Chemical characterization and quantification of the brown algal storage compound laminarin

In search of a meaningful stress indicator for Fucus vesiculosus we found that the often used quantitative determination procedures for the polysaccharide laminarin (beta-1,3-glucan) result in different kind of problems, uncertainties and limitations. This chemical long-term storage form of carbon enables perennial brown algae in seasonally fluctuating ecosystems to uncouple growth from photosynthesis. Because of this high ecological relevance a reliable and precise method for determination and quantification of laminarin is needed. Therefore, a simple, cold water extraction method coupled to a new quantitative liquid chromatography-mass spectrometrical method (LC-MS) was developed. Laminarin was determined in nine out of twelve brown algal species, and its expected typical molar mass distribution of 2000-7000 Da was confirmed. Furthermore, laminarin consisted of a complex mixture of different chemical forms, since fifteen chemical laminarin species with distinct molecular weights were measured in nine species of brown algae. Laminarin concentrations in the algal tissues ranged from 0.03 to 0.86% dry weight (DW). The direct chemical characterization and quantification of laminarin by LC-MS represents a powerful method to verify the biochemical and ecological importance of laminarin for brown algae. Single individuals of Laminaria hyperborea, L. digitata, Saccharina latissima, F. serratus, F. vesiculosus, F. spiralis, Himanthalia elongata, Cystoseira tamariscifolia, Pelvetia canaliculata, Ascophyllum nodosum, Halidrys siliquosa and Dictyota dichotoma were collected in fall (18.11.2013) during spring low tide from the shore of Finavarra, Co. Clare, west coast of Ireland (53° 09' 25'' N, 09° 06' 58'' W). After sampling, the different algae were immediately transported to the lab, lyophilized and sent to the University of Rostock. Laminarin was extracted with cold ultrapure water from the algal samples. Before extraction they were ground to < 1 mm grain size with an analytical mill (Ika MF 10 Basic). The algal material (approx. 1.5 g DW) was extracted in ultrapure water (8 mL) on a shaker (250 rpm) for 5 h. After the addition of surplus ultrapure water (4 mL) and shaking manually, 1 mL of the sample was filter centrifuged (45 µm) at 14,000 rpm (Hettich Mikro 22 R). The slightly viscous supernatant was free of suspended material and converted into a microvial (300 µL) for further analysis. The extracts were analyzed using liquid chromatography-mass spectrometry (LC-MS) analysis (LTQ Velos Pro ion trap spectrometer with Accela HPLC, Thermo Scientific). Laminarin species were separated on a KinetexTM column (2.6 µm C18, 150 x 3 mm). The mobile phase was 90 % ultrapure water and 10 % acetonitrile, run isocratically at a flow rate of 0.2 mL min-1. MS was working in ESI negative ion mode in a mass range of 100 - 4000 amu. Glucose contents were determined after extraction using high-performance liquid chromatography (HPLC). Extracted samples were analyzed in an HPLC (SmartLine, Knauer GmbH) equipped with a SUPELCOGELTM Ca column (30 x 7,8 mm without preColumn) and RI-detector (S2300 PDA S2800). Water was used as eluent at a flow rate of 0.8 mL min-1 at 75 °C. Glucose was quantified by comparison of the retention time and peak area with standard solutions using ChromGate software. Mannitol was extracted from three subsamples of 10-20 mg powdered alga material (L. hyperborea, L. digitata, S. latissima, F. serratus, F. vesiculosus, F. spiralis, H. elongata, P. canaliculata, A. nodosum, H. siliquosa) and quantified, following the HPLC method described by Karsten et al. (1991). For analyzing carbon and nitrogen contents, dried algal material was ground to powder and three subsamples of 2 mg from each alga thalli were loaded and packed into tin cartridges (6×6×12 mm). The packages were combusted at 950 °C and the absolute contents of C and N were automatically quantified in an elemental analyzer (Elementar Vario EL III, Germany) using acetanilide as standard according to Verardo et al. (1990).

Carbonate content and d13C record of a Jordan Cretaceous outcrop (Section GM3 (Ghawr Al Mazar (Ghor al Mazrar))

An evaluation of the global synchronicity and duration of "3rd-order" sea-level fluctuations during the Cretaceous greenhouse has been hampered by poor constraints on potential climatic and tectonic drivers, and limitations of geochronology and chronostratigraphic correlation. To provide insight into the nature of such sea-level fluctuations, here we present a new Late Cretaceous record from the Jordanian Levant Platform, comprising a detailed physical-, bio-, chemo- and sequence stratigraphy. Carbonate content of these strata reflects overall sequence stratigraphic development, and demonstrates a dramatic 3rd-order-scale cycle that is also apparent in the d°C record. Updated radioisotopic constraints and astrochronologic testing provide support for the inference of an ~1 million year long sea-level oscillation associated with this 3rd-order cycle, which likely reflects a long-period obliquity (1.2 Myr) control on eustasy and stratigraphic sequence development, linked to the global carbon cycle. The observation of cyclic sea-level fluctuations on this time scale suggests sustained global modulation of continental fresh-water-storage. The hypothesized link between astronomical forcing and sea-level forms a baseline approach in the global correlation of sequence boundaries.

Radionuclide analyses, and sedimentation and accumulation rates of sediment core PS2319-1

There is increasing evidence indicating that syndepositional redistribution of sediment on the seafloor by bottom currents is common and can significantly affect sediment mass accumulation rates. Notwithstanding its common incidence, this process (generally referred to as sediment focusing) is often difficult to recognize. If redistribution is near synchronous to deposition, the stratigraphy of the sediment is not disturbed and sediment focusing can easily be overlooked. Ignoring it, however, can lead to serious misinterpretations of sedimentary fluxes, particularly when past changes in export flux from the overlying water are inferred. In many instances, this problem can be resolved, at least for sediments deposited during the late Quaternary, by normalizing to the flux of 230Th scavenged from seawater, which is nearly constant and equivalent to the known rate of production of 230Th from the decay of dissolved 234U. We review the principle, advantages and limitations of this method. Notwithstanding its limitations, it is clear that 230Th normalization does provide a means of achieving more accurate interpretations of sedimentary fluxes and eliminates the risk of serious misinterpretations of sediment mass accumulation rates.