植物酸性转化酶基因及其表达调控

酸性转化酶是蔗糖代谢的关键酶,在植物体中具有重要的生理作用。近十几年来,许多植物酸性转化酶基因已经克隆,其基因表达调控的研究也取得了很大的进展。本文综述了植物酸性转化酶﹑ 基因及其蛋白结构、基因表达的器官和发育特异性以及糖、受伤、病原胁迫和激素对基因表达的调节和蛋白抑制因子对酶活性的影响,并讨论了当前在该研究领域存在的问题。     

荔枝果实酸性转化酶LcSAI生物信息学和表达分析

酸性转化酶作为蔗糖代谢中的关键酶,在还原糖积累型荔枝中表达及酶活性显著高于蔗糖积累型荔枝。为了全面了解酸性转化酶的生物学特征,该文通过运用生物信息学方法对荔枝果实酸性转化酶LcSAI基因的基本理化性质、蛋白二级结构、亲水性/疏水性、跨膜结构域、信号肽、磷酸化位点、保守结构域、系统进化进行系统的分析;利用qRT-PCR技术对LcSAI在'妃子笑'不同组织和果实不同发育阶段进行表达分析。结果表明:(1)荔枝果实酸性转化酶为定位于液泡的亲水性不稳定蛋白,无信号肽,其蛋白的二级结构主要有无规则卷曲和延伸链构件,并散布于整个蛋白;(2)在N-端含有一个跨膜区,包含两个保守结构域,位于N-端的Pfam DUF3357结构域和Glyco_32结构域,属于糖基水解酶基因家族32超家族;(3)系统进化分析结果显示LcSAI与龙眼酸性转化酶基因同源,不同组织间LcSAI表达水平为雄花根嫩茎种子嫩叶雌花果皮老叶,果实不同发育阶段LcSAI表达具有特异性。该研究为深入研究LcSAI果实酸性转化酶基因调控蔗糖代谢途径机理提供数据依据。     

植物苹果酸和柠檬酸通道蛋白基因的研究进展

铝毒是酸性土壤中抑制植物生长和减少作物产量的主要因素。近年来研究表明植物主要通过根部有机酸通道蛋白将小分子有机酸阴离子转运到细胞膜外来缓解铝毒。本文综述了植物中编码铝诱导的苹果酸转运蛋白和多药及毒性复合物的排出转运蛋白两种耐铝基因,并从基因克隆、蛋白质同源性比较、基因表达调控、基因的功能和应用以及预测耐铝基因作用模式等方面进行了阐述;同时对这些耐铝基因的应用前景进行了展望。     

苹果液泡酸性转化酶基因片段的克隆及序列分析

在分析植物可溶性转化酶基因的保守区序列基础上,设计了一对PCR引物,以苹果(辽伏)基因组DNA为模板,采用PCR方法扩增出长约743bp的DNA片段,克隆入pUCm—T载体,测序结果表明,所获得的片段推测为苹果酸性转化酶基因的片段,该基因片段长度为743bp,是开放阅读框的一部分,无内含子。其序列已在GenBank中登记(登记号为AF433644)。在Gen-Bank中进行同源性检索,结果表明该成员编码的氨基酸与植物可溶性酸性转化酶的氨基酸同源性较高,与温州蜜柑(GenBank AF014925)、胡萝卜(X67163)、甜菜(GenBank X81796)和绿豆(GenBank D10265)液泡酸性转化酶基因编码蛋白质的氨基酸分别具有80％、80％、78％和77％的同源性。而与定位于细胞壁的酸性转化酶(不溶性)同源性较低,由此可推测出研究获得的苹果转化酶基因可能定于液泡。该基因片段的获得对于深入研究苹果蔗糖代谢以及果实发育和品质形成具有重要的意义。     

一种适用于酸性转化酶蛋白定量的酶联免疫吸附法

利用纯化的苹果果实可溶性酸性转化酶蛋白及其多克隆抗体,建立了一套适合于果实酸性转化酶蛋白定量测定的酶联免疫吸附方法(ELISA).以ABA对苹果、葡萄果实酸性转化酶蛋白量的调节为例,从方法学的角度对Western blotting和ELISA进行了比较,显示出ELISA具有定量分析的优势以及操作简便、灵敏度高的特点.     

小桐子转化酶基因家族的生物信息学分析及表达特性

转化酶是植物蔗糖代谢与抗逆性形成的关键酶。基于小桐子基因组数据,鉴定到16个小桐子转化酶基因,并对其系统进化、基因结构、理化性质、保守基序及表达特性进行了分析。结果表明,小桐子转化酶基因家族聚类为3个亚家族,中性/碱性转化酶α亚组与酸性转化酶基因包含6或7个外显子,β亚组包含4或5个外显子。同时,酸性转化酶N端都包含信号肽序列,且都鉴定到GH32家族保守的-NDPNG/A-与-MWECV/P-基序。qRT-PCR表达分析显示,JcC-INVD基因在小桐子子叶中表达量最高,而在根中表达量较低,且在叶中属于低温诱导基因。     

拟南芥中CBF对COR基因表达的调控1

拟南芥中COR基因的启动子含有CRT/DRE元件,转录激活因子CBF蛋白与之结合,促进COR基因的表达.CBF基因超表达不需要低温刺激就能促进COR基因的表达而增加植物的抗寒性,显示其调节COR基因表达的能力.作者对CBF基因和CBF蛋白的特性、CBF基因表达与植物抗寒性之间的关系以及CBF调节COR基因表达的机理进行了介绍.     

基因表达调控中的核因子作用

利用病毒和动物系统对基因表达调控进行了广泛和深入的研究,发现了顺式作用调节序列,鉴定了序列专一的DNA结合蛋白,DNA与蛋白质相互识别、结合及蛋白质与蛋白质相互作用中起作用的蛋白质结构域,并且对调节蛋白基因的克隆和序列进行了分析．基因表达调控领域又由于植物基因调控机制取得的发展而得到了补充,文章着重介绍植物基因中的DNA与蛋白质间的作用;植物调节蛋白基因的分离;这一领域的今后研究方向及展望．     

植物转化酶的种类、特性与功能

介绍了植物转化酶的种类、生化特性以及生理功能的研究现状,着重讨论酸性转化酶在参与光合作用调节、韧皮部卸载、贮藏器官糖的组成、细胞渗透调节以及细胞对胁迫的响应中的作用,同时对今后这一领域的研究前景提出了一些看法.     

蔗糖转化酶在高等植物生长发育及胁迫响应中的功能研究进展

随着分子生物学和测序技术的发展,植物蔗糖转化酶基因的克隆、表达调控及其功能方面的研究取得了长足的进展。综述了近年来蔗糖转化酶在植物生长发育、以及转化酶介导的植物对生物与非生物胁迫等过程中的重要作用,并对该领域今后的研究前景进行了展望。     

转化酶在高等植物蔗糖代谢中的作用研究进展

蔗糖转化酶在高等植物蔗糖代谢中起着关键的作用。研究表明 ,转化酶参与植物的生长、器官建成、糖分运输、韧皮部卸载及调节库组织糖分构成及水平。近年来关于该酶的生化特性、基因表达与调控以及结构与功能等的研究取得了重要进展。本文介绍了转化酶在植物体内的种类、分布、分子结构特点、生理作用及分子生物学研究进展。     

转化酶在高等植物蔗糖代谢中的作用研究进展

蔗糖转化酶在高等植物蔗糖代谢中起着关键的作用。研究表明,转化酶参与植物的生长、器官建成、糖分运输、韧皮部卸载及调节库组织糖分构成及水平。近年来关于该酶的生化特性、基因表达与调控以及结构与功能等的研究取得了重要进展。本文介绍了转化酶在植物体内的种类、分布、分子结构特点、生理作用及分子生物学研究进展。     

Genome-Wide Identification of the Invertase Gene Family in Populus

Invertase plays a crucial role in carbohydrate partitioning and plant development as it catalyses the irreversible hydrolysis of sucrose into glucose and fructose. The invertase family in plants is composed of two sub-families: acid invertases, which are targeted to the cell wall and vacuole; and neutral/alkaline invertases, which function in the cytosol. In this study, 5 cell wall invertase genes (PtCWINV1-5), 3 vacuolar invertase genes (PtVINV1-3) and 16 neutral/alkaline invertase genes (PtNINV1-16) were identified in the Populus genome and found to be distributed on 14 chromosomes. A comprehensive analysis of poplar invertase genes was performed, including structures, chromosome location, phylogeny, evolutionary pattern and expression profiles. Phylogenetic analysis indicated that the two sub-families were both divided into two clades. Segmental duplication is contributed to neutral/alkaline sub-family expansion. Furthermore, the Populus invertase genes displayed differential expression in roots, stems, leaves, leaf buds and in response to salt/cold stress and pathogen infection. In addition, the analysis of enzyme activity and sugar content revealed that invertase genes play key roles in the sucrose metabolism of various tissues and organs in poplar. This work lays the foundation for future functional analysis of the invertase genes in Populus and other woody perennials.     

Quantitative analysis of the regulation scheme of invertase expression in Saccharomyces cerevisiae.

In Saccharomyces cerevisiae, the expression of invertase, which is the hydrolyzing enzyme of sucrose, is controlled by the presence of monosaccharides, such as glucose and fructose, and referred to as carbon catabolite repression. To date, efforts have been made to identify the mechanism by which cells sense extracellular monosaccharide concentrations and trigger the genes involved in the repression pathway. The aim of the present work was to quantitatively investigate the cellular regulation of invertase expression in the wild-type strain S. cerevisiae CEN.PK113-7D during batch growth containing mixed sugar substrates under different initial conditions. Because of the high frequency and accurate online analysis of multiple components, a tight control of invertase expression could be observed, and threshold concentrations of the monosaccharides for derepression could be determined to 0.5 gl(-1) for glucose and 2 gl(-1) for fructose. Also, the existence of a hitherto undescribed regulatory state, in which cells regulate invertase expression very precisely and operate over long periods at monosaccharide concentrations lower than the above thresholds, could be demonstrated. All experimental observations could be summarized in a formulation of the cellular regulation scheme of invertase expression. A simple kinetic model could show that the regulation scheme explains the observed behavior very well. Additionally, the model was able to explain consequences of the regulation on the global metabolism.     

Post-translational inhibitory regulation of acid invertase induced by fructose and glucose in developing apple fruit

Acid invertase (EC 3.2.1.26) is one of the key enzymes involved in the carbohydrate sink-organ development and the sink strength modulation in crops. The experiment conducted with 'Starkrimson' apple (Malus domestica Borkh) fruit showed that, during the fruit development, the activity of acid invertase gradually declined concomitantly with the progressive accumulation of fructose, glucose and sucrose, while Western blotting assay of acid invertase detected a 30 ku peptide of which the immuno-signal intensity increased during the fruit development. The im-muno-localization via immunogold electron microscopy showed that, on the one hand, acid invertase was mainly located on the flesh cell wall with numbers of the immunosignals present in the vacuole at the late stage of fruit development; and on the other hand, the amount of acid invertase increased during fruit development, which was consistent with the results of Western blotting. The in vivo pre-incubation of fruit discs with soluble sugars showed that