细胞分裂素结合蛋白的研究进展

迄今为止,已从多种植物中分离到细胞分裂素结合蛋白（ＣＢＰｓ）,它们可能在细胞分裂素的信号转导、体内运输及代谢中起作用。根据现有研究结果认为,大多数ＣＴＫｓ受体可能位于膜上,通过与Ｇ－蛋白耦联的信号转导系统或双组分信号转导系统完成ＣＴＫｓ信号的跨膜转导。少数ＣＴＫｓ受体可能位于细胞质中,与胞内ＣＴＫｓ结合后进入细胞核,直接调节基因的表达。本文综述了近年来对ＣＢＰｓ的研究进展,分析了ＣＴＫｓ受体的可能     

细胞分裂素信号转导研究进展

对应用突变体研究细胞分裂素信号转导、细胞分裂素受体以及参与细胞分裂素信号转导相关的蛋白质作了简要介绍 ;并且对细胞分裂素信号途径进行了推测 :细胞分裂素被受体 (CKI1、IBC6或者GCR1 )接受后 ,经传导系统转化形成特定的信息 ,一方面可能调节基因的表达 ,从而可能调节受体水平 ,导致细胞对细胞分裂素浓度应答范围发生改变 ,另外 ,基因表达使细胞产生相关的生理反应 ;另一方面形成的特定信息可能激活MAPK级联途径 (mitogen_activatedproteinkinasecascade) ,导致细胞产生相关的生理反应。     

细胞分裂素信号转导研究进展

对应用突变体研究细胞分裂素信号转导、细胞分裂素受体以及参与细胞分裂素信号转导相关的蛋白质作了简要介绍;并且对细胞分裂素信号途径进行了推测：细胞分裂素被受体（CKI1、IBC6或者GCR1）接受后,经传导系统转化形成特定的信息,一方面可能调节基因的表达,从而可能调节受体水平,导致细胞对细胞分裂素浓度应答范围发生改变,另外,基因表达使细胞产生相关的生理反应;另一方面形成的特定信息可能激活MAPK级联途径（mitogen-activated protein kinase cascade）,导致细胞产生相关的生理反应。     

细胞分裂素信号转导分子机制

细胞分裂素受体家族与细菌二元组分系统的感受器组氨酸激酶具有同源性,证实下游事件与传统的磷酸转运作用具有相似性.借助于AHP蛋白的瞬间转运作用,细胞分裂素信号通过定位在细胞膜的类组氨酸激酶受体传到细胞核内,AHP蛋白使B型ARR活化,随后B型ARR激活A型ARR或其它靶基因的转录,逐步形成从质膜接受部位到激活核内基因表达的细胞分裂素信号转导模式.     

亚种间杂交稻根系伤流液与地上部细胞分裂素的变化

本文以水稻亚种间杂交组合Ⅱ优2070及恢复系2070、Ⅱ优419及恢复系中419为材料．应用HPLC和ELISA法测定灌浆期间根系伤流液中根源细胞分裂素（CTKs）种类和数量以及稻叶和籽粒中细胞分裂素组分含量的变化．研究表明在亚种杂交稻及其恢复系灌浆起始阶段,玉米素（Z）占总CTKs的比例高达62．8％-89．1％,是根系伤流液中细胞分裂素的主要成分．而二氢玉米素（diHZ）和二氢玉米素核苷（diHZR）则在灌浆后期明显上升,这种变化动态与II优2070和2070剑叶中ZRs（Z＋ZR）、diHZRs（diHZ＋diHZR）的波动变化是相符合的。与根系伤流液的主要细胞分裂素组成不同,IPAs在叶片和籽粒中占总CTKs量的比例最高。说明在灌浆期间．根系伤流液中CTKs的种类及其活性存在着变化,这种变化可能与细胞分裂素代谢酶活性变化及其相关酶基因表达的不同有关。讨论了强、弱势粒的CTKs组分的变化与籽粒结实的可能关系。     

亚种间杂交稻根系伤流液与地上部细胞分裂素的变化

本文以水稻亚种间杂交组合Ⅱ优2070及恢复系2070、Ⅱ优419及恢复系中419为材料．应用HPLC和ELISA法测定灌浆期间根系伤流液中根源细胞分裂素(CTKs)种类和数量以及稻叶和籽粒中细胞分裂素组分含量的变化,研究表明在亚种杂交稻及其恢复系灌浆起始阶段,玉米素(Z) 占总CTKs的比例高达62．8％-89．1％,是根系伤流液中细胞分裂素的主要成分,而二氢玉米素(di- HZ)和二氢玉米素核苷(diHZR)则在灌浆后期明显上升．这种变化动态与Ⅱ优2070和2070剑叶中ZRs(Z ZR)、diHZRs(diHZ diHZR)的波动变化是相符合的。与根系伤流液的主要细胞分裂素组成不同,IPAs在叶片和籽粒中占总CTKs量的比例最高。说明在灌浆期间,根系伤流液中CTKs的种类及其活性存在着变化,这种变化可能与细胞分裂素代谢酶活性变化及其相关酶基因表达的不同有关。讨论了强、弱势粒的CTKs组分的变化与籽粒结实的可能关系。     

细胞分裂素: 代谢、信号转导、交叉反应与农艺性状改良

在高等植物中, 细胞分裂素通过对细胞分裂与分化的调节而广泛参与了对植物生长发育的调控。在过去的10余年, 利用模式植物拟南芥的研究, 在阐明细胞分裂素的代谢、转运与信号转导等方面取得了重要的进展。同时, 关于细胞分裂素与其它信号途径之间存在的广泛交叉反应也受到了人们的注意。根据我们现有的知识, 细胞分裂素信号转导是通过磷酸基团在一个双元组分系统之间的系列传递而完成的, 该过程被称之为“磷酸接力传递”(phosphorelay)。细胞分裂素与其它信号途径的互作可能也主要是通过双元组分系统链接的。双元组分系统中目前已知的主要信号元件不仅表现出功能冗余性, 同时在调控特定的植物生长发育过程时也具有特异性。本文在对细胞分裂素的代谢与转运过程简要评述的基础上,对其信号转导以及与其它信号途径间交叉反应的研究进展进行重点讨论, 并展望细胞分裂素研究对重要农业性状改良的意义。     

细胞分裂素:代谢、信号转导、交叉反应与农艺性状改良

在高等植物中,细胞分裂素通过对细胞分裂与分化的调节而广泛参与了对植物生长发育的调控。在过去的10余年,利用模式植物拟南芥的研究,在阐明细胞分裂素的代谢、转运与信号转导等方面取得了重要的进展。同时,关于细胞分裂素与其它信号途径之间存在的广泛交叉反应也受到了人们的注意。根据我们现有的知识,细胞分裂素信号转导是通过磷酸基团在一个双元组分系统之间的系列传递而完成的,该过程被称之为“磷酸接力传递”（phosphorelay）。细胞分裂素与其它信号途径的互作可能也主要是通过双元组分系统链接的。双元组分系统中目前已知的主要信号元件不仅表现出功能冗余性,同时在调控特定的植物生长发育过程时也具有特异性。本文在对细胞分裂素的代谢与转运过程简要评述的基础上,对其信号转导以及与其它信号途径间交叉反应的研究进展进行重点讨论,并展望细胞分裂素研究对重要农业性状改良的意义。     

细胞分裂素对植物基因表达的调节

细胞分裂素能显著改变植物基因转录的水平,在转录水平上或转录后水平上调控植物基因的表达。蛋白质磷酸化在细胞分裂素的信号转导过程中起着重要的作用,但迄今为止细胞分裂素的顺式作用元件和反式作用因子尚未有报道。文章最后就细胞分裂素对植物基因表达调节研究作了展望。     

植物双组分信号系统

植物中存在着一种双组分信号系统,该系统由组氨酸蛋白激酶,HPt蛋白和反应调节器组成,它参与乙烯,细胞分裂素等的信号转导,并且与光敏色素信号产生有关。     

Detection of membrane-bound cytokinin-binding proteins in Arabidopsis thaliana cells.

In order to isolate cytokinin-binding proteins (CBPs), we have developed new affinity probes constituted of a cytokinin such as zeatin riboside ([9R]Z) conjugated to a carrier protein. These probes were used for detecting CBPs in an ELISA procedure. The efficiency of the cytokinin conjugate in detecting CBPs was controlled with protein model: proteins having an affinity for cytokinin such as the monoclonal anti-[9R]Z antibodies did bind the cytokinin conjugate whereas proteins unable to bind cytokinin such as bovine serum albumin did not. Using these new affinity probes, we showed that CBPs are present in the membrane fraction of in vitro cultured Arabidopsis thaliana cells. The nature of the protein at the detected binding sites was demonstrated by submitting the microsomal proteins to a proteolytic treatment, which was found to eradicate the binding. Free biologically active cytokinins or monoclonal anti-[9R]Z antibodies inhibited the binding, thus showing the specificity of the interaction. The detected CBPs were partially solubilized from the membranes with potassium chloride, indicating their peripheral membrane location. The separation by anion exchange chromatography of solubilized microsomal proteins revealed the existence of two different CBPs. They were present at higher levels in cells during the exponential growth phase.     

Analysis of protein interactions within the cytokinin-signaling pathway of Arabidopsis thaliana

The signal of the plant hormone cytokinin is perceived by membrane-located sensor histidine kinases and transduced by other members of the plant two-component system. In Arabidopsis thaliana, 28 two-component system proteins (phosphotransmitters and response regulators) act downstream of three receptors, transmitting the signal from the membrane to the nucleus and modulating the cellular response. Although the principal signaling mechanism has been elucidated, redundancy in the system has made it difficult to understand which of the many components interact to control the downstream biological processes. Here, we present a large-scale interaction study comprising most members of the Arabidopsis cytokinin signaling pathway. Using the yeast two-hybrid system, we detected 42 new interactions, of which more than 90% were confirmed by in vitro coaffinity purification. There are distinct patterns of interaction between protein families, but only a few interactions between proteins of the same family. An interaction map of this signaling pathway shows the Arabidopsis histidine phosphotransfer proteins as hubs, which interact with members from all other protein families, mostly in a redundant fashion. Domain-mapping experiments revealed the interaction domains of the proteins of this pathway. Analyses of Arabidopsis histidine phosphotransfer protein 5 mutant proteins showed that the presence of the canonical phospho-accepting histidine residue is not required for the interactions. Interaction of A-type response regulators with Arabidopsis histidine phosphotransfer proteins but not with B-type response regulators suggests that their known activity in feedback regulation may be realized by interfering at the level of Arabidopsis histidine phosphotransfer protein-mediated signaling. This study contributes to our understanding of the protein interactions of the cytokinin-signaling system and provides a framework for further functional studies in planta.     

Cross talk between gibberellin and cytokinin: the Arabidopsis GA response inhibitor SPINDLY plays a positive role in cytokinin signaling

SPINDLY (SPY) is a negative regulator of gibberellin (GA) responses; however, spy mutants exhibit various phenotypic alterations not found in GA-treated plants. Assaying for additional roles for SPY revealed that spy mutants are resistant to exogenously applied cytokinin. GA also repressed the effects of cytokinin, suggesting that there is cross talk between the two hormone-response pathways, which may involve SPY function. Two spy alleles showing severe (spy-4) and mild (spy-3) GA-associated phenotypes exhibited similar resistance to cytokinin, suggesting that SPY enhances cytokinin responses and inhibits GA signaling through distinct mechanisms. GA and spy repressed numerous cytokinin responses, from seedling development to senescence, indicating that cross talk occurs early in the cytokinin-signaling pathway. Because GA3 and spy-4 inhibited induction of the cytokinin primary-response gene, type-A Arabidopsis response regulator 5, SPY may interact with and modify elements from the phosphorelay cascade of the cytokinin signal transduction pathway. Cytokinin, on the other hand, had no effect on GA biosynthesis or responses. Our results demonstrate that SPY acts as both a repressor of GA responses and a positive regulator of cytokinin signaling. Hence, SPY may play a central role in the regulation of GA/cytokinin cross talk during plant development.     

The cytokinin receptors of Arabidopsis are located mainly to the endoplasmic reticulum

The plant hormone cytokinin is perceived by membrane-located sensor histidine kinases. Arabidopsis (Arabidopsis thaliana) possesses three cytokinin receptors: ARABIDOPSIS HISTIDINE KINASE2 (AHK2), AHK3, and CYTOKININ RESPONSE1/AHK4. The current model predicts perception of the cytokinin signal at the plasma membrane. However, cytokinin-binding studies with membrane fractions separated by two-phase partitioning showed that in the wild type, as well as in mutants retaining only single cytokinin receptors, the major part of specific cytokinin binding was associated with endomembranes. Leaf epidermal cells of tobacco (Nicotiana benthamiana) expressing receptor-green fluorescent protein fusion proteins and bimolecular fluorescence complementation analysis showed strong fluorescence of the endoplasmic reticulum (ER) network for all three receptors. Furthermore, separation of the microsomal fraction of Arabidopsis plants expressing Myc-tagged AHK2 and AHK3 receptors by sucrose gradient centrifugation followed by immunoblotting displayed the Mg2?-dependent density shift typical of ER membrane proteins. Cytokinin-binding assays, fluorescent fusion proteins, and biochemical fractionation all showed that the large majority of cytokinin receptors are localized to the ER, suggesting a central role of this compartment in cytokinin signaling. A modified model for cytokinin signaling is proposed.     

Novel signals for plant development

Classic signal molecules such as auxin, cytokinin, gibberellins, abscisic acid and more recently brassinosteroids have been extensively studied in the context of their role in morphogenetic processes in plants. In the past five years, it has become apparent that there are novel signaling molecules, such as N-acylethanolamides, alkamides, glutamate and nitric oxide, that might play important roles in the regulation of morphogenetic and adaptive processes. There is information pointing out that these molecules might be involved in diverse processes, including seed germination, pathogenesis, modulation of plant architecture and response to abiotic factors. In animals, alkamides and N-acylethanolamides act as endogenous signaling molecules that activate cannabinoid receptors, which are coupled to signal transduction cascades involving glutamate and nitric oxide. Hence, there is a possibility that cannabinoid signaling represents an evolutionary conserved pathway that modulates cellular and physiological processes in eukaryotes.