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Publications - Cell and Metabolic Biology

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Publications

Ahkami, A. H.; Lischewski, S.; Haensch, K.-T.; Porfirova, S.; Hofmann, J.; Rolletschek, H.; Melzer, M.; Franken, P.; Hause, B.; Druege, U.; Hajirezaei, M. R.; Molecular physiology of adventitious root formation in Petunia hybrida cuttings: involvement of wound response and primary metabolism New Phytol. 181, 613-625, (2009) DOI: 10.1111/j.1469-8137.2008.02704.x

Adventitious root formation (ARF) in the model plant Petunia hybrida cv. Mitchell has been analysed in terms of anatomy, gene expression, enzymatic activities and levels of metabolites. This study focuses on the involvement of wound response and primary metabolism.Microscopic techniques were complemented with targeted transcript, enzyme and metabolite profiling using real time polymerase chain reaction (PCR), Northern blot, enzymatic assays, chromatography and mass spectrometry.Three days after severance from the stock plants, first meristematic cells appeared which further developed into root primordia and finally adventitious roots. Excision of cuttings led to a fast and transient increase in the wound‐hormone jasmonic acid, followed by the expression of jasmonate‐regulated genes such as cell wall invertase. Analysis of soluble and insoluble carbohydrates showed a continuous accumulation during ARF. A broad metabolite profiling revealed a strong increase in organic acids and resynthesis of essential amino acids.Substantial changes in enzyme activities and metabolite levels indicate that specific enzymes and metabolites might play a crucial role during ARF. Three metabolic phases could be defined: (i) sink establishment phase characterized by apoplastic unloading of sucrose and being probably mediated by jasmonates; (ii) recovery phase; and (iii) maintenance phase, in which a symplastic unloading occurs.
Publications

Mrosk, C.; Forner, S.; Hause, G.; Küster, H.; Kopka, J.; Hause, B.; Composite Medicago truncatula plants harbouring Agrobacterium rhizogenes-transformed roots reveal normal mycorrhization by Glomus intraradices J. Exp. Bot. 60, 3797-3807, (2009) DOI: 10.1093/jxb/erp220

Composite plants consisting of a wild-type shoot and a transgenic root are frequently used for functional genomics in legume research. Although transformation of roots using Agrobacterium rhizogenes leads to morphologically normal roots, the question arises as to whether such roots interact with arbuscular mycorrhizal (AM) fungi in the same way as wild-type roots. To address this question, roots transformed with a vector containing the fluorescence marker DsRed were used to analyse AM in terms of mycorrhization rate, morphology of fungal and plant subcellular structures, as well as transcript and secondary metabolite accumulations. Mycorrhization rate, appearance, and developmental stages of arbuscules were identical in both types of roots. Using Mt16kOLI1Plus microarrays, transcript profiling of mycorrhizal roots showed that 222 and 73 genes exhibited at least a 2-fold induction and less than half of the expression, respectively, most of them described as AM regulated in the same direction in wild-type roots. To verify this, typical AM marker genes were analysed by quantitative reverse transcription-PCR and revealed equal transcript accumulation in transgenic and wild-type roots. Regarding secondary metabolites, several isoflavonoids and apocarotenoids, all known to accumulate in mycorrhizal wild-type roots, have been found to be up-regulated in mycorrhizal in comparison with non-mycorrhizal transgenic roots. This set of data revealed a substantial similarity in mycorrhization of transgenic and wild-type roots of Medicago truncatula, validating the use of composite plants for studying AM-related effects.
Publications

Lukačin, R.; Matern, U.; Hehmann, M.; Specker, S.; Vogt, T.; Corrigendum to “Cations modulate the substrate specificity of bifunctional class I O-methyltransferase from Ammi majus” [FEBS Lett. 577 (2004) 367-370] FEBS Lett. 583, 855-855, (2009) DOI: 10.1016/j.febslet.2009.01.050

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Publications

Hohnjec, N.; Lenz, F.; Fehlberg, V.; Vieweg, M. F.; Baier, M. C.; Hause, B.; Küster, H.; The Signal Peptide of the Medicago truncatula Modular Nodulin MtNOD25 Operates as an Address Label for the Specific Targeting of Proteins to Nitrogen-Fixing Symbiosomes Mol. Plant Microbe Interact. 22, 63-72, (2009) DOI: 10.1094/MPMI-22-1-0063

The nodule-specific MtNOD25 gene of the model legume Medicago truncatula encodes a modular nodulin composed of different repetitive modules flanked by distinct N- and C-termini. Although similarities are low with respect to all repetitive modules, both the N-terminal signal peptide (SP) and the C-terminus are highly conserved in modular nodulins from different legumes. On the cellular level, MtNOD25 is only transcribed in the infected cells of root nodules, and this activation is mediated by a 299-bp minimal promoter containing an organ-specific element. By expressing mGFP6 translational fusions in transgenic nodules, we show that MtNOD25 proteins are exclusively translocated to the symbiosomes of infected cells. This specific targeting only requires an N-terminal MtNOD25 SP that is highly conserved across a family of legume-specific symbiosome proteins. Our finding sheds light on one possible mechanism for the delivery of host proteins to the symbiosomes of infected root nodule cells and, in addition, defines a short molecular address label of only 24 amino acids whose N-terminal presence is sufficient to translocate proteins across the peribacteroid membrane.
Publications

Hause, B.; Schaarschmidt, S.; The role of jasmonates in mutualistic symbioses between plants and soil-born microorganisms Phytochemistry 70, 1589-1599, (2009) DOI: 10.1016/j.phytochem.2009.07.003

Many plants are able to develop mutualistic interactions with arbuscular mycorrhizal fungi and/or nitrogen-fixing bacteria. Whereas the former is widely distributed among most of the land plants, the latter is restricted to species of ten plant families, including the legumes. The establishment of both associations is based on mutual recognition and a high degree of coordination at the morphological and physiological level. This requires the activity of a number of signals, including jasmonates. Here, recent knowledge on the putative roles of jasmonates in both mutualistic symbioses will be reviewed. Firstly, the action of jasmonates will be discussed in terms of the initial signal exchange between symbionts and in the resulting plant signaling cascade common for nodulation and mycorrhization. Secondly, the putative role of jasmonates in the autoregulation of the endosymbioses will be outlined. Finally, aspects of function of jasmonates in the fully established symbioses will be presented. Various processes will be discussed that are possibly mediated by jasmonates, including the redox status of nodules and the carbohydrate partitioning of mycorrhizal roots.
Publications

Grunwald, U.; Guo, W.; Fischer, K.; Isayenkov, S.; Ludwig-Müller, J.; Hause, B.; Yan, X.; Küster, H.; Franken, P.; Overlapping expression patterns and differential transcript levels of phosphate transporter genes in arbuscular mycorrhizal, Pi-fertilised and phytohormone-treated Medicago truncatula roots Planta 229, 1023-1034, (2009) DOI: 10.1007/s00425-008-0877-z

A microarray carrying 5,648 probes of Medicago truncatula root-expressed genes was screened in order to identify those that are specifically regulated by the arbuscular mycorrhizal (AM) fungus Gigaspora rosea, by Pi fertilisation or by the phytohormones abscisic acid and jasmonic acid. Amongst the identified genes, 21% showed a common induction and 31% a common repression between roots fertilised with Pi or inoculated with the AM fungus G. rosea, while there was no obvious overlap in the expression patterns between mycorrhizal and phytohormone-treated roots. Expression patterns were further studied by comparing the results with published data obtained from roots colonised by the AM fungi Glomus mosseae and Glomus intraradices, but only very few genes were identified as being commonly regulated by all three AM fungi. Analysis of Pi concentrations in plants colonised by either of the three AM fungi revealed that this could be due to the higher Pi levels in plants inoculated by G. rosea compared with the other two fungi, explaining that numerous genes are commonly regulated by the interaction with G. rosea and by phosphate. Differential gene expression in roots inoculated with the three AM fungi was further studied by expression analyses of six genes from the phosphate transporter gene family in M. truncatula. While MtPT4 was induced by all three fungi, the other five genes showed different degrees of repression mirroring the functional differences in phosphate nutrition by G. rosea, G. mosseae and G. intraradices.
Publications

Floss, D. S.; Walter, M. H.; Role of carotenoid cleavage dioxygenase 1 (CCD1) in apocarotenoid biogenesis revisited Plant Signal Behav. 4, 172-175, (2009) DOI: 10.4161/psb.4.3.7840

Oxidative tailoring of C40 carotenoids by double bond-specific cleavage enzymes (carotenoid cleavage dioxygenases, CCDs) gives rise to various apocarotenoids. AtCCD1 generating C13 and C14 apocarotenoids and orthologous enzymes in other plants are the only CCDs acting in the cytosol, while the hitherto presumed C40 substrate is localized in the plastid. A new model for CCD1 action arising from a RNAi-mediated CCD1 gene silencing study in mycorrhizal hairy roots of Medicago truncatula may solve this contradiction. This approach unexpectedly resulted in the accumulation of C27 apocarotenoids but not C40 carotenoids suggesting C27 as the main substrates for CCD1 in planta. It further implies a consecutive two-step cleavage process, in which another CCD performs the primary cleavage of C40 to C27 in the plastid followed by C27 export and further cleavage by CCD1 in the cytosol. We compare the specificities and subcellular locations of the various CCDs and propose the plastidial CCD7 to be the first player in mycorrhizal apocarotenoid biogenesis.
Publications

Fellenberg, C.; Böttcher, C.; Vogt, T.; Phenylpropanoid polyamine conjugate biosynthesis in Arabidopsis thaliana flower buds Phytochemistry 70, 1392-1400, (2009) DOI: 10.1016/j.phytochem.2009.08.010

Phenylpropanoid polyamine conjugates have been identified in flowers of many plant species. Their presence in Arabidopsis thaliana has only been recently established in flower buds and pollen grains. Annotation and location of a cation-dependent O-methyltransferase AtTSM1 specifically to the tapetum of young flower buds enabled the subsequent identification of several genes with a putative role in phenylpropanoid polyamine conjugate biosynthesis. Based on the analysis of several A. thaliana knockout mutants, a biosynthetic pathway of these conjugates is proposed, which involves two methylation steps catalyzed by different cation-dependent O-methyltransferases, a cytochrome P450 (CYP98A8) catalyzed hydroxylation, and a conjugating acyl transfer performed by a BAHD-like, hydroxycinnamoyl (HC)-transferase. LC/MS based metabolite profiling of the cyp98A8 knockout line identified new feruloyl- and 4-coumaroylspermidine conjugates in the corresponding flowers consistent with a role of this gene in the hydroxylation of these conjugates. A pattern of minor amounts of bis- and tris-acylspermidine conjugates, likely the products of additional HC-transferases were identified in wild type as well as in the mutant lines. Transcript suppression of the genes early in the pathway was observed in knockout or RNAi-lines of the genes encoding late enzymatic steps. The implication of these findings for spermidine conjugate biosynthesis in flower buds of A. thaliana is discussed.
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