@phdthesis{Czechowski2005, author = {Czechowski, Tomasz}, title = {Nitrogen signalling in Arabidopsis thaliana}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:517-opus-5445}, school = {Universit{\"a}t Potsdam}, year = {2005}, abstract = {Nitrogen is an essential macronutrient for plants and nitrogen fertilizers are indispensable for modern agriculture. Unfortunately, we know too little about how plants regulate their use of soil nitrogen, to maximize fertilizers-N use by crops and pastures. This project took a dual approach, involving forward and reverse genetics, to identify N-regulators in plants, which may prove useful in the future to improve nitrogen-use efficiency in agriculture. To identify nitrogen-regulated transcription factor genes in Arabidopsis that may control N-use efficiency we developed a unique resource for qRT-PCR measurements on all Arabidpsis transcription factor genes. Using closely spaced, gene-specific primer pairs and SYBR® Green to monitor amplification of double-stranded DNA, transcript levels of 83\% of all target genes could be measured in roots or shoots of young Arabidopsis wild-type plants. Only 4\% of reactions produced non-specific PCR products, and 13\% of TF transcripts were undetectable in these organs. Measurements of transcript abundance were quantitative over six orders of magnitude, with a detection limit equivalent to one transcript molecule in 1000 cells. Transcript levels for different TF genes ranged between 0.001-100 copies per cell. Real-time RT-PCR revealed 26 root-specific and 39 shoot-specific TF genes, most of which have not been identified as organ-specific previously. An enlarged and improved version of the TF qRT-PCR platform contains now primer pairs for 2256 Arabidopsis TF genes, representing 53 gene families and sub-families arrayed on six 384-well plates. Set-up of real-time PCR reactions is now fully robotized. One researcher is able to measure expression of all 2256 TF genes in a single biological sample in a just one working day. The Arabidopsis qRT-PCT platform was successfully used to identify 37 TF genes which transcriptionaly responded at the transcriptional level to N-deprivation or to nitrate per se. Most of these genes have not been characterized previously. Further selection of TF genes based on the responses of selected candidates to other macronutrients and abiotic stresses allowed to distinguish between TFs regulated (i) specifically by nitrogen (29 genes) (ii) regulated by general macronutrient or by salt and osmotic stress (6 genes), and (iii) responding to all major macronutrients and to abiotic stresses. Most of the N-regulated TF genes were also regulated by carbon. Further characterization of sixteen selected TF genes, revealed: (i) lack of transcriptional response to organic nitrogen, (ii) two major types of kinetics of induction by nitrate, (iii) specific responses for the majority of the genes to nitrate but not downstream products of nitrate assimilation. All sixteen TF genes were cloned into binary vectors for constitutive and ethanol inducible over expression, and the first generation of transgenic plants were obtained for almost all of them. Some of the plants constitutively over expressing TF genes under control of the 35S promoter revealed visible phenotypes in T1 generation. Homozygous T-DNA knock out lines were also obtained for many of the candidate TF genes. So far, one knock out line revealed a visible phenotype: retardation of flowering time. A forward genetic approach using an Arabidopsis ATNRT2.1 promoter : Luciferase reporter line, resulted in identification of eleven EMS mutant reporter lines affected in induction of ATNRT2.1 expression by nitrate. These lines could by divided in the following classes according to expression of other genes involved in primary nitrogen and carbon metabolism: (i) lines affected exclusively in nitrate transport, (ii) those affected in nitrate transport, acquisition, but also in glycolysis and oxidative pentose pathway, (iii) mutants affected moderately in nitrate transport, oxidative pentose pathway and glycolysis but not in primary nitrate assimilation. Thus, several different N-regulatory genes may have been mutated in this set of mutants. Map-based cloning has begun to identify the genes affected in these mutants.}, subject = {Stickstoff}, language = {en} } @phdthesis{Dreyer2005, author = {Dreyer, Ingo}, title = {Biophysikalische und molekulare Grundlagen der Regulation des Kaliumtransports in Pflanzen}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:517-opus-7708}, school = {Universit{\"a}t Potsdam}, year = {2005}, abstract = {Kaliumionen (K+) sind die am h{\"a}ufigsten vorkommenden anorganischen Kationen in Pflanzen. Gemessen am Trockengewicht kann ihr Anteil bis zu 10\% ausmachen. Kaliumionen {\"u}bernehmen wichtige Funktionen in verschiedenen Prozessen in der Pflanze. So sind sie z.B. essentiell f{\"u}r das Wachstum und f{\"u}r den Stoffwechsel. Viele wichtige Enzyme arbeiten optimal bei einer K+ Konzentration im Bereich von 100 mM. Aus diesem Grund halten Pflanzenzellen in ihren Kompartimenten, die am Stoffwechsel beteiligt sind, eine kontrollierte Kaliumkonzentration von etwa 100 mM aufrecht. Die Aufnahme von Kaliumionen aus dem Erdreich und deren Transport innerhalb der Pflanze und innerhalb einer Pflanzenzelle wird durch verschiedene Kaliumtransportproteine erm{\"o}glicht. Die Aufrechterhaltung einer stabilen K+ Konzentration ist jedoch nur m{\"o}glich, wenn die Aktivit{\"a}t dieser Transportproteine einer strikten Kontrolle unterliegt. Die Prozesse, die die Transportproteine regulieren, sind bis heute nur ansatzweise verstanden. Detailliertere Kenntnisse auf diesem Gebiet sind aber von zentraler Bedeutung f{\"u}r das Verst{\"a}ndnis der Integration der Transportproteine in das komplexe System des pflanzlichen Organismus. In dieser Habilitationsschrift werden eigene Publikationen zusammenfassend dargestellt, in denen die Untersuchungen verschiedener Regulationsmechanismen pflanzlicher Kaliumkan{\"a}le beschrieben werden. Diese Untersuchungen umfassen ein Spektrum aus verschiedenen proteinbiochemischen, biophysikalischen und pflanzenphysiologischen Analysen. Um die Regulationsmechanismen grundlegend zu verstehen, werden zum einen ihre strukturellen und molekularen Besonderheiten untersucht. Zum anderen werden die biophysikalischen und reaktionskinetischen Zusammenh{\"a}nge der Regulationsmechanismen analysiert. Die gewonnenen Erkenntnisse erlauben eine neue, detailliertere Interpretation der physiologischen Rolle der Kaliumtransportproteine in der Pflanze.}, subject = {Kaliumion}, language = {de} } @phdthesis{Dolniak2005, author = {Dolniak, Blazej}, title = {Functional characterisation of NIC2, a member of the MATE family from Arabidopsis thaliana (L.) Heynh.}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:517-opus-5372}, school = {Universit{\"a}t Potsdam}, year = {2005}, abstract = {The multidrug and toxic compounds extrusion (MATE) family includes hundreds of functionally uncharacterised proteins from bacteria and all eukaryotic kingdoms except the animal kingdom, that function as drug/toxin::Na+ or H+ antiporters. In Arabidopsis thaliana the MATE family comprises 56 members, one of which is NIC2 (Novel Ion Carrier 2). Using heterologous expression systems including Escherichia coli and Saccharomyces cerevisiae, and the homologous expression system of Arabidopsis thaliana, the functional characterisation of NIC2 was performed. It has been demonstrated that NIC2 confers resistance of E. coli towards the chemically diverse compounds such as tetraethylammonium chloride (TEACl), tetramethylammonium chloride (TMACl) and a toxic analogue of indole-3-acetic acid, 5-fluoro-indole-acetic acid (F-IAA). Therefore, NIC2 may be able to transport a broad range of drug and toxic compounds. In wild-type yeast the expression of NIC2 increased the tolerance towards lithium and sodium, but not towards potassium and calcium. In A. thaliana, the overexpression of NIC2 led to strong phenotypic changes. Under normal growth condtions overexpression caused an extremely bushy phenotype with no apical dominance but an enhanced number of lateral flowering shoots. The amount of rossette leaves and flowers with accompanying siliques were also much higher than in wild-type plants and the senescence occurred earlier in the transgenic plants. In contrast, RNA interference (RNAi) used to silence NIC2 expression, induced early flower stalk development and flowering compared with wild-type plants. In additon, the main flower stalks were not able to grow vertically, but instead had a strong tendency to bend towards the ground. While NIC2 RNAi seedlings produced many lateral roots outgrowing from the primary root and the root-shoot junction, NIC2 overexpression seedlings displayed longer primary roots that were characterised by a 2 to 4 h delay in the gravitropic response. In addition, these lines exhibited an enhanced resistance to exogenously applied auxins, i.e. indole-3-acetic acid (IAA) and indole-3-butyric acid (IBA) when compared with the wild-type roots. Based on these results, it is suggested that the NIC2 overexpression and NIC2 RNAi phenotypes were due to decreased or increased levels of auxin, respectively. The ProNIC2:GUS fusion gene revealed that NIC2 is expressed in the stele of the elongation zone, in the lateral root cap, in new lateral root primordia, and in pericycle cells of the root system. In the vascular tissue of rosette leaves and inflorescence stems, the expression was observed in the xylem parenchyma cells, while in siliques it was also in vascular tissue, but as well in the dehiscence and abscission zones. The organ- and tissue-specific expression sites of NIC2 correlate with the sites of auxin action in mature Arabidopsis plants. Further experiments using ProNIC2:GUS indicated that NIC2 is an auxin-inducible gene. Additionally, during the gravitropic response when an endogenous auxin gradient across the root tip forms, the GUS activity pattern of the ProNIC2:GUS fusion gene markedly changed at the upper side of the root tip, while at the lower side stayed unchanged. Finally, at the subcellular level NIC2-GFP fusion protein localised in the peroxisomes of Nicotana tabacum BY2 protoplasts. Considering the experimental results, it is proposed that the hypothetical function of NIC2 is the efflux transport which takes part in the auxin homeostasis in plant tissues probably by removing auxin conjugates from the cytoplasm into peroxisomes.}, subject = {Ackerschmalwand}, language = {en} } @phdthesis{GomezPorras2005, author = {G{\´o}mez-Porras, Judith Lucia}, title = {In silico identification of genes regulated by abscisic acid in Arabidopsis thaliana (L.) Heynh.}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:517-opus-7401}, school = {Universit{\"a}t Potsdam}, year = {2005}, abstract = {Abscisic acid (ABA) is a major plant hormone that plays an important role during plant growth and development. During vegetative growth ABA mediates (in part) responses to various environmental stresses such as cold, drought and high salinity. The response triggered by ABA includes changes in the transcript level of genes involved in stress tolerance. The aim of this project was the In silico identification of genes putatively regulated by ABA in A. thaliana. In silico predictions were combined with experimental data in order to evaluate the reliability of computational predictions. Taking advantage of the genome sequence of A. thaliana publicly available since 2000, 1 kb upstream sequences were screened for combinations of cis-elements known to be involved in the regulation of ABA-responsive genes. It was found that around 10 to 20 percent of the genes of A. thaliana might be regulated by ABA. Further analyses of the predictions revealed that certain combinations of cis-elements that confer ABA-responsiveness were significantly over-represented compared with results in random sequences and with random expectations. In addition, it was observed that other combinations that confer ABA-responsiveness in monocotyledonous species might not be functional in A. thaliana. It is proposed that ABA-responsive genes in A. thaliana show pairs of ABRE (abscisic acid responsive element) with MYB binding sites, DRE (dehydration responsive element) or with itself. The analysis of the distances between pairs of cis-elements suggested that pairs of ABREs are bound by homodimers of ABRE binding proteins. In contrast, pairs between MYB binding sites and ABRE, or DRE and ABRE showed a distance between cis-elements that suggested that the binding proteins interact through protein complexes and not directly. The comparison of computational predictions with experimental data confirmed that the regulatory mechanisms leading to the induction or repression of genes by ABA is very incompletely understood. It became evident that besides the cis-elements proposed in this study to be present in ABA-responsive genes, other known and unknown cis-elements might play an important role in the transcriptional regulation of ABA-responsive genes. For example, auxin-related cis elements, or the cis-elements recognized by the NAM-family of transcription factors (Non-Apical meristem). This work documents the use of computational and experimental approaches to analyse possible interactions between cis-elements involved in the regulation of ABA-responsive genes. The computational predictions allowed the distinction between putatively relevant combinations of cis-elements from irrelevant combinations of cis-elements in ABA-responsive genes. The comparison with experimental data allowed to identify certain cis-elements that have not been previously associated to the ABA-mediated transcriptional regulation, but that might be present in ABA-responsive genes (e.g. auxin responsive elements). Moreover, the efforts to unravel the gene regulatory network associated with the ABA-signalling pathway revealed that NAM-transcription factors and their corresponding binding sequences are important components of this network.}, subject = {Bioinformatik}, language = {en} } @phdthesis{Blacha2009, author = {Blacha, Anna Maria}, title = {Investigating the role of regulatory genes in heterosis for superior growth and biomass production in Arabidopsis thaliana}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:517-opus-46146}, school = {Universit{\"a}t Potsdam}, year = {2009}, abstract = {'Heterosis' is a term used in genetics and breeding referring to hybrid vigour or the superiority of hybrids over their parents in terms of traits such as size, growth rate, biomass, fertility, yield, nutrient content, disease resistance or tolerance to abiotic and abiotic stress. Parental plants which are two different inbred (pure) lines that have desired traits are crossed to obtain hybrids. Maximum heterosis is observed in the first generation (F1) of crosses. Heterosis has been utilised in plant and animal breeding programs for at least 90 years: by the end of the 21st century, 65\% of worldwide maize production was hybrid-based. Generally, it is believed that an understanding of the molecular basis of heterosis will allow the creation of new superior genotypes which could either be used directly as F1 hybrids or form the basis for the future breeding selection programmes. Two selected accessions of a research model plant Arabidopsis thaliana (thale cress) were crossed to obtain hybrids. These typically exhibited a 60-80\% increase of biomass when compared to the average weight of both parents. This PhD project focused on investigating the role of selected regulatory genes given their potentially key involvement in heterosis. In the first part of the project, the most appropriate developmental stage for this heterosis study was determined by metabolite level measurements and growth observations in parents and hybrids. At the selected stage, around 60 candidate regulatory genes (i.e. differentially expressed in hybrids when compared to parents) were identified. Of these, the majority were transcription factors, genes that coordinate the expression of other genes. Subsequent expression analyses of the candidate genes in biomass-heterotic hybrids of other Arabidopsis accessions revealed a differential expression in a gene subset, highlighting their relevance for heterosis. Moreover, a fraction of the candidate regulatory genes were found within DNA regions closely linked to the genes that underlie the biomass or growth heterosis. Additional analyses to validate the role of selected candidate regulatory genes in heterosis appeared insufficient to establish their role in heterosis. This uncovered a need for using novel approaches as discussed in the thesis. Taken together, the work provided an insight into studies on the molecular mechanisms underlying heterosis. Although studies on heterosis date back to more than one hundred years, this project as many others revealed that more investigations will be needed to uncover this phenomenon.}, language = {en} } @phdthesis{Arvidsson2010, author = {Arvidsson, Samuel Janne}, title = {Identification of growth-related tonoplast proteins in Arabidopsis thaliana}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:517-opus-52408}, school = {Universit{\"a}t Potsdam}, year = {2010}, abstract = {In a very simplified view, the plant leaf growth can be reduced to two processes, cell division and cell expansion, accompanied by expansion of their surrounding cell walls. The vacuole, as being the largest compartment of the plant cell, plays a major role in controlling the water balance of the plant. This is achieved by regulating the osmotic pressure, through import and export of solutes over the vacuolar membrane (the tonoplast) and by controlling the water channels, the aquaporins. Together with the control of cell wall relaxation, vacuolar osmotic pressure regulation is thought to play an important role in cell expansion, directly by providing cell volume and indirectly by providing ion and pH homestasis for the cytosoplasm. In this thesis the role of tonoplast protein coding genes in cell expansion in the model plant Arabidopsis thaliana is studied and genes which play a putative role in growth are identified. Since there is, to date, no clearly identified protein localization signal for the tonoplast, there is no possibility to perform genome-wide prediction of proteins localized to this compartment. Thus, a series of recent proteomic studies of the tonoplast were used to compile a list of cross-membrane tonoplast protein coding genes (117 genes), and other growth-related genes from notably the growth regulating factor (GRF) and expansin families were included (26 genes). For these genes a platform for high-throughput reverse transcription quantitative real time polymerase chain reaction (RT-qPCR) was developed by selecting specific primer pairs. To this end, a software tool (called QuantPrime, see http://www.quantprime.de) was developed that automatically designs such primers and tests their specificity in silico against whole transcriptomes and genomes, to avoid cross-hybridizations causing unspecific amplification. The RT-qPCR platform was used in an expression study in order to identify candidate growth related genes. Here, a growth-associative spatio-temporal leaf sampling strategy was used, targeting growing regions at high expansion developmental stages and comparing them to samples taken from non-expanding regions or stages of low expansion. Candidate growth related genes were identified after applying a template-based scoring analysis on the expression data, ranking the genes according to their association with leaf expansion. To analyze the functional involvement of these genes in leaf growth on a macroscopic scale, knockout mutants of the candidate growth related genes were screened for growth phenotypes. To this end, a system for non-invasive automated leaf growth phenotyping was established, based on a commercially available image capture and analysis system. A software package was developed for detailed developmental stage annotation of the images captured with the system, and an analysis pipeline was constructed for automated data pre-processing and statistical testing, including modeling and graph generation, for various growth-related phenotypes. Using this system, 24 knockout mutant lines were analyzed, and significant growth phenotypes were found for five different genes.}, language = {en} } @phdthesis{Ivakov2011, author = {Ivakov, Alexander}, title = {Metabolic interactions in leaf development in Arabidopsis thaliana}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:517-opus-59730}, school = {Universit{\"a}t Potsdam}, year = {2011}, abstract = {Das Wachstum und {\"U}berleben von Pflanzen basiert auf der Photosynthese in den Bl{\"a}ttern. Diese beinhaltet die Aufnahme von Kohlenstoffdioxid aus der Atmosph{\"a}re und das simultane Einfangen von Lichtenergie zur Bildung organischer Molek{\"u}le. Diese werden nach dem Eintritt in den Metabolismus in viele andere Komponenten umgewandelt, welche die Grundlage f{\"u}r die Zunahme der Biomasse bilden. Bl{\"a}tter sind Organe, die auf die Fixierung von Kohlenstoffdioxid spezialisiert sind. Die Funktionen der Bl{\"a}tter beinhalten vor allem die Optimierung und Feinregulierung vieler Prozesse, um eine effektive Nutzung von Ressourcen und eine maximale Photosynthese zu gew{\"a}hrleisten. Es ist bekannt, dass sich die Morphologie der Bl{\"a}tter den Wachstumsbedingungen der Pflanze anpasst und eine wichtige Rolle bei der Optimierung der Photosynthese spielt. Trotzdem ist die Regulation dieser Art der Anpassung bisher nicht verstanden. Die allgemeine Zielsetzung dieser vorliegenden Arbeit ist das Verst{\"a}ndnis wie das Wachstum und die Morphologie der Bl{\"a}tter im Modellorganismus Arabidopsis thaliana reguliert werden. Besondere Aufmerksamkeit wurde hierbei der M{\"o}glichkeit geschenkt, dass es interne metabolische Signale in der Pflanze geben k{\"o}nnte, die das Wachstum und die Entwicklung von Bl{\"a}ttern beeinflussen. Um diese Fragestellung zu untersuchen, muss das Wachstum und die Entwicklung von Bl{\"a}ttern oberhalb des Levels des einzelnen Organs und im Kontext der gesamten Pflanze betrachtet werden, weil Bl{\"a}tter nicht eigenst{\"a}ndig wachsen, sondern von Ressourcen und regulatorischen Einfl{\"u}ssen der ganzen Pflanze abh{\"a}ngig sind. Aufgrund der Komplexit{\"a}t dieser Fragestellung wurden drei komplement{\"a}re Ans{\"a}tze durchgef{\"u}hrt. Im ersten und spezifischsten Ansatz wurde untersucht ob eine flussabw{\"a}rts liegende Komponente des Zucker-Signalwegs, Trehalose-6-Phosphat (Tre-6-P), das Blattwachstum und die Blattentwicklung beinflussen kann. Um diese Frage zu beantworten wurden transgene Arabidopsis-Linien mit einem gest{\"o}rten Gehalt von Tre-6-P durch die Expression von bakteriellen Proteinen die in dem metabolismus von trehalose beteiligt sind. Die Pflanzen-Linien wurden unter Standard-Bendingungen in Erde angebaut und ihr Metabolismus und ihre Blattmorphologie untersucht. Diese Experimente f{\"u}hrten auch zu einem unerwarteten Projekt hinsichtlich einer m{\"o}glichen Rolle von Tre-6-P in der Regulation der Stomata. In einem zweiten, allgemeineren Ansatz wurde untersucht, ob {\"A}nderungen im Zucker-Gehalt der Pflanzen die Morphogenese der Bl{\"a}tter als Antwort auf Licht beeinflussen. Dazu wurden eine Reihe von Mutanten, die im Zentralmetabolismus beeintr{\"a}chtigt sind, in derselben Lichtbedingung angezogen und bez{\"u}glich ihrer Blattmorphologie analysiert. In einem dritten noch allgemeineren Ansatz wurde die nat{\"u}rliche Variation von morphologischen Auspr{\"a}gungen der Bl{\"a}tter und Rosette anhand von wilden Arabidopsis {\"O}kotypen untersucht, um zu verstehen wie sich die Blattmorphologie auf die Blattfunktion und das gesamte Pflanzenwachstum auswirkt und wie unterschiedliche Eigenschaften miteinander verkn{\"u}pft sind. Das Verh{\"a}ltnis der Blattanzahl zum Gesamtwachstum der Pflanze und Blattgr{\"o}ße wurde gesondert weiter untersucht durch eine Normalisierung der Blattanzahl auf das Frischgewicht der Rosette, um den Parameter „leafing Intensity" abzusch{\"a}tzen. Leafing Intensity integrierte Blattanzahl, Blattgr{\"o}ße und gesamtes Rosettenwachstum in einer Reihe von Kompromiss-Interaktionen, die in einem Wachstumsvorteil resultieren, wenn Pflanzen weniger, aber gr{\"o}ßere Bl{\"a}tter pro Einheit Biomasse ausbilden. Dies f{\"u}hrte zu einem theoretischen Ansatz in dem ein einfaches allometrisch mathematisches Modell konstruiert wurde, um Blattanzahl, Blattgr{\"o}ße und Pflanzenwachstum im Kontext der gesamten Pflanze Arabidopsis zu verkn{\"u}pfen.}, language = {en} } @article{RuprechtMutwilSaxeetal.2011, author = {Ruprecht, Colin and Mutwil, Marek and Saxe, Friederike and Eder, Michaela and Nikoloski, Zoran and Persson, Staffan}, title = {Large-scale co-expression approach to dissect secondary cell wall formation across plant species}, series = {Frontiers in plant science}, volume = {2}, journal = {Frontiers in plant science}, publisher = {Frontiers Research Foundation}, address = {Lausanne}, issn = {1664-462X}, doi = {10.3389/fpls.2011.00023}, pages = {13}, year = {2011}, abstract = {Plant cell walls are complex composites largely consisting of carbohydrate-based polymers, and are generally divided into primary and secondary walls based on content and characteristics. Cellulose microfibrils constitute a major component of both primary and secondary cell walls and are synthesized at the plasma membrane by cellulose synthase (CESA) complexes. Several studies in Arabidopsis have demonstrated the power of co-expression analyses to identify new genes associated with secondary wall cellulose biosynthesis. However, across-species comparative co-expression analyses remain largely unexplored. Here, we compared co-expressed gene vicinity networks of primary and secondary wall CESAsin Arabidopsis, barley, rice, poplar, soybean, Medicago, and wheat, and identified gene families that are consistently co-regulated with cellulose biosynthesis. In addition to the expected polysaccharide acting enzymes, we also found many gene families associated with cytoskeleton, signaling, transcriptional regulation, oxidation, and protein degradation. Based on these analyses, we selected and biochemically analyzed T-DNA insertion lines corresponding to approximately twenty genes from gene families that re-occur in the co-expressed gene vicinity networks of secondary wall CESAs across the seven species. We developed a statistical pipeline using principal component analysis and optimal clustering based on silhouette width to analyze sugar profiles. One of the mutants, corresponding to a pinoresinol reductase gene, displayed disturbed xylem morphology and held lower levels of lignin molecules. We propose that this type of large-scale co-expression approach, coupled with statistical analysis of the cell wall contents, will be useful to facilitate rapid knowledge transfer across plant species.}, language = {en} } @article{RocchettiSharmaWulfetangeetal.2012, author = {Rocchetti, Alessandra and Sharma, Tripti and Wulfetange, Camilla and Scholz-Starke, Joachim and Grippa, Alexandra and Carpaneto, Armando and Dreyer, Ingo and Vitale, Alessandro and Czempinski, Katrin and Pedrazzini, Emanuela}, title = {The putative K+ channel subunit AtKCO3 forms stable dimers in arabidopsis}, series = {Frontiers in plant science}, volume = {3}, journal = {Frontiers in plant science}, publisher = {Frontiers Research Foundation}, address = {Lausanne}, issn = {1664-462X}, doi = {10.3389/fpls.2012.00251}, pages = {13}, year = {2012}, abstract = {The permeation pore of K+ channels is formed by four copies of the pore domain. AtKCO3 is the only putative voltage-independent K+ channel subunit of Arabidopsis thaliana with a single pore domain. KCO3-like proteins recently emerged in evolution and, to date, have been found only in the genus Arabidopsis (A. thaliana and A. lyrata). We show that the absence of KCO3 does not cause marked changes in growth under various conditions. Only under osmotic stress we observed reduced root growth of the kco3-1 null-allele line. This phenotype was complemented by expressing a KCO3 mutant with an inactive pore, indicating that the function of KCO3 under osmotic stress does not depend on its direct ability to transport ions. Constitutively overexpressed AtKCO3 or AtKCO3::G FP are efficiently sorted to the tonoplast indicating that the protein is approved by the endoplasmic reticulum quality control. However, vacuoles isolated from transgenic plants do not have significant alterations in current density. Consistently, both AtKCO3 and AtKCO3::GFP are detected as homodimers upon velocity gradient centrifugation, an assembly state that would not allow for activity. We conclude that if AtKCO3 ever functions as a K+ channel, active tetramers are held by particularly weak interactions, are formed only in unknown specific conditions and may require partner proteins.}, language = {en} } @article{SakurabaBalazadehTanakaetal.2012, author = {Sakuraba, Yasuhito and Balazadeh, Salma and Tanaka, Ryouichi and M{\"u}ller-R{\"o}ber, Bernd and Tanaka, Ayumi}, title = {Overproduction of Chl b retards senescence through transcriptional reprogramming in arabidopsis}, series = {Plant \& cell physiology}, volume = {53}, journal = {Plant \& cell physiology}, number = {3}, publisher = {Oxford Univ. Press}, address = {Oxford}, issn = {0032-0781}, doi = {10.1093/pcp/pcs006}, pages = {505 -- 517}, year = {2012}, abstract = {Leaf senescence is a developmentally and environmentally regulated process which includes global changes in gene expression. Using Arabidopsis as a model, we modified Chl arrangement in photosystems by overexpressing the catalytic domain (the C domain) of chlorophyllide a oxygenase (CAO) fused with the linker domain (the B domain) of CAO and green fluorescent protein (GFP). In these plants (referred to as the BCG plants for the B and C domains of CAO and GFP), the Chl a/b ratio was drastically decreased and Chl b was incorporated into core antenna complexes. The BCG plants exhibited a significant delay of both developmental and dark-induced leaf senescence. The photosynthetic apparatus, CO2 fixation enzymes and the chloroplast structure were lost in wild-type plants during senescence, while BCG plants retained them longer than the wild type. Large-scale quantitative real-time PCR analyses of 1,880 transcription factor (TF) genes showed that 241 TFs are differentially expressed between BCG plants and wild-type plants at senescence, similar to 40\% of which are known senescence-associated genes (SAGs). Expression profiling also revealed the down-regulation of a large number of additional non-TF SAGs. In contrast, genes involved in photosynthesis were up-regulated, while those encoding Chl degradation enzymes were down-regulated in BCG plants. These results demonstrate that alteration of pigment composition in the photosynthetic apparatus retards senescence through transcriptional reprogramming.}, language = {en} }