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.     

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.     

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.     

Physiological development of brooded larvae from two pocilloporid corals in Taiwan

In southern Taiwan, brooded larvae of Pocillopora damicornis and Seriatopora caliendrum are released year-round in synchrony with new moons, and each larval release occurs over multiple days. Using P. damicornis and S. caliendrum as a model system, we describe within-brood variation in larval phenotypes and test for release-day effects that influence larval performance in the pelagic phase. Research was conducted in 2010 using larvae from corals collected in June and July from Nanwan Bay (21°56.179??N, 120°44.85??E). In June, larval phenotypes of both species were characterized immediately following release, and their competency to settle assessed. In July, larvae of P. damicornis were collected on 3?days over the peak release period and incubated for 7?C11?days at 28.0?°C and 320???mol?quanta?m^?2?s^?1; their phenotypes and settlement competency were measured every 2?days. P. damicornis larvae released close to peak release were 1.6 times larger in size, contained twice the number of Symbiodinium larva^?1, and were 44?% more likely to settle in the first 24?h than larvae released early in the brood. In addition, peak-release larvae respired at a lower rate than larvae released late in the brood. Similarly, S. caliendrum larvae released close to peak release were 1.4 times larger in size and were 33?% more likely to settle in the first 5?h than larvae released early in the brood. In July, P. damicornis larvae differed between early (2?days prior to peak), peak, and late (2?days after peak) release. Protein content of early-release larvae was lower than peak- and late-release larvae, and this difference persisted throughout the development. Further, release day affected the way larval respiration varied throughout development. By showing that brooded coral larvae differ between release days and display maternal effects influencing performance in the swimming phase, our results suggest that pocilloporid corals utilize bet-hedging to increase reproductive success.     

Mode of reproduction,recruitment, and genetic subdivision in the brooding sponge<Emphasis Type="Italic"> Haliclona</Emphasis> sp.

Sponges display a variety of reproductive strategies that have the potential to influence population genetic structure. Histological examination of ten reproductive individuals of the Western Australian sponge Haliclona sp. showed that this species broods embryonic larvae that are potentially limited in dispersal capabilities. Because sponges have the potential to propagate in a number of modes, allozyme electrophoresis was used to assess the relative importance of asexual and sexual reproduction to recruitment, and to quantify genetic subdivision over different spatial scales. Tissue samples from 227 sponges were collected from reefs within two areas 400 km apart: Hamelin Bay and Rottnest Island. Contrary to expectations for highly clonal populations, genotypic diversity within sites was high, no linkage disequilibrium was found, and there was no evidence of genotypic clustering within reefs. There was no genetic evidence that asexual reproduction is important for the maintenance of populations. Genetic comparisons were consistent with mixing of sexually produced recruits within reefs, on a scale up to a few hundred metres, but significant genetic subdivision between reefs (F_ST=0.069 at Hamelin Bay, 0.130 at Rottnest Island) indicated that water gaps of several hundred metres are effective at preventing dispersal. Subdivision between the two areas, separated by 400 km, was moderately greater (F_ST=0.142) than within, but the same alleles were predominant in the two areas. These genetic patterns are consistent with limited dispersal capabilities of brooded larvae.Communicated by G.F. Humphrey, Sydney     

Polymorphism in soft coral larvae revealed by amplified fragment-length polymorphism (AFLP) markers

The dioecious Red Sea soft coral Parerythropodium fulvum fulvum breeds its nonsymbiotic planula larvae on the surface of female colonies for less than a week. After completing their development, larvae crawl and settle near maternal colonies. Here we study the genetic polymorphism of developing larvae by the use of amplified fragment-length polymorphism markers. Four reproductive colonies from shallow water populations (two from a dense population and two from a less densely populated area 100 m away) were chosen, and ten larvae were randomly collected from each colony. DNA was analyzed by using three different primer combinations producing 61, 63, 63 polymorphic markers, respectively. All larvae exhibited different banding patterns from one another, illustrating the prominent role of sexual reproduction for the production of larvae. Nei's mean genetic distances for all 12 possible pair-wise combinations for larval origins revealed, in most cases, that sister larvae are genetically closer than larvae from different colonies and that larvae may be grouped into three statistical clusters in accordance with colony origin and population studied. The usefulness of molecular methodologies in coral population genetics is discussed. Received: 26 March 1999 / Accepted: 19 October 1999     

Fissiparity and population genetics of Coscinasterias calamaria

The effect of asexual reproduction on the population genetics of the fissiparous seastar Coscinasterias calamaria was examined at Rottnest Island and the adjacent mainland, Western Australia, in 1985. Field samples and laboratory observations on growth rate and regeneration showed that fission is common at all 14 sites examined. Electrophoretic analysis of six polymorphic enzymes revealed low genotypic diversity and strong genetic disequilibrium at each site, confirming the highly clonal structure of local populations. Striking variation in clonal composition over distances as short as 50 m emphasizes the very localized subdivision of these populations. In the combined sample from all sites, however, the expected range of multilocus genotypes was found in the proportions expected from random mixing of sexually produced larvae, confirming that clonal diversity results from sexual reproduction. Local genotypic diversity was not correlated with fission, emphasizing the difficulty of determining the short-term roles of asexual and larval recruitment in the maintenance of populations. Subtidal populations, however, appear to have both lower incidence of fission and lower larval recruitment than do intertidal populations.     

Reef coral reproduction in the eastern Pacific: Costa Rica,Panama, and Galapagos Islands (Ecuador)

Reproductive ecology of two major reef-building corals in the eastern Pacific [Pocillopora damicornis (Linnaeus) andPocillopora elegans Dana] was investigated between 1984 and 1990 in Costa Rica, Panama (Gulf of Chiriqui and Gulf of Panama), and the Galapagos Islands (Ecuador) following the 1982–83 El Niño disturbance. Mature spermatocytes and oocytes were found in both species and were usually present in the same polyp in nearly equal ratios. Numerous zooxanthellae were observed in mature, unfertilized oocytes. Although spawning in field populations was not observed, it is likely that both pocilloporids are simultaneous hermaphroditic spawners, as evidenced by the disappearance of mature gametes after full moon. This is in contrast with most known pocilloporid corals that brood and release planula larvae. Corals were reproductively most active in the thermally stable environments of Costa Rica and the Gulf of Chiriqui (Panama) where 32 to 90% of all colonies contained gametes. In the moderately varying thermal conditions in the Galapagos Islands, 16 to 40% of colonies contained gametes, and in the pronounced seasonal upwelling environment of the Pearl Islands (Panama) only 6 to 18% of colonies contained gametes. Year-round reproduction occurred in Costa Rica and the Gulf of Chiriqui, whereas reproduction was confined to warm periods in the seasonally varying environments of the Galapagos Islands and the Gulf of Panama. Pocilloporid corals in Costa Rica and the Gulf of Chiriqui demonstrated lunar spawning activity, with mature gametes present a few days before and after full moon. Some limited spawning may have occurred also at new moon. While frequent gamete maturation has been demonstrated in this study, the relatively low rates of larval recruitment occurring on eastern Pacific coral reefs disturbed by the 1982–83 El Niño suggest that the recovery of important frame-building corals could be greatly prolonged.     

Evidence of amictic reproduction in the brittle-star Ophiomyxa brevirima

Ophiomyxa brevirima is a direct-developing species of brittle-star found around the coast of New Zealand. The mode of reproduction of this ophiuroid was studied using isozyme electrophoresis of mother and brooded offspring. Multi-locus assessment of the genotypes of 35 adult females and their brooded offspring was undertaken. A minimum of 45% of broods assessed showed evidence of amictic reproduction, with all offspring in these broods possessing genotypes that were identical to the genotype of their heterozygous mother. In most other broods there was evidence of segregation. These observations are interpreted as evidence of both sexual and asexual reproduction in this ophiuroid species. Received: 19 November 1996 / Accepted: 28 January 1997     

Genetic subdivision in the subtidal,clonal sea anemone Anthothoe albocincta

Anthothoe albocincta, a common subtidal anemone along south-eastern Australia, reproduces both sexually through broadcast spawning and clonally through fission. Clones may be distinguished both by their electrophoretic genotypes and the colour of their tentacles and oral discs. Local populations typically consist of many, dense clonal aggregations. However, some clones appear to have locally extensive distributions, forming a series of separate aggregations. The capacity of clones to disperse among local populations is unknown. In this study we used an electrophoretic survey at six allozyme loci to quantify levels of variation among samples from each of 13 local populations and four geographic regions sampled between April 1992 and June 1993. These data revealed that populations of A. albocincta, separated by up to 930 km, were at least moderately subdivided. Levels of geographic variation were high and the average F _ST value (standardised genetic variation) was 0.27 based on genotypes of all individual polyps. This value reflects substantial variation both within (F _PR =0.13) and among (F _RT =0.16) regions. Estimates of gene flow among both neighbouring populations and regions are therefore low (N _e m=1.7 and 1.3, respectively). UPGMA (unweighted pair-group method using arithmetic averages) dendrograms suggest that a genetic discontinuity occurs at the very south-east corner of Australia, paralleling reports for two other south-eastern Australian marine invertebrates. In addition, our analyses and theoretical predictions imply that localised proliferation of clonal genotypes may have caused us to underestimate the potential importance of gene flow via larval dispersal. Moreover, the abundance and vast geographic range of this species suggests that widespread dispersal does occur. Collections from three populations covering the peroid December 1992 to June 1993 were examined by crude dissection along with histological sectioning, and showed A. albocincta to be dioecious, with unisexual clones. Eggs within the ovaries of six females sampled over a 3 mo period were small (96±4 m) and similar to those of related species that produce planktotrophic larvae. In contrast, we found no evidence that clones were dispersed (shared) among neighbouring local populations. An average of only 6% of six-locus genotypes were common to pairs of local populations separated by up to 125 km, this being equal to the percentage expected through sexual reproduction alone. In addition, the percentage of shared genotypes did not decline with increasing geographic separation. These data imply that although asexual reproduction may be used to maintain local populations, the sexual production of genotypically diverse larvae is the primary source of widely dispersed colonists and hence of new clones.     

Spatial autocorrelation analysis of small-scale genetic structure in a clonal soft coral with limited larval dispersal

The philopatric larval dispesal and small effective population sizes characteristic of many clonal species should promote the development of significant small-scale genetic structure within populations as a result of isolation-by-distance. We used spatial autocorrelation statistics to detect genetic structure, arising from both clonal reproduction and philopatric dispersal of sexual propagules, for five allozyme loci within populations of the soft coral Alcyonium sp. In a population on Tatoosh Island, Washington, USA, sampled in 1991/1992, we found significant positive spatial autocorrelation at all loci among individuals separated by <40 cm, reflecting the presence of significant smallscale genetic structure due to associations among clonemates. For 4 of 5 loci, however, we detected no significant spatial autocorrelation among the different clones within this population over distances of 1 to 40 m. Analysis of soft-coral populations from six additional, topographically diverse sites in the north-east Pacific also did not reveal significant spatial autocorrelation among clones at any loci. This general lack of spatial autocorrelation of genotypes among clones suggests that significant small-scale genetic structure has not arisen in populations of Alcyonium sp. as a consequence of isolation-by-distance.     

Influence of contrasting larval developmental types upon the population-genetic structure of cheilostome bryozoans

For many sedentary or sessile benthic marine invertebrates the planktonic duration of the larval stage is believed to be a key determinant of the magnitude of genetic differences between populations. An obvious dichotomy in dispersal potential exists within cheilostome bryozoans that develop from either (1) a cyphonautes larva that spends several weeks in the plankton or (2) a brooded coronate larva that settles soon after release from the adult colony. This study characterises the pattern of variation at allozyme loci among British populations of four species of bryozoan—two species with cyphonautes and two with coronate larvae. There is some variation in the estimates of genetic differentiation among similarly separated populations that may be a consequence of non-equilibrium genetic conditions arising from sporadic migration, possibly through dispersal by rafting on macroalgae by mature colonies. Despite this, however, both the level of genetic differentiation between populations and the pattern of migrant exchange correlate with the larval developmental mode. Bryozoan species that brood coronate larvae show higher levels of genetic heterogeneity between populations and significant isolation by distance genetic structure while, by contrast, distance has little or no effect upon the amount of genetic differentiation among populations of bryozoans with cyphonautes larvae. For cheilostome bryozoans, therefore, it appears that genetic differentiation between populations is directly associated with the type of larval development. These data are discussed also with respect to levels of gene diversity and the geological pattern of cheilostome bryozoan species diversity.     

DNA fingerprints of a gorgonian coral: a method for detecting clonal structure in a vegetative species

Clonal reproduction, a common life history strategy among sessile marine invertebrates, can lead to high local abundances of one to a few genotypes in a population. Analysis of the clonal structure of such populations can provide insight into the ecological and evolutionary history of the population, but requires markers that can identify individual genets. Forensic and demographic studies have demonstrated that DNA fingerprinting can provide markers that are unique for an individual genotype. We have generated DNA fingerprints for over 70 colonies of the clonal gorgonian, Plexaura A (Plexaura sp. A) collected from June 1990 through July 1991 in the San Blas Islands, Panama. DNA fingerprints within a singic individual were identical and fingerprinting resolved multiple genotypes within and among reefs. On one reef in the San Blas Islands, Panama, 59% of the colonies sampled were of one genotype and this genotype was not found on any other sampled reefs. A previous study using tissue grafts identified 13 putative clones on these reefs, while DNA fingerprints of the same colonies differentiated 17 genotypes. The present study demonstrates the utility of DNA fingerprinting for distinguishing clones and for identifying clonal structure of marine invertebrate populations.     

The relative contribution of dinoflagellate photosynthesis and stored lipids to the survivorship of symbiotic larvae of the reef-building corals

The long-distance dispersal of larvae provides important linkages between populations of reef-building corals and is a critical part of coral biology. Some coral planulae have symbiotic dinoflagellates (Symbiodinium spp.) that probably provide energy in addition to the lipids provisioned within the egg. However, our understanding of the influence of symbionts on the energy metabolism and survivorship of planulae remains limited. This study examines the relative roles of symbiotic dinoflagellate photosynthesis and stored lipid content in the survivorship of the developing stages of the corals Pocillopora damicornis and Montipora digitata. We found that survivorship decreased under dark conditions (i.e. no photosynthetic activity) for P. damicornis and M. digitata at 31 and 22 days after release/spawning, respectively. The lipid content of P. damicornis and M. digitata planulae showed a significant decrease, at a higher rate, under dark conditions, when compared with light conditions. When converted to energy equivalents, the available energy provided by the depletion of lipids could account for 41.9 and 84.7% of larval metabolism for P. damicornis (by day 31) and 38.4 and 90.1% for M. digitata (by day 21) under light and dark conditions, respectively. This finding indicates that not all energy requirements of the larvae are met by lipids: energy is also sourced from the photosynthetic activities of the symbiotic dinoflagellates within these larvae, especially under light conditions. In addition, the amounts of three main lipid classes (wax esters, triglycerides, and phospholipids) decreased throughout the experiment in the planulae of both species, with the wax ester content decreasing more rapidly under dark conditions than under light conditions. The observations that the planulae of both species derive considerable amounts of energy from wax esters, and that symbiotic dinoflagellates enable larvae to use their stores at lower rates, suggested that symbiotic dinoflagellates have the potential to extend larval life under light conditions.     

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.     

Population structure of the Spanish sardine Sardinella aurita: natural morphological variation in a genetically homogeneous population

The importance of larval dispersal in determining the distribution and abundance of benthic marine organisms is well recognized; however, the contribution of post-larval dispersal has not been measured. I compared the dispersal of swimming larvae with that of rafting colonies in a population of compound ascidians, Botrylloides sp., living attached to leaves of the eelgrass Zostera marina in Tomales Bay, California, USA in 1990–1992. Colonies rafting on broken eelgrass traveled over 200 times farther and had comparable recruitment success relative to swimming larvae. The recruitment of rafting colonies into new habitats was facilitated by the ability of thesecolonial animals to grow asexually onto surrounding substrata. Rafting colonies brooded larvae that were subsequently relased after the colony settled into a new habitat. These results suggest that colonization of new habitats can occur by post-larvae as well as larvae, and that long-range dispersal by species with short-lived larvae may occur by post-larval rafting.     

Sexual reproduction of the photosymbiotic ascidian <Emphasis Type="Italic">Diplosoma virens</Emphasis> in the Ryukyu Archipelago,Japan: vertical transmission,seasonal change,and possible impact of parasitic copepods

Diplosoma virens is a colonial ascidian hosting prokaryotic algae Prochloron sp. in the common cloacal cavity of the colonies and is sometimes parasitized by notodelphyid copepods. In ascidian–Prochloron symbiosis, it is generally known that the host larvae acquire the algal symbionts from their mother colonies to maintain the symbiosis. A histological study of the sexually mature colonies of D. virens showed that the algal symbionts attach to pre-hatching larvae on the rastrum (plant rake) projected from the postero-dorsal part of the larval trunk, and then the rastrum is packed in the posterior half of the larval trunk that will become a cloacal cavity after metamorphosis. This process is the same as that of D. simile. Monthly sampling of D. virens colonies showed that they have embryos in summer in Ryukyus, situated near the northern-most limit of the coral reefs in the West Pacific. While the frequencies of copepod parasitism were variable among the populations, the colonies from a highly parasitized population had a significantly smaller number of eggs/embryos per zooid than the colonies from the less parasitized populations. The parasites probably have an inhibitory impact on the sexual reproduction of the host colonies.Communicated by T. Ikeda, Hakodate     

Regulation of ovary activation in worker honey-bees (Apis mellifera): larval signal production and adult response thresholds differ between anarchistic and wild-type bees

One-day-old anarchistic (selected for successful worker reproduction) and wild-type honey-bee workers were introduced into queenright colonies of honey-bees of two treatments. In treatment 1, all eggs and larvae were offspring of queens from an anarchistic line. In treatment 2, all eggs and larvae were offspring of wild-type queens. In both treatments, adult workers were wild type. This experimental arrangement was used to test the importance of larval genotype on ovary activation in young adult workers. After 12 days, the introduced bees were dissected to determine the frequency of ovary activation. In those colonies provided with wild-type brood, 0% of introduced wild-type bees and 16% of anarchistic bees had activated ovaries. In those colonies provided with anarchistic brood, 13% of introduced wild-type bees and 41% of anarchistic bees had activated ovaries. These results strongly support the hypothesis that selection for high levels of worker reproduction in anarchistic stocks has reduced the amount or composition of brood pheromones produced by larvae that normally signal workers to refrain from reproduction. They also suggest that anarchistic workers have a higher threshold for these signals than wild-type bees.