Other language confidence: 0.7514720156747902
Here, we traced the abundance of 27 polysaccharide epitopes in dissolved and particulate organic matter along a three month diatom bloom period in the North Sea. We used a bioanalytic approach based on carbohydrate microarrays and monoclonal antibodies (mAbs). Details describing the data: Carbohydrate microarray data show the relative polysaccharide abundance (antibody signal intensity) detected in samples harvested during a spring phytoplankton bloom period (21 sampling dates) in the North Sea (54˚11.3'N, 7˚54.0'E). Samples include high molecular weight dissolved organic matter (HMWDOM) and particulate organic matter (POM). Polysaccharides from all samples were sequentially extracted with the solvents H2O, 50 mM EDTA pH 7.5 and 4 M NaOH with 0.1% w/v NaBH4. A library of identical microarrays was created, each populated with the same time-series of extracted polysaccharides. These microarrays were then individually incubated with polysaccharide-specific mAbs or carbohydrate binding modules (CBMs) as probes, which specifically bind to a single polysaccharide epitope. The binding of probes to polysaccharide epitopes on the microarrays was detected using a secondary antibody coupled to alkaline phosphatase, which converts its substrate into a coloured product, the amount of which correlates with polysaccharide concentration. Antibody signal intensity was quantified and the highest signal value in the data set for HMWDOM and for POM was set to 100 and all other values were normalised accordingly. The temporal dynamics but not the absolute number should be compared between HMWDOM and POM pools as they required independent normalisation since their sampling was different. A cut-off of 5 arbitrary units was applied and all probe profiles where in at least one date an antibody positive signal (value ≥ 5) was detected are included in the data set. The epitope recognised by each probe is shown at the top of each column and the name of the corresponding mAb or CBM is depicted in parentheses. Size fractions correspond to: POM 10 µm, over 10 µm; POM 3 µm, between 10 and 3 µm; POM 0.2 µm, between 3 and 0.2 µm; HMWDOM, between 0.2 µm and 1 kDa. HG, homogalacturonan; DE, degree of esterification; AGP, arabinogalactan protein; GlcA, glucuronic acid. This microarray data set reveals the temporal dynamics of 27 polysaccharide epitopes in HMWDOM and POM.
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.
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.
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.
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.
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.
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.
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.
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.
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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