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Species discrimination of plant roots by Fourier transform infrared (FTIR) spectroscopy

Comprehension of belowground competition between plant species is a central part in understanding the complex interactions in intercropped agricultural systems, between crops and weeds as well as in natural ecosystems. So far, no simple and rapid method for species discrimination of roots in the soil exists. We will be developing a method for root discrimination of various species based on Fourier Transform Infrared (FTIR)-Attenuated Total Reflexion (ATR) Spectroscopy and expanding its application to the field. The absorbance patterns of FTIR-ATR spectra represent the chemical sample composition like an individual fingerprint. By means of multivariate methods, spectra will be grouped according to spectral and chemical similarity in order to achieve species discrimination. We will investigate pea and oat roots as well as maize and barnyard grass roots using various cultivars/proveniences grown in the greenhouse. Pea and oat are recommendable species for intercropping to achieve superior grain and protein yields in an environmentally sustainable manner. To evaluate the effects of intercropping on root distribution in the field, root segments will be measured directly at the soil profile wall using a mobile FTIR spectrometer. By extracting the main root compounds (lipids, proteins, carbohydrates) and recording their FTIR-ATR spectra as references, we will elucidate the chemical basis of species-specific differences.

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.

Konsequenzen der Massenvermehrung phytophager Insekten für Ökosystem-Funktionen auf verschiedenen Zeitskalen in Kiefernwäldern Nordost-Deutschlands; Auswirkungen von Massenvermehrungen phytophager Insekten auf biogeochemische Prozesse und mikrobiologische Aktivitäten in Baumkronen und Böden

Insektenkalamitäten können Menge und chemische Zusammensetzung von gelöster und partikulärer organischer Substanz (DOM, POM) innerhalb des Transfers zwischen Baumkronen und Boden verändern. Dies kann mikrobielle Aktivitäten in der Phyllosphäre und im Boden beeinflussen, was zu veränderten C und N Umsätzen führt. Projektziel ist, die C und N Verbindung zwischen Kronenraum und Boden in 60-jährigen Kiefernwäldern (Pinus silvestris L.) unter Insektenbefall zu untersuchen. Um die Hypothese zu testen, dass Massenvermehrung von herbivoren Insekten den C und N Umsatz in Kiefernwäldern steigert, wird (1) der Eintrag quantifiziert: DOM und POM Flüsse vom Kronenraum in den Boden, (2) Mechanismen bewertet: Effekte durch leicht- und schwerabbaubare Verbindungen in DOM und POM (Phenole, Lipide, Kohlenhydrate, Proteine, freie Aminosäuren) auf Kronen- und Bodenmikroorganismen (mikrobielle Biomasse, Enzymaktivitäten), sowie biogeochemische Prozesse (C-Mineralisierung) im Boden und (3) Konsequenzen quantifiziert: Treibhausgasemissionen (THG) und flüchtige organische Verbindungen (VOCs) vom Boden. Veränderte C und N Pfade werden über neu entwickelte Algorithmen modelliert, um langfristige Auswirkungen auf ökosystemarer Ebene abzuschätzen. Damit wird der Kurzschluss zwischen erhöhter DOM und POM Produktion im Kronenraum durch Herbivore einerseits, mit C und N Einträgen im Boden und Umsatzprozesse andererseits analysiert und modelliert.

Wirkungen wasserlöslicher organischer Substanzen auf die Stabilisierung und den Abbau organischer Bodensubstanz

Mikrobielle Umsetzungsprozesse im Boden verlaufen fast ausschließlich unter Beteiligung einer gelösten Phase, da alle lebenden Zellen von einem Wasserfilm umgeben sind, durch den Substrate hindurchdiffundieren müssen, oder über den Exoenzyme und andere Exsudate abgegeben werden. Bei der Mineralisierung organischer Substanzen kommt daher der gelösten organischen Substanz (DOM) als Substrat für Mikroorganismen eine entscheidende Rolle zu. In dem Vorhaben wird der Frage nachgegangen, ob bestimmte streu- und wurzelbürtige DOM-Komponenten wie Kohlenhydrate oder Phenole darüberhinaus die mikrobielle Aktivität in einem Maße fördern oder hemmen können, daß von ihnen Auswirkungen auf den Abbau oder die Stabilisierung der organischen Bodensubstanz auftreten können. Zur Untersuchung solcher 'Priming Effekte' sollen umfangreiche Inkubationsversuche durchgeführt werden, bei denen die Wirkung unterschiedlicher gelöster 14C-markierter Einzelverbindungen und von DOM-Lösungen unterschiedlicher 13C-Signatur auf die Mineralisierung von Modellsubstanzen und der organischen Substanz verschiedener Bodenproben ermittelt wird. Ein daraus berechneter Priming Index gibt Auskunft darüber, inwieweit es durch die zugesetzten DOM-Lösungen zu einem verstärkten oder vermindertem Abbau der organischen Bodensubstanz kommt

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.

Photosynthese bei der Weinrebe

Einbau und Verteilung von Kohlenstoff unter verschiedenen Umweltsbedingungen werden am Beispiel der Weinrebe untersucht. Besonderes Augenmerk wird dabei auf die Bildung bzw. Remobilisierung von Transportkohlenhydraten nach Befall durch pilzliche Parasiten (Mehltau) oder nach Schaedigung des Photosyntheseapparates (z.B. Hagelschlag, Toxineinwirkung) gerichtet.

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.

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.

Microbial cell abundance quantified via DAPI-cell counting 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.

Physiological data of Pacific oyster Crassostrea (Magallana) gigas after exposure to intermittent hypoxia and the combination with (fluctuating) elevated temperature

Organisms in intertidal zones experience fluctuations in environmental stressors such as hypoxia and temperature. These stressors and their fluctuations often appear in combination. Combination of stressors can have different effects compared to single stressors. In this study, we investigate the physiological effects of intermittent hypoxia in combination with different temperature regimes on the Pacific Oyster Crassostrea (Magallana) gigas. The oysters were exposed to hypoxic cycles (12h hypoxia by emersion/12h submersion) at normal (15°C), elevated (30°C) or fluctuating (15°C submersion/30°C emersion) temperature for 10 days. After the last submersion phase, the gills and digestive gland were sampled. We measured markers for bioenergetics and redox-balance in the gills and digestive gland using colorimetric methods as well as a set of metabolites (predominantly amino acids, osmolytes, anaerobic end products and energetic metabolites) in the gills using LC-MS/MS. Oysters kept submerged for up to 10 days were used as controls.

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