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Imaging Unit

Nowadays, the elucidation of molecular and biochemical processes requires its combination with investigation of cells – their physiological properties, their structure, the organelles they contain, interactions with their environment, their life cycle, division and death. For centuries, progress in biological research has been connected to the development of tools and equipment that allow new insights into living matter. The invention of and improvements in optical systems were very important because exceeding the limits of the optical resolution of the human eye delivered new insights into tissues, cells and subcellular compartments on the one hand and cellular processes on the other.

The Imaging Unit of the IPB aims to support all research groups in their use of cell biological methods. Currently, at least 13 research groups of the institute are using this unit.

The unit provides:

  • Coordinated supervision and maintenance of equipment
  • Optimal training of coworkers
  • Maximal use of IPB investments
  • Updating of equipment according to state-of-the-art and to requirements of actual research

The working principle of this unit is:

  • Unit headed by one scientist (Prof. Dr. Bettina Hause) and supported by one technician (Hagen Stellmach)
  • Equipment is located decentralized, but its maintenance is carried out centralized
  • For all cell biological methods advice, training and help is provided, extensive experiments, however, have to be done by the co-workers themselves

Devices and materials:

 

Microscopes:

Several stereo microscopes
(Zeiss and Nikon)

Multipurpose MacroMicroSystem equipped with epi-fluorescence: AZ100 (Nikon) with camera (one in each Dept. MSV and SEB)

Lightsheet Microscope
 

Lightsheet Z1 (Zeiss)                                                                                                                                               

Epi-fluorescence microscopes

Axioplan 2 (Zeiss) with differential interference contrast (DIC) device and ApoTome to obtain optical sections, with two cameras (AxioCam MRm and AxioCam MRc5)

AxioImager (Zeiss) with differential interference contrast (DIC) device and ApoTome to obtain optical sections, with two cameras (AxioCam MRm and AxioCam MRc5)

Confocal Laser Scanning Microscope

LSM780 (Zeiss) with Airyscan

LSM700 (Zeiss)


Microtomes

Rotary microtomes to perform semi-thin sections (Microm und Leica)

Vibrating microtome (Vibratome VT1000S, Leica) (Dept. SZB)

Cryo-Microtom CM1950 (Leica)                                                                      

Miscellaneous

InsituPro VSi (Intavis) for automated in situ detection (Dept. SZB)

Micromanipulator (Eppendorf)

Laser Capture Microdissection                                                                    

More devices

  • Organelle-marker: Vectors and transgenic lines of Arabidopsis (Nelson et al., 2007)
  • Wave-marker: Vectors and transgenic lines of Arabidopsis (Geldner et al., 2009)









Methods established:

  • Fixiation, embedding and sectioning of plant materials
  • Laser-Micro-Dissection
  • Immuno labelling
  • in situ-hybridisation
  • light microscopy including fluorescence microscopy
  • confocal laser scanning microscopy
  • Determination of protein interactions via FRET and BiFC (Split-YFP)

Publications by Tag: Cell Biology

Sort by: sort ascending Year Type of publication

Displaying results 1 to 10 of 16.

Publications

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

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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 <em>Medicago truncatula</em>modular nodulin MtNOD25 operates as an address label for the specific targeting of proteins to nitrogen-fixing symbiosomes Mol. Plant Microbe In 22, 63-72, (2009)

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Publications

Bethke, G.; Unthan, T.; Uhrig, J.F.; Pöschl, Y.; Gust, A.A.; Scheel, D.; Lee, J. Flg22 regulates the release of an ethylene response factor substrate from MAP kinase 6 in <em>Arabidopsis thaliana</em> via ethylene signaling Proc Natl Acad Sci U S A 106, 8067-72, (2009)

Mitogen-activated protein kinase (MAPK)mediated responses are in part regulated by the repertoire of MAPK substrates, which is still poorly elucidated in plants. Here, the in vivo enzymesubstrate interaction of the Arabidopsis thaliana MAP kinase, MPK6, with an ethylene response factor (ERF104) is shown by fluorescence resonance energy transfer. The interaction was rapidly lost in response to flagellin-derived flg22 peptide. This complex disruption requires not only MPK6 activity, which also affects ERF104 stability via phosphorylation, but also ethylene signaling. The latter points to a novel role of ethylene in substrate release, presumably allowing the liberated ERF104 to access target genes. Microarray data show enrichment of GCC motifs in the promoters of ERF104up-regulated genes, many of which are stress related. ERF104 is a vital regulator of basal immunity, as altered expression in both erf104 and overexpressors led to more growth inhibition by flg22 and enhanced susceptibility to a non-adapted bacterial pathogen.
Publications

Teutschbein, J.; Gross, W.; Nimtz, M.; Milkowski, C.; Hause, B.; Strack, D. Identification and localization of a lipase-like acyltransferase in phenylpropanoid metabolism of tomato (<em>Solanum lycopersicum</em>) J. Biol. Chem 285, 38374-38381 , (2010)

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Publications

Breuillin, F.; Hajirezaei, M.-R.; Ahkami, A.; Favre, P.; Druege, U.; Hause, B.; Bucher, M.; Kretzschmar, T.; Bossolini, E.; Kuhlemeier, C.; Martinoia, E.; Franken, P.; Scholz, U.; Reinhardt, D. Phosphate systemically inhibits development of arbuscular mycorrhiza in <em>Petunia hybrida</em> and represses genes involved in mycorrhizal functioning Plant J 64, 1002-1017 , (2010)

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Publications

Zdyb, A.; Demchenko, K.; Heumann, J.; Mrosk, C.; Grzeganek, P.; Göbel, C.; Feussner, I.; Pawlowski, K.; Hause, B. Jasmonate biosynthesis in legume and actinorhizal nodules New Phytol 189, 568-579 , (2011)

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Publications

Helber, N.; Wippel, K.; Sauer, N.; Schaarschmidt, S.; Hause, B.; Requena, N. A versatile monosaccharide transporter that operates in the arbuscular mycorrhizal fungus <i>Glomus sp</i> is crucial for the symbiotic relationship with plants. Plant Cell 23, 3812-3823, (2011)

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Publications

Mielke, K.; Forner, S.; Kramell, R.; Conrad, U.; Hause, B. Cell-specific visualization of jasmonates in wounded tomato and Arabidopsis leaves using jasmonate-specific antibodies New Phytol 190, 1069-1080, (2011)

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Publications

Goetz, S.; Hellwege, A.; Stenzel, I.; Kutter, C.; Hauptmann, V.; Forner, S.; Mc Caig, B.; Hause, G.; Miersch, O.; Wasternack, C.; Hause, B. Role of cis-12-oxo-phytodienoic acid in tomato embryo development. Plant Physiol 158 (4), 1715-1727, (2012)

Oxylipins including jasmonates are signaling compounds in plant growth, development, and responses to biotic and abiotic stresses. In Arabidopsis (Arabidopsis thaliana) most mutants affected in jasmonic acid (JA) biosynthesis and signaling are male sterile, whereas the JA-insensitive tomato (Solanum lycopersicum) mutant jai1 is female sterile. The diminished seed formation in jai1 together with the ovule-specific accumulation of the JA biosynthesis enzyme allene oxide cyclase (AOC), which correlates with elevated levels of JAs, suggest a role of oxylipins in tomato flower/seed development. Here, we show that 35S::SlAOC-RNAi lines with strongly reduced AOC in ovules exhibited reduced seed set similarly to the jai1 plants. Investigation of embryo development of wild-type tomato plants showed preferential occurrence of AOC promoter activity and AOC protein accumulation in the developing seed coat and the embryo, whereas 12-oxo-phytodienoic acid (OPDA) was the dominant oxylipin occurring nearly exclusively in the seed coat tissues. The OPDA- and JA-deficient mutant spr2 was delayed in embryo development and showed an increased programmed cell death in the developing seed coat and endosperm. In contrast, the mutant acx1a, which accumulates preferentially OPDA and residual amount of JA, developed embryos similar to the wild type, suggesting a role of OPDA in embryo development. Activity of the residual amount of JA in the acx1a mutant is highly improbable since the known reproductive phenotype of the JA-insensitive mutant jai1 could be rescued by wound-induced formation of OPDA. These data suggest a role of OPDA or an OPDA-related compound for proper embryo development possibly by regulating carbohydrate supply and detoxification.
Publications

Stenzel, I.; Otto, M.; Delker, C.; Kirmse, N.; Schmidt, D.; Miersch, O.; Hause, B.; Wasternack, C. ALLENE OXIDE CYCLASE (AOC) gene family members of <em>Arabidopsis thaliana</em>: tissue- and organ-specific promoter activities and in vivo heteromerization* J Exp Bot. 63, 6125-6138, (2012)

Jasmonates are important signals in plant stress responses and plant development. An essential step in the biosynthesis of jasmonic acid (JA) is catalysed by ALLENE OXIDE CYCLASE (AOC) which establishes the naturally occurring enantiomeric structure of jasmonates. In Arabidopsis thaliana, four genes encode four functional AOC polypeptides (AOC1, AOC2, AOC3, and AOC4) raising the question of functional redundancy or diversification. Analysis of transcript accumulation revealed an organ-specific expression pattern, whereas detailed inspection of transgenic lines expressing the GUS reporter gene under the control of individual AOC promoters showed partially redundant promoter activities during development: (i) In fully developed leaves, promoter activities of AOC1, AOC2, and AOC3 appeared throughout all leaf tissue, but AOC4 promoter activity was vascular bundle-specific; (ii) only AOC3 and AOC4 showed promoter activities in roots; and (iii) partially specific promoter activities were found for AOC1 and AOC4 in flower development. In situ hybridization of flower stalks confirmed the GUS activity data. Characterization of single and double AOC loss-of-function mutants further corroborates the hypothesis of functional redundancies among individual AOCs due to a lack of phenotypes indicative of JA deficiency (e.g. male sterility). To elucidate whether redundant AOC expression might contribute to regulation on AOC activity level, protein interaction studies using bimolecular fluorescence complementation (BiFC) were performed and showed that all AOCs can interact among each other. The data suggest a putative regulatory mechanism of temporal and spatial fine-tuning in JA formation by differential expression and via possible heteromerization of the four AOCs.

This page was last modified on 14.11.2018.

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