High Magnesium Calcite and Dolomite composition carbonate in Amphiroa (Lithophyllaceae, Corallinales, Rhodophyta): further documentation of elevated Mg in Corallinales with climate change implications.

Species of the calcified, articulate coralline Amphiroa are key components of many shallow marine ecosystems. Understanding their mineral composition is important as their susceptibility to dissolution, due to ocean acidification, may vary with mineral composition. We studied the distribution of Mg-calcite, very high magnesium calcite (VHMC), and dolomite within Amphiroa species to elucidate their mineral properties and susceptibility to dissolution. Results revealed that the asymmetrical X-ray diffraction (XRD) pattern typical of Amphiroa globally represents high levels of VHMC and dolomite composition carbonate. The dolomite seems most likely to be disordered, but higher resolution XRD is required for confirmation. The calcified long sides of medullary cells have predominantly VHMC/dolomite and the corners have bands of VHMC/dolomite. Epithallial cell walls are low Mg-calcite, and cortical cells are low Mg-calcite with bands of VHMC. VHMC/dolomite is more stable than Mg-calcite, and this may provide a competitive advantage for Amphiroa species as seawater pH declines. Further work is required to determine the metabolic controls on VHMC/dolomite mineral formation.

Coralline algal calcification: A morphological and process-based understanding.

RESEARCH PURPOSE AND FINDINGS: Coralline algae are key biological substrates of many carbonate systems globally. Their capacity to build enduring crusts that underpin the formation of tropical reefs, rhodolith beds and other benthic substrate is dependent on the formation of a calcified thallus. However, this important process of skeletal carbonate formation is not well understood. We undertook a study of cellular carbonate features to develop a model for calcification. We describe two types of cell wall calcification; 1) calcified primary cell wall (PCW) in the thin-walled elongate cells such as central medullary cells in articulated corallines and hypothallial cells in crustose coralline algae (CCA), 2) calcified secondary cell wall (SCW) with radial Mg-calcite crystals in thicker-walled rounded cortical cells of articulated corallines and perithallial cells of CCA. The distinctive banding found in many rhodoliths is the regular transition from PCW-only cells to SCW cells. Within the cell walls there can be bands of elevated Mg with Mg content of a few mol% higher than radial Mg-calcite (M-type), ranging up to dolomite composition (D-type). MODEL FOR CALCIFICATION: We propose the following three-step model for calcification. 1) A thin (< 0.5 µm) PCW forms and is filled with a mineralising fluid of organic compounds and seawater. Nanometer-scale Mg-calcite grains precipitate on the organic structures within the PCW. 2) Crystalline cellulose microfibrils (CMF) are extruded perpendicularly from the cellulose synthase complexes (CSC) in the plasmalemma to form the SCW. 3) The CMF soaks in the mineralising fluid as it extrudes and becomes calcified, retaining the perpendicular form, thus building the radial calcite. In Clathromorphum, SCW formation lags PCW creating a zone of weakness resulting in a split in the sub-surface crust. All calcification seems likely to be a bioinduced rather than controlled process. These findings are a substantial step forward in understanding how corallines calcify.

The role of chitin-rich skeletal organic matrix on the crystallization of calcium carbonate in the crustose coralline alga Leptophytum foecundum.

The organic matrix (OM) contained in marine calcifiers has a key role in the regulation of crystal deposition, such as crystalline structure, initiation of mineralization, inhibition, and biological/environmental control. However, the functional properties of the chitin-rich skeletal organic matrix on the biological aspect of crystallization in crustose coralline algae have not yet been investigated. Hence, the characterization of organic matrices in the biomineralization process of this species was studied to understand the functions of these key components for structural formation and mineralization of calcium carbonate crystals. We purified skeletal organic matrix proteins from this species and explored how these components are involved in the mineralization of calcium carbonate crystals and environmental control. Intriguingly, the analytical investigation of the skeletal OM revealed the presence of chitin in the crustose coralline alga Leptophytum foecundum. The sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis of the OM revealed a high molecular mass protein as 300-kDa. Analysis of glycosylation activity exposed two strong glycoproteins as 300-kDa and 240-kDa. Our study of the biominerals of live collected specimens found that in addition to Mg-calcite up to 30% aragonite were present in the skeleton. Our experiment demonstrated that the chitin-rich skeletal OM of coralline algae plays a key role in the biocalcification process by enabling the formation of Mg-calcite. In addition, this OM did not inhibit the formation of aragonite suggesting there is an as yet unidentified process in the living coralline that prevents the formation of aragonite in the living skeletal cell walls.

Presence of skeletal banding in a reef-building tropical crustose coralline alga.

The presence of banding in the skeleton of coralline algae has been reported in many species, primarily from temperate and polar regions. Similar to tree rings, skeletal banding can provide information on growth rate, age, and longevity; as well as records of past environmental conditions and the coralline alga's growth responses to such changes. The aim of this study was to explore the presence and characterise the nature of banding in the tropical coralline alga Porolithon onkodes, an abundant and key reef-building species on the Great Barrier Reef (GBR) Australia, and the Indo-Pacific in general. To achieve this we employed various methods including X-ray diffraction (XRD) to determine seasonal mol% magnesium (Mg), mineralogy mapping to investigate changes in dominant mineral phases, scanning electron microscopy-electron dispersive spectroscopy (SEM-EDS), and micro-computed tomography (micro-CT) scanning to examine changes in cell size and density banding, and UV light to examine reproductive (conceptacle) banding. Seasonal variation in the Mg content of the skeleton did occur and followed previously recorded variations with the highest mol% MgCO3 in summer and lowest in winter, confirming the positive relationship between seawater temperature and mol% MgCO3. Rows of conceptacles viewed under UV light provided easily distinguishable bands that could be used to measure vertical growth rate (1.4 mm year-1) and age of the organism. Micro-CT scanning showed obvious banding patterns in relation to skeletal density, and mineralogical mapping revealed patterns of banding created by changes in Mg content. Thus, we present new evidence for seasonal banding patterns in the tropical coralline alga P. onkodes. This banding in the P. onkodes skeleton can provide valuable information into the present and past life history of this important reef-building species, and is essential to assess and predict the response of these organisms to future climate and environmental changes.

Multiple phases of mg-calcite in crustose coralline algae suggest caution for temperature proxy and ocean acidification assessment: lessons from the ultrastructure and biomineralization in Phymatolithon (Rhodophyta, Corallinales)1.

Magnesium content, strongly correlated with temperature, has been developed as a climate archive for the late Holocene without considering anatomical controls on Mg content. In this paper, we explore the ultrastructure and cellular scale Mg-content variations within four species of North Atlantic crust-forming Phymatolithon. The cell wall has radial grains of Mg-calcite, whereas the interfilament (middle lamella) has grains aligned parallel to the filament axis. The proportion of interfilament and cell wall carbonate varies by tissue and species. Three distinct primary phases of Mg-calcite were identified: interfilament Mg-calcite (mean 8.9 mol% MgCO3 ), perithallial cell walls Mg-calcite (mean 13.4 mol% MgCO3 ), and hypothallium Mg-calcite (mean 17.1 mol% MgCO3 ). Magnesium content for the bulk crust, an average of all phases present, showed a strongly correlated (R2  = 0.975) increase of 0.31 mol% MgCO3 per °C. Of concern for climate reconstructions is the potential for false warming signals from undetected postgrazing wound repair carbonate that is substantially enriched in Mg, unrelated to temperature. Within a single crust, Mg-content of component carbonates ranged from 8 to 20 mol% MgCO3 , representing theoretical thermodynamic stabilities from aragonite-equivalent to unstable higher-Mg-calcite. It is unlikely that existing current predictions of ocean acidification impact on coralline algae, based on saturation states calculated using average Mg contents, provide an environmentally relevant estimate.

Discovery of the mineral brucite (magnesium hydroxide) in the tropical calcifying alga Polystrata dura (Peyssonneliales, Rhodophyta).

Red algae of the family Peyssonneliaceae typically form thin crusts impregnated with aragonite. Here, we report the first discovery of brucite in a thick red algal crust (~1 cm) formed by the peyssonnelioid species Polystrata dura from Papua New Guinea. Cells of P. dura were found to be infilled by the magnesium-rich mineral brucite [Mg(OH)2 ]; minor amounts of magnesite and calcite were also detected. We propose that cell infill may be associated with the development of thick (> ~5 mm) calcified red algal crusts, integral components of tropical biotic reefs. If brucite infill within the P. dura crust enhances resistance to dissolution similarly to crustose coralline algae that infill with dolomite, then these crusts would be more resilient to future ocean acidification than crusts without infill.

Greenhouse conditions induce mineralogical changes and dolomite accumulation in coralline algae on tropical reefs.

Human-induced ocean acidification and warming alter seawater carbonate chemistry reducing the calcification of reef-building crustose coralline algae (CCA), which has implications for reef stability. However, due to the presence of multiple carbonate minerals with different solubilities in seawater, the algal mineralogical responses to changes in carbonate chemistry are poorly understood. Here we demonstrate a 200% increase in dolomite concentration in living CCA under greenhouse conditions of high pCO2 (1,225 µatm) and warming (30 °C). Aragonite, in contrast, increases with lower pCO2 (296 µatm) and low temperature (28 °C). Mineral changes in the surface pigmented skeleton are minor and dolomite and aragonite formation largely occurs in the white crust beneath. Dissolution of high-Mg-calcite and particularly the erosive activities of endolithic algae living inside skeletons play key roles in concentrating dolomite in greenhouse treatments. As oceans acidify and warm in the future, the relative abundance of dolomite in CCA will increase.

A short-term in situ CO2 enrichment experiment on Heron Island (GBR).

Ocean acidification poses multiple challenges for coral reefs on molecular to ecological scales, yet previous experimental studies of the impact of projected CO2 concentrations have mostly been done in aquarium systems with corals removed from their natural ecosystem and placed under artificial light and seawater conditions. The Coral-Proto Free Ocean Carbon Enrichment System (CP-FOCE) uses a network of sensors to monitor conditions within each flume and maintain experimental pH as an offset from environmental pH using feedback control on the injection of low pH seawater. Carbonate chemistry conditions maintained in the -0.06 and -0.22 pH offset treatments were significantly different than environmental conditions. The results from this short-term experiment suggest that the CP-FOCE is an important new experimental system to study in situ impacts of ocean acidification on coral reef ecosystems.