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Schwerpunktprogramm (SPP) 1158: Antarctic Research with Comparable Investigations in Arctic Sea Ice Areas; Bereich Infrastruktur - Antarktisforschung mit vergleichenden Untersuchungen in arktischen Eisgebieten, Kombinierte Effekte von Temperatur und Ressourcenverfügbarkeit auf den Abbau von organischem Material durch Antarktisches Bakterioplankton

Die globale Erwärmung führt zu neuen Bedrohungen in den Ozeanen, da die steigende Temperatur Kaskadeneffekte in biogeochemischen Kreisläufen und Nahrungsnetzen auslösen kann. Das Scientific Committee on Antarctic Research (SCAR) hat ein besseres Verständnis von potentiellen Effekten des Klimawandels auf die physikalische und biologische CO2-Aufnahme des Südozeans als dringende Fragestellung der Antarktisforschung identifiziert. Bakterien sind die Hauptproduzenten von CO2 und wirken so der biologischen Zehrung von CO2 durch die Primärproduktion entgegen. In Antarktischen marinen Systemen sind die niedrige Temperatur und die geringe Verfügbarkeit von labilem organischem Material Hauptfaktoren, die Wachstum und Aktivität von Bakterien begrenzen. Temperatur und Ressourcen-Verfügbarkeit für Bakterien werden sich durch den Klimawandel in Verbindung mit Eisschmelze und Folgen für die Primärproduktion jedoch erheblich verändern. Laborexperimente mit Batch-Kulturen haben gezeigt, dass Temperatureffekte auf bakterielles Wachstum nahe der minimalen Wachstumstemperatur ein hohes Potential haben mit der Konzentration von organischen Substraten zu interagieren. Durch diese Interaktionen waren Temperatureffekte auf bakterielles Wachstum überproportional stark, wenn Substrate verfügbar wurden. Die Relevanz dieses synergistischen Effektes für die bakterielle Produktion und die damit verbundene Freisetzung von CO2 in natürlichen Gemeinschaften ist jedoch unklar. Dieses Projekt beabsichtigt einzelne und kombinierte Effekte von Temperatur und Verfügbarkeit organischen Materials auf Antarktisches Bakterioplankton zu testen. Zu diesem Zweck werden Aktivierungsenergien von extrazellulären Enzymen, Substrataufnahme und Produktion in bakteriellen Gemeinschaften des Weddell Meeres bestimmt. Die Ratenmessungen im Weddell Meer werden mit der Analyse von organischem Material kombiniert, um Temperatureffekte auf Flüsse von labilem und semilabilem organischen Kohlenstoff abzuschätzen. Mit Hilfe der Ergebnisse werden der natürliche Bereich der Temperatursensitivität und die Modulation durch abiotische und biotische Faktoren bestimmt. In Experimenten an Bord werden kombinierte Effekte von Erwärmung und Substratzugabe auf die Zusammensetzung der bakteriellen Gemeinschaft, auf Muster der Genexpression und auf den Umsatz des organischen Materials getestet, um Veränderungen in der Gemeinschaft in Bezug zu Veränderungen ihrer Funktionen zu setzen. Es werden zudem Chemostat-Experimente mit Isolaten aus dem Südozean durchgeführt, um Temperatureffekte auf Wachstumseffizienzen und die chemische Zusammensetzung bakterieller Biomasse zu quantifizieren. Eine bessere Bestimmung von bakterieller Remineralisierung und ihrer Abhängigkeit von Temperatur und Substratkonzentration ist notwendig um biogeochemische Modelle besser zu parametrisieren, die den zukünftigen marinen Kohlenstoffkreislauf und den Austausch von CO2 zwischen Ozean und Atmosphäre in einem sich veränderndem Klima projizieren.

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.

Particulate fucoidan concentrations 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.

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).

Specific antibody analysis 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.

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.

MEPHYSTO: Combining population dynamics and drought related ecophysiology in the regional forest model TreeMig

The project is part of the COST action FP0603 Forest models for research and decision support in sustainable forest management (http://www.cost.esf.org/index.php?id=143&action number=FP0603) which aims at extending the scope of forest models from growth only to population dynamics and ecophysiology. Rationale: For sustainable forest management over large areas and for simulating different forest functions especially under changing conditions, different aspects of the system forest' must be modelled jointly: ecophysiological/biogeochemical processes, population dynamics, spatial interactions, and horizontal/vertical species stand structure. We develop a forest model with a stand-size grain suitable to be applied on large areas for assessment of, e.g., climate change or management effects on forest functions. This is achieved by merging and if necessary up- and down-scaling model functions of ecophysiological and population dynamical processes contained in existing models (single tree physiology, local scale ecophysiological, empirical forest growth, spatio-temporal forest landscape, and dynamic global vegetation models). Drought is predicted to occur more frequently with climate change, thus the main focus is on drought and the mechanisms how it affects the trees. Research questions: What are the mechanisms by which drought affects trees? Which is the best (sufficiently accurate and efficient) way to model and simulate these mechanisms? How can population dynamics and ecophysiology be combined in a landscape scale model concerning - allocation of water and carbohydrates to trees and organs? - spatial heterogeneity of soil water and trees? Methods: The project builds on the climate-driven forest landscape model TreeMig (Lischke et al., 2006). Process descriptions from various existing models are compiled, evaluated and included into TreeMig. This involves a thorough scaling of process formulations. Drought effects, involving soil water balance, stomata regulation, photosynthesis, CO2 fertilization effects, allocation of carbohydrates, dynamics of reserve pools and the relationship between these and regeneration, growth and mortality are studied in literature and other models and included into MEPHYSTO.

Dissolved 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.

Incubation set-up during 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.

BAM1 antibody (fucoidan) analysis 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.

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