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EXO modifies sucrose and trehalose responses and connects the extracellular carbon status to growth
(2013)
Plants have the capacity to adapt growth to changing environmental conditions. This implies the modulation of metabolism according to the availability of carbon (C). Particular interest in the response to the C availability is based on the increasing atmospheric levels of CO2. Several regulatory pathways that link the C status to growth have emerged. The extracellular EXO protein is essential for cell expansion and promotes shoot and root growth. Homologous proteins were identified in evolutionarily distant green plants. We show here that the EXO protein connects growth with C responses. The exo mutant displayed altered responses to exogenous sucrose supplemented to the growth medium. Impaired growth of the mutant in synthetic medium was associated with the accumulation of starch and anthocyanins, altered expression of sugar-responsive genes, and increased abscisic acid levels. Thus, EXO modulates several responses related to the C availability. Growth retardation on medium supplemented with 2-deoxy-glucose, mannose, and palatinose was similar to the wildtype. Trehalose feeding stimulated root growth and shoot biomass production of exoplants where as it inhibited growth of the wildtype. The phenotypic features of the exo mutant suggest that apoplastic processes coordinate growth and C responses.
The ability of some chemical compounds to cause oxidative stress offers a fast and convenient way to study the responses of plants to reactive oxygen species (ROS). In order to unveil potential novel genetic players of the ROS-regulatory network, a population of similar to 2,000 randomly selected Arabidopsis thaliana T-DNA insertion mutants was screened for ROS sensitivity/resistance by growing seedlings on agar medium supplemented with stress-inducing concentrations of the superoxide-eliciting herbicide methyl viologen or the catalase inhibitor 3-amino-triazole. A semi-robotic setup was used to capture and analyze images of the chemically treated seedlings which helped interpret the screening results by providing quantitative information on seedling area and healthy-to-chlorotic tissue ratios for data verification. A ROS-related phenotype was confirmed in three of the initially selected 33 mutant candidates, which carry T-DNA insertions in genes encoding a Ring/Ubox superfamily protein, ABI5 binding protein 1 (AFP1), previously reported to be involved in ABA signaling, and a protein of unknown function, respectively. In addition, we identified six mutants, most of which have not been described yet, that are related to growth or chloroplast development and show defects in a ROS-independent manner. Thus, semi-automated image capturing and phenotyping applied on publically available T-DNA insertion collections adds a simple means for discovering novel mutants in complex physiological processes and identifying the genes involved.
Translatome and metabolome effects triggered by gibberellins during rosette growth in Arabidopsis
(2012)
Although gibberellins (GAs) are well known for their growth control function, little is known about their effects on primary metabolism. Here the modulation of gene expression and metabolic adjustment in response to changes in plant (Arabidopsis thaliana) growth imposed on varying the gibberellin regime were evaluated. Polysomal mRNA populations were profiled following treatment of plants with paclobutrazol (PAC), an inhibitor of GA biosynthesis, and gibberellic acid (GA(3)) to monitor translational regulation of mRNAs globally. Gibberellin levels did not affect levels of carbohydrates in plants treated with PAC and/or GA(3). However, the tricarboxylic acid cycle intermediates malate and fumarate, two alternative carbon storage molecules, accumulated upon PAC treatment. Moreover, an increase in nitrate and in the levels of the amino acids was observed in plants grown under a low GA regime. Only minor changes in amino acid levels were detected in plants treated with GA(3) alone, or PAC plus GA(3). Comparison of the molecular changes at the transcript and metabolite levels demonstrated that a low GA level mainly affects growth by uncoupling growth from carbon availability. These observations, together with the translatome changes, reveal an interaction between energy metabolism and GA-mediated control of growth to coordinate cell wall extension, secondary metabolism, and lipid metabolism.
All's well that ends well
(2012)
The transition from cell proliferation to cell expansion is critical for determining leaf size. Andriankaja et al. (2012) demonstrate that in leaves of dicotyledonous plants, a basal proliferation zone is maintained for several days before abruptly disappearing, and that chloroplast differentiation is required to trigger the onset of cell expansion.
To gain a deeper understanding of the mechanisms behind biomass accumulation, it is important to study plant growth behavior. Manually phenotyping large sets of plants requires important human resources and expertise and is typically not feasible for detection of weak growth phenotypes. Here, we established an automated growth phenotyping pipeline for Arabidopsis thaliana to aid researchers in comparing growth behaviors of different genotypes.
The analysis pipeline includes automated image analysis of two-dimensional digital plant images and evaluation of manually annotated information of growth stages. It employs linear mixed-effects models to quantify genotype effects on total rosette area and relative leaf growth rate (RLGR) and ANOVAs to quantify effects on developmental times.
Using the system, a single researcher can phenotype up to 7000 plants d(-1). Technical variance is very low (typically < 2%). We show quantitative results for the growth-impaired starch-excessmutant sex4-3 and the growth-enhancedmutant grf9.
We show that recordings of environmental and developmental variables reduce noise levels in the phenotyping datasets significantly and that careful examination of predictor variables (such as d after sowing or germination) is crucial to avoid exaggerations of recorded phenotypes and thus biased conclusions.
Die vorliegende Dissertation behandelt die Ökologie von Cnidium dubium (Schkuhr) Thell. (Sumpf-Brenndolde), Gratiola officinalis L. (Gottes-Gnadenkraut) und Juncus atratus Krocker (Schwarze Binse), drei gefährdeten Arten, die als sogenannte Stromtalpflanzen in Mitteleuropa in ihrem Vorkommen eng an die Flussauen gebunden sind. Die Arbeit basiert auf verschiedenen Simulationsexperimenten und Feldstudien in der Unteren Havelniederung, einem „Feuchtgebiet von internationaler Bedeutung“. Sie behandelt Themenkomplexe wie das Samenbankverhalten, die Samenkeimung, die Stickstofflimitierung, die Konkurrenzkraft, das Verhalten der Pflanzen nach einer Sommertrockenheit und nach einer Winter/Frühjahrsüberflutung. Ferner widmet sie sich der Populationsbiologie der Arten und dem Verhalten der Pflanzen nach besonderen Störungsereignissen wie Mahd, Herbivorie und der Sommerflut 2002. Der Leser erfährt, wie die Pflanzen in verschiedenen Lebensphasen auf die auentypische Umwelt reagieren und erhält umfassende Einblicke in physiologische Mechanismen, die der Anpassung an die typischen Bedingungen einer mitteleuropäischen Flussaue dienen. Eine Interpretation der Ergebnisse zeigt auf, welche der spezifischen Eigenschaften zur Gefährdung der drei Stromtalarten beitragen. Die Arbeit ist für den Arten-, Biotop- und Landschaftsschutz interessant. Darüber hinaus bietet sie zahlreiche Anknüpfungspunkte zur ökophysiologischen Grundlagenforschung. Die verstärkte Nutzung physiologischer Methoden bei der Klärung ökologischer Fragestellungen wird angeregt.
In-situ Wachstumsuntersuchungen beim reaktiven Anlassen von Cu, In Schichten in elementarem Schwefel
(2003)
In dieser Arbeit wurde das reaktive Anlassen von dünnen Kupfer-Indium-Schichten in elementarem Schwefel mittels energiedispersiver Röntgenbeugung untersucht. Durch die simultane Aufnahme der Röntgenspektren und der Messung der diffusen Reflexion von Laserlicht der Wellenlänge 635 nm an der Oberfläche der Probe während des Schichtwachstums von CuInS<SUB>2</SUB> konnte eine Methode zur Prozesskontrolle für ein Herstellungsverfahren von CuInS<SUB>2</SUB> etabliert werden. Die Bildung von CuInS<SUB>2</SUB> aus Kupfer-Indium-Vorläuferschichten wird dominiert von Umwandlungen der intermetallischen Phasen. CuInS<SUB>2</SUB> wächst innerhalb der Aufheizperiode ab einer Temperatur von ca. 200°C aus der Phase Cu11In9. Jedoch zerfällt letztere metallische Phase in Cu16In9 und flüssiges Indium bei einer Temperatur von ca. 310°C. Das flüssige Indium reagiert im Falle von Kupferarmut mit dem Schwefel und führt zu einem zusätzlichen Reaktionspfad über InS zu CuIn5S8. Unter Präparationsbedingungen mit Kupferüberschuss wird das Indium in einer intermetallischen Phase gebunden.Erstmals konnte die Phase Digenite bei Temperaturen über 240°C beobachtet werden. Beim Abkühlen auf Raumtemperatur wandelt sich diese Phase unter dem Verbrauch von Schwefel in Covellite um.Für Proben mit Kupferüberschuss konnte eine Wachstumskinetik proportional zur Temperatur beobachtet werden. Dieses Verhalten wurde durch eine stress-induzierte Diffusion als dominierenden Reaktionsmechanismus interpretiert. Dabei werden während der Bildung von CuInS<SUB>2</SUB> durch unterschiedliche Ausdehnungen der metallischen und sulfidischen Schichten eine Spannung in der CuInS<SUB>2</SUB>-Schicht induziert, die nach Überschreiten einer Grenzspannung zu Rissen in der CuInS2-Schicht führt. Entlang dieser Risse findet ein schneller Transport der Metalle zur Oberfläche, wo diese mit dem Schwefel reagieren können, statt. Die Risse heilen durch die Bildung neuen Sulfids wieder aus.