Publications (13)69.32 Total impact
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Article: Analysis of cytokinin mutants and regulation of cytokinin metabolic genes reveals important regulatory roles of cytokinins in drought, salt and abscisic acid responses, and abscisic acid biosynthesis.
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ABSTRACT: Cytokinins (CKs) regulate plant growth and development via a complex network of CK signaling. Here, we perform functional analyses with CK-deficient plants to provide direct evidence that CKs negatively regulate salt and drought stress signaling. All CK-deficient plants with reduced levels of various CKs exhibited a strong stress-tolerant phenotype that was associated with increased cell membrane integrity and abscisic acid (ABA) hypersensitivity rather than stomatal density and ABA-mediated stomatal closure. Expression of the Arabidopsis thaliana ISOPENTENYL-TRANSFERASE genes involved in the biosynthesis of bioactive CKs and the majority of the Arabidopsis CYTOKININ OXIDASES/DEHYDROGENASES genes was repressed by stress and ABA treatments, leading to a decrease in biologically active CK contents. These results demonstrate a novel mechanism for survival under abiotic stress conditions via the homeostatic regulation of steady state CK levels. Additionally, under normal conditions, although CK deficiency increased the sensitivity of plants to exogenous ABA, it caused a downregulation of key ABA biosynthetic genes, leading to a significant reduction in endogenous ABA levels in CK-deficient plants relative to the wild type. Taken together, this study provides direct evidence that mutual regulation mechanisms exist between the CK and ABA metabolism and signals underlying different processes regulating plant adaptation to stressors as well as plant growth and development.The Plant Cell 06/2011; 23(6):2169-83. · 8.99 Impact Factor -
Article: Combining enhanced root and shoot growth reveals cross talk between pathways that control plant organ size in Arabidopsis.
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ABSTRACT: Functionally distinct Arabidopsis (Arabidopsis thaliana) genes that positively affect root or shoot growth when ectopically expressed were combined to explore the feasibility of enhanced biomass production. Enhanced root growth resulting from cytokinin deficiency was obtained by overexpressing CYTOKININ OXIDASE/DEHYDROGENASE3 (CKX3) under the control of the root-specific PYK10 promoter. Plants harboring the PYK10-CKX3 construct were crossed with four different transgenic lines showing enhanced leaf growth. For all combinations, the phenotypic traits of the individual lines could be combined, resulting in an overall growth increase. Unexpectedly, three out of four combinations had more than additive effects. Both leaf and root growth were synergistically enhanced in plants ectopically expressing CKX3 and BRASSINOSTEROID INSENSITIVE1, indicating cross talk between cytokinins and brassinosteroids. In agreement, treatment of PYK10-CKX3 plants with brassinolide resulted in a dramatic increase in lateral root growth that could not be observed in wild-type plants. Coexpression of CKX3 and the GROWTH-REGULATING FACTOR5 (GRF5) antagonized the effects of GRF5 overexpression, revealing an interplay between cytokinins and GRF5 during leaf cell proliferation. The combined overexpression of CKX3 and GIBBERELLIN 20-OXIDASE1 led to a synergistic increase in leaf growth, suggesting an antagonistic growth control by cytokinins and gibberellins. Only additive effects on root and shoot growth were visible in plants ectopically expressing both CKX3 and ARABIDOPSIS VACUOLAR PYROPHOSPHATASE1, hinting at an independent action mode. Our results show new interactions and contribute to the molecular and physiological understanding of biomass production at the whole plant level.Plant physiology 01/2011; 155(3):1339-52. · 6.53 Impact Factor -
Article: Root-specific reduction of cytokinin causes enhanced root growth, drought tolerance, and leaf mineral enrichment in Arabidopsis and tobacco.
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ABSTRACT: Optimizing root system architecture can overcome yield limitations in crop plants caused by water or nutrient shortages. Classic breeding approaches are difficult because the trait is governed by many genes and is difficult to score. We generated transgenic Arabidopsis thaliana and tobacco (Nicotiana tabacum) plants with enhanced root-specific degradation of the hormone cytokinin, a negative regulator of root growth. These transgenic plants form a larger root system, whereas growth and development of the shoot are similar. Elongation of the primary root, root branching, and root biomass formation were increased by up to 60% in transgenic lines, increasing the root-to-shoot ratio. We thus demonstrated that a single dominant gene could regulate a complex trait, root growth. Moreover, we showed that cytokinin regulates root growth in a largely organ-autonomous fashion that is consistent with its dual role as a hormone with both paracrine and long-distance activities. Transgenic plants had a higher survival rate after severe drought treatment. The accumulation of several elements, including S, P, Mn, Mg, Zn, as well as Cd from a contaminated soil, was significantly increased in shoots. Under conditions of sulfur or magnesium deficiency, leaf chlorophyll content was less affected in transgenic plants, demonstrating the physiological relevance of shoot element accumulation. Our approach might contribute to improve drought tolerance, nutrient efficiency, and nutrient content of crop plants.The Plant Cell 12/2010; 22(12):3905-20. · 8.99 Impact Factor -
Article: Cytokinin action in plant development.
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ABSTRACT: Cytokinin regulates many important aspects of plant development in aerial and subterranean organs. The hormone is part of an intrinsic genetic network controlling organ development and growth in these two distinct environments that plants have to cope with. Cytokinin also mediates the responses to variable extrinsic factors, such as light conditions in the shoot and availability of nutrients and water in the root, and has a role in the response to biotic and abiotic stress. Together, these activities contribute to the fine-tuning of quantitative growth regulation in plants. We review recent progress in understanding the cytokinin system and its links to the regulatory pathways that respond to internal and external signals.Current opinion in plant biology 10/2009; 12(5):527-38. · 10.33 Impact Factor -
Article: The purine derivative PI-55 blocks cytokinin action via receptor inhibition.
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ABSTRACT: One of several potential approaches to study mechanisms of action of biologically active compounds is to develop their agonists and antagonists. In the present study, we report the identification of the first known molecule antagonizing the activity of the plant hormone cytokinin at the receptor level. This compound, 6-(2-hydroxy-3-methylbenzylamino)purine, designated PI-55 in the present study, is structurally closely related to cytokinin 6-benzylaminopurine, but substitutions at specific positions of the aromatic side chain strongly diminished its cytokinin activity and conferred antagonistic properties. PI-55 competitively inhibited the binding of the natural ligand trans-zeatin to the Arabidopsis cytokinin receptors cytokinin response 1 (CRE1)/Arabidopsis histidine kinase (AHK) 4 and AHK3 and repressed induction of the cytokinin response gene ARR5:GUS. Genetic analysis revealed that CRE1/AHK4 is the primary target of PI-55. Cytokinin bioassays also demonstrated the anticytokinin effect of PI-55 in several other species. Furthermore, we show that PI-55 accelerated the germination of Arabidopsis seeds and promoted the root growth and formation of lateral roots, thus phenocopying the known consequences of a lowered cytokinin status and demonstrating its potential to inhibit cytokinin perception in planta. PI-55 is the first example for the targeted development of a cytokinin antagonist and represents an initial step for the preparation of cytokinin antagonists with broad activity and reduced agonistic properties.FEBS Journal 12/2008; 276(1):244-53. · 3.79 Impact Factor -
Article: Cytokinin deficiency causes distinct changes of sink and source parameters in tobacco shoots and roots.
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ABSTRACT: Cytokinin deficiency causes pleiotropic developmental changes such as reduced shoot and increased root growth. It was investigated whether cytokinin-deficient tobacco plants, which overproduce different cytokinin oxidase/dehydrogenase enzymes, show changes in different sink and source parameters, which could be causally related to the establishment of the cytokinin deficiency syndrome. Ultrastructural analysis revealed distinct changes in differentiating shoot tissues, including an increased vacuolation and an earlier differentiation of plastids, which showed partially disorganized thylakoid structures later in development. A comparison of the ploidy levels revealed an increased population of cells with a 4C DNA content during early stages of leaf development, indicating an inhibited progression from G2 to mitosis. To compare physiological characteristics of sink leaves, source leaves and roots of wild-type and cytokinin-deficient plants, several photosynthetic parameters, content of soluble sugars, starch and adenylates, as well as activities of enzymes of carbon assimilation and dissimilation were determined. Leaves of cytokinin-deficient plants contained less chlorophyll and non-photochemical quenching of young leaves was increased. However, absorption rate, photosynthetic capacity (F(v)/F(m) and J(CO2 max)) and efficiency (Phi CO(2 app)), as well as the content of soluble sugars, were not strongly altered in source leaves, indicating that chlorophyll is not limiting for photoassimilation and suggesting that source strength did not restrict shoot growth. By contrast, shoot sink tissues showed drastically reduced contents of soluble sugars, decreased activities of vacuolar invertases, and a reduced ATP content. These results strongly support a function of cytokinin in regulating shoot sink strength and its reduction may be a cause of the altered shoot phenotype. Roots of cytokinin-deficient plants contained less sugar compared with wild-type. However, this did not negatively affect glycolysis, ATP content, or root development. It is suggested that cytokinin-mediated regulation of the sink strength differs between roots and shoots.Journal of Experimental Botany 02/2008; 59(10):2659-72. · 5.36 Impact Factor -
Chapter: Cytokinin Metabolism and Signal Transduction
11/2007: pages 93 - 123; , ISBN: 9780470988800 -
Article: A linear programming approach to max-sum problem: a review.
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ABSTRACT: The max-sum labeling problem, defined as maximizing a sum of binary (i.e., pairwise) functions of discrete variables, is a general NP-hard optimization problem with many applications, such as computing the MAP configuration of a Markov random field. We review a not widely known approach to the problem, developed by Ukrainian researchers Schlesinger et al. in 1976, and show how it contributes to recent results, most importantly, those on the convex combination of trees and tree-reweighted max-product. In particular, we review Schlesinger et al.'s upper bound on the max-sum criterion, its minimization by equivalent transformations, its relation to the constraint satisfaction problem, the fact that this minimization is dual to a linear programming relaxation of the original problem, and the three kinds of consistency necessary for optimality of the upper bound. We revisit problems with Boolean variables and supermodular problems. We describe two algorithms for decreasing the upper bound. We present an example application for structural image analysis.IEEE Transactions on Pattern Analysis and Machine Intelligence 08/2007; 29(7):1165-79. · 4.91 Impact Factor -
Article: Cytokinin oxidase/dehydrogenase genes in barley and wheat: cloning and heterologous expression.
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ABSTRACT: The cloning of two novel genes that encode cytokinin oxidase/dehydrogenase (CKX) in barley is described in this work. Transformation of both genes into Arabidopsis and tobacco showed that at least one of the genes codes for a functional enzyme, as its expression caused a cytokinin-deficient phenotype in the heterologous host plants. Additional cloning of two gene fragments, and an in silico search in the public expressed sequence tag clone databases, revealed the presence of at least 13 more members of the CKX gene family in barley and wheat. The expression of three selected barley genes was analyzed by RT-PCR and found to be organ-specific with peak expression in mature kernels. One barley CKX (HvCKX2) was characterized in detail after heterologous expression in tobacco. Interestingly, this enzyme shows a pH optimum at 4.5 and a preference for cytokinin ribosides as substrates, which may indicate its vacuolar targeting. Different substrate specificities, and the pH profiles of cytokinin-degrading enzymes extracted from different barley tissues, are also presented.European Journal of Biochemistry 11/2004; 271(20):3990-4002. · 3.58 Impact Factor -
Article: Cytokinin-deficient transgenic Arabidopsis plants show multiple developmental alterations indicating opposite functions of cytokinins in the regulation of shoot and root meristem activity.
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ABSTRACT: Cytokinins are hormones that regulate cell division and development. As a result of a lack of specific mutants and biochemical tools, it has not been possible to study the consequences of cytokinin deficiency. Cytokinin-deficient plants are expected to yield information about processes in which cytokinins are limiting and that, therefore, they might regulate. We have engineered transgenic Arabidopsis plants that overexpress individually six different members of the cytokinin oxidase/dehydrogenase (AtCKX) gene family and have undertaken a detailed phenotypic analysis. Transgenic plants had increased cytokinin breakdown (30 to 45% of wild-type cytokinin content) and reduced expression of the cytokinin reporter gene ARR5:GUS (beta-glucuronidase). Cytokinin deficiency resulted in diminished activity of the vegetative and floral shoot apical meristems and leaf primordia, indicating an absolute requirement for the hormone. By contrast, cytokinins are negative regulators of root growth and lateral root formation. We show that the increased growth of the primary root is linked to an enhanced meristematic cell number, suggesting that cytokinins control the exit of cells from the root meristem. Different AtCKX-green fluorescent protein fusion proteins were localized to the vacuoles or the endoplasmic reticulum and possibly to the extracellular space, indicating that subcellular compartmentation plays an important role in cytokinin biology. Analyses of promoter:GUS fusion genes showed differential expression of AtCKX genes during plant development, the activity being confined predominantly to zones of active growth. Our results are consistent with the hypothesis that cytokinins have central, but opposite, regulatory functions in root and shoot meristems and indicate that a fine-tuned control of catabolism plays an important role in ensuring the proper regulation of cytokinin functions.The Plant Cell 12/2003; 15(11):2532-50. · 8.99 Impact Factor -
Article: Structure and function of cytokinin oxidase/dehydrogenase genes of maize, rice, Arabidopsis and other species.
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ABSTRACT: Cytokinin oxidases/dehydrogenases (CKX) catalyze the irreversible degradation of the cytokinins isopentenyladenine, zeatin, and their ribosides in a single enzymatic step by oxidative side chain cleavage. To date the sequences of 17 fully annotated CKX genes are known, including two prokaryotic genes. The CKX gene families of Arabidopsis thaliana and rice comprise seven and at least ten members, respectively. The main features of CKX genes and proteins are summarized in this review. Individual proteins differ in their catalytic properties, their subcellular localization and their expression domains. The evolutionary development of cytokinin-catabolizing gene families and the individual properties of their members indicate an important role for the fine-tuned control of catabolism to assure proper regulation of cytokinin functions. The use of CKX genes as a tool in studies of cytokinin biology and biotechnological applications is discussed.Journal of Plant Research 07/2003; 116(3):241-52. · 1.75 Impact Factor -
Article: New cytokinin metabolites in IPT transgenic Arabidopsis thaliana plants.
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ABSTRACT: Two novel cytokinin metabolites were identified in transgenic Arabidopsis thaliana (L.) Heynh. plants containing the bacterial IPT gene under the transcriptional control of a heat-regulated promoter. After cyclic heat-shock treatment, the endogenous cytokinin concentrations were elevated up to 100-fold compared to the wild-type plants. More then 20 different cytokinin metabolites were found, with zeatin-type cytokinins being the most abundant. The metabolic inactivation of these compounds occurred predominantly through N-glucosylation. No significant accumulation of isopentenyladenine-type cytokinins, and only a small increase in dihydrozeatin metabolites, was observed. Subsequent studies of the abundant, unidentified conjugates revealed the presence of zeatin and dihydrozeatin diglucoside conjugates. Structural analysis, utilizing electrospray-liquid tandem mass spectrometry, identified these as a zeatin-O-glucoside-9-glucoside and dihydrozeatin-O-glucoside-9-glucoside, respectively. A third unknown metabolite, was tentatively identified as a phosphorylated form of zeatin-9-glucoside. The biological activity of these compounds in three cytokinin bioassays was low. A comparison of the cytokinin pattern in transgenic and wild-type plants indicates that these specific metabolites accumulate as a consequence of enhanced cytokinin biosynthesis, and are probably involved in the homeostatic mechanisms that control endogenous cytokinin levels.Physiologia Plantarum 06/2003; 118(1):127-137. · 3.11 Impact Factor -
Article: Cytokinin oxidase/cytokinin dehydrogenase assay: optimized procedures and applications.
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ABSTRACT: Spectrophotometric methods for determining the activity of cytokinin oxidase/cytokinin dehydrogenase (EC 1.5.99.12) were developed and optimized. A sensitive end-point method based on a combination of the electron acceptor 2,6-dichlorophenolindophenol and Schiff base formation of the reaction product with 4-aminophenol under acidic conditions can be applied to crude cell and tissue extracts. The assay was also adapted for other substrates than N6-(2-isopentenyl)adenine, such as zeatin and the aromatic cytokinins, although an enzyme which degrades the latter compounds has not yet been identified. The second novel method is an initial rate method based on the coupled redox reaction of phenazine methosulfate and 3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide resulting in the formation of a formazan dye. This method can be used for kinetic studies with purified enzyme and is entirely substrate independent.Analytical Biochemistry 08/2002; 306(1):1-7. · 3.00 Impact Factor
Top co-authors
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Institutions
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2008–2010
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Freie Universität Berlin
Berlin, Land Berlin, Germany
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2007
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Rothamsted Research
Harpenden, ENG, United Kingdom -
Czech Technical University in Prague
- Department of Cybernetics and Robotics
Praha, Hlavni mesto Praha, Czech Republic
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