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Joachim Fisahn

Publications and source records attributed to Joachim Fisahn.

5 recordsLinked to original sources

An optical multifrequency phase-modulation method using microbeads for measuring intracellular oxygen concentrations in plants.

A technique has been developed to measure absolute intracellular oxygen concentrations in green plants. Oxygen-sensitive phosphorescent microbeads were injected into the cells and an optical multifrequency phase-modulation technique was used to discriminate the sensor signal from the strong autofluorescence of the plant tissue. The method was established using photosynthesis-competent cells of the giant algae Chara corallina L., and was validated by application to various cell types of other plant species.

Chara↗

Cell-specific protein profiling in Arabidopsis thaliana trichomes: identification of trichome-located proteins involved in sulfur metabolism and detoxification.

Metabolite, protein, and transcript analysis at the cellular level gives unparalleled insight into the complex roles tissues play in the plant system. However, while capillary electrophoresis and PCR amplification strategies make the profiling of metabolites and transcripts in specific cell types possible, the profiling of proteins in small samples represents a bottleneck. Here for the first time protein profiling has been achieved in a specific plant cell type: The application of specific cell sampling and shotgun peptide sequencing (nano LC/MS/MS) resulted in the identification of 63 unique proteins from pooled Arabidopsis trichome cells. A complete S-adenosylmethionine pathway cluster, two S-adenosylmethionine synthase isoforms, a glutathione S-conjugate translocator and other proteins involved in sulfur metabolism and detoxification are shown to be present in these cells, in agreement with previous work done at the level of trichome transcript analysis. The technology described here brings the simultaneous identification and localization of physiologically relevant cellular proteins within reach.

Arabidopsis↗

Analysis of the transient increase in cytosolic Ca2+ during the action potential of higher plants with high temporal resolution: requirement of Ca2+ transients for induction of jasmonic acid biosynthesis and PINII gene expression.

Plants respond to various abiotic stimuli by activation and propagation of fast electrical signals, action potentials. To resolve the temporal increase in cytosolic Ca(2)(+) during the action potentials of higher plants, we regenerated transgenic potato plants that expressed the Ca(2)(+) photoprotein apoaequorin. These genetically engineered potato plants were used for simultaneous measurements of transient changes in the membrane potential and the Ca(2)(+) luminescence triggered by heat-induced action potentials. High temporal resolution for recording of the fast transient electrical and light signals was accomplished by a sampling rate of 1 kHz. Upon elicitation by heat the membrane potential depolarization preceded the rise of cytosolic Ca(2)(+) by 50-100 ms. Several Ca(2)(+) channel blockers were tested to inhibit the rise in cytosolic Ca(2)(+). Treatment of plants with Ruthenium Red blocked the elevation in cytosolic Ca(2)(+) that was associated with heat-stimulated action potentials. Furthermore, action potentials have been demonstrated to stimulate jasmonic acid biosynthesis and PINII gene expression. Therefore, we measured jasmonic acid and PINII gene expression levels subsequent to action potential initiation by a short heating pulse. As expected, jasmonic acid biosynthesis and PINII gene expression were induced by action potentials. Pretreatment of potato plants with Ruthenium Red inhibited induction of jasmonic acid biosynthesis and PINII gene expression that was generally triggered by heat-activated action potentials.

Action Potentials↗

Adjustment of diurnal starch turnover to short days: depletion of sugar during the night leads to a temporary inhibition of carbohydrate utilization, accumulation of sugars and post-translational activation of ADP-glucose pyrophosphorylase in the following light period.

A larger proportion of the fixed carbon is retained as starch in the leaf in short days, providing a larger store to support metabolism and carbon export during the long night. The mechanisms that facilitate this adjustment of the sink-source balance are unknown. Starchless pgm mutants were analysed to discover responses that are triggered when diurnal starch turnover is disturbed. Sugars accumulated to high levels during the day, and fell to very low levels by the middle of the night. Sugars rose rapidly in the roots and rosette after illumination, and decreased later in the light period. Global transcript profiling revealed only small differences between pgm and Col0 at the end of the day but large differences at the end of the night, when pgm resembled Col0 after a 4-6 h prolongation of the night and many genes required for biosynthesis and growth were repressed [Plant J. 37 (2004) 914]. It is concluded that transient sugar depletion at the end of the night inhibits carbon utilization at the start of the ensuing light period. A second set of experiments investigated the stimulation of starch synthesis in response to short days in wild-type Col0. In short days, sugars were very low in the roots and rosette at the end of the dark period, and after illumination accumulated rapidly in both organs to levels that were higher than in long days. The response resembles pgm, except that carbohydrate accumulated in the leaf as starch instead of sugars. A similar response was found after transfer from long to short days. Inclusion of sugar in the rooting medium attenuated the stimulation of starch synthesis. Post-translational activation of ADP-glucose pyrophosphorylase (AGPase) was increased in pgm, and in Col0 in short days. It is concluded that starch synthesis is stimulated in short day conditions because sugar depletion at the end of the night triggers a temporary inhibition of growth and carbohydrate utilization in the first part of the light period, leading to transient accumulation of sugar and activation of AGPase.

Arabidopsis↗