脱落酸与Em基因的表达调控

En基因的表达受ABA诱导,干旱和盐胁通过增加ABA含量或改变植物细胞对ABA的敏感性而诱导Em基因的表达。植物Em基因启动存在3个功能区：5′远端AT富集区通过影响转录调节表达量,作用类似于非专一性增强子;ABA应答元件ABRE在ABA存在的情况下与转录因子EmBP-1相互作用能显著增强Em基因的表达;5′UTR可能通过转录后调控而影响最终表达水平。     

非编码序列对仙台病毒HN基因表达的调控作用

仙台病毒的血凝素神经氨酸酶 (HN)蛋白在COS 7细胞中得到了表达 .结果表明 ,SRα启动子驱动HN基因表达的活性高于鸡 β肌动蛋白启动子 .而且 ,HN基因的 5′非编码序列能促进其表达 .Northern杂交证明 ,高表达是由HN mRNA转录引起 .为研究HN基因 5′编码序列对其转录的调控作用 ,用位点专一性突变分别以角蛋白基因和细胞色素P45 0基因的 5′非编码序列替代HN基因的 5′非编码序列 ,并分别缺失HN基因 3′非编码序列 ,构建了一系列表达载体 .以氯霉素乙酰转移酶(CAT)基因为报道基因 ,用S1酶对HN mRNA转录量进行定量分析 .实验证明 ,HN基因 5′非编码序列能非特异性提高HN mRNA的转录 ,3′非编码序列对其转录也有某种特殊的调控作用 .     

植物体内干旱信号的传递与基因表达

干旱是严重影响植物生长发育的重要环境胁迫因子之一。干旱能影响植物的水分状态,使植物缺水遭受伤害。近年来,相继从拟南芥等植物中克隆出了一些受干旱诱导的基因,如蛋白激酶基因、光合基因、渗透调节基因、功能蛋白基因（如LEA基因）等。干旱等胁迫信号经历一系列的传递过程,最后诱导这些特定基因的表达。在植物体中,可能存在依赖ABA型和不依赖ABA型两条干旱信号的传递途径。近年来从高等植物中分离出一系列调控干旱相关基因表达的转录因子,通过转录因子之间以及与其它相关蛋白之间的相互作用,激活或抑制干旱等胁迫因子诱导的基因表达。     

脱落酸激素诱导拟南芥幼苗中花青素的合成

脱落酸(abscisic acid,ABA)激素是一类重要的生长调节物质,参与调控植物的多种生理过程。花青素(anthocyanins)是植物次生代谢产生的类黄酮化合物,对植物的生长发育和逆境胁迫响应有重要作用。该文以拟南芥(Arabidopsis thaliana)为研究对象,探讨ABA信号对花青素生物合成的调控功能和作用机制。结果表明:外源施加ABA显著提高野生型幼苗茎尖中花青素的积累。相一致的是,ABA能诱导某些与花青素合成相关的转录因子及合成酶基因的表达。遗传学分析发现,ABA诱导花青素合成部分依赖于MBW复合体中的核心转录因子,如TTG1、TT8及MYB75等。初步机制研究揭示,ABA信号途径中的bZIP类转录因子ABI5能与TTG1、TT8及MYB75等相互作用形成蛋白复合物。综上结果认为,ABA信号诱导拟南芥幼苗中花青素的积累,并可能通过ABI5与MBW复合体协同作用调控花青素的合成。     

内源ABA信号水平动态调控的分子机制

从逆境信号感知、ABA合成的触发到ABA水平的动态调控, 是细胞内重要的逆境信号传导途径, 相对于应答ABA的下游信号事件, 该领域研究滞后。研究显示, 根系中ZEP、限速酶NCED、AtRGS1等合成酶基因及ABA2基因响应胁迫反应上调ABA信号水平。而7′-, 8′-, 9′-hydroxylase和糖基转移酶基因受逆境诱导激活, 负调节ABA的积累。同时, 提高的内源ABA信号水平能激活合成酶基因和代谢酶基因的表达。此外, 基因表达和源库动力学分析显示, 叶片ABA动态库的维持依赖根源ABA的持续供应。值得一提的是, miRNA与ABA信号起源及动态水平维持有关。进一步的代谢动力学分析揭示, ABA信号水平受合成酶基因和代谢酶基因表达的协同控制, 多因素共同参与内源ABA信号水平的动态调控。     

病原菌诱导型启动子顺式作用元件及其互作的转录因子

启动子是位于基因5′端上游的一段DNA序列, 负责调控基因的转录。与植物抗病相关基因的启动子区含有能针对病原菌胁迫做出应答的顺式作用元件, 这些顺式作用元件通过与转录因子特异性结合, 进而增强抗病基因的转录表达, 提高植物的抗病性。该文主要综述了病原菌诱导型启动子相关顺式作用元件及与这些元件互作的转录因子, 特别对一类特殊的转录因子--病原菌TAL效应子与植物靶基因启动子之间的相互作用机制进行了阐述, 并对其应用前景进行了展望。     

植物冷驯化的分子机理研究进展

植物冷驯化是一个非常复杂的过程,包括植物将感受到的低温信号转变成生化信号,以激活冷诱导基因的启动子,刺激特定的mRNA的转录,并在特定的组织中合成冷驯化蛋白.冷驯化蛋白通过增强膜脂流动性和阻止胞间冰晶形成等方式,以保护细胞免受低温伤害.冷驯化基因的表达以转录后调控为主,也有转录调控.某些冷诱导基因也可受ABA或其他环境胁迫(如高温、干旱、高盐、脱水等)诱导表达.     

玉米bHLH类转录因子ABP7的启动子克隆及其活性分析

bHLH类转录因子广泛存在于植物中,在花的形态建成、光和激素信号转导等方面发挥着重要作用,但其表达调控机理尚待深入研究。从玉米中克隆了bHLH类转录因子ABP7,初步研究结果显示它可能参与了籽粒形成、激素应答等多个信号转导途径的调控。为进一步阐明ABP7基因上游调控途径,克隆了ABP7的5′侧翼序列,利用原生质体瞬时转化体系验证了其启动子活性,并对其顺式作用元件进行了分析。结果显示,位于ABP7基因起始密码子上游长为2 266 bp的DNA序列具有启动子活性,能够启动报告基因的表达,并且存在许多潜在的与GA、IAA和ABA激素信号转导和逆境胁迫响应相关的元件,为进一步鉴定ABP7基因发挥作用的信号途径、上游调控因子和分子机理以及ABP7基因在玉米生长发育和逆境应答中的功能奠定了基础。     

植物启动子的诱导模序

启动子是位于基因 5′端并负责该基因转录的DNA序列。诱导性启动子中有一些对伤害、真菌侵染、紫外线照射、激素等做出应答反应的顺式作用元件 ,被称为模序。已在植物启动子中鉴定出许多与诱导表达相关的模序 ,如伤害诱导模序 ,真菌诱导模序 ,植物激素诱导模序和光诱导模序等。这些模序作为启动子的顺式元件对各种诱导因子做出反应、调控基因的表达。     

普通小麦TaNAC1基因的表达及在拟南芥中的遗传转化

NAC类转录因子是植物特有的转录因子家族,在调节植物生长发育及逆境胁迫应答反应中起着重要作用。本文从普通小麦幼叶中获得了一个编码NAC结构域的转录因子基因,命名为Ta NAC1;氨基酸序列分析表明,Ta NAC1具有典型的NAC类转录因子所具有的五个亚结构域,隶属于NAC类转录因子的ATAF亚类;亚细胞定位实验表明,Ta NAC1蛋白在细胞核内表达;转录水平上,Ta NAC1基因的表达受到PEG、ABA、低温及高盐等非生物胁迫条件的诱导;将Ta NAC1转化拟南芥后,与野生型比较发现,Ta NAC1基因的过量表达会使转基因植株出现叶片发育畸形且生长缓慢,植株矮化及茎部融合等表型,表明Ta NAC1基因可能在参与小麦叶片及茎的发育中起着重要的调控作用。     

Analysis of an osmotically regulated pathogenesis-related osmotin gene promoter

Osmotin is a small (24 kDa), basic, pathogenesis-related protein, that accumulates during adaptation of tobacco (Nicotiana tabacum) cells to osmotic stress. There are more than 10 inducers that activate the osmotin gene in various plant tissues. The osmotin promoter contains several sequences bearing a high degree of similarity to ABRE, as-1 and E-8 cis element sequences. Gel retardation studies indicated the presence of at least two regions in the osmotin promoter that show specific interactions with nuclear factors isolated from cultured cells or leaves. The abundance of these binding factors increased in response to salt, ABA and ethylene. Nuclear factors protected a 35 bp sequence of the promoter from DNase I digestion. Different 5 deletions of the osmotin promoter cloned into a promoter-less GUS-NOS plasmid (pBI 201) were used in transient expression studies with a Biolistic gun. The transient expression studies revealed the presence of three distinct regions in the osmotin promoter. The promoter sequence from –108 to –248 bp is absolutely required for reporter gene activity, followed by a long stretch (up to –1052) of enhancer-like sequence and then a sequence upstream of –1052, which appears to contain negative elements. The responses to ABA, ethylene, salt, desiccation and wounding appear to be associated with the –248 bp sequence of the promoter. This region also contains a putative ABRE (CACTGTG) core element. Activation of the osmotin gene by various inducers is discussed in view of antifungal activity of the osmotin protein.     

Functional dissection of an abscisic acid (ABA)-inducible gene reveals two independent ABA-responsive complexes each containing a G-box and a novel cis-acting element.

To elucidate the mechanism by which abscisic acid (ABA) regulates gene expression, the promoter of the barley ABA-responsive HVA22 gene has been analyzed by both loss- and gain-of-function studies. Previous reports indicate that G-box sequences, which are present in genes responding to a variety of environmental and physiological cues, are involved in ABA response. However, our data suggest that G-box sequences are necessary but not sufficient for ABA response. Instead, an ABA response complex consisting of a G-box, namely, ABRE3 (GCCACGTACA), and a novel coupling element, CE1 (TGCCACCGG), is sufficient for high-level ABA induction, and replacement of either of these sequences abolishes ABA responsiveness. We suggest that the interaction between G-box sequences, such as ABRE3 in the HVA22 gene, and CE-type sequences determines the specificity in ABA-regulated gene expression. Our results also demonstrate that the ABA response complex is the minimal promoter unit governing high-level ABA induction; four copies of this 49-bp-long complex linked to a minimal promoter can confer more than 100-fold ABA-induced gene expression. In addition to ABA response complex 1, composed of ABRE3 and CE1, the HVA22 promoter contains another ABA response complex. The ABA responsiveness of this ABA response complex 2 relies on the interaction of G-box (ABRE2; CGCACGTGTC) with another yet unidentified coupling element. These two complexes contribute incrementally to the expression level of HVA22 in response to ABA.     

Cis-elements and trans-factors that regulate expression of the maize Cat1 antioxidant gene in response to ABA and osmotic stress: H2O2 is the likely intermediary signaling molecule for the response

The mechanisms by which the maize antioxidant Cat1 gene responds to abscisic acid (ABA) and osmotic stress have been investigated. Results show that during late embryogenesis, Cat1 expression in vivo is independent of endogenous ABA levels. However, exogenously applied ABA significantly enhances Cat1 expression. Transient assays using particle bombardment show that the proximal ABRE2 element on the Cat1 promoter is responsible for the induction of Cat1 expression by ABA. We further show that ABA induces the expression of Cat1 via the interaction between ABRE2 and one of its binding proteins, CBF1 (Cat1 binding factor 1). Using ABA-deficient mutant embryos, we show that osmotic stress induces Cat1 expression through two alternate signal transduction pathways: an ABA signaling pathway leading to the interaction between the ABRE2 motif and CBF1, and a pathway via the interaction of ABRE2 and CBF2 (Cat1 binding factor 2) that is independent of ABA. The data presented clearly suggest that hydrogen peroxide (H2O2) plays an important intermediary role in the ABA signal transduction pathway leading to the induction of the Cat1 gene.     

Interaction between two cis-acting elements,ABRE and DRE,in ABA-dependent expression of Arabidopsis rd29A gene in response to dehydration and high-salinity stresses

Many abiotic stress-inducible genes contain two cis-acting elements, namely a dehydration-responsive element (DRE; TACCGACAT) and an ABA-responsive element (ABRE; ACGTGG/TC), in their promoter regions. We precisely analyzed the 120 bp promoter region (-174 to -55) of the Arabidopsis rd29A gene whose expression is induced by dehydration, high-salinity, low-temperature, and abscisic acid (ABA) treatments and whose 120 bp promoter region contains the DRE, DRE/CRT-core motif (A/GCCGAC), and ABRE sequences. Deletion and base substitution analyses of this region showed that the DRE-core motif functions as DRE and that the DRE/DRE-core motif could be a coupling element of ABRE. Gel mobility shift assays revealed that DRE-binding proteins (DREB1s/CBFs and DREB2s) bind to both DRE and the DRE-core motif and that ABRE-binding proteins (AREBs/ABFs) bind to ABRE in the 120 bp promoter region. In addition, transactivation experiments using Arabidopsis leaf protoplasts showed that DREBs and AREBs cumulatively transactivate the expression of a GUS reporter gene fused to the 120 bp promoter region of rd29A. These results indicate that DRE and ABRE are interdependent in the ABA-responsive expression of the rd29A gene in response to ABA in Arabidopsis.     

Sequence analysis of a functional member of the Em gene family from wheat.

We report the complete sequence of one functional member of the Em gene family whose expression in wheat embryos is regulated by a complex set of environmental and developmental controls, including the phytohormone abscisic acid (ABA). The Em coding region contains one short intron, and there is an inverted repeat in the transcribed 3'-flanking region. A 646 bp fragment from the 5' promoter, which was previously shown to direct ABA-regulated expression in transformed tobacco tissue and rice cells, is characterized by: (1) three stretches of between 33 and 73 nucleotides of A/T rich (greater than 86%) boxes, (2) one copy of an eight bp palindrome (CATGCATG) which is identical to the RY repeat found in the 5' promoters of many legume genes expressed during embryo development, (3) 15 copies of a six bp repeat (PuCACGPy), found primarily in the 5' region, and (4) two sequences in the ABA-response region, CGAGCAG and a CACGT motif, both of which are conserved in 5' non-coding regions of other plant genes that are expressed in response to ABA and/or in embryos. These sequence comparisons are discussed in relation to the regulation of Em gene expression and other ABA-regulated genes.