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Algae were maintained in a mesocosm setting for calibration purposes using light and temperature conditions reflecting the natural environment at their site of collection before being frozen at −30°C.Freezing was done to prevent contamination by bacteria or fungi prior to analysis.
Frozen specimens were thawed immediately prior to processing.Coralline algae are among the calcifying organisms that appear to be the most sensitive to ocean acidification formation in the algal skeletons and the mechanism of growth, especially regarding the key components of organic molecules in the biomineralization process.Therefore, we studied the coralline alga Clathromorphum compactum to understand the skeletal structural system and explore the functional molecules in the biomineralization process by which the skeletal minerals form in this species (see Fig. Figure 1: Schematic approach in understanding the dynamic mechanism of calcification, purification of the organic matrix components and the nucleation, growth, and orientation of mineral crystals by the chitin and collagen matrices at different stages of mineralization in the algal skeletons. compactum were live collected in 2008 at 10 m depth near Quirpon, Newfoundland, Canada (51°35.135′ 55°25.490′), and Bay Bulls, Newfoundland, Canada (47°18.496′ 52°47.354′).Thus, it is crucial to obtain a detailed understanding of their calcification process.We here extracted calcified skeletal organic matrix components including soluble and insoluble fractions from the widely-distributed Subarctic and Arctic coralline alga Clathromorphum compactum.For this study only portions of the algal skeleton that had formed prior to field collection were analyzed in order to avoid sampling skeletal material formed during the mesocosm experiment.
To avoid contamination by other associated tissues (e.g., fungus, bacteria etc.) and unwanted substances, the preparation of skeletons from the algal body must be performed carefully.
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Interest in calcifying coralline algae has been increasing over the past years due to the discovery of extensive coralline algal dominated ecosystems in Arctic and Subarctic latitudes, their projected sensitivity to ocean acidification and their utility as palaeoenvironmental proxies.
The lyophilized skeletal organic matrix fractions showed comparatively high concentrations of soluble and insoluble organic matrices comprising 0.9% and 4.5% of skeletal weight, respectively.
This is significantly higher than in other skeletal marine calcifiers.
Attenuated Total Reflection-Fourier Transform Infrared Spectroscopy (ATR-FTIR) and X-Ray Diffraction (XRD) results indicate that chitin is present in the skeletal organic matrices of C. This polymer exhibits similar hierarchical structural organizations with collagen present in the matrix and serves as a template for nucleation and controls the location and orientation of mineral phases.