Revisiting the connectivity puzzle of the common coral Pocillopora damicornis

Understanding levels of connectivity among scleractinian coral populations over a range of temporal and spatial scales is vital for managing tropical coral reef ecosystems. Here, we use multilocus microsatellite genotypes to assess the spatial genetic structure of two molecular operational taxonomic units (MOTUs, types α and β) of the widespread coral Pocillopora damicornis on the Great Barrier Reef (GBR) and infer the extent of connectivity on spatial scales spanning from local habitat types to latitudinal sectors of the GBR. We found high genetic similarities over large spatial scales spanning > 1000 km from the northern to the southern GBR, but also strong genetic differentiation at local scales in both MOTUs. The presence of a considerable number of first‐generation migrants within the populations sampled (12% and 27% for types α and β, respectively) suggests that genetic differentiation over small spatial scales is probably a consequence of stochastic recruitment from different genetic pools into recently opened up spaces on the reef, for example, following major disturbance events. We explain high genetic similarity among populations over hundreds of kilometres by long competency periods of brooded zooxanthellate larvae and multiple larval release events each year, combined with strong longshore currents typical along the GBR. The lack of genetic evidence for predominantly clonal reproduction in adult populations of P. damicornis, which broods predominantly asexually produced larvae, further undermines the paradigm that brooded larvae settle close to parent colonies shortly after the release.     

Contact reactions between young colonies of the coral Pocillopora damicornis

 Newly settled larvae (primary polyps) or young colonies of the coral Pocillopora damicornis were brought into contact at various periods after planulation to examine isogeneic and allogeneic responses. While young colonies derived from the same colony always fused, those derived from different colonies showed either fusion, nonfusion, or incompatible fusion. Tissues were continuous in incompatibly fused pairs, but a white zone, without zooxanthellae, was observed at the interface. The skeleton was also continuous but a groove with skeletal spines on both sides was observed under the white zone. Polyps originating near the white zone later disappeared or were partially resorbed. After 2–8 months, several incompatibly fused pairs became separated by a skeletal ridge, or by a narrow zone of skeleton without living tissue. Incompatible fusion appears to be a distinct histoincompatible response which later transforms into nonfusion. The period between planulation and initial contact of colonies did not affect the outcomes of the contact experiments. Accepted: 31 January 1996     

Experimental fragmentation reduces sexual reproductive output by the reef-building coral Pocillopora damicornis

Natural and anthropogenic disturbances may fragment stony reef corals, but few quantitative data exist on the impacts of skeletal fragmentation on sexual reproduction in corals. We experimentally fragmented colonies of the branching coral Pocillopora damicornis and determined the number and size of planula larvae released during one lunar reproductive cycle. Partially fragmented colonies significantly delayed both the onset and peak period of planula release compared with intact control colonies. Most fragments removed from the corals died within 11–18 days, and released few planulae. The total number of planulae released per coral colony varied exponentially with remaining tissue volume, and was significantly lower in damaged versus undamaged colonies. However, the number of planulae produced per unit tissue volume, and planula size, did not vary with damage treatment. We conclude that even partial fragmentation of P. damicornis colonies (<25% of tissue removed) decreases their larval output by reducing reproductive tissue volume. Repeated breakage of corals, such as caused by intensive diving tourism or frequent storms, may lead to substantially reduced sexual reproduction. Therefore, reef management should limit human activities that fracture stony corals and lead to decreases in colony size and reproductive output. Accepted: 2 February 2000     

Mortality and growth of juvenile coral Pocillopora damicornis (Linnaeus)

Early mortality in cohortsof the coral Pocillopora damicornis (Linnaeus) was monitored under experimental conditions on a reef in order to evaluate effects of sedimentation, grazing, predation and competition. Corals that settled in dishes in the laboratory were placed on the reef flat about 3 days after metamorphosis. Six different conditions were tested in each series of experiments: orientation of dishes (upward, vertical and downward) by with and without protection against potential grazers by covering the dish with a net. Survival of juvenile corals on both protected and unprotected dishes facing upward was lower than in the vertical or downward direction. Under the vertical facing and protected conditions, algal growth was more intensive and algae trapped sediment; mortality of juvenile corals by algae and sedimentation increased gradually. In the uprotected and vertical conditions, algal growth on the surface was removed constantly by grazing invertebrates and fishes and the juvenile corals were removed or killed as well. On the downward facing dishes, survivorship of juveniles was relatively high in both protected and unprotected conditions. This habitat attracted many sessile animals that killed some juveniles by competition. Thus, juvenile corals survived better in experimentally manipulated microhabitats not affected by direct sedimentation, not exposed to direct grazing activity, and not occupied by rapidly growing filamentous algae.     

Biomineralization in newly settled recruits of the scleractinian coral Pocillopora damicornis

Calcium carbonate biomineralization of scleractinian coral recruits is fundamental to the construction of reefs and their survival under stress from global and local environmental change. Establishing a baseline for how normal, healthy coral recruits initiate skeletal formation is, therefore, warranted. Here, we present a thorough, multiscale, microscopic and spectroscopic investigation of skeletal elements deposited by Pocillopora damicornis recruits, from 12 h to 22 days after settlement in aquarium on a flat substrate. Six growth stages are defined, primarily based on appearance and morphology of successively deposited skeletal structures, with the following average formation time‐scales: A (<24 h), B (24–36 h), C (36–48 h), D (48–72 h), E (72–96 h), and F (>10 days). Raman and energy dispersive X‐ray spectroscopy indicate the presence of calcite among the earliest components of the basal plate, which consist of micrometer‐sized, rod‐shaped crystals with rhomboidal habit. All later CaCO₃ skeletal structures are composed exclusively of aragonite. High‐resolution scanning electron microscopy reveals that, externally, all CaCO₃ deposits consist of <100 nm granular units. Fusiform, dumbbell‐like, and semispherulitic structures, 25–35 µm in longest dimension, occur only during the earliest stages (Stages A–C), with morphologies similar to structures formed abiotically or induced by organics in in vitro carbonate crystallization experiments. All other skeletal structures of the basal plate are composed of vertically extending lamellar bundles of granules. From Stage D, straight fibrils, 40–45 nm in width and presumably of organic composition, form bridges between these aragonitic bundles emerging from the growing front of fusing skeletal structures. Our results show a clear evolution in the coral polyp biomineralization process as the carbonate structures develop toward those characterizing the adult skeleton. J. Morphol. 275:1349–1365, 2014. © 2014 Wiley Periodicals, Inc.     

Genetic differentiation amongst populations of the coral Pocillopora damicornis off southwestern Australia

An electrophoretic study of genetic differentiation amongst local populations of the reef-coral Pocillopora damicornis was used to group coral heads into units defined as the area of effective dispersal of a clone, and termed colonies. For reefs off southwestern Australia, colonies were usually under a few hundred metres in extent. Although most new recruits within a colony were derived asexually, sexually produced propagules acted to connect populations outside the boundaries of a colony. Such connections were weak, and allelic frequencies varied considerably over a few kilometres. The primary agent of genetic differentiation was suggested to be the small effective population size resulting from the asexual proliferation of a few genotypes at any site. The effective number of genotypes per colony was approximately six. Asexual reproduction appears also to limit gene flow and accentuate selection in this species.     

Ten polymorphic STR loci in the cosmopolitan reef coral, Pocillopora damicornis

We report the development of 10 polymorphic molecular markers containing short tandem repeats in the cosmopolitan reef-building coral, Pocillopora damicornis, an important model species for coral health, physiology, ecology, and genetics. The availability of polymorphic DNA markers in P. damicornis can act as impetus for investigations into inheritance and population genetics, as well as novel investigations into host-symbiont ecology and evolution. Coral bleaching and gene flow studies performed with these markers can have direct conservation implications.     

Temperature-regulated bleaching and lysis of the coral Pocillopora damicornis by the novel pathogen Vibrio coralliilyticus

Coral bleaching is the disruption of symbioses between coral animals and their photosynthetic microalgal endosymbionts (zooxanthellae). It has been suggested that large-scale bleaching episodes are linked to global warming. The data presented here demonstrate that Vibrio coralliilyticus is an etiological agent of bleaching of the coral Pocillopora damicornis. This bacterium was present at high levels in bleached P. damicornis but absent from healthy corals. The bacterium was isolated in pure culture, characterized microbiologically, and shown to cause bleaching when it was inoculated onto healthy corals at 25 degrees C. The pathogen was reisolated from the diseased tissues of the infected corals. The zooxanthella concentration in the bacterium-bleached corals was less than 12% of the zooxanthella concentration in healthy corals. When P. damicornis was infected with V. coralliilyticus at higher temperatures (27 and 29 degrees C), the corals lysed within 2 weeks, indicating that the seawater temperature is a critical environmental parameter in determining the outcome of infection. A large increase in the level of the extracellular protease activity of V. coralliilyticus occurred at the same temperature range (24 to 28 degrees C) as the transition from bleaching to lysis of the corals. We suggest that bleaching of P. damicornis results from an attack on the algae, whereas bacterium-induced lysis and death are promoted by bacterial extracellular proteases. The data presented here support the bacterial hypothesis of coral bleaching.     

Assessing hidden species diversity in the coral Pocillopora damicornis from Eastern Australia

The incredible range of morphological plasticity present in scleractinian corals has confused the taxonomy of the group, prompting the introduction of “ecomorphs” to explain the observed correlation between local environmental conditions and phenotypic variation. Pocillopora damicornis (Linnaeus, 1758) represents one of the best known examples of eco-phenotypic variation in scleractinian corals with a variety of forms and reproductive strategies reported across its global distribution range. Here, we reconstruct genealogical relationships of P. damicornis colonies collected from thirteen locations along the East Australian coast to examine the relationship between genetic and phenotypic diversity in this species. Haplotype networks computed from two mitochondrial DNA regions (CR, ORF) indicate that the range of morphotypes observed within this taxon fall into at least five genetically distinct mitochondrial lineages. Nuclear (HSP70, ITS2) haplowebs on the other hand recover sharp genetic discontinuities among three of the morphological groups. We conclude that P. damicornis from Eastern Australia constitutes a cryptic species complex. The misinterpretation of taxonomical units within P. damicornis may well explain its perceived variation in the ecology, biology and life history across its range.     

Linear extension rate is independent of colony size in the coral Pocillopora damicornis

Measurements of the skeletal extension rate of branches of the reef coral Pocillopora damicornis showed that the linear extension rate is independent of colony size for colonies from 1.9 to 19 cm in diameter. Analysis of existing data from Western Australia, Samoa, the Great Barrier Reef and Hawaii supports the finding that linear extension is not related to colony size in this species.     

Influence of water flow on skeletal isotopic composition in the coral Pocillopora damicornis

Fragments of branching Pocillopora damicornis coral colonies were grown in experimental flumes under two water flow regimes. Colony size and buoyant weight increased most rapidly in the fast-flow regime. Branch tips from the upper and outer parts of the colonies showed the lowest and most consistent skeletal oxygen isotope ratios. Flow regime had little influence on the lowest oxygen isotope ratios, which were at least 3.5‰ lighter than the calculated oxygen isotopic equilibrium. These “kinetic” isotope effects are comparable to those observed in Porites corals. Relatively more branch tips showed extreme ¹⁸O depletions under low-flow conditions, and among small coral colonies. Isotopic variability was greater among branch tips from the lower and inner parts of the colonies and at high flow. Skeletal oxygen and carbon isotope ratios generally showed positive correlations. Despite the particularly large offsets from isotopic equilibrium, the isotopically lightest branches showed the greatest isotopic consistency and therefore would make the best isotopic thermometers. Isotopic variability within the colony may provide an indication of flow regime.     

Ontogenetic changes in the capacity of the coral Pocillopora damicornis to originate branches

Most colonial corals vary intraspecifically in growth forms, and the diversity in branching morphology is especially striking. While the effects of environmental factors on growth forms have been studied, the genetic control of coral branching patterns has received little attention. The discovery of ontogenetic changes in the capacity to originate branching would set the stage for studies of how branch formation is genetically controlled. During experiments investigating contact reactions in the coral Pocillopora damicornis, we observed that young colonies derived from settled planulae and colonies regenerated from adult branch tips assumed different growth forms. Young colonies formed at least one branch from the central region of the colony, while colonies regenerated from adult branch tips (3-5 mm long) did not form branches during the 9-month observation period. This pattern was invariable, regardless of the types and outcomes of the contact experiments or the orientation of the branch tips. However, some fragments taken from 1- or 2-year-old colonies formed branches. This suggests that the rate of branch formation in P. damicornis colonies decreases with age. These findings will facilitate investigations of the mechanism of coral branch formation at the molecular level.     

Endosymbiotic copepods may feed on zooxanthellae from their coral host, Pocillopora damicornis

The Xarifiidae is one of the most common families of endosymbiotic copepods that live in close association with scleractinian corals. Previous studies on xarifiids primarily focused on their taxonomy and morphology, while their influence on corals is still unknown. In this study, we collected a total of 1,579 individuals belonging to 6 species of xarifiids from 360 colonies of Pocillopora damicornis at Nanwan Bay, southern Taiwan from July 2007 to May 2008. Furthermore, using optical and electron microscopic observations, we examined the gut contents of Xarifia fissilis, the most abundant species of the Xarifiidae that we collected. We found that the gut of X. fissilis was characterized by a reddish-brown color due to the presence of numerous unicellular algae with diameters of 5–10 μm. TEM observations indicated that the unicellular algae possessed typical characteristics of Symbiodinium including a peripheral chloroplast, stalked pyrenoids, starch sheaths, mesokaryotic nuclei, amphiesmas, an accumulation body, and mitochondria. After starving the isolated X. fissilis in the light and dark (light intensity: 140 μmol photon m⁻² s⁻¹; photoperiod: 12 h light/12 h dark) for 2 weeks, fluorescence was clearly visible in its gut and fecal pellets under fluorescent microscopic observations. The cultivation experiment supports the hypothesis that the unicellular algae were beneficial to the survival of X. fissilis under light conditions, possibly through transferring photosynthates to the hosts. These results suggest that X. fissilis may consume and retain unicellular algae for further photosynthesis.     

Oxidative DNA damage induced by iron chloride in the larvae of the lace coral Pocillopora damicornis

Biochemical and molecular biomarkers tools are utilized as early warning signatures of contaminant exposure to target and non-target organisms. The objective of this study was to investigate the sublethal effects of iron chloride to the larvae of the lace coral Pocillopora damicornis by measuring a suit of oxidative-stress biomarkers. The larvae were exposed to a range of sublethal concentrations of iron chloride (0.01, 0.1, 1, 10, and 100 ppm) for seven days. With reference to oxidative stress biomarkers, the no-observed effect concentration (NOEC) and the lowest observed effect concentration (LOEC) of iron chloride were observed to be 0.01 and 100 ppm respectively. At the end of the seventh day the antioxidant status of the larvae was evaluated by the levels of glutathione (GSH), glutathione peroxidase (GPX), glutathione reductase (GR), and glutathione-S-transferase (GST), in both experimental and control groups. For the quantification of cellular oxidative damage, lipid peroxidation (LPO) activity was determined in the same and the extent of DNA damage was assessed by the expression of DNA apurinic/apyrimidinic (AP) sites. Iron chloride exhibited a concentration-dependent inhibition of GSH and GPX and induction of GR, GST, LPO, and DNA-AP sites in the P. damicornis larvae when compared to the control group. The oxidative stress biomarkers of the larvae exposed to 0.1, 1, and 10 ppm of iron chloride did not show any significant overall differences when compared to the control group. However the activities of LPO, GSH, GPX, GR, GST and DNA-AP in the larval group exposed to 100 ppm of iron chloride exhibited statistically significant (P=0.002, 0.003, 0.002, 0.002, 0.005 and 0.007) differences when compared to the control group. The research results indicated that iron chloride in concentrations at the 100 ppm level caused oxidative stress in the P. damicornis larvae.     

Population genetics of an ecosystem-defining reef coral Pocillopora damicornis in the Tropical Eastern Pacific

Background

Coral reefs in the Tropical Eastern Pacific (TEP) are amongst the most peripheral and geographically isolated in the world. This isolation has shaped the biology of TEP organisms and lead to the formation of numerous endemic species. For example, the coral Pocillopora damicornis is a minor reef-builder elsewhere in the Indo-West Pacific, but is the dominant reef-building coral in the TEP, where it forms large, mono-specific stands, covering many hectares of reef. Moreover, TEP P. damicornis reproduces by broadcast spawning, while it broods mostly parthenogenetic larvae throughout the rest of the Indo-West Pacific. Population genetic surveys for P. damicornis from across its Indo-Pacific range indicate that gene flow (i.e. larval dispersal) is generally limited over hundreds of kilometers or less. Little is known about the population genetic structure and the dispersal potential of P. damicornis in the TEP.

Methodology

Using multilocus microsatellite data, we analyzed the population structure of TEP P. damicornis among and within nine reefs and test for significant genetic structure across three geographically and ecologically distinct regions in Panama.

Principal Findings/Conclusions

We detected significant levels of population genetic structure (global R_ST = 0.162), indicating restricted gene flow (i.e. larvae dispersal), both among the three regions (R_RT = 0.081) as well as within regions (R_SR = 0.089). Limited gene flow across a distinct environmental cline, like the regional upwelling gradient in Panama, indicates a significant potential for differential adaptation and population differentiation. Individual reefs were characterized by unexpectedly high genet diversity (avg. 94%), relatively high inbreeding coefficients (global F_IS = 0.183), and localized spatial genetic structure among individuals (i.e. unique genets) over 10 m intervals. These findings suggest that gene flow is limited in TEP P. damicornis populations, particularly among regions, but even over meter scales within populations.     

Diversity of bacteria associated with the coral Pocillopora damicornis from the Great Barrier Reef

The microbial community associated with the reef building coral Pocillopora damicornis located on the Great Barrier Reef was investigated using culture-independent molecular microbial techniques. The microbial communities of three separate coral colonies were assessed using clone library construction alongside restriction fragment length polymorphism and phylogenetic analysis. Diversity was also investigated spatially across six replicate samples within each single coral colony using 16S rDNA and rpoB-DGGE analysis. Clone libraries demonstrated that the majority of retrieved sequences from coral tissue slurry libraries affiliated with gamma-Proteobacteria. This contrasted with clone libraries of seawater and coral mucus, which were dominated by alpha-Proteobacteria. A number of retrieved clone sequences were conserved between coral colonies; a result consistent with previous studies suggesting a specific microbe-coral association. rpoB-DGGE patterns of replicate tissue slurry samples underestimated microbial diversity, but demonstrated that fingerprints were identical within the same coral. These fingerprints were also conserved across coral colonies. The 16S rDNA-DGGE patterns of replicate tissue slurry samples were more complex, although non-metric multidimensional scaling (nMDS) analysis showed groupings of these banding patterns indicating that some bacterial diversity was uniform within a coral colony. Sequence data retrieved from DGGE analysis support clone library data in that the majority of affiliations were within the gamma-Proteobacteria. Many sequences retrieved also affiliated closely with sequences derived from previous studies of microbial diversity of healthy corals in the Caribbean. Clones showing high 16S rDNA sequence identity to both Vibrio shiloi and Vibrio coralliilyticus were retrieved, suggesting that these may be opportunist pathogens. Comparisons of retrieved microbial diversity between two different sampling methods, a syringe extracted coral mucus sample and an airbrushed coral tissue slurry sample were also investigated. Non-metric multidimensional scaling of clone library data highlighted that clone diversity retrieved from a coral mucus library more closely reflected the diversity of surrounding seawater than a corresponding coral tissue clone library.     

Response to higher sediment loads by Pocillopora damicornis planulae

Planulae collected from the reef coral Pocillopora damicornis were exposed to four different levels of sediment (0, 10, 100, 1000 mg l^-1) for 14 days under two contrasting water agitation levels (aerated vs. non-aerated). There was no significant difference in larval settlement among the varius sediment treatments and between agitation levels used in this study. However, a significant number of settled planulae underwent reversed metamorphosis (polyp bail-out) in the 100 and 1000 mg l^-1 treatments of both water aeration regimes indicating conditions were not appropriate for continued development and growth. Data suggested that successful recruitment may be limited in areas with greater than normal sediment loads.     

Survival of tissue balls from the coral Pocillopora damicornis L. exposed to cryoprotectant solutions

In this study, the tolerance of tissue balls (TBs, 100–300 μm in diameter) from the coral Pocillopora damicornis produced using mechanical excision to exposure to cryoprotectant (CPA) solutions was tested. TBs were treated for 20 min at room temperature with solutions of ethylene glycol (EG), methanol (Met), glycerol (Gly) or dimethyl sulfoxide (Me₂SO) at concentrations between 1.0 and 4.5 M. Two parameters were used to evaluate the survival of TBs following CPA treatment. The Undamaged Duration of Tissue Balls (expressed in h) corresponded to the time period during which the membrane surface of TBs remained smooth and their motility was preserved. Tissue Ball Regression (expressed in μm/h) corresponded to the size reduction of TBs over time. TBs tolerated exposure to all CPAs tested at the three lower concentrations employed (1.0 M, 1.5 M and 2.0 M). No survival was achieved following exposure to a 4.5 M CPA solution. At concentrations of 3.0 and 4.0 M, higher Undamaged Duration of Tissue Balls and lower Tissue Ball Regression were obtained following treatment with EG compared to the other three CPAs. Our experiments show that TBs constitute a good experimental material to evaluate CPA toxicity on corals using large numbers of samples. Performing preliminary experiments with TBs may allow reducing the number of tests carried out with less easily available coral forms such as planulae, thereby preserving larval stocks.     

Mixed asexual and sexual reproduction in the Indo‐Pacific reef coral Pocillopora damicornis

Pocillopora damicornis is one of the best studied reef‐building corals, yet it's somewhat unique reproductive strategy remains poorly understood. Genetic studies indicate that P. damicornis larvae are produced almost exclusively parthenogenetically, and yet population genetic surveys suggest frequent sexual reproduction. Using microsatellite data from over 580 larvae from 13 colonies, we demonstrate that P. damicornis displays a mixed reproductive strategy where sexual and asexual larvae are produced simultaneously within the same colony. The majority of larvae were parthenogenetic (94%), but most colonies (10 of the 13) produced a subset of their larvae sexually. Logistic regression indicates that the proportion of sexual larvae varied significantly with colony size, cycle day, and calendar day. In particular, the decrease in sexual larvae with colony size suggests that the mixed reproductive strategy changes across the life of the coral. This unique shift in reproductive strategy leads to increasingly asexual replications of successful genotypes, which (in contrast to exclusive parthenogens) have already contributed to the recombinant gene pool.