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Menge, Zusammensetzung und Umsetzung der organischen Substanz im Unterboden

Das Wissen über die Menge, Zusammensetzung und Umsetzung der organischen Substanz in Böden der gemäßigten Breiten beschränkt sich bis auf wenige Ausnahmen auf die Oberböden (A-Horizonte und Auflagen). Hier finden sich die höchsten Konzentrationen der organischen Substanz. Jüngere Inventurarbeiten haben nun gezeigt, dass auch im Unterboden (B- und Cv-Horizonte) beträchtliche Mengen an organischer Substanz, allerdings in niedrigen Konzentrationen vorliegen. Ziel des geplanten Vorhabens ist es, (1) die Menge der organischen Substanz im Unterboden zu erfassen, (2) ihre Zusammensetzung und Herkunft zu bestimmen und (3) ihre Umsetzbarkeit zu erfassen. Daraus sollen Rückschlüsse auf die Stabilisierungsmechanismen der organischen Substanz im Unterboden gezogen werden. Nach einer Inventur der Bodenprofile an den SPP-Standorten (C-Gehalte, 14C-Alter) erfolgt die Erfassung der Zusammensetzung der organischen Substanz mittels Festkörper-13C-NMR-Spektroskopie. Die Zusammensetzung der Lipid-, Polysaccharid- und Ligninfraktion soll Hinweise auf die Herkunft der stabilisierten organischen Substanz differenziert nach oberirdischen, unterirdischen Pflanzenrückständen und mikrobiellen Resten geben. Abbauversuche unter kontrollierten Bedingungen im Labor und die Erfassung des 14C-Alters des freigesetzten CO2 sollen Aufschluß über die Umsetzbarkeit des 'jungen' und 'alten' C im Unterboden geben. Dabei werden jeweils die Profile über die gesamte Entwicklungstiefe betrachtet, um die Unterbodenhorizonte in Bezug zu den Oberböden und zu den Ergebnissen anderer AG im SPP zu setzen. Darauf aufbauend können dann in den nächsten Phasen des SPP die Eigenschaften der organischen Substanz im Unterboden und die Regulation der C-Umsetzungen im Unterboden untersucht werden.

Grundlage von Trockentoleranz in hoeheren Pflanzen

Das Ziel unserer Untersuchungen ist es, molekulare Mechanismen aufzuklaeren, die zur Trockentoleranz bei hoeheren Pflanzen fuehren. Dazu untersuchen wir als Modellsystem die Wiederauferstehungspflanze C. plantagineum. Diese Pflanze zeichnet sich durch eine extreme Trockentoleranz aus. Wir haben mehrere Gene isoliert, die waehrend des Trockenstresses induziert werden. Es wird untersucht, inwieweit diese Genprodukte zur Trockentoleranz beitragen. Die Gene koennen in drei Gruppen eingeteilt werden: Lea-(late anbryogenesis abundant) Gene, Gene, die fuer Produkte des Kohlenhydratstoffwechsels kodieren, sowie regulatorische Gene.

Biofilme, Makromoleküle und organische Restsubstanzen als Matrizen bei der Bildung von Organo- und Biomineralen - Geobiologische Faktoren bei der Evolution der Biomineralisation

Durch vergleichende Analyse von Organo- und Biomineralen aus evolutionsbiologisch zunehmend komplexeren Systemen - von Organofilmen (Ooide) über Biofilme zu Poriferen - sollen systemspezifische Wechselwirkungen zwischen Makromolekülen und Mineralphasen sowie Steuerungsmechanismen der Mineralbildung aufgezeigt werden. Dazu werden aus verschiedenen Habitaten (Hartwasserseen, Salzseen, Sodaseen, Meerwasser) makromolekulare Überzüge (Organofilme), polysaccharidreiche phototrophe und heterotrophe Biofilme sowie proteinreiche heterotrophe BiofilmMetazoen-Gemeinschaften (Riffhöhlen) untersucht. Ausgehend von der hydrochemischen Charakterisierung der Habitate, wird eine biochemische Charakterisierung der primären organischen Substanzen und Matrix sowie der Restsubstanzen in den assoziierten Mineralisaten durchgeführt. Eine strukturelle und mikrobiologische Analyse der beteiligten Organo- und Biofilme folgt (histochemische Färbungen, Applikation von Oligonukleotidsonden zur in situ Identifikation nicht-phototropher Bakterien - FISH). In kontrollierten Experimenten wird mittels kultivierter Mikroorganismen, Labor-Biofilme und extrahierter organischer Substanzen eine Fällung induziert. Die aus den Fallbeispielen abgeleiteten Steuerungsmechanismen der Mineralisation werden unter dem Mikroskop u.a. mit Ionen- und pH-sensitive Fluorochromen zur qualitativen Messung von chemischen Mikrogradienten und durch elektronenoptische Charakterisierung der Fällungsprodukte verifiziert. Ein Schwerpunkt liegt dabei auf der Produktion und dem Abbau Ca2+-adsorbierender extrazellulärer polymerer Substanzen (EPS), die in Organo- und Biofilmen bezüglich Nukleation, Fällung und Gefügebildung von entscheidender Bedeutung sind und Voraussetzungen für eine enzymatisch gesteuerte Biomineralisation darstellen.

Particulate organic carbon measurements during 24-days of incubations in mesocosm experiments with brown algae

Six mesocosm experiments with specimens of Fucales or Laminariales were conducted across six georegions (3 mesocosms with brown algae, 3 mesocosms without brown algae). Incubations lasted 24 days, followed by a year-long monitoring of incubation water. During the first 12 days, brown algae were maintained in mesocosms adjacent to control mesocosms, with 1 L of water sampled every second day. Half of the mesocosm water was replaced with fresh seawater after each sampling. Environmental conditions and primary productivity of specimens was recorded during the incubation. After 12 days, specimens were removed and incubation continued for another 12 days, maintaing the same sampling routine. At the end of the 24 day- incubation period, long-term monitoring was set-up with 6-10L of incubation water in two different conditions: one exposed to a controlled light cycle at 20°C, the second set in darkness at 4°C with added nutrients (40 µM NO3- and 3µM PO43-). Additional water samples were collected along transects extending from near-shore brown algae poplulations. Water samples were filtered over pre-combusted GFF filters (450°C, 4.5h), and both the filtrate and filters were analysed for dissolved organic carbon (DOC), particulate organic carbon (POC). Fucoidan was quantified in dissolved (>1kDa) fraction and surface active fraction (SAF) (> 1kDa and negative charged fraction purified with anion exchange chromatography) fractions through monosaccharide quantification after acid-hydrolysis (100°C, 24h) using HPAEC-PAD, according to Engel and Händel, 2011. Intact polysaccharides were detected using structure-sensitive monoclonal antibodies (Torode et al., 2015; Vidal-Melgosa et al., 2021). Microbial cells were quantified using DAPI-cell staining and counting. Semi-quantitative measurements of particulate fucoidan were performed via acid hydrolysis of GFF filter pieces, followed by monosaccharide analysis via HPAEC-PAD. Sedimented particles to bottom of mesocosms were scooped out on day 24 for monosaccharide analysis and BAM1 antibody binding specific to fucoidan.

Total carbohydrates quantified in sediment cores from coastal vegetated ecosystems

50-cm deep sediment cores were taken in saltmarsh, seagrass, mangroves and unvegetated areas around the German Bight, Malaysia and Columbia in 2022 and 2023. Up to 3 points per ecosystem were sampled along a transect, in total 93 cores were analysed. Carbohydrates were sequentially extracted using MilliQ-water and 0.3 M EDTA for later analyses. The total carbohydrate content was assessed using the phenol-sulfuric acid assay (Dubois et al., 1956). Briefly, 100 µL of resuspended samples or extracts were mixed with 100 µL of 5% phenol solution, followed by the addition of 500 µL of concentrated sulfuric acid. The reaction mixture was incubated at room temperature for 10 minutes, then further incubated at 30°C for 20 minutes. Absorbance at 490 nm was measured using a Spectramax Id3 plate reader (Molecular Devices) and quantified against a glucose standard curve.

Polysaccharides quantified in sediment cores from coastal vegetated ecosystems

50-cm deep sediment cores were taken in saltmarsh, seagrass, mangroves and unvegetated areas around the German Bight, Malaysia and Columbia in 2022 and 2023. Up to 3 points per ecosystem were sampled along a transect, in total 93 cores were analysed. Carbohydrates were sequentially extracted using MilliQ-water and 0.3 M EDTA for later analyses. Polysaccharides were screened using microarray analysis following the method described by Vidal-Melgosa et al. (2022). Briefly, sediment extracts from MilliQ-water and EDTA were combined in equal volumes, and 30 µL of the mixture was transferred into wells of 384-microwell plates. Two consecutive two-fold dilutions were performed using a printing buffer (55.2% glycerol, 44% water, 0.8% Triton X-100). The plates were then centrifuged at 3,500 × g for 10 minutes at 15 °C. Each microarray was individually probed with a monoclonal antibody (mAb), and binding was detected using a secondary antibody conjugated to alkaline phosphatase. In the presence of its substrate, this reaction produced a colorimetric signal. Developed arrays were scanned at 2400 dots per inch, and binding signal intensity was quantified using Array-Pro Analyzer 6.3 software (Media Cybernetics).

Monosaccharides quantified in sediment cores from coastal vegetated ecosystems

50-cm deep sediment cores were taken in saltmarsh, seagrass, mangroves and unvegetated areas around the German Bight, Malaysia and Columbia in 2022 and 2023. Up to 3 points per ecosystem were sampled along a transect, in total 93 cores were analysed. Carbohydrates were sequentially extracted using MilliQ-water and 0.3 M EDTA for later analyses. Extracts were acid hydrolysed (1 M HCl, 24 h, 100°C) and monosaccharides were analysed using anion exchange chromatography with pulsed amperometric detection (HPAEC-PAD), according to Engel et al., 2011. Briefly, sample analysis was performed using a Dionex ICS-5000+ system with a CarboPac PA10 analytical column (2 × 250 mm) and a CarboPac PA10 guard column (2 × 50 mm). Neutral and amino sugars were separated under isocratic conditions with 18 mM NaOH, while acidic monosaccharides were separated using a gradient up to 200 mM NaCH₃COO.

Polysaccharide fucoidan BAM1 and arabinogalactan-protein glycan JIM13 antibody binding in sediment cores from coastal vegetated ecosystems

50-cm deep sediment cores were taken in saltmarsh, seagrass, mangroves and unvegetated areas around the German Bight, Malaysia and Columbia in 2022 and 2023. Up to 3 points per ecosystem were sampled along a transect, in total 93 cores were analysed. Carbohydrates were sequentially extracted using MilliQ-water and 0.3 M EDTA for later analyses. For more specific analysis enzyme-linked immunosorbent assay (ELISA) was used for detection of fucoidan and arabinogalactan-protein glycan as described in the following studies (Vidal-Melgosa et al., 2021 and Cornuault et al., 2014). In short, 100 µL of sediment extracts were added to a pre-coated 96-well plate and incubated overnight at 4°C. The signal was developed using primary antibodies, BAM1 (fucoidan) and JIM13 (arabinogalactan-protein glycan), diluted 1:10 in skim milk PBS solution, followed by anti-rat antibody at a 1:1000 dilution in the same solution. Absorbance was measured at 450 nm using a Spectramax Id3 plate reader (Molecular Devices).

Chlorophyll a, extracellular polymeric substance concentration and 16S rRNA gene copy numbers in saltmarsh sediments in response to a storm surge

Samples were taken to study the effect of storm surges on ecosystem functioning of salt marsh microbial communities. Sediment samples were collected from experimental salt marsh islands located in the back-barrier tidal flats of Spiekeroog Island, German North Sea (53°45′N, 7°43′E). The islands consist of three elevation zones (0.7 m, 1.0 m, and 1.3 m above mean sea level), corresponding to pioneer zone, lower salt marsh, and upper salt marsh. Six islands were sampled (three initially bare; three transplanted with lower salt marsh sediment and vegetation). Sampling was conducted in September 2022 (pre-disturbance), March 2023 (post-winter storm surges), and August 2023 (recovery phase). Surface sediments (upper 2 cm) were collected using syringe cores. Pooled samples were analyzed for chlorophyll a as a proxy for microphytobenthos biomass using ethanol extraction and spectrophotometric pigment analysis. Extracellular polymeric substances (EPS) were quantified using EDTA extraction followed by phenol–sulfuric acid carbohydrate analysis. DNA was extracted from sediment subsamples using a Qiagen PowerSoil kit. Prokaryotic abundance was estimated by quantitative PCR targeting the 16S rRNA gene (primers 519F/907R), using an Escherichia coli 16S rRNA gene standard curve. The dataset includes chlorophyll a concentrations (µg g⁻¹ dry sediment), EPS carbohydrate concentrations, and prokaryotic 16S rRNA gene copy numbers for all sampling times, elevations, and treatments.

Chlorophyll a, extracellular polymeric substance concentration and 16S rRNA gene copy numbers in sediments in response to a marine heatwave

This dataset originates from a laboratory mesocosm experiment investigating the effects of simulated marine heatwaves (MHWs) on benthic microbial communities from sublittoral sediments of Spiekeroog (Wadden Sea, Germany). Surface sediments were sieved (1 mm) and incubated under controlled light and temperature conditions before exposure to three temperature treatments (19 °C control, 23 °C, and 25 °C). Following a gradual warming phase, MHW conditions were maintained for six days, followed by a recovery phase at ambient temperature. Sediment samples were collected repeatedly throughout the experiment. Surface sediments (upper 2 cm) were collected using syringe cores. Pooled samples were analyzed for chlorophyll a as a proxy for microphytobenthos biomass using ethanol extraction and spectrophotometric pigment analysis. Extracellular polymeric substances (EPS) were quantified using EDTA extraction followed by phenol–sulfuric acid carbohydrate analysis. DNA was extracted from sediment subsamples using a Qiagen PowerSoil kit. Prokaryotic abundance was estimated by quantitative PCR targeting the 16S rRNA gene (primers 519F/907R), using an Escherichia coli 16S rRNA gene standard curve. The dataset includes chlorophyll a concentrations (µg g⁻¹ dry sediment), EPS carbohydrate concentrations, and prokaryotic 16S rRNA gene copy numbers for all sampling times, elevations, and treatments.

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