μM), mannose (92 μM), galactose (145 μM), fructose (224 μM), xylose (1.0 mM), and fucose (3.7 mM) at pH 5.6 at a depolarized membrane potential of -40 mV. The results presented here suggest PvSTP13.1 contributes to retrieval of hexoses from the apoplasmic space in source leaves and coats of developing seeds.
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  • 文章类型: Journal Article
    植物特异性VQ蛋白在植物生长发育调控中具有重要作用,以及在植物非生物胁迫反应中。它们的作用已经在模式植物拟南芥中得到了很好的确立;然而,马铃薯VQ蛋白的功能尚未得到充分研究。VQ蛋白核心区含有短的FxxhVQxhTG氨基酸基序序列。在这项研究中,对马铃薯VQ31蛋白进行了克隆和功能鉴定。StVQ31的完整开放阅读框(ORF)大小为672bp,编码223个氨基酸。亚细胞定位分析显示StVQ31位于细胞核中。与野生型(WT)植物相比,过表达StVQ31的转基因拟南芥植物表现出增强的耐盐性。正如根长增加所证明的那样,发芽率,盐度胁迫下的叶绿素含量。转基因植物的耐受性增加与渗透势(脯氨酸和可溶性糖)增加有关。MDA积累减少,总蛋白质含量降低,和提高膜的完整性。这些结果表明StVQ31过表达增强了植物的渗透潜能以维持正常的细胞生长。与WT相比,转基因植物的抗氧化酶活性显着增加,减少细胞膜损伤。此外,实时荧光定量PCR分析表明,StVQ31调控盐胁迫应答相关基因的表达,包括ERD,LEA4-5、At2g38905和AtNCED3。这些发现表明,StVQ31显著影响渗透和抗氧化细胞稳态,从而提高耐盐性。
    Plant-specific VQ proteins have crucial functions in the regulation of plant growth and development, as well as in plant abiotic stress responses. Their roles have been well established in the model plant Arabidopsis thaliana; however, the functions of the potato VQ proteins have not been adequately investigated. The VQ protein core region contains a short FxxhVQxhTG amino acid motif sequence. In this study, the VQ31 protein from potato was cloned and functionally characterized. The complete open reading frame (ORF) size of StVQ31 is 672 bp, encoding 223 amino acids. Subcellular localization analysis revealed that StVQ31 is located in the nucleus. Transgenic Arabidopsis plants overexpressing StVQ31 exhibited enhanced salt tolerance compared to wild-type (WT) plants, as evidenced by increased root length, germination rate, and chlorophyll content under salinity stress. The increased tolerance of transgenic plants was associated with increased osmotic potential (proline and soluble sugars), decreased MDA accumulation, decreased total protein content, and improved membrane integrity. These results implied that StVQ31 overexpression enhanced the osmotic potential of the plants to maintain normal cell growth. Compared to the WT, the transgenic plants exhibited a notable increase in antioxidant enzyme activities, reducing cell membrane damage. Furthermore, the real-time fluorescence quantitative PCR analysis demonstrated that StVQ31 regulated the expression of genes associated with the response to salt stress, including ERD, LEA4-5, At2g38905, and AtNCED3. These findings suggest that StVQ31 significantly impacts osmotic and antioxidant cellular homeostasis, thereby enhancing salt tolerance.
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  • 文章类型: Journal Article
    GATA转录因子(TF)在许多植物物种的各种生物过程中的调节作用已被广泛研究。GATATFs在调节非生物胁迫耐受性中的功能和分子机制尚未在普通豆类中进行研究。本研究分析了在不同非生物和植物激素胁迫下,寻常型假单胞菌基因组中GATA基因家族的功能特性。在寻常型疟原虫基因组中系统地研究了GATA基因家族,并鉴定了31个PvGATATFs。该研究发现,31个PvGATA基因中有18个发生了重复事件,强调基因复制在GATA基因扩增中的作用。所有的PvGATA基因被分为四个重要的亚家族,每个亚家族有8、3、6和13个成员(亚家族I,II,III,andIV),分别。所有的PvGATA蛋白序列都包含一个GATA结构域,但亚家族II成员有其他结构域,如CCT和tify.在PvGATA中预测了总共799个启动子顺式调节元件(CREs)。此外,我们使用qRT-PCR研究了五个PvGATA基因在非生物条件下在普通豆根中的表达谱。结果表明,PvGATA01/10/25/28可能在调节植物对盐和干旱胁迫的抗性中起重要作用,并可能参与植物激素介导的胁迫信号通路。选择PvGATA28用于过表达,并使用农杆菌介导的转化克隆入N.benthamiana。转基因系受到非生物胁迫,结果表明,与野生型对应物相比,转基因品系对胁迫条件具有显着的耐受性。种子萌发试验表明,与野生型品系相比,转基因品系的休眠时间延长。本研究全面分析了PvGATA基因家族,为今后研究GATATFs在普通豆科植物非生物胁迫耐受性中的作用奠定了基础。
    The GATA transcription factors (TFs) have been extensively studied for its regulatory role in various biological processes in many plant species. The functional and molecular mechanism of GATA TFs in regulating tolerance to abiotic stress has not yet been studied in the common bean. This study analyzed the functional identity of the GATA gene family in the P. vulgaris genome under different abiotic and phytohormonal stress. The GATA gene family was systematically investigated in the P. vulgaris genome, and 31 PvGATA TFs were identified. The study found that 18 out of 31 PvGATA genes had undergone duplication events, emphasizing the role of gene duplication in GATA gene expansion. All the PvGATA genes were classified into four significant subfamilies, with 8, 3, 6, and 13 members in each subfamily (subfamilies I, II, III, and IV), respectively. All PvGATA protein sequences contained a single GATA domain, but subfamily II members had additional domains such as CCT and tify. A total of 799 promoter cis-regulatory elements (CREs) were predicted in the PvGATAs. Additionally, we used qRT-PCR to investigate the expression profiles of five PvGATA genes in the common bean roots under abiotic conditions. The results suggest that PvGATA01/10/25/28 may play crucial roles in regulating plant resistance against salt and drought stress and may be involved in phytohormone-mediated stress signaling pathways. PvGATA28 was selected for overexpression and cloned into N. benthamiana using Agrobacterium-mediated transformation. Transgenic lines were subjected to abiotic stress, and results showed a significant tolerance of transgenic lines to stress conditions compared to wild-type counterparts. The seed germination assay suggested an extended dormancy of transgenic lines compared to wild-type lines. This study provides a comprehensive analysis of the PvGATA gene family, which can serve as a foundation for future research on the function of GATA TFs in abiotic stress tolerance in common bean plants.
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