赤霉素关键蛋白DELLA对植物雄蕊发育研究进展

在植物生长发育过程中,DELLA蛋白作为响应赤霉素（GA）信号通路负调控因子,其家族成员在植物雄蕊组织中均有表达,对植物生长发育起着重要作用.通过近几年研究发现,无论是拟南芥还是水稻,DELLA基因的突变都会导致植株的雄性不育.该文综述了DELLA蛋白基本特征、参与赤霉素信号通路的调控,以及在雄蕊发育中的作用,为今后的研究提供一定的理论基础.     

海岛棉DELLA蛋白基因GbGAI2的克隆及其在胚珠中表达分析

赤霉素(Gibberellins,GA)在棉花纤维的发育过程中起重要作用,DELLA蛋白是GA信号通路中的起关键作用的负调控因子。本研究采用同源克隆方法,获得了海岛棉的DELLA蛋白同源基因GbGAI2。序列比对分析结果表明,GbGAI2编码的氨基酸序列具有DELLA蛋白的典型结构域,并与陆地棉DELLA蛋白GhGAI2氨基酸有90.13%相同。半定量RT-PCR分析结果表明,GbGAI2在1-5DPA和23DPA棉胚中的表达量较高,说明该基因可能在海岛棉棉纤维发育的起始阶段和棉纤维次生壁加厚阶段起作用。     

棉花DELLA蛋白GhGAI2b基因的克隆和功能初步分析

DELLA蛋白是赤霉素信号途径的负调节因子,在棉花纤维发育中有着重要作用。本研究采用同源克隆方法,从棉花中克隆DELLA蛋白Gh GAI2b基因,构建p BP35S:Gh GAI2b和p BP35S:Ghgai2b植物过表达载体,通过农杆菌介导的滴花法转化Col野生型拟南芥。结果表明,棉花DELLA蛋白Gh GAI2b包含了DELLA蛋白家族中所有的典型保守区域;转基因拟南芥表现出莲座叶半径变短,植株矮化,花序紧凑,花器官发育迟缓等生长发育受抑制表型。说明棉花DELLA蛋白Gh GAI2b基因可能参与GA信号途径抑制植物生长发育。     

2013-26 科技要闻

揭示植物激素作用新机制华中师范大学杨光富教授研究组在深入分析GID1-GA(GA,赤霉素;GID1,GIBBERELLIN INSENSITIVE DWARF1)复合体晶体结构的基础上,通过对GID1蛋白的构象进行微秒级的分子动力学模拟,首次发现了赤霉素与受体GID1蛋白结合的新通道,并提出了"非变构学说"的赤霉素作用新机制,即赤霉素在通过新通道与GID1蛋白结合时并不介导其氮端结构域发生构象变化,而是通过稳定GID1与DELLA蛋白之间的氢键而介导DELLA蛋     

用CRISPR/Cas9系统构建原位表达dXPR1-GFP的果蝇品系

XPR1是最近鉴定出的一个原发性家族性脑钙化症(primary familial brian calcification,PFBC)的致病基因,它在果蝇体内的同源基因是CG10483(dXPR1).使用CRISPR/Cas9系统,构建原位表达dXPR1-GFP的果蝇品系,在CG10483终止密码子之前插入绿色荧光蛋白(green fluorescent protein,GFP)的编码序列,使果蝇表达dXPR1-GFP融合蛋白.该融合蛋白既可以被CG10483编码蛋白的抗体和GFP的抗体识别又能在激发光下发出绿色荧光,能为dXPR1编码蛋白提供准确的定位,从而为研究XPR1基因的功能提供方便.     

NAC1启动子驱动GFP表达的组织特征及对生长素和赤霉素的反应

将拟南芥中转录因子基因NAC1上游调控区1kb克隆到pRD420质粒，构建由NAC1启动子驱动绿色荧光蛋白基因（GFP）表达的植物表达载体，并以此载体转化烟草，对阳性转基因植株研究表明，GFP在根部有较高水平的表达.进一步用生长素、生长素拮抗剂NPA和赤霉素处理，荧光强度的变化表明：该调控区受生长素作用的同时也受赤霉素诱导.初步说明NAC1基因不仅是一个生长素反应基因，同时可能也是一个赤霉素反应基因.     

NACl启动子驱动GFP表达的组织特征及对生长素和赤霉素的反应

将拟南芥中转录因子基因NACl上游调控区1kb克隆到pRD420质粒，构建由NACl启动子驱动绿色荧光蛋白基因（GFP）表达的植物表达载体，并以此载体转化烟草，对阳性转基因植株研究表明，GFP在根部有较高水平的表达．进一步用生长素、生长素拮抗剂NPA和赤霉素处理，荧光强度的变化表明：该调控区受生长素作用的同时也受赤霉素诱导．初步说明NACl基因不仅是一个生长素反应基因，同时可能也是一个赤霉素反应基因．     

P因子介导mir-2基因簇原位过表达调节Yki的活性分析

Hippo信号通路在动物器官大小发育和肿瘤发生过程中发挥关键性作用.为了发掘新的Hippo信号通路活性调节因子,通过P因子插入介导基因原位过表达,在果蝇表型修饰筛选中鉴定了1株可以显著增强Hippo信号通路下游效应因子Yki活性的果蝇株MS004.反向PCR定位显示,MS004中P因子位于2号染色体左臂蛋白编码基因spitz的内含子区域;进一步的遗传分析确定,MS004果蝇株通过诱导位于spitz内含子区域的mir-2基因簇原位过表达增强Yki的活性.     

海岛棉ERF族转录因子EREB6基因克隆特征分析

以电子克隆,同源扩增和RACE技术相结合的方法从海岛棉中克隆出了一条新基因,命名为EREB6,它包含一个669 bp长的开放阅读框,预期编码222个氨基酸长的蛋白质.序列分析发现预期蛋白含有一段核定位信号序列,GFP融合蛋白瞬时表达实验证明该蛋白定位于细胞核;含有一个保守的ERF域,属于ERF族;同源分析表明该转录因子...     

用CRISPR/CAS9系统构建原位表达dPit:GFP的果蝇品系

SLC20A2基因是Ⅲ型纳磷转运蛋白2(PiT2)的编码基因,PiT2可将无机磷从细胞外运送至细胞内.SLC20A2基因突变导致PiT2不能正常地将无机磷运送到细胞内,血液中的无机磷含量升高,磷和钙离子结合形成沉淀,堵塞血管.SLC20A2基因在果蝇中的同源基因是CG42575(dpit).本实验使用CRISPR/CAS9系统,构建原位表达dPiT:GFP的果蝇品系,将绿色荧光蛋白(green fluorescent protein,GFP)编码序列插入dpit基因末端,使果蝇表达dPiT:GFP融合蛋白.该融合蛋白可以被dPiT及GFP的抗体同时识别,通过荧光直接观察dPiT:GFP的位置,从而准确定位dpit,进而为dpit基因的功能研究提供基础.     

Auxin promotes Arabidopsis root growth by modulating gibberellin response

The growth of plant organs is influenced by a stream of the phytohormone auxin that flows from the shoot apex to the tip of the root. However, until now it has not been known how auxin regulates the cell proliferation and enlargement that characterizes organ growth. Here we show that auxin controls the growth of roots by modulating cellular responses to the phytohormone gibberellin (GA). GA promotes the growth of plants by opposing the effects of nuclear DELLA protein growth repressors, one of which is Arabidopsis RGA (for repressor of gal-3). GA opposes the action of several DELLA proteins by destabilizing them, reducing both the concentration of detectable DELLA proteins and their growth-restraining effects. We also show that auxin is necessary for GA-mediated control of root growth, and that attenuation of auxin transport or signalling delays the GA-induced disappearance of RGA from root cell nuclei. Our observations indicate that the shoot apex exerts long-distance control on the growth of plant organs through the effect of auxin on GA-mediated DELLA protein destabilization.     

Expression pattern of GASA,downstream genes of DELLA,in Arabidopsis

Separation and functional research of related components involved in gibberellins （GAs） signaling are important to clarify the mechanism of GA functioning. Research on the downstream components of DELLA, the key factor of the GA signaling pathway, is limited at present. GASA （GA-Stimulated in Arabidopsis） family contains 15 genes usually regulated by GA in Arabidopsis thaliana. All GASA proteins have a cleavable signal peptide in N terminus and a conserved GASA domain including 12 cysteines in C terminus. RT-PCR analysis revealed that the expression of GASA4 and GASA6 were down-regulated, but GASA 1 and GASA9 were up-regulated in the DELLA mutants, gai-t6 and rga-24, as well as the double mutant, consisting with the results that GASA4 and GASA6 were induced, but GASA1 and GASA9 were inhibited by exogenous GA3. In addition, the expression patterns of other GASA genes were regulated by GA and ABA, separately or cooperatively. Most of GASA genes were expressed in roots, stems, leaves, flowers and developing siliques. GUS gene driven by the promoters of GASA6, GASA7, GASA8, GASA9, GASAIO, GASA11 and GASA12were used as reporters and it was found that all GASA genes expressed in the growing and differentiating organs and abscission zones, suggesting the role of these genes in cell growth and differentiation. This study provided an important basis for functional study of the GASA gene family in the GA and ABA signaling pathway.     

GIBBERELLIN INSENSITIVE DWARF1 encodes a soluble receptor for gibberellin

Gibberellins (GAs) are phytohormones that are essential for many developmental processes in plants. It has been postulated that plants have both membrane-bound and soluble GA receptors; however, no GA receptors have yet been identified. Here we report the isolation and characterization of a new GA-insensitive dwarf mutant of rice, gid1. The GID1 gene encodes an unknown protein with similarity to the hormone-sensitive lipases, and we observed preferential localization of a GID1-green fluorescent protein (GFP) signal in nuclei. Recombinant glutathione S-transferase (GST)-GID1 had a high affinity only for biologically active GAs, whereas mutated GST-GID1 corresponding to three gid1 alleles had no GA-binding affinity. The dissociation constant for GA4 was estimated to be around 10(-7) M, enough to account for the GA dependency of shoot elongation. Moreover, GID1 bound to SLR1, a rice DELLA protein, in a GA-dependent manner in yeast cells. GID1 overexpression resulted in a GA-hypersensitive phenotype. Together, our results indicate that GID1 is a soluble receptor mediating GA signalling in rice.     

植物生长调节剂对翅果油树坐果率的影响

本文研究了不同植物生长调节剂对翅果油树坐果率及有机物含量的影响．结果表明：在光照充足的条件下,喷施植物生长调节剂能减少翅果油树的落花落果现象,提高坐果率．其中增幅最大处理为：400mg·L^-1 GA3和300mg·L^-1 GA3＋100mg·L^-16-BA;此外,喷施50mg·L^-1 NAA和200mg·L^-1 GA3～400mg·L^-1还可使果实中的可溶性糖与蛋白质含量增加．     

DELLA基因家族在被子植物中的系统演化关系

基于被子植物各主要分支代表类群的DELLA氨基酸序列，开展它们在被子植物中演化关系的系统发育分析.DELLA基因家族在双子叶植物早期演化阶段经历了一次复制事件，形成2大分支，每支都包含相应的蔷薇类和菊类；单子叶植物没有经历早期的复制事件，所有的DELLA基因聚在一起形成单一的支系.另外，证实了在被子植物的双子叶植物中存在第3个DELLA基因支系.     

植物激素-赤霉素(GA)细胞信号转导机制

植物激素赤霉素(GA)参与植物种子萌发、茎伸长、开花、结果等多个生长发育过程.研究GA细胞信号转导的分子机制对进一步阐明其生物学功能具有重要的意义.GA合成突变体和细胞信号转导突变体的研究表明,GAI及其同功蛋白具有受体功能,GA被受体识别后进一步与DELLA蛋白作用,从而解除DELLA蛋白对植物生长的抑制作用.文章主要综述这些分子具体的作用模式.