Gene expression of Pocillopora damicornis coral larvae in response to acidification and ocean warming.

OBJECTIVES: The endosymbiosis with Symbiodiniaceae is key to the ecological success of reef-building corals. However, climate change is threatening to destabilize this symbiosis on a global scale. Most studies looking into the response of corals to heat stress and ocean acidification focus on coral colonies. As such, our knowledge of symbiotic interactions and stress response in other stages of the coral lifecycle remains limited. Establishing transcriptomic resources for coral larvae under stress can thus provide a foundation for understanding the genomic basis of symbiosis, and its susceptibility to climate change. Here, we present a gene expression dataset generated from larvae of the coral Pocillopora damicornis in response to exposure to acidification and elevated temperature conditions below the bleaching threshold of the symbiosis. DATA DESCRIPTION: This dataset is comprised of 16 samples (30 larvae per sample) collected from four treatments (Control, High pCO2, High Temperature, and Combined pCO2 and Temperature treatments). Freshly collected larvae were exposed to treatment conditions for five days, providing valuable insights into gene expression in this vulnerable stage of the lifecycle. In combination with previously published datasets, this transcriptomic resource will facilitate the in-depth investigation of the effects of ocean acidification and elevated temperature on coral larvae and its implication for symbiosis.

New branched Porolithon species (Corallinales, Rhodophyta) from the Great Barrier Reef, Coral Sea, and Lord Howe Island.

Porolithon is one of the most ecologically important genera of tropical and subtropical crustose (non-geniculate) coralline algae growing abundantly along the shallow margins of coral reefs and functioning to cement reef frameworks. Thalli of branched, fruticose Porolithon specimens from the Indo-Pacific Ocean traditionally have been called P. gardineri, while massive, columnar forms have been called P. craspedium. Sequence comparisons of the rbcL gene both from type specimens of P. gardineri and P. craspedium and from field-collected specimens demonstrate that neither species is present in east Australia and instead resolve into four unique genetic lineages. Porolithon howensis sp. nov. forms columnar protuberances and loosely attached margins and occurs predominantly at Lord Howe Island; P. lobulatum sp. nov. has fruticose to clavate forms and free margins that are lobed and occurs in the Coral Sea and on the Great Barrier Reef (GBR); P. parvulum sp. nov. has short (<2 cm), unbranched protuberances and attached margins and is restricted to the central and southern GBR; and P. pinnaculum sp. nov. has a mountain-like, columnar morphology and occurs on oceanic Coral Sea reefs. A rbcL gene sequence of the isotype of P. castellum demonstrates it is a different species from other columnar species. In addition to the diagnostic rbcL and psbA marker sequences, the four new species may be distinguished by a combination of features including thallus growth form, margin shape (attached or unattached), and medullary system (coaxial or plumose). Porolithon species, because of their ecological importance and sensitivity to ocean acidification, need urgent documentation of their taxonomic diversity.

Seasonal upwelling conditions promote growth and calcification in reef-building coralline algae.

Crustose coralline algae (CCA) are important components of reef ecology contributing to reef framework construction. However, little is known about how seasonal upwelling systems influence growth and calcification of tropical CCA. We assessed marginal and vertical growth and net calcification rates of two dominant but morphologically different reef-building CCA, Porolithon antillarum and Lithophyllum cf. kaiseri, in a shallow coral reef of the Colombian Caribbean during upwelling and non-upwelling seasons. Growth and calcification rates varied seasonally with higher values during the upwelling compared to the non-upwelling (rainy) season. Annual vertical growth showed rates of 4.48 ± 1.58 and 4.31 ± 2.17 mm · y-1 , net calcification using crust growth estimates of 0.75 ± 0.30 g and 0.68 ± 0.60 g CaCO3 · cm-2 · y-1 and net calcification using the buoyant weight method of 1.49 ± 0.57 and 0.52 ± 0.11 g CaCO3 · cm-2 · y-1 in P. antillarum and L. kaiseri, respectively. Seawater temperature was inversely related with growth and calcification; however, complex oceanographic interactions between temperature and resource availability (e.g., light, nutrients, and CO2 ) are proposed to modulate CCA vital rates. Although CCA calcification rates are comparable to hard corals, CCA vertical accretion is much lower, suggesting that the main contribution of CCA to reef construction is via cementation processes. These results provide baseline data on CCA in the region and generate useful information for monitoring the impacts of environmental changes on tropical upwelling environments.

Iron limitation of kelp growth may prevent ocean afforestation.

Carbon dioxide removal (CDR) and emissions reduction are essential to alleviate climate change. Ocean macroalgal afforestation (OMA) is a CDR method already undergoing field trials where nearshore kelps, on rafts, are purposefully grown offshore at scale. Dissolved iron (dFe) supply often limits oceanic phytoplankton growth, however this potentially rate-limiting factor is being overlooked in OMA discussions. Here, we determine the limiting dFe concentrations for growth and key physiological functions of a representative kelp species, Macrocystis pyrifera, considered as a promising candidate for OMA. dFe additions to oceanic seawater ranging 0.01-20.2 nM Fe' ‒ Fe' being the sum of dissolved inorganic Fe(III) species ‒ result in impaired physiological functions and kelp mortality. Kelp growth cannot be sustained at oceanic dFe concentrations, which are 1000-fold lower than required by M. pyrifera. OMA may require additional perturbation of offshore waters via dFe fertilisation.

Strong seasonal patterns of DOC release by a temperate seaweed community: Implications for the coastal ocean carbon cycle.

Release of dissolved organic carbon (DOC) by seaweed underpins the microbial food web and is crucial for the coastal ocean carbon cycle. However, we know relatively little of seasonal DOC release patterns in temperate regions of the southern hemisphere. Strong seasonal changes in inorganic nitrogen availability, irradiance, and temperature regulate the growth of seaweeds on temperate reefs and influence DOC release. We seasonally surveyed and sampled seaweed at Coal Point, Tasmania, over 1 year. Dominant species with or without carbon dioxide (CO2 ) concentrating mechanisms (CCMs) were collected for laboratory experiments to determine seasonal rates of DOC release. During spring and summer, substantial DOC release (10.06-33.54 µmol C · g DW-1 · h-1 ) was observed for all species, between 3 and 27 times greater than during autumn and winter. Our results suggest that inorganic carbon (Ci ) uptake strategy does not regulate DOC release. Seasonal patterns of DOC release were likely a result of photosynthetic overflow during periods of high gross photosynthesis indicated by variations in tissue C:N ratios. For each season, we calculated a reef-scale net DOC release for seaweed at Coal Point of 7.84-12.9 g C · m-2 · d-1 in spring and summer, which was ~16 times greater than in autumn and winter (0.2-1.0 g C · m-2 · d-1 ). Phyllospora comosa, which dominated the biomass, contributed the most DOC to the coastal ocean, up to ~14 times more than Ecklonia radiata and the understory assemblage combined. Reef-scale DOC release was driven by seasonal changes in seaweed physiology rather than seaweed biomass.

Settlement cue selectivity by larvae of the destructive crown-of-thorns starfish.

Population irruptions of crown-of-thorns starfish (COTS) cause extensive degradation of coral reefs, threatening the structure and function of these important ecosystems. For population irruptions to initiate and spread, large numbers of planktonic larvae have to successfully transition into their benthic life-history stage (i.e. settlement), whereby larval behaviour and the presence of settlement cues may shape spatial patterns of recruitment and adult densities. Our results demonstrate that a wide range of coralline algae species induce COTS larvae to settle; however, the capacity to promote settlement success varied manyfold among algal species, ranging from greater than 90% in Melyvonnea cf. madagascariensis to less than 2% in Lithophyllum cf. kotschyanum and two Porolithon species at 24 h. Because many coralline algae species that promote high settlement success are prevalent in shallow reef habitats, our findings challenge the hypothesis that COTS larvae predominantly settle in deep water. Considering both larval behaviour and algal ecology, this study highlights the ecological significance of coralline algae communities in driving recruitment patterns of COTS. More specifically, the local abundance of highly inductive coralline algae (especially, Melyvonnea cf. madagascariensis) may explain some of the marked spatial heterogeneity of COTS populations and the incidence of population irruptions.

Cell wall organic matrix composition and biomineralization across reef-building coralline algae under global change.

Crustose coralline algae (CCA) are one of the most important benthic substrate consolidators on coral reefs through their ability to deposit calcium carbonate on an organic matrix in their cell walls. Discrete polysaccharides have been recognized for their role in biomineralization, yet little is known about the carbohydrate composition of organic matrices across CCA taxa and whether they have the capacity to modulate their organic matrix constituents amidst environmental change, particularly the threats of ocean acidification (OA) and warming. We simulated elevated pCO2 and temperature (IPCC RCP 8.5) and subjected four mid-shelf Great Barrier Reef species of CCA to 2 months of experimentation. To assess the variability in surficial monosaccharide composition and biomineralization across species and treatments, we determined the monosaccharide composition of the polysaccharides present in the cell walls of surficial algal tissue and quantified calcification. Our results revealed dissimilarity among species' monosaccharide constituents, which suggests that organic matrices are composed of different polysaccharides across CCA taxa. We also observed that species differentially modulate composition in response to ocean acidification and warming. Our findings suggest that both variability in composition and ability to modulate monosaccharide abundance may play a crucial role in surficial biomineralization dynamics under the stress of OA and global warming.

Transcriptomic stability or lability explains sensitivity to climate stressors in coralline algae.

BACKGROUND: Crustose coralline algae (CCA) are calcifying red macroalgae that play important ecological roles including stabilisation of reef frameworks and provision of settlement cues for a range of marine invertebrates. Previous research into the responses of CCA to ocean warming (OW) and ocean acidification (OA) have found magnitude of effect to be species-specific. Response to OW and OA could be linked to divergent underlying molecular processes across species. RESULTS: Here we show Sporolithon durum, a species that exhibits low sensitivity to climate stressors, had little change in metabolic performance and did not significantly alter the expression of any genes when exposed to temperature and pH perturbations. In contrast, Porolithon onkodes, a major coral reef builder, reduced photosynthetic rates and had a labile transcriptomic response with over 400 significantly differentially expressed genes, with differential regulation of genes relating to physiological processes such as carbon acquisition and metabolism. The differential gene expression detected in P. onkodes implicates possible key metabolic pathways, including the pentose phosphate pathway, in the stress response of this species. CONCLUSIONS: We suggest S. durum is more resistant to OW and OA than P. onkodes, which demonstrated a high sensitivity to climate stressors and may have limited ability for acclimatisation. Understanding changes in gene expression in relation to physiological processes of CCA could help us understand and predict how different species will respond to, and persist in, future ocean conditions predicted for 2100.

Phymatolithopsis gen. nov. (Hapalidiales, Corallinophycidae, Rhodophyta) based on molecular and morpho-anatomical evidence.

A multigene (psbA, rbcL, 18S rDNA) molecular phylogeny of the genus Phymatolithon showed a polyphyletic grouping of two monophyletic clades within the Hapalidiales. DNA sequence data integrated with morpho-anatomical comparisons of type material and of recently collected specimens were used to establish Phymatolithopsis gen. nov. with three species, P. prolixa comb. nov., the generitype, P. repanda comb. nov. and P. donghaensis sp. nov. Phymatolithopsis is sister to Mesophyllum and occurs in a clade distinct from Phymatolithon and boreal species currently assigned to Lithothamnion. Morpho-anatomically, Phymatolithopsis is comprised of species that are non-geniculate and encrusting, bear epithallial cells with rounded walls (not flared), subepithallial initials that are usually as short as or shorter than their immediate inward derivatives, conceptacle primordia from all stages forming superficially directly from subepithallial initials, mature carposporangial conceptacles with a discontinuous fusion cell, gonimoblast filaments that develop at the margins of the fusion cell around the periphery of the carposporangial conceptacle chambers, and multiporate tetra/bisporangial conceptacles. Phymatolithopsis can be distinguished from Phymatolithon by the origin of its conceptacle primordia, which are initiated superficially, directly from the layer of subepithallial initials below the epithallial cells and the distribution of gonimoblast filaments in carposporangial conceptacles, that are at the margins of the fusion cells.

Changes in physiological performance and protein expression in the larvae of the coral Pocillopora damicornis and their symbionts in response to elevated temperature and acidification.

Climate change causes ocean warming and acidification, which threaten coral reef ecosystems. Ocean warming and acidification cause bleaching and mortality, and decrease calcification in adult corals, leading to changes in the composition of coral communities; however, their interactive effects on coral larvae are not comprehensively understood. To examine the underlying molecular mechanisms of larval responses to elevated temperature and pCO2, we examined the physiological performance and protein expression profiles of Pocillopora damicornis at two temperatures (29 and 33 °C) and pCO2 levels (500 and 1000 µatm) for 5 d. Extensive physiological and proteomic changes were observed in coral larvae. The results indicated a significant decrease in net photosynthesis (PNET) and autotrophic capability (PNET/RD) of larvae exposed to elevated temperature but a marked increase in PNET and PNET/RD of larvae exposed to high pCO2 levels. Elevated temperature significantly reduced endosymbiont densities by 70% and photochemical efficiency, indicating that warming impaired host-symbiont symbiosis. Expression of photosynthesis-related proteins, the photosystem (PS) I reaction center subunits IV and XI as well as oxygen-evolving enhancer 1, was downregulated at higher temperatures in symbionts, whereas expression of the PS I iron­sulfur center protein was increased under high pCO2 conditions. Furthermore, expression of phosphoribulokinase (involved in the Calvin cycle) and phosphoenolpyruvate carboxylase (related to the C4 pathway) was downregulated in symbionts under thermal stress; this finding suggests reduced carbon fixation at high temperatures. The abundance of carbonic anhydrase-associated proteins, which are predicted to exert biochemical roles in dissolved inorganic carbon transport in larvae, was reduced in coral host and symbionts at high temperatures. These results elucidate potential mechanisms underlying the responses of coral larvae exposed to elevated temperature and acidification and suggest an important role of symbionts in the response to warming and acidification.

Rate and fate of dissolved organic carbon release by seaweeds: A missing link in the coastal ocean carbon cycle.

Dissolved organic carbon (DOC) release by seaweeds (marine macroalgae) is a critical component of the coastal ocean biogeochemical carbon cycle but is an aspect of seaweed carbon physiology that we know relatively little about. Seaweed-derived DOC is found throughout coastal ecosystems and supports multiple food web linkages. Here, we discuss the mechanisms of DOC release by seaweeds and group them into passive (leakage, requires no energy) and active release (exudation, requires energy) with particular focus on the photosynthetic "overflow" hypothesis. The release of DOC from seaweeds was first studied in the 1960s, but subsequent studies use a range of units hindering evaluation: we convert published values to a common unit (µmol C · g DW-1 · h-1 ) allowing comparisons between seaweed phyla, functional groups, biogeographic region, and an assessment of the environmental regulation of DOC production. The range of DOC release rates by seaweeds from each phylum under ambient environmental conditions was 0-266.44 µmol C · g DW-1 · h-1 (Chlorophyta), 0-89.92 µmol C · g DW-1 · h-1 (Ochrophyta), and 0-41.28 µmol C · g DW-1 · h-1 (Rhodophyta). DOC release rates increased under environmental factors such as desiccation, high irradiance, non-optimal temperatures, altered salinity, and elevated dissolved carbon dioxide (CO2 ) concentrations. Importantly, DOC release was highest by seaweeds that were desiccated (<90 times greater DOC release compared to ambient). We discuss the impact of future ocean scenarios (ocean acidification, seawater warming, altered irradiance) on DOC release rates by seaweeds, the role of seaweed-derived DOC in carbon sequestration models, and how they inform future research directions.

Global declines in coral reef calcium carbonate production under ocean acidification and warming.

Ocean warming and acidification threaten the future growth of coral reefs. This is because the calcifying coral reef taxa that construct the calcium carbonate frameworks and cement the reef together are highly sensitive to ocean warming and acidification. However, the global-scale effects of ocean warming and acidification on rates of coral reef net carbonate production remain poorly constrained despite a wealth of studies assessing their effects on the calcification of individual organisms. Here, we present global estimates of projected future changes in coral reef net carbonate production under ocean warming and acidification. We apply a meta-analysis of responses of coral reef taxa calcification and bioerosion rates to predicted changes in coral cover driven by climate change to estimate the net carbonate production rates of 183 reefs worldwide by 2050 and 2100. We forecast mean global reef net carbonate production under representative concentration pathways (RCP) 2.6, 4.5, and 8.5 will decline by 76, 149, and 156%, respectively, by 2100. While 63% of reefs are projected to continue to accrete by 2100 under RCP2.6, 94% will be eroding by 2050 under RCP8.5, and no reefs will continue to accrete at rates matching projected sea level rise under RCP4.5 or 8.5 by 2100. Projected reduced coral cover due to bleaching events predominately drives these declines rather than the direct physiological impacts of ocean warming and acidification on calcification or bioerosion. Presently degraded reefs were also more sensitive in our analysis. These findings highlight the low likelihood that the world's coral reefs will maintain their functional roles without near-term stabilization of atmospheric CO2 emissions.

Evaluating bloom potential of the green-tide forming alga Ulva ohnoi under ocean acidification and warming.

The occurrence of green-tides, whose bloom potential may be increased by various human activities and biogeochemical process, results in enormous economic losses and ecosystem collapse. In this study, we investigated the ecophysiology of the subtropical green-tide forming alga, Ulva ohnoi complex (hereafter: U. ohnoi), under simulated future ocean conditions in order to predict its bloom potential using photosynthesis and growth measurements, and stable isotope analyses. Our mesocosm system included four experimental conditions that simulated the individual and combined effects of elevated CO2 and temperature, namely control (450 µatm CO2 & 20 °C), acidification (900 µatm CO2 & 20 °C), warming (450 µatm CO2 & 25 °C), and greenhouse (900 µatm CO2 & 25 °C). Photosynthetic electron transport rates (rETR) increased significantly under acidification conditions, but net photosynthesis and growth were not affected. In contrast, rETR, net photosynthesis, and growth all decreased significantly under elevated temperature conditions (i.e. both warming and greenhouse). These results represent the imbalance of energy metabolism between electron transport and O2 production that may be expected under ocean acidification conditions. This imbalance appears to be related to carbon and nitrogen assimilation by U. ohnoi. In particular, 13C and 15N discrimination data suggest U. ohnoi prefers CO2 and NH4+ over HCO3- and NO3- as sources of carbon and nitrogen, respectively, and this results in increased N content in the thallus under ocean acidification conditions. Together, our results suggest a trade-off in which the bloom potential of U. ohnoi could increase under ocean acidification due to greater N accumulation and through the saving of energy during carbon and nitrogen metabolism, but that elevated temperatures could decrease U. ohnoi's bloom potential through a decrease in photosynthesis and growth.

Corallinapetrales and Corallinapetraceae: A new order and family of coralline red algae including Corallinapetra gabrielii comb. nov.

The coralline algal genus Corallinapetra is currently monospecific and was established on the species Corallinapetra novaezelandiae, known from a single collection from north-eastern New Zealand. On the basis of multi-gene phylogenetic analyses, Corallinapetra has been resolved apart from all currently recognized families and orders within the Corallinophycidae. We analyzed DNA sequence data from the holotype of Lithothamnion gabrielii, which has been considered a heterotypic synonym of L. muelleri, and an unidentified sample collected from Stewart Island in New Zealand, using psbA, rbcL, and COI-5P genes. We also observed detailed morpho-anatomical characters with light and scanning electron microscopy. Our phylogenetic analyses showed that L. gabrielii and the sample from New Zealand belonged to the same clade as Corallinapetra, distinct from other families and orders in the Corallinophycidae. Members of this clade are distinguishable from other families and orders in the Corallinophycidae by possessing sporangia that are surrounded by remnant sterile filaments that are weakly calcified in mature multiporate sporangial conceptacles that produce zonately divided tetrasporangia. Therefore, we propose that Corallinapetra be placed in its own family, Corallinapetraceae and order, Corallinapetrales, and that L. gabrielii should be assigned to Corallinapetra, as C. gabrielii, to reflect their phylogenetic relationships. We also obtained a partial rbcL sequence data from the lectotype of L. muelleri, the generitype of Lithothamnion. Comparison of the L. muelleri type sequence with L. gabrielii unambiguously demonstrated that these two species are not conspecific, and confirm the placement of L. muelleri within the Hapalidiales.

Inorganic carbon uptake strategies in coralline algae: Plasticity across evolutionary lineages under ocean acidification and warming.

Dissolved inorganic carbon (DIC) assimilation is essential to the reef-building capacity of crustose coralline algae (CCA). Little is known, however, about the DIC uptake strategies and their potential plasticity under ongoing ocean acidification (OA) and warming. The persistence of CCA lineages throughout historical oscillations of pCO2 and temperature suggests that evolutionary history may play a role in selecting for adaptive traits. We evaluated the effects of pCO2 and temperature on the plasticity of DIC uptake strategies and associated energetic consequences in reef-building CCA from different evolutionary lineages. We simulated past, present, moderate (IPCC RCP 6.0) and high pCO2 (RCP 8.5) and present and high (RCP 8.5) temperature conditions and quantified stable carbon isotope fractionation (13ε), organic carbon content, growth and photochemical efficiency. All investigated CCA species possess CO2-concentrating mechanisms (CCMs) and assimilate CO2 via diffusion to varying degrees. Under OA and warming, CCA either increased or maintained CCM capacity, which was associated with overall neutral effects on metabolic performance. More basal taxa, Sporolithales and Hapalidiales, had greater capacity for diffusive CO2 use than Corallinales. We suggest that CCMs are an adaptation that supports a robust carbon physiology and are likely responsible for the endurance of CCA in historically changing oceans.

Global warming offsets the ecophysiological stress of ocean acidification on temperate crustose coralline algae.

Dramatic increases in the release of anthropogenic CO2 and global temperatures have resulted in alterations to seawater carbonate chemistry and metabolisms of marine organisms. There has been recent interest in the effects of these stressors on crustose coralline algae (CCA) because photosynthesis and calcification are influenced by all components of carbonate chemistry. To examine this, a mesocosm experiment was conducted to evaluate photosynthesis, calcification and growth in the temperate CCA Chamberlainium sp. under acidification (doubled CO2), warming (+5 °C), and greenhouse (doubled CO2 and +5 °C) conditions compared to present-day conditions. After 47 days of acclimation to these conditions, productivity was lowest under acidification, although photochemical properties were improved, while respiration was highest under warming. Likewise, growth was lowest under acidification, but this negative response was offset by elevated temperature under greenhouse. Together, these results suggest that warming offsets the negative effects of acidification by creating more suitable conditions for photosynthesis and growth.

Plasticity of adult coralline algae to prolonged increased temperature and pCO2 exposure but reduced survival in their first generation.

Crustose coralline algae (CCA) are vital to coral reefs worldwide, providing structural integrity and inducing the settlement of important invertebrate larvae. CCA are known to be impacted by changes in their environment, both during early development and adulthood. However, long-term studies on either life history stage are lacking in the literature, therefore not allowing time to explore the acclimatory or potential adaptive responses of CCA to future global change scenarios. Here, we exposed a widely distributed, slow growing, species of CCA, Sporolithon cf. durum, to elevated temperature and pCO2 for five months and their first set of offspring (F1) for eleven weeks. Survival, reproductive output, and metabolic rate were measured in adult S. cf. durum, and survival and growth were measured in the F1 generation. Adult S. cf. durum experienced 0% mortality across treatments and reduced their O2 production after five months exposure to global stressors, indicating a possible expression of plasticity. In contrast, the combined stressors of elevated temperature and pCO2 resulted in 50% higher mortality and 61% lower growth on germlings. On the other hand, under the independent elevated pCO2 treatment, germling growth was higher than all other treatments. These results show the robustness and plasticity of S. cf. durum adults, indicating the potential for them to acclimate to increased temperature and pCO2. However, the germlings of this species are highly sensitive to global stressors and this could negatively impact this species in future oceans, and ultimately the structure and stability of coral reefs.

CO2 Enrichment Stimulates Dissolved Organic Carbon Release in Coral Reef Macroalgae.

Dissolved organic carbon (DOC) released by macroalgae is important in the context of coral reef degradation as it contributes to coral mortality by promoting bacterial metabolism on the coral surface. Using experimental carbon dioxide (CO2 ) manipulations in outdoor flow-through tanks, we found that seawater CO2 enrichment enhances daily net DOC release in a range of macroalgal species in the Great Barrier Reef (Australia). There was, however, large variability in DOC release among species, light and dark conditions, and CO2 exposure times. Under light conditions, DOC release in the red macroalga Amansia was 15 times higher under high CO2 conditions compared to ambient CO2 , however, CO2 enhancement did not affect DOC production in the other species. Results from the night incubations were more consistent as three of the four species (Amansia, Lobophora, and Sargassum) enhanced DOC release when enriched with CO2 . DOC fluxes shifted from production in the 1-d incubations to consumption in the 19-d experiment under light conditions, suggesting an important role of bacteria in DOC balances. The results suggest that rising CO2 (and ocean acidification) will continue to intensify space competition in favor of the macroalgae, potentially exacerbating reef degradation and ecological phase shifts from coral to macroalgal dominance.

De novo transcriptome assembly for four species of crustose coralline algae and analysis of unique orthologous genes.

Crustose coralline algae (CCA) are calcifying red macroalgae that reef build in their own right and perform essential ecosystem functions on coral reefs worldwide. Despite their importance, limited genetic information exists for this algal group. De novo transcriptomes were compiled for four species of common tropical CCA using RNA-seq. Sequencing generated between 66 and 87 million raw reads. Transcriptomes were assembled, redundant contigs removed, and remaining contigs were annotated using Trinotate. Protein orthology analysis was conducted between CCA species and two noncalcifying red algae species from NCBI that have published genomes and transcriptomes, and 978 orthologous protein groups were found to be uniquely shared amongst CCA. Functional enrichment analysis of these 'CCA-specific' proteins showed a higher than expected number of sequences from categories relating to regulation of biological and cellular processes, such as actin related proteins, heat shock proteins, and adhesion proteins. Some proteins found within these enriched categories, i.e. actin and GH18, have been implicated in calcification in other taxa, and are thus candidates for involvement in CCA calcification. This study provides the first comprehensive investigation of gene content in these species, offering insights not only into the evolution of coralline algae but also of the Rhodophyta more broadly.

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.