Genetic engineering of cytokinins and their application to agriculture

Cytokinins are master regulators of plant growth and development. They are involved in the regulation of many important physiological and metabolic processes. Recent progress in cytokinin research at the molecular level, including identification of related genes and cytokinin receptors, plus elucidation of signal transduction, has greatly increased our understanding of cytokinin actions. Although still in its infant stage, molecular breeding of crops with altered cytokinin metabolism, when combined with the transgenic approach, has shown very promising potential for application to agriculture. In this review we briefly introduce recent progress in cytokinin molecular biology, discuss applications of cytokinin genetic engineering to agriculture, and present implications and future research directions.     

Cytokinin signaling

Cytokinins influence many aspects of plant growth and development. The current model for cytokinin signaling is a multi-step phosphorelay similar to the prokaryotic two-component systems that are used in responses to environmental stimuli. Recently, progress has been made in improving our understanding of the molecular mechanism that underlies cytokinin signaling. Molecular and genetic analyses of loss-of-function mutants indicate that the two-component elements that are involved in cytokinin signaling have redundant and overlapping functions. These elements regulate both the shoot and root meristems, are required for the development of fertile flowers, and modulate the response to varying nutrient levels.     

Degradation of cytokinins by cytokinin oxidases in plants

The degradation metabolism of cytokinins is an important process that controls the levels of cytokinin active forms and their distribution in plant tissues. It appears to be due, in large part, to the activity of a specific enzyme, cytokinin oxidase. This review attempts to collate the limited information available about this enzyme and introduce new facts, obtained in our laboratory, concerning the mechanism of degradation of cytokinins bearing unsaturated isoprene side chains. However, complete clarification of the effects of cytokinin oxidase on cytokinin regulation and its molecular and biochemical properties will be dependent upon the purification of the protein with cytokinin oxidase activity to homogeneity and progress in the development of requisite molecular probes.     

The discovery of cytokinin receptors and biosynthesis of cytokinins: A true story

The year 2001 saw an amazing progress in cytokinin studies. Ten years ago, cytokinin receptor genes and genes encoding cytokinin biosynthetic enzymes together with the corresponding proteins were identified in plants. These studies elucidated the molecular mechanism of cytokinin effects on the expression of cytokinin responsive genes and ultimately established the endogenous synthesis of cytokinins in plant cells, justifying their membership among plant hormones. The paper describes in short the edifying and sometimes paradoxical story of these fundamental and captivating discoveries.     

Rapid alteration of the phosphoproteome in the moss Physcomitrella patens after cytokinin treatment

Cytokinin hormones are crucial regulators of a large number of processes in plant development. Recently, significant progress has been made toward the elucidation of the molecular details of cytokinin that has led to a model for signal transduction involving a phosphorylation cascade. However, the current knowledge of cytokinin action remains largely unknown and does not explain the different roles of this hormone. To gain further insights into this aspect of cytokinin action and the inducible phosphorelay, we have produced the first large-scale map of a phosphoproteome in the moss Physcomitrella patens. Using a protocol that we recently published (Heintz, D.; et al. Electrophoresis 2004, 25, 1149-1159) that combines IMAC, MALDI-TOF-MS, and LC-MS/MS, a total of 172 phosphopeptide sequences were obtained by a peptide de novo sequencing strategy. Specific P. patens EST and raw genomic databases were interrogated, and protein homology searches resulted in the identification of 112 proteins that were then classified into functional categories. In addition, the temporal dynamics of the phosphoproteome in response to cytokinin stimulation was studied at 2, 4, 6, and 15 min after hormone addition. We identified 13 proteins that were not previously known targets of cytokinin action. Among the responsive proteins, some were involved in metabolism, and several proteins of unknown function were also identified. We have mapped the time course of their activation in response to cytokinin and discussed their hypothetical biological significance. Deciphering these early induced phosphorylation events has shown that the cytokinin effect can be rapid (few minutes), and the duration of this effect can be variable. Also phosphorylation events can be differentially regulated. Taken together our proteomic study provides an enriched look of the multistep phosphorelay system mediating cytokinin response and suggests the existence of a multidirectional interaction between cytokinin and numerous other pathways.     

细胞分裂素在植物抗逆和延衰中的作用

本文综述了细胞分裂素类物质的种类、分布和在植物抗水分胁迫、低温冷害、病虫害等方面的作用以及在延缓果实、叶片、切花等衰老中的效果,讨论了其生理机制、细胞分裂素与其它植物激素的相互关系,并提出了有关细胞分裂素类物质作用机理中值得深入研究的若干问题,如嘌呤型与苯基脲型细胞分裂素的作用特点,细胞分裂素与生长素、脱落酸的协调作用和拮抗作用细胞分裂素的从头合成途径和tＲＮＡ途径等。     

细胞分裂素在植物抗逆和延衰中的作用

本文综述了细胞分裂素类物质的种类、分布和在植物抗水分胁迫、低温冷害、病虫害等方面的作用以及在延缓果实、叶片、切花等衰老中的效果,讨论了其生理机制、细胞分裂素与其它植物激素的相互关系,并提出了有关细胞分裂素类物质作用机理中值得深入研究的若干问题,如嘌呤型与苯基脲型细胞分裂素的作用特点,细胞分裂素与生长素、脱落酸的协调作用和拮抗作用,细胞分裂素的从头合成途径和tRNA途径等。     

Cytokinin biosynthesis and perception

Cytokinin has been considered to be a master regulator of plant growth and development, but only in the past several years has substantial progress been made uncovering the roles of cytokinins at various developmental stages. Recent studies on key metabolic enzymes and signaling components have contributed to understanding the basic mechanism of biosynthesis and perception of cytokinin within a whole plant body. The initial products of de novo cytokinin biosynthesis in higher plants and Agrobacterium are different, and the regulatory systems in biosynthesis and homeostasis are finely controlled and appear to be important in communicating nutrient signals to morphogenetic responses. The cytokinin receptors have largely overlapping, but still specific, functions in diverse cytokinin responses. In this review, we will specifically emphasize the biosynthesis of isoprenoid cytokinins and perception of cytokinin signals in Arabidopsis.     

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。     

Cytokinin signal perception and transduction

In the past few years, enormous progress has been made in understanding cytokinin perception and signalling. Three cytokinin receptor proteins, which are hybrid histidine kinases, have been identified in Arabidopsis. These receptors may transduce signals in a quantitative rheostat-like fashion, thus permitting long-lasting and continuously variable signalling that is directly dependent on the hormone concentration. Evidence has been provided that downstream signalling is transmitted through a His-to-Asp phospho-relay involving phosphotransmitter and response regulator proteins, typical of two-component systems. On the basis of mutant analysis, protein-protein interaction studies and target gene identification, a cellular network is emerging that links cytokinin activity to both developmental and physiological processes.     

Cytokinin oxidase: Biochemical features and physiological significance

The catabolism of cytokinin in plant tissues appears to be due, in large part, to the activity of a specific enzyme, cytokinin oxidase. This enzyme catalyses the oxidation of cytokinin substrates bearing unsaturated isoprenoid side chains, using molecular oxygen as the oxidant. In general, substrate specificity is highly conserved and cytokinin substrates bearing saturated or cyclic side chains do not serve as substrates for most cytokinin oxidases tested to date. Despite variation in molecular properties of the enzyme from a number of higher plants, oxygen is always required for the reaction. Cytokinin oxidases from several sources have been shown to be glycosylated. Cytokinin oxidase activity appears to be universally inhibited by cytokinin-active urea derivatives. Auxin has been reported to act as an allosteric regulator which increases activity of the enzyme.
Cytokinin oxidase activity is subject to tight regulation. Levels of the enzyme are controlled by a mechanism sensitive to cytokinin supply. The up-regulation of cytokinin oxidase expression in response to exogenous application of cytokinin suggests that the metabolic fate of exogenously applied cytokinins may not accurately mimic that of the endogenous compounds.
Cytokinin oxidase is believed to be a copper-containing amine oxidase (EC 1.4.3.6). Considerable evidence strongly supports a common mechanism for amine oxidases. It is possible that advances in understanding of other amine oxidases could be extrapolated to increase our understanding of cytokinin oxidase at the molecular level. This is discussed with reference to what is currently known about the catalytic mechanism of the enzyme. The possibility of pyrroloquinoline quinone, or a closely related compound, as a redox cofactor of cytokinin oxidase is considered, as are the implications of the glycosylated nature of the enzyme for its regulation and compartmentalisation within the cell.     

Properties, functions and evolution of cytokinin receptors

The discovery of cytokinin receptors of Arabidopsis thaliana ten years ago was a milestone in plant hormone research. Since then, research has yielded insights into the biochemical properties and functions of these sensor histidine kinases. Their affinities to both trans-zeatin and isopentenyladenine are in the low nM range. Cytokinin ribosides, cis-zeatin and thidiazuron were established as compounds with genuine cytokinin activity and the first cytokinin antagonists were identified. Numerous functions of cytokinin receptors in plant development, as well as in the plant's responses to the environment, have been elucidated and are summarized. Finally, we address the question how the receptors have evolved during plant evolution.     

Comparison of phytohormone signaling mechanisms

Plant hormones are crucial signaling molecules that coordinate all aspects of plant growth, development and defense. A great deal of attention has been attracted from biologists to study the molecular mechanisms for perception and signal transduction of plant hormones during the last two decades. Tremendous progress has been made in identifying receptors and key signaling components of plant hormones. The holistic picture of hormone signaling pathways is extremely complicated, this review will give a general overview of perception and signal transduction mechanisms of auxin, gibberellin, cytokinin, abscisic acid, ethylene, brassinosteroid, and jasmonate.     

飞蝗研究进展

飞蝗作为农业重大害虫,其分布遍及欧洲、亚洲、非洲和澳洲。全世界共有10个亚种,我国有3个亚种。飞蝗不但分布广,而且数量多、发生频率高,危害极为严重。作者在查阅了国内外近20年文献资料的基础上,结合我国蝗害情况,分别从形态、组织、生理、生化、遗传、毒力及分子系统学等方面综述了国内外飞蝗的研究进展,以期对我国飞蝗的基础及应用研究提供参考。