Members of the genus Sorbus are the only endemic tree species that occur in Czechia. They are important components of endangered plant communities. Their natural regeneration is usually problematic because of their mode of reproduction and because they can survive in rare populations with small numbers of individuals. The aim of this study was to develop a successful micropropagation protocol for selected Sorbus species, of which two are endemic (S. gemella and S. omissa) and two are hybrid (S. × abscondita and S. × kitaibeliana). We found significant differences in shoot induction and rooting ability between the Sorbus species under study. With the exception of S. × abscondita, N6-benzyladenine had a significantly greater effect on shoot regeneration, both in terms of shoot number and total shoot length, than meta-topolin. Root induction was key to the successful micropropagation of the Sorbus species studied. Our results show that four Sorbus species can be successfully rooted under ex vitro conditions, without a rooting powder treatment in a steamed peat-perlite substrate. Auxin-untreated microcuttings of S. gemella, S. × kitaibeliana and S. omissa, but not S. × abscondita, rooted better than ones treated with indole-3-butyric acid. This is the first time a micropropagation protocol for S. omissa, S. × abscondita and S. × kitaibeliana has been published.

• Keywords
• Sorbus, endemic species, ex vitro rooting, micropropagation, substrate,
• Publication type
• Journal Article MeSH

Cytokinins are a class of phytohormones, signalling molecules specific to plants. They act as regulators of diverse physiological processes in complex signalling pathways. It is necessary for plants to continuously regulate cytokinin distribution among different organs, tissues, cells, and compartments. Such regulatory mechanisms include cytokinin biosynthesis, metabolic conversions and degradation, as well as cytokinin membrane transport. In our review, we aim to provide a thorough picture of the latter. We begin by summarizing cytokinin structures and physicochemical properties. Then, we revise the elementary thermodynamic and kinetic aspects of cytokinin membrane transport. Next, we review which membrane-bound carrier proteins and protein families recognize cytokinins as their substrates. Namely, we discuss the families of "equilibrative nucleoside transporters" and "purine permeases", which translocate diverse purine-related compounds, and proteins AtPUP14, AtABCG14, AtAZG1, and AtAZG2, which are specific to cytokinins. We also address long-distance cytokinin transport. Putting all these pieces together, we finally discuss cytokinin distribution as a net result of these processes, diverse in their physicochemical nature but acting together to promote plant fitness.

• Keywords
• ABCG14, AZG1, AZG2, PUP14, cytokinin distribution, cytokinin hydrophobicity, cytokinin transport, membrane transport,
• MeSH
• Arabidopsis metabolism   MeSH
• Biological Transport MeSH
• Cell Membrane metabolism   MeSH
• Cytokinins metabolism   MeSH
• Homeostasis MeSH
• Hydrophobic and Hydrophilic Interactions MeSH
• Kinetics MeSH
• Plant Roots metabolism   MeSH
• Membrane Transport Proteins metabolism   MeSH
• Arabidopsis Proteins genetics   MeSH
• Gene Expression Regulation, Plant MeSH
• Plant Growth Regulators metabolism   MeSH
• Signal Transduction physiology   MeSH
• Thermodynamics MeSH
• Plant Shoots metabolism   MeSH
• Publication type
• Journal Article MeSH
• Review MeSH
• Names of Substances
• Cytokinins MeSH
• Membrane Transport Proteins MeSH
• Arabidopsis Proteins MeSH
• Plant Growth Regulators MeSH

Cytokinins are plant hormones, derivatives of adenine with a side chain at the N6-position. They are involved in many physiological processes. While the metabolism of trans-zeatin and isopentenyladenine, which are considered to be highly active cytokinins, has been extensively studied, there are others with less obvious functions, such as cis-zeatin, dihydrozeatin, and aromatic cytokinins, which have been comparatively neglected. To help explain this duality, we present a novel hypothesis metaphorically comparing various cytokinin forms, enzymes of CK metabolism, and their signalling and transporter functions to the comics superheroes Hulk and Deadpool. Hulk is a powerful but short-lived creation, whilst Deadpool presents a more subtle and enduring force. With this dual framework in mind, this review compares different cytokinin metabolites, and their biosynthesis, translocation, and sensing to illustrate the different mechanisms behind the two CK strategies. This is put together and applied to a plant developmental scale and, beyond plants, to interactions with organisms of other kingdoms, to highlight where future study can benefit the understanding of plant fitness and productivity.

• Keywords
• Hulk/Deadpool, aromatic cytokinins, cis-zeatin, cytokinin biosynthesis, cytokinin oxidase/dehydrogenase, cytokinin signalling, cytokinin transport, cytokinins, isopentenyl transferase,
• MeSH
• Arabidopsis metabolism   MeSH
• Models, Biological MeSH
• Biological Transport MeSH
• Biological Assay MeSH
• Cytokinins metabolism   MeSH
• Plant Physiological Phenomena * MeSH
• Glycosylation MeSH
• Hydrolysis MeSH
• Kinetics MeSH
• Kinetin metabolism   MeSH
• Oxidoreductases metabolism   MeSH
• Plant Growth Regulators metabolism   MeSH
• Plants metabolism   MeSH
• Signal Transduction * MeSH
• Protein Binding MeSH
• Zeatin analogs & derivatives   MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Review MeSH
• Names of Substances
• cytokinin oxidase MeSH Browser
• Cytokinins MeSH
• dihydrozeatin MeSH Browser
• Kinetin MeSH
• Oxidoreductases MeSH
• Plant Growth Regulators MeSH
• Zeatin MeSH

Cytokinins (CKs) are a class of phytohormones affecting many aspects of plant growth and development. In the complex process of CK homeostasis in plants, N-glucosylation represents one of the essential metabolic pathways. Its products, CK N7- and N9-glucosides, have been largely overlooked in the past as irreversible and inactive CK products lacking any relevant physiological impact. In this work, we report a widespread distribution of CK N-glucosides across the plant kingdom proceeding from evolutionary older to younger plants with different proportions between N7- and N9-glucosides in the total CK pool. We show dramatic changes in their profiles as well as in expression levels of the UGT76C1 and UGT76C2 genes during Arabidopsis ontogenesis. We also demonstrate specific physiological effects of CK N-glucosides in CK bioassays including their antisenescent activities, inhibitory effects on root development, and activation of the CK signaling pathway visualized by the CK-responsive YFP reporter line, TCSv2::3XVENUS. Last but not least, we present the considerable impact of CK N7- and N9-glucosides on the expression of CK-related genes in maize and their stimulatory effects on CK oxidase/dehydrogenase activity in oats. Our findings revise the apparent irreversibility and inactivity of CK N7- and N9-glucosides and indicate their involvement in CK evolution while suggesting their unique function(s) in plants.

• Keywords
• Arabidopsis, N-glucosides, Zea mays, [2-3H]tZ9G, cis-zeatin, cytokinin, cytokinin oxidase/dehydrogenase, oat, senescence, trans-zeatin, β-D-glucosidase Zm-p60.1,
• MeSH
• Arabidopsis genetics   metabolism   MeSH
• Cytokinins genetics   MeSH
• Glucosides genetics   MeSH
• Glucosyltransferases genetics   MeSH
• Zea mays genetics   metabolism   MeSH
• Evolution, Molecular * MeSH
• Oxidoreductases genetics   MeSH
• Gene Expression Regulation, Plant genetics   MeSH
• Plant Growth Regulators genetics   metabolism   MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Cytokinins MeSH
• Glucosides MeSH
• Glucosyltransferases MeSH
• Oxidoreductases MeSH
• Plant Growth Regulators MeSH

Leaf senescence, accompanied by chlorophyll breakdown, chloroplast degradation and inhibition of photosynthesis, can be suppressed by an exogenous application of cytokinins. Two aromatic cytokinin arabinosides (6-benzylamino-9-β-d-arabinofuranosylpurines; BAPAs), 3-hydroxy- (3OHBAPA) and 3-methoxy- (3MeOBAPA) derivatives, have recently been found to possess high anti-senescence activity. Interestingly, their effect on the maintenance of chlorophyll content and maximal quantum yield of photosystem II (PSII) in detached dark-adapted leaves differed quantitatively in wheat (Triticum aestivum L. cv. Aranka) and Arabidopsis (Arabidopsisthaliana L. (Col-0)). In this work, we have found that the anti-senescence effects of 3OHBAPA and 3MeOBAPA in wheat and Arabidopsis also differ in other parameters, including the maintenance of carotenoid content and chloroplasts, rate of reduction of primary electron acceptor of PSII (QA) as well as electron transport behind QA, and partitioning of absorbed light energy in light-adapted leaves. In wheat, 3OHBAPA had a higher protective effect than 3MeOBAPA, whereas in Arabidopsis, 3MeOBAPA was the more efficient derivative. We have found that the different anti-senescent activity of 3OHBAPA and 3MeOBAPA was coupled to different ethylene production in the treated leaves: the lower the ethylene production, the higher the anti-senescence activity. 3OHBAPA and 3MeOBAPA also efficiently protected the senescing leaves of wheat and Arabidopsis against oxidative damage induced by both H2O2 and high-light treatment, which could also be connected with the low level of ethylene production.

• Keywords
• Arabidopsis, chlorophyll fluorescence, cytokinin derivative, ethylene, oxidative stress, photosystem II, phytohormone, senescence, wheat,
• MeSH
• Arabidopsis drug effects   growth & development   metabolism   MeSH
• Cytokinins pharmacology   MeSH
• Ethylenes metabolism   MeSH
• Photosynthesis MeSH
• Plant Leaves drug effects   growth & development   metabolism   MeSH
• Triticum drug effects   growth & development   metabolism   MeSH
• Plant Growth Regulators pharmacology   MeSH
• Cellular Senescence * MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Cytokinins MeSH
• ethylene MeSH Browser
• Ethylenes MeSH
• Plant Growth Regulators MeSH

Cytokinins and their sugar or non-sugar conjugates are very active growth-promoting factors in plants, although they occur at very low concentrations. These compounds have been identified in numerous plant species. This review predominantly focuses on 9-substituted adenine-based cytokinin conjugates, both artificial and endogenous, sugar and non-sugar, and their roles in plants. Acquired information about their biological activities, interconversions, and metabolism improves understanding of their mechanisms of action and functions in planta. Although a number of 9-substituted cytokinins occur endogenously, many have also been prepared in laboratories to facilitate the clarification of their physiological roles and the determination of their biological properties. Here, we chart advances in knowledge of 9-substituted cytokinin conjugates from their discovery to current understanding and reciprocal interactions between biological properties and associated structural motifs. Current organic chemistry enables preparation of derivatives with better biological properties, such as improved anti-senescence, strong cell division stimulation, shoot forming, or more persistent stress tolerance compared to endogenous or canonical cytokinins. Many artificial cytokinin conjugates stimulate higher mass production than naturally occurring cytokinins, improve rooting, or simply have high stability or bioavailability. Thus, knowledge of the biosynthesis, metabolism, and activity of 9-substituted cytokinins in various plant species extends the scope for exploiting both natural and artificially prepared cytokinins in plant biotechnology, tissue culture, and agriculture.

• Keywords
• D-arabinoside, cytokinin nucleosides, cytokinin sugar conjugates, disaccharides, glucoside, meta-topolin, plant biotechnology, plant tissue culture, riboside, zeatin,
• MeSH
• Adenine chemistry   metabolism   MeSH
• Cytokinins biosynthesis   chemistry   metabolism   MeSH
• Molecular Structure MeSH
• Plants chemistry   metabolism   MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Review MeSH
• Names of Substances
• Adenine MeSH
• Cytokinins MeSH

A number of scientific reports published to date contain data on endogenous levels of various phytohormones in potato (Solanum tuberosum L.) but a complete cytokinin profile of potato tissues, that would include data on all particular molecular forms of cytokinin, has still been missing. In this work, endogenous levels of all analytically detectable isoprenoid cytokinins, as well as the auxin indole-3-acetic acid (IAA), and abscisic acid (ABA) have been determined in shoots and roots of 30 day old in vitro grown potato (cv. Désirée). The results presented here are generally similar to other data reported for in vitro grown potato plants, whereas greenhouse-grown plants typically contain lower levels of ABA, possibly indicating that in vitro grown potato is exposed to chronic stress. Cytokinin N-glucosides, particularly N7-glucosides, are the dominant cytokinin forms in both shoots and roots of potato, whereas nucleobases, as the bioactive forms of cytokinins, comprise a low proportion of cytokinin levels in tissues of potato. Differences in phytohormone composition between shoots and roots of potato suggest specific patterns of transport and/or differences in tissue-specific metabolism of plant hormones. These results represent a contribution to understanding the hormonomics of potato, a crop species of extraordinary economic importance.

• MeSH
• Cytokinins analysis   metabolism   MeSH
• Stress, Physiological MeSH
• Plant Roots metabolism   MeSH
• Abscisic Acid analysis   metabolism   MeSH
• Indoleacetic Acids analysis   metabolism   MeSH
• Plant Growth Regulators analysis   metabolism   MeSH
• Solanum tuberosum growth & development   metabolism   MeSH
• Tandem Mass Spectrometry MeSH
• Plant Shoots metabolism   MeSH
• Chromatography, High Pressure Liquid MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Cytokinins MeSH
• indoleacetic acid MeSH Browser
• Abscisic Acid MeSH
• Indoleacetic Acids MeSH
• Plant Growth Regulators MeSH

Cytokinins (CKs) and their metabolites and derivatives are essential for cell division, plant growth regulation and development. They are typically found at minute concentrations in plant tissues containing very complicated biological matrices. Therefore, defined standards labelled with stable isotopes are required for precise metabolic profiling and quantification of CKs, as well as in vivo elucidation of CK biosynthesis in various plant species. In this work, 11 [15N]-labelled C6-purine derivatives were prepared, among them 5 aromatic (4, 5, 6, 7, 8) and 3 isoprenoid (9, 10, 11) CKs. Compared to current methods, optimized syntheses of 6-amino-9H-[15N5]-purine (adenine) and 6-chloro-9H-[15N4]-purine (6-chloropurine) were performed to achieve more effective, selective and generally easier approaches. The chemical identity and purity of prepared compounds were confirmed by physico-chemical analyses (TLC; HRMS; HPLC-MS; 1H, 13C, 15N NMR). The presented approach is applicable for the synthesis of any other desired [15N4]-labelled C6-substituted purine derivatives.

• Keywords
• 15N-labelled, cytokinin, purine, synthesis,
• Publication type
• Journal Article MeSH

Stress-induced senescence is a global agro-economic problem. Cytokinins are considered to be key plant anti-senescence hormones, but despite this practical function their use in agriculture is limited because cytokinins also inhibit root growth and development. We explored new cytokinin analogs by synthesizing a series of 1,2,3-thiadiazol-5-yl urea derivatives. The most potent compound, 1-(2-methoxy-ethyl)-3-1,2,3-thiadiazol-5-yl urea (ASES - Anti-Senescence Substance), strongly inhibited dark-induced senescence in leaves of wheat (Triticum aestivum L.) and Arabidopsis thaliana. The inhibitory effect of ASES on wheat leaf senescence was, to the best of our knowledge, the strongest of any known natural or synthetic compound. In vivo, ASES also improved the salt tolerance of young wheat plants. Interestingly, ASES did not affect root development of wheat and Arabidopsis, and molecular and classical cytokinin bioassays demonstrated that ASES exhibits very low cytokinin activity. A proteomic analysis of the ASES-treated leaves further revealed that the senescence-induced degradation of photosystem II had been very effectively blocked. Taken together, our results including data from cytokinin content analysis demonstrate that ASES delays leaf senescence by mechanism(s) different from those of cytokinins and, more effectively. No such substance has yet been described in the literature, which makes ASES an interesting tool for research of photosynthesis regulation. Its simple synthesis and high efficiency predetermine ASES to become also a potent plant stress protectant in biotechnology and agricultural industries.

• Keywords
• ASES, cytokinin, photosystem II, senescence, stress, thidiazuron, wheat,
• Publication type
• Journal Article MeSH

BACKGROUND: When applied to a nutrition solution or agar media, the non-substituted aromatic cytokinins caused thickening and shortening of the primary root, had an inhibitory effect on lateral root branching, and even showed some negative effects on development of the aerial part at as low as a 10 nanomolar concentration. Novel analogues of aromatic cytokinins ranking among topolins substituted on N9-atom of adenine by tetrahydropyranyl or 4-chlorobutyl group have been prepared and tested in standardized cytokinin bioassays [1]. Those showing comparable activities with N(6)-benzylaminopurine were further tested in planta. METHODOLOGY/PRINCIPAL FINDINGS: The main aim of the study was to explain molecular mechanism of function of novel cytokinin derivatives on plant development. Precise quantification of cytokinin content and profiling of genes involved in cytokinin metabolism and perception in treated plants revealed several aspects of different action of m-methoxytopolin base and its substituted derivative on plant development. In contrast to standard cytokinins, N9- tetrahydropyranyl derivative of m-topolin and its methoxy-counterpart showed the negative effects on root development only at three orders of magnitude higher concentrations. Moreover, the methoxy-derivative demonstrates a positive effect on lateral root branching and leaf emerging in a nanomolar range of concentrations, in comparison with untreated plants. CONCLUSIONS/SIGNIFICANCE: Tetrahydropyranyl substitution at N9-position of cytokinin purine ring significantly enhances acropetal transport of a given cytokinins. Together with the methoxy-substitution, impedes accumulation of non-active cytokinin glucoside forms in roots, allows gradual release of the active base, and has a significant effect on the distribution and amount of endogenous isoprenoid cytokinins in different plant tissues. The utilization of novel aromatic cytokinin derivatives can distinctively improve expected hormonal effects in plant propagation techniques in the future.

• MeSH
• Enzyme Activation MeSH
• Arabidopsis drug effects   genetics   growth & development   metabolism   MeSH
• Biomass MeSH
• Cytokinins chemistry   pharmacology   MeSH
• Plant Roots chemistry   drug effects   growth & development   MeSH
• Zea mays drug effects   genetics   growth & development   metabolism   MeSH
• Plant Leaves chemistry   drug effects   growth & development   MeSH
• Oxidoreductases metabolism   MeSH
• Cell Proliferation drug effects   MeSH
• Gene Expression Regulation, Plant MeSH
• Plant Growth Regulators chemistry   pharmacology   MeSH
• Seedlings drug effects   MeSH
• Gene Expression Profiling MeSH
• Xylem chemistry   metabolism   MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Cytokinins MeSH
• Oxidoreductases MeSH
• Plant Growth Regulators MeSH