Asexual production of planulae in the coral Pocillopora damicornis

The reproduction of scleractinian corals through planular larvae has traditionally been viewed as a strictly sexual process. Here, the results of an electrophoretic study of a ubiquitous Indo-Pacific coral, Pocillopora damicornis, show an exact inheritance of parental genotypes by brooded planulae, demonstrating the existence of an asexual mode of production of planular larvae. Comparisons of the genetical structure of a number of populations with structures predicted for sexual reproduction suggest that, although there is probably also a sexual form of reproduction, asexually produced planulae can be of major importance in the maintenance of populations of this species.     

Genetical structure within populations of the coral Pocillopora damicornis

Genetic variation of allozymes within populations of Pocillopora damicornis from southwestern Australia was consistent with a primary role of local asexual proliferation of clones in population maintenance. Populations were composed typically of two to four multilocus genotypes accounting for 40 to 80% of individuals, with the remainder assigned to genotypes occasionally in twos or threes but more commonly singly. In the three populations where recruitment was examined genetically, 84% of all first-year recruits was assigned to clones represented in the population's resident adults. The majority of these recruits came from the most highly-replicated of the adult clones. The observed genotypic diversity was, on average, about half that calculated to occur for the same allelic frequencies in a sexually-reproducing population with free recombination. Despite the prevalence of asexual reproduction, both through planulae and fragments, the existence of a sexual mode of reproduction was inferred from the high level of variation produced by pooling populations, the existence of novel genotypes and the concordance of clonal gene frequencies at many sites with the predictions of Hardy-Weinberg equilibria.     

Ecological and physiological differences between two colour morphs of the coral Pocillopora damicornis

Pocillopora damicornis (Linnaeus) is a ubiquitous branching coral found throughout the Indo-Pacific region. Like many other species of coral, P. damicornis displays a large range of morphologies. At One Tree Island, it occurs as two distinct morphs that are easily distinguished by the presence or absence of pink pigmentation. The two colour morphs of P. damicornis were found to differ in their distribution and abundance in the One Tree Island Lagoon. The brown morph was more abudant than the pink morph in the shallows (<1 m),whereas the pink morph was more abundant at deeper sites (>3 m). The two morphs also differed physiologically. The brown morph tended to have a greater calcification rate than the pink morph, regardless of environmental conditions. However, the difference in the calcification rate between the two morphs became non-significant under shaded conditions (5% full sunlight), indicating some degree of physiological plasticity of the morphs. The pink colour in P. damicornis was due to a hydrophilic pigment with a major peak absorbance at 560 nm. The expression of pink pigment had both genetic and phenotypic components. The brown morph has a reduced genetic capacity to express the pigment relative to the pink morph. On the other hand, pigment expression could be induced by light in the pink morph. Although genetic differences ultimately determine the differences between the two morphs of P. damicornis, the extent of pigment expression is under some degree of environmental influence.     

Intra-colonial variability in light acclimation of zooxanthellae in coral tissues of Pocillopora damicornis

We investigated heterogeneity of light acclimation of photosynthesis in sun- and shade-adapted coenosarc and polyp tissues of Pocillopora damicornis. The zooxanthellar community within P. damicornis colonies at Heron Island is genetically uniform, yet they showed a large degree of plasticity in their photo-physiological acclimation linked to light microclimates characterised by fibre-optic microprobes. Microscale scalar irradiance measurements showed higher absorption in polyp than coenosarc tissues and higher absorption in the more densely pigmented shade-adapted polyps than in sun-adapted polyps. The combination of an O₂ microelectrode with a fibre-optic microprobe (combined sensor diameter 50–100 μm) enabled parallel measurements of O₂ concentration, gross photosynthesis rate and photosystem II (PSII) quantum yield at the coral surface under steady-state conditions as a function of increasing irradiances. Lower O₂ levels at the tissue surface and higher compensation irradiance indicated a higher respiration activity in sun-adapted polyp tissue as compared to shade-adapted polyps. Shade-adapted coenosarc and polyp tissues exhibited lower maxima of relative electron transport rates (rETR_max) (84±15 and 41±10, respectively) than sun-adapted coenosarc and polyp tissues (136±14 and 77±13, respectively). Shade-adapted tissues showed stronger decrease of rETR at high scalar irradiances as compared to sun-adapted tissues. The relationship between the relative PSII electron transport and the rate of gross photosynthesis, as well as O₂ concentration, was non-linear in sun-adapted tissues over the entire irradiance range, whereas for shade-adapted tissues the relationship became non-linear at medium to high scalar irradiances >200 μmol photons m⁻² s⁻¹. This suggests that rETR measurements should be used with caution in corals as a proxy for photosynthesis rates. The apparently high rates of photosynthesis (oxygen evolution rates) suggest that there must be a considerable electron transport rate through the photosystems that is not observed by the rETR measurements. This may be accounted for by vertical heterogeneity of zooxanthellae in the tissue and the operation of an alternative electron pathway such as cyclic electron flow around PSII.     

Evidence for broadcast spawning as well as brooding in the scleractinian coral Pocillopora damicornis

The reproductive activity of 88 colonies of Pocillopora damicornis at Rottnest Island, Western Australia, was examined over 12 mo (20 August 1988 to 20 August 1989). Larvae were found close to the three new moons of February, March and April, 1989, but in large numbers only during March; 67% of all corals produced larvae. There was no obvious connection between gametogenesis and the production of brooded larvae; however, those corals which did not produce, larvae also did not produce ova but did produce, sperm, and must be considered male colonies. All colonies which produced ova went on to produce planulae. Spermatogenesis occurred throughout the year, but oogenesis occurred between September 1988 and April 1989. Dramatic declines in the percentage of polyps containing eggs and sperm occurred in early February and early April 1989. On these dates the amount of lipid in the tissue of the corals decreased, which suggests that the decline in the percentage of polyps with sperm and eggs was not due to the gametes being resorbed by the polyps. Therefore, P. damicornis at Rottnest Island appears to be both a brooder and a broadcaster with asexual reproduction through brooded larvae and sexual reproduction through spawning of gametes. This accounts for earlier evidence in the literature that some input from sexual reproduction had been occurring in the populations of P. damicornis at Rottnest Island. There is no direct evidence for spawning, as broadcasting of gametes has not been observed, but the indirect evidence provides a strong argument for the existence of this dual reproduction.     

Effect of temperature and temperature adaptation on calcification rate in the hermatypic coral Pocillopora damicornis

Using ⁴⁵Ca incorporation into the coral skeleton as a measure of calcification rate, the effect of temperature on clacification rate was studied in the hermatypic coral Pocillopora damicornis. Both immediate and long-term (adaptation) effects were investigated. Temperature has a marked effect on rate — an effect that varies depending on the temperature history of the coral (i.e., temperature adaptation occurs). P. damicornis showed both 27° and 31°C temperature optima, one or the other being dominant depending on the natural water temperature to which the coral was adapted. The two optimum temperatures may indicate two isoenzymes or two alternate metabolic pathways involved in the calcification process.     

Energetics,competency, and long-distance dispersal of planula larvae of the coral Pocillopora damicornis

Pocillopora damicornis (Linnaeus) were collected from Enewetak Atoll, Marshall Islands, in 1980–1981) and Kaneohe Bay, Hawaii, in 1982. Their planula larvae contained 17% protein, 70% lipid, and 13% carbohydrate by dry weight. Calculations based on stored energy reserves and daily metabolic expenditure indicate that planulae could survive approximately 100 d and still settle successfully. Competency experiments demonstrated that larvae settled and metamorphosed after 103 d. This period of time is sufficient to allow immigration of larvae from the Central Pacific to the eastern Pacific, and supports the hypothesis of long-distance dispersal of larvae for the origin of present eastern Pacific populations of P. damicornis.     

Availability of two forms of dissolved nitrogen to the coral Pocillopora damicornis and its symbiotic zooxanthellae

The relative contribution of dissolved nitrogen (ammonium and dissolved free amino acids DFAAs) to the nitrogen budget of the reef-building coral Pocillopora damicornis was assessed for colonies growing on control and ammonium-enriched reefs at One Tree Island (southern Great Barrier Reef) during the ENCORE (Enrichment of Nutrient on Coral Reef; 1993 to 1996) project. P. damicornis acquired ammonium at rates of between 5.1 and 91.8 nmol N cm⁻² h⁻¹ which were not affected by nutrient treatment except in the case of one morph. In this case, uptake rates decreased from 80.5 to 42.8 nmol cm⁻² h⁻¹ (P < 0.05) on exposure to elevated ammonium over 12 mo. The presence or absence of light during measurement did not influence the uptake of ammonium ions. Nitrogen budgets revealed that the uptake of ammonium from concentrations of 0.11 to 0.13 μM could completely satisfy the demand of growing P. damicornis for new nitrogen. P. damicornis also took up DFAAs at rates ranging from 4.9 to 9.8 nmol N cm⁻² h⁻¹. These rates were higher in the dark than in the light (9.0 vs 5.1 nmol m⁻² h⁻¹, P < 0.001). Uptake rates were highest for the amino acids serine, arginine and alanine, and lowest for tyrosine. DFAA concentrations within the ENCORE microatolls that received ammonium were undetectable, whereas they ranged up to 100 nM within the control microatolls. The contribution of DFAAs to the nitrogen budget of P. damicornis constituted only a small fraction of the nitrogen potentially contributed by ammonium under field conditions. Even at the highest field concentrations measured during this study, DFAAs could contribute only ≃11.3% of the nitrogen demand of P.␣damicornis. This contribution, however, may be an important source of nitrogen when other sources such as ammonium are scarce or during periods when high concentrations of DFAAs become sporadically available (e.g. cell breakage during fish-grazing). Received: 22 April 1998 / Accepted: 3 November 1998     

Night irradiance and synchronization of lunar release of planula larvae in the reef coral Pocillopora damicornis

Pocillopora damicornis (Linnaeus), which is known to release planula larvae on a monthly cycle, was grown in full daytime solar irradiance, but with four treatments of night irradiance: (1) natural night irradiance, (2) shifted-phase (total darkness during nights of full moon with artificial irradiance at lunar intensity on nights of new moon), (3) constant full moon (full lunar irradiance every night), and (4) constant new moon (total darkness every night). The reproductive cycle of the corals held in the shifted-phase treatment moved out of synchrony with the cycle of corals exposed to a natural lunar cycle of night irradiance. Two previously described types of P. damicornis were tested. The Type Y normally start releasing larvae at full moon, with peak production at third quarter. In the shifted-phase treatment they began releasing planulae at new moon (artificial full moon), with peak production at first quarter. The Type B corals, that normally start releasing planulae at new moon with peak production at first quarter, began to release planulae at full moon (artificial new moon), with peak production at third quarter. Populations of corals grown either in the constant full moon or constant new moon treatment quickly lost synchronization of monthly larva production, although production of planulae continued. Thus spawning is synchronized by night irradiance.Contribution No. 702 of the Hawaii Institute of Marine Biology     

Influence of size on primary production in the reef coral Pocillopora damicornis and the macroalga Acanthophora spicifera

Size influences the photosynthesis-irradiance (P-I) relationship in colonies of the branched reef-coral Pocillopora damicornis and in intact plants of the branched redmacroalga Acanthophora spicifera. The light saturation constant is proportional to size. Maximum net rate of oxygen production (net photosynthesis) per colony and nocturnal dark oxygen-uptake rate per colony (respiration) increase with increasing size, but the latter increases at a much lower rate. Therefore, the photosynthesis to respiration ratio increases with increasing canopy size. Large increases in chlorophyll per unit reef area also accompany increase in size. The initial slope (alpha) of the chlorophyll-specific P-I curve and assimilation number are inversely related to size. Integrated daytime oxygen production increases with size more rapidly than nighttime oxygen consumption. Consequently, net primary production of an entire colony or plant (or rate per unit area of reef) increases with increasing size of the canopy. Production efficiency also increases with size. The coral is rigid, symmetrical and highly organized. Chlorophyll distribution is more stratified in comparison to the macroalga. The coral shows higher photosynthetic efficiency, as would be expected according to the stratified production model of Odum et al. (1958). This research was conducted on specimens from Kaneohe Bay, Oahu, Hawaii, USA in 1981.     

The production of sexual and asexual larvae within single broods of the scleractinian coral, Pocillopora damicornis

The genotypic compositions of two populations of the brooding coral, Pocillopora damicornis, were studied by exhaustive sampling of adult colonies on a fine scale in southern Taiwan. In addition, 100 larvae were randomly selected among more than 1,000 brooded larvae collected from a single broodparent within each population. Using 7 polymorphic microsatellite markers, both populations were found to be highly clonal, and 7 clonal lineages were characterized. One clonal lineage (C1) dominated both study areas and comprises 54.9% of all colonies sampled, while any of the other 6 clonal lineages represented no more than 5%. Among the 100 larvae randomly selected for genotyping from each broodparent, the extent of clonal reproduction was high, and only 29 and 6 larvae, respectively, were found to be genotypically different from their broodparent. Among the 35 genotypically distinct larvae, 33 were thought to be derived from sexual reproduction, and 2 were assumed to be clonal propagules that had undergone somatic mutations. Two genotypically identical larvae were also found in one of the 2 sexually derived larva arrays, indicating the possible existence of polyembryony. The high proportions of clonality in both adult colonies and brooded larvae suggest that asexually produced larvae might significantly contribute to the recruitment of local populations. The dense clonal population of P. damicornis in the study area favors the quick recolonization view of clonal propagules after disturbances.     

Genetic variation and clonal structure in the scleractinian coral Pocillopora damicornis in the Ryukyu Archipelago, southern Japan

Samples of the scleractinian coral Pocillopora damicornis were collected from six sites located around four islands in the Ryukyu Archipelago, southern Japan, and subjected to allozyme electrophoresis. Seven polymorphic loci were examined for their allelic patterns. The ratio of observed to expected genotypic diversity (0.30 < G _o :G _e  < 0.64), the ratio of the observed number of genotypes to the number of individuals (0.47 < N _g :N _i  < 0.75), and deviations from Hardy–Weinberg equilibrium indicated that asexual reproduction plays a major role in the maintenance of established populations. However, populations were not completely dominated by a single or a few clones, and most clones were represented by only a few individual samples. The high frequency of typhoons in the region suggests that, in P. damicornis, fragmentation caused through occasional exposure to powerful waves is a major mode of asexual reproduction, but asexual production of planulae may also be contributing to the maintenance of populations. A significant genetic differentiation (F ST) was found between the six populations examined (0.027 < F ST < 0.092, average F ST = 0.056). The moderate gene flow is discussed according to characteristics of the larval stage of the species, and to circulation patterns in the region. Received: 7 August 1998 / Accepted: 18 May 1999     

Variation in the genetic composition of coral (Pocillopora damicornis and Acropora palifera) populations from different reef habitats

The genetic structure of populations of the corals Pocillopora damicornis and Acropora palifera was examined in three habitats at One Tree Island during March and April 1993, using electrophoretically detectable variation at six allozyme loci. There were significant genetic differences among populations of P. damicornis within each of the reef crest, lagoon and microatoll habitats. The level of differentiation among populations was similar in each of the habitats. Differences between populations of P. damicornis from lagoon and microatolls were no greater than that within habitats, but genetic differentiation of these from crest populations was much higher. There was no difference in the genetic composition of A. palifera populations within or between the lagoon and microatolls, the only habitats where this species was found. Both coral species had observed:expected (G _O:G_E) genotypic diversity rations >0.80, indicating predominantly sexual reproduction. These data, the high genotype diversity and general conformance of genotype frequencies to those expected under conditions of Hardy-Weinberg, suggested panmixis at each site. The high degree of sexual reproduction in the P. damicornis populations is unusual for a species where asexual reproduction has been the dominant mode of reproduction reported to date. Gene flow in both species was considerable between the lagoon and the closed microatolls. The genetic differences between populations of P. damicornis in these habitats and the reef crest may reflect the relative isolation of all populations within the closed One Tree Lagoon from those outside. However, local currents appear to offer effective means of dispersal between the habitats, suggesting that the genetic differences result from natural selection in the different environments within One Tree Lagoon and the reef crest.     

The physiological and molecular responses of larvae from the reef-building coral Pocillopora damicornis exposed to near-future increases in temperature and pCO2

Given the threats of greenhouse gas emissions and a changing climate to marine ecosystems, there is an urgent need to better understand the response of not only adult corals, which are particularly sensitive to environmental changes, but also their larvae, whose mechanisms of acclimation to both temperature increases and ocean acidification are not well understood. Brooded larvae from the reef coral Pocillopora damicornis collected from Nanwan Bay, Southern Taiwan, were exposed to ambient or elevated temperature (25 or 29 °C) and pCO₂ (415 or 635 μatm) in a factorial experiment for 9 days, and a variety of physiological and molecular parameters were measured. Respiration and rubisco protein expression decreased in larvae exposed to elevated temperature, while those incubated at high pCO₂ were larger in size. Collectively, these findings highlight the complex metabolic and molecular responses of this life history stage and the need to integrate our understanding across multiple levels of biological organization. Our results also suggest that for this pocilloporid larval life stage, the impacts of elevated temperature are likely a greater threat under near-future predictions for climate change than ocean acidification.     

Patterns of formation and the ultrastructure of the larval skeleton of Pocillopora damicornis

The growth and development of the tissues and skeleton of settled larvae of the reef coral Pocillopora damicornis (Linnaeus), collected in December 1983 from Ko Phuket, Thailand, were investigated using light microscopy, scanning electron microscopy and transmission electron microscopy. The rate of development of larval skeletons was very variable, preventing the chronological sequencing of skeletal growth. However, four growth stages in the development of a complete larval skeleton from first settlement were identificd: Stage 1, deposition of the first elements of the basal plate upon settlement; Stage 2, completion of basal plate, and deposition of skeletal spines and ridges in positions corresponding to the septal cycles; Stage 3, formation of the corallite wall and septal and costal cycles; Stage 4, the complete larval skeleton which represented the maximum growth attained eight days after settlement. The configuration of the larval tissues, particularly the aboral ectoderm, mirrored the four developmental stages. The deposition of the larval skeleton was correlated with the metamorphosis of the aboral ectoderm from a columnar to a squamous morphology. The basal plate of the larval skeleton had two layers of crystals orientated perpendicular to each other. The architecture of the complete larval skeleton is described and compared to that of the adult skeleton of P. damicornis. The results are discussed with respect to previous concepts of the formation of the larval skeleton of scleractinian corals and coral calcification.     

Genetic diversity,clonality and connectivity in the scleractinian coral Pocillopora damicornis: a multi-scale analysis in an insular,fragmented reef system

Clonality and genetic structure of the coral Pocillopora damicornis sensu lato were assessed using five microsatellites in 12 populations from four islands of the Society Archipelago (French Polynesia) sampled in June 2008. The 427 analysed specimens fell into 132 multilocus genotypes (MLGs), suggesting that asexual reproduction plays an important role in the maintenance of these populations. A haploweb analysis of ITS2 sequences of each MLG was consistent with all of them being conspecific. Genetic differentiation was detected both between and within islands, but when a single sample per MLG was included in the analyses, the populations turned out to be nearly panmictic. These observations provide further evidence of the marked variability in reproductive strategies and genetic structure of P. damicornis throughout its geographic range; comparison with results previously obtained for the congeneric species Pocillopora meandrina underlines the importance of life history traits in shaping the genetic structure of coral populations.     

Studies on reef corals. I. Skeleton formation by newly settled planula larva of Pocillopora damicornis

Initiation of coral-skeleton formation was studied in the reef-coral Pocillopora damicornis Lamarck. Observations were made on sequential skeletal growth stages of newly settled planula larvae during the first 22 days following settling onto glass microscope slides. Techniques used include phase light microscopy, scanning and transmission electron microscopy, and powder X-ray and selected area electron micro-diffraction. Formation of the skeleton is initiated immediately on settling of the larva. The primary calcareous elements are of two types — flattened spherulitic platelets, and smaller rod-like granules. Rudimentary primary septa are clearly defined within 6 h after settling. Fusion of the primary calcareous elements results in the formation of the larval basal disc within 48 to 72 h. With transmission electron microscopy, this basal disc is found to differ from subsequent adult calcification in (1) considerably lesser degree of mineralization, (2) smaller crystal size, (3) more random orientation of the crystals, and (4) the presence of trace amounts of calcite in addition to aragonite. The basal disc with its septal rudiments constitutes a true larval skeleton, differing in morphology, micro-architecture, and crystal type from the fibrous growth characterizing the adult skeleton.Contribution 419, Hawaii Institute of Marine Biology, University of Hawaii.     

Effects of nitrogen and phosphorus enrichement on in vivo symbiotic zooxanthellae of Pocillopora damicornis

The reef coral Pocillopora damicornis (Linnaeus) was grown for 8 wk in four nutrient treatments: control, consisting of ambient, unfiltered Kaneohe Bay seawater [dissolved inorganic nitrogen (DIN, 1.0 M) and dissolved inorganic phosphate (DIP, 0.3 M)]; nitrogen enrichment (15 M DIN as ammonium); phosphorus enrichment (1.2 M DIP as inorganic phosphate); and 15 M DIN+1.2 M DIP. Analyses of zooxanthellae for C, N, P and chlorophyll a after the 8 wk experiment indicated that DIN enrichment increased the cellular chlorophyll a and excess nitrogen fraction of the algae, but did not affect C cell^-1. DIP enrichment decreased both C and P cell^-1, but the decrease was proportionally less for C cell^-1. the response of cellular P to both DIN and DIP enrichment appeared to be in the same direction and could not be explained as a primary effect of external nutrient enrichment. The observed response of cellular P might be a consequence of in situ CO₂ limitation. DIN enrichment could increase the CO₂ (aq) demand by increasing the net production per unit area. DIP enrichment could slow down calcification, thus decreasing the availability of CO₂ (aq) in the coral tissue.Hawaii Institute of Marine Biology Contribution No. 920     

Studies on reef corals. III. Fine structural changes of calicoblast cells in Pocillopora damicornis during settling and calcification

The site of reef-coral calcification has been studied in the branching coral Pocillopora damicornis Lamarck. Electron microscopy and X-ray microprobe analysis were performed on the calicoblast epidermis of newly settled larval stages and of adult coral. During settling, the heterogeneous columnar cell composition of the planktonic larva epidermis is replaced by a simple epithelium consisting of a single cell type, the calicoblast cell. Metamorphosis appears tightly linked to settling, with cell changes occurring within hours after attachment, and is marked by the appearance of a new secretory cell. The calicoblast cell of the adult coral is extremely flattened, and interdigitates extensively with adjacent calicoblast cells. This cell possesses a featureless plasma membrane lacking microvilli or flagella. It characteristically contains large membrane-bound vesicles with homogeneously fine granular contents. Preliminary microprobe analysis indicated a higher calcium content in these vesicles than in surrounding tissue; however, not in concentrations suggesting calcium-carbonate precipitation. They may represent sites of organic matrix synthesis. The calicoblast epidermis is separated from the underlying coral skeleton by a narrow gap. This gap appeared devoid of substructure, either organic or inorganic. The coral soft tissues are attached to the skeleton by mesogleal attachment processes, the desmoidal processes. These consist of a complex fibrous network originating in the mesoglea, and inserting onto the skeleton via specialized attachment regions consisting of electron-opaque membranous plaques. Skeletogenesis in reef-corals probably occurs extra-cellularly, external to the calicoblast epidermis, by simple overgrowth of the skeleton.     

Studies on reef corals. II. Fine structure of planktonic planula larva of Pocillopora damicornis,with emphasis on the aboral epidermis

Tissue and cellular organization of the planula larva of the reef-coral Pocillopora damicornis are detailed ultrastructurally. Emphasis is placed on fine structure of the aboral (=presumptive calicoblast) epidermis. The pre-settling exploratory behavior of the coral larva is discussed. The aboral epidermis consists of a tall columnar epithelium, made up of a heterogeneous cell population-nematocytes, secretory and mucus cells, and flagellated cells. Basally, the columnar epithelium is modified into complexly interdigitating cellular processes, appearing as a discrete histological layer. A-type granules (Jha and Mackie, 1967) indicate neurosecretory activity. The flagellated cell contains a peculiarly comlex flagellar apparatus, including a new intracellular structure, an electron-opaque fibrous cup. It extends across the entire width of the flagellated cell apically, enveloping the flagellar basal bodies and paired transverse accessory rootlets. The flagellum main basal rootlet passes freely through an opening in the base of the rootlet cup. The flagellated cell is here viewed as a sensory cell, as suggested by the corona of microvilli encircling the flagellum, the bundles of microvilli, and extensive areas of tight junctions with adjacent cells. There is no evidence in the gastrodermis or epidermis of the planktonic larva for pre-settling accumulation of calcareous skeletal material. Vesicles containing a highly oriented fibrous material may represent sites of skeletal organic-matrix precusor buildup.Contribution no. 451. Hawaii Institute of Marine Biology, Kaneohe, Hawaii.