Sexual reproduction of three coral species from the Mexican South Pacific

Our understanding of the reproductive biology of corals from the Mexican southeastern Pacific is limited, and consequently, the role of reproduction in structuring coral communities is unclear. As a first attempt to understand the importance of sexual reproduction in structuring and maintaining of the coral communities from this region, we documented the reproductive cycles over 2 years (2003–2004) in three main reef-building corals species in the region. Pocillopora damicornis was shown to be hermaphroditic with asynchronous gamete development observed only in 2004; P. gigantea was characterized as both gonochoric and cosexually hermaphroditic. Absence of mature gametes was documented in both species, and an observation may be attributed to the 2003 El Niño Southern Oscillation event, which may have inhibited reproductive maturation via thermal stress. Porites panamensis was gonochoric with asynchronous development, and planulae were generally brooded. The presence of mature gametes and planulae in P. panamensis polyps suggests that this species is an important contributor to local and, likely, regional recruitment of this species. Further research should seek to identify important source populations for these coral recruits and document the exchange of larvae between coral populations of the Mexican Pacific.     

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.     

Quantification of sexual reproduction in the marine benthic hydroid Campanularia everta

Campanularia everta is an epiphytic hydroid that may form dense populations on the macroalga Halimeda tuna. The main objects of this study were to quantify sexual reporduction and estimate sexual reproductive output of this hydroid. Sexual reproduction occurred from mid-October to mid-December 1991 along the Spanish Mediterranean coast. During this period, male and female colonies formed gonangia. Four oocytes mature in each female gonangium, and embryonic development starts probably after internal fertilization occurs. Planulae complete their development within a mucus sheath attached to the top of the gonotheca (acrocyst). Eight successive gonangia with a life-span of 1 wk each were formed over a 2 mo period. The fertile period was characterized by high initial production of gonangia followed by a progressive decline resulting from a decrease both in the number of fertile colonies and in the gonangia density of fertile colonies. Annual production was estimated at 42000 gonangia m^-2, representing 83000 oocytes m^-2. The high fertilization rates observed (77 to 100%) yielded a minimum production of 64000 planulae m^-2. Reproduction in C. everta is characterized by: (1) a high number of larvae produced m^-2; (2) formation and gradual release of larvae throughout the sexual reproduction period; (3) direct formation of planulae with no intermediate medusa stage; (4) low dispersive ability of the planula. All these mechanisms are part of a reproductive strategy designed to ensure the permanence of the population in its habitat.     

A floating mid-water coral nursery as larval dispersion hub: testing an idea

The global decline in reef health has prompted the need for effective management methodologies, including the development of active restoration measures. One such approach is the ‘gardening concept’ that involves use of underwater nurseries where coral fragments are farmed before their transplantation into denuded reefs. Here we document enhanced sexual reproduction in colonies of the coral Stylophora pistillata cultured in mid-water floating nursery situated in nutrient enriched water, near the fish farms in Eilat, Red Sea. We found that after 2 years of nursery, the average number of oocytes per polyp in farmed colonies was ca. 35% higher than in corresponding naturally growing colonies. Small branches in the nursery developed gravid colonies that released equal (or more) numbers of planula larvae as compared to similar size, 5-year old naturally growing colonies. These nursery-borne planulae possessed more zooxanthellae and contained more chlorophyll per larva. While settled and metamorphosed in equal rates compared to planulae originated from reef-grown colonies, the nursery borne planulae developed faster growing young colonies. We estimate that a coral nursery could generate, during the reproductive season, tens of millions of planula larvae and therefore should be regarded as a ‘larval dispersion hub’ that can be used as a management tool for natural recruitment enhancement.     

Embryogenesis and acquisition of algal symbionts by planulae of Xenia umbellata (Octocorallia: Alcyonacea)

Early embryogenesis of the internally brooding soft coral Xenia umbellata and acquisition of algal symbionts in the course of its planular ontogenesis have been examined by scanning and transmission electron microscopy and by light microscopy. The endoderm of adult X. umbellata harbours symbionts mainly in the tentacles and in the peripheral solenia system. The colonies are gonochoric brooders. Algal symbionts were never found in the sperm sacs, and were only rarely found in the follicular tissue enclosing the oocytes. Fertilized eggs pass into endodermal brood pouches where embryogenesis occurs. Cleavage is holoblastic and leads to formation of a solid blastula. Algal symbionts are conspicuously embedded in the parental mesoglea that coats the young embryo, most probably transmitted by surface adherence. At a further stage, this integument disappears and the algae reside extracellularly among the cells of the newly-formed blastula. After subsequent cell proliferation developing planulae possess an inner mass of yolk-laden cells that contain numerous symbiotic algae. Gradually the yolk disintegrates, leaving a cavity enclosed by ectoderm, a thin mesoglea and an inner endoderm with intracellular symbionts. The mature planulae have already been provided with numerous intracellular symbionts by the time they are expelled from the brood pouches. The markedly early symbiont acquisition by the embryos of X. umbellata may help support their developmental requirements in the course of planular ontogenesis.     

Allozyme electrophoresis demonstrates that the scleractinian coral Montipora digitata is two species

Allozyme electrophoresis at five loci was used to clarify the species status of two morphs of the scleractinian coral Montipora digitata (Dana, 1846), using specimens collected from three locations off the Queensland coast between October 1991 and April 1993. The two morphs, occurring sympatrically, were distinguished by one fixed gene difference at Locus LT-2 and frequency differences at three other loci, indicating reproductive isolation between them. Populations of both morphs were generally in Hardy-Weinberg equilibrium, suggesting that sexual reproduction does occur. Despite sampling designed to reduce the chance of collecting clonemates, the genetic diversity ratio (G _O:G_E) indicated that asexual reproduction also occurs. This is in agreement with the known modes of reproduction for this species. There was significant genetic differentiation between populations of one of the morphs of M. digitata. This could have been influenced by greater degrees of asexual reproduction in this morph, selection within sites, or variation in larval survival and recruitment patterns. Relatively low F _st (differentiation among populations) values found for a broadcast spawning species during this study are in accordance with the hypothesis that brooded planulae are adapted for rapid settlement, whereas planulae produced by broadcast spawners are adapted for widespread dispersal.     

Sexual and asexual production of planulae in reef corals

Electrophoresis was used to provide genetic evidence of the mode of production of brooded planulae in each of four species of scleractinian coral collected from the central region of the Great Barrier Reef during September, October and November 1984. Comparisons were made of the multi-locus genotypes of planulae and their broodparents for two ahermatypic (non zooxanthellate) species,Tubastraea diaphana andT. coccinea and two hermatypic (zooxanthellate) species,Acropora palifera andSeriatopora hystrix. For both ahermatypic species, all planulae were found to be genetically identical to their broodparents, including 91 planulae which were heterozygous for at least one locus. These results are consistent with asexual (ameiotic) reproduction. In contrast, non-parental genotypes were detected in the majority of hermatypic broods, which is consistent with expectations for sexual reproduction with at least some outcrossing. These data confirmed that brooded planulae may be produced both sexually and asexually and countered the suggestion that electrophoretic studies of hermatypic corals may be weakened by the contaminating effect of the enzymes of their symbiotic xooxanthellae.Contribution No. 306 of the Australian Institute of Marine Science     

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.     

An AFLP-based test of clonality in widespread, putatively asexual populations of Porphyra umbilicalis (Rhodophyta) in the Northwest Atlantic with an in silico analysis for bacterial contamination

Asexual reproduction often predominates in populations of species experiencing range expansion. Locally selected genotypes can lead to the establishment of clonal populations that accumulate genetic diversity through time. Sexual reproduction has never been observed in extensive field and culture studies of the red alga Porphyra umbilicalis from Maine, USA, even though sexual reproduction predominates in this species in the eastern Atlantic (Europe). This suggests that Maine populations are indeed asexual and might consist of one or only a few genetic clones; we have tested this using AFLPs. Individuals were sampled at two sites in Maine (Cobscook Bay [n?=?25], Schoodic Point [n?=?26]) and compared to sexual individuals from England (Sidmouth [n?=?17]). The AFLP analysis determined that individuals at two sites in Maine containing putative asexuals were not strictly clonal; however, two multilocus lineages were sampled more than once. Two genetic clones, one at each Maine site, were comprised of 6 individuals each; the 39 additional Maine individuals had distinctive AFLP genotypes. However, when the individuals from Maine were compared with a known sexual population from Sidmouth, England, much greater genetic diversity was found within the sexual population in England. Finally, we examine how preparation of field-collected material for AFLP investigations can affect the inclusion of non-target DNA and demonstrate an in silico approach for removing some cryptic contaminants from analysis.     

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.     

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.     

Reproductive biology of the deep-sea octocoral Drifa glomerata in the Northwest Atlantic

The present study examined the mode and timing of reproduction of poorly understood deep-water octocorals and the environmental factors that may influence their reproductive patterns. Data on reproductive characteristics of the octocoral Drifa glomerata (Alcyonacea: Nephtheidae) collected between 2004 and 2007 at ca. 100–330 m depth off Newfoundland and Labrador (eastern Canada) were compared among years, months and depth ranges. No male gonad was ever observed during the study. The ratio of fertile colonies possessing large pinkish polyps with oocytes/planulae was >50% throughout the year. The number of brooded planula larvae within a single fertile polyp varied between 1 and 10 for a total of approximately 40–3,000 in a whole colony. The size of oocytes and/or planulae was consistently greater in the polyps than in the branchlets, indicating that the development pathway of oocytes to planulae is from the branchlets to the polyps. Although larval production seemed to persist year round, the onset of major planulation events occurred in December–January of two consecutive years, when large mature planulae were released in correlation with the first increase of photoperiod and maximum temperature at 150 m. A second peak in planulation between April and early June followed the phytoplankton bloom. Seasonal trends were more apparent in colonies from <200 m, and the planula index varied among sampling depths and years. Larval release in a live colony under laboratory conditions occurred between January and June 2008, closely following predictions based on field samples.     

Distribution and zoogeography of the marine cladoceran Podon schmackeri in the northwestern Pacific

Embryology of the hermaphroditic and dimorphic alcyonacean Heteroxenia fuscescens has been examined by scanning electron microscopy and light microcopy. Initial embryonic development in this octocoral occurs while the embryos migrate freely inside the anthocodiae and the tentacles. Immature planulae are extruded externally into intersiphonozooid spaces, where they mature. All stages of planular morphogenesis, from egg to planula, occur while the embryo is coated by the original egg mesogleal coat derived from the parent colony. Hatching from this mesogleal coat occurs as late as immediately prior to planulation. H. fuscescens demonstrates highly specialized brood care involving the retention of embryos in internal polyp cavities as well as in external spaces. This highly specialized brood care, coupled with the embryo coating, may provide better protection for the embryo and greater fecundity for the colony.     

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.     

Gonad development and the planulae of the temperate Australian soft coral Capnella gaboensis

Capnella gaboensis Verseveldt, 1977 was sampled at four sites in Sydney Harbour, during 1981–1984. This soft coral has an annual cycle of gonad development, with gonad number reaching a peak in May several weeks prior to spawning, and gonad size reaching a peak in May–June at spawning. The gonads develop during the warm months, and colonies spawn their gametes in late autumn and early winter. Gonad development is neither synchronous within colonies nor within populations, possibly reflecting the protracted nature of spawning. The histology of the developing oocytes and spermaries is described in detail C. gaboensis is a surface-brooder. The planulae are similar in structure to the larvae of other octocorals. The larvae are benthic, settling quickly upon suitable substratum, metamorphosing into polyps with mouths, tentacles and spicules, approximately one week after settling.     

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.     

Genetic structure of fissiparous populations of Holothuria (Halodeima) atra on the Great Barrier Reef

Allozyme variation at five polymorphic loci was surveyed in a total of 311 individuals of the sea cucumber Holothuria (Halodeima) atra (Jäger, 1833) collected from two nearshore and two midshelf populations in the Great Barrier Reef in November 1996. Strong deviations in genotype frequencies from those expected under Hardy–Weinberg equilibrium, particularly a large number of heterozygote excesses, confirmed the occurrence of asexual reproduction. Females and males differed significantly in genotype frequencies as a result of differences in either the amount of fission in each sex or in the mortality of asexually produced recruits. The estimated maximum sexual input (number of sexually produced individuals: sample size = N*:N_i) to the two nearshore reefs (38 to 67%) was low relative to that to the two midshelf reefs (74 to 87%). The three ratios and G _o :G _e , N _go :N _i , N*:N _i , (where G _o = observed genotypic diversity, G _e = expected genotypic diversity, N _go = number of genotypes) considered to be indicators of the extent of asexual reproduction, showed a consistent trend in the degree of asexual reproduction similar to that derived from the number of regenerating individuals observed in the populations for which data were available. F-statistic analyses of clonal gene frequencies demonstrated that all populations received sexual recruits from the same gene pool. There was evidence of restricted sexual recruitment to the Fantome population, suggesting that asexual reproduction was dominant only in areas where sexual recruitment was limited by other factors.     

Queen life-span and total reproductive success are positively associated in the ant Cardiocondyla cf. kagutsuchi

While reproduction and longevity are negatively correlated in a large number of animals, this fundamental trade-off appears to be absent from social insect queens. Here, we examine the distribution of the life-spans of queens and their mates and the age-trajectory of reproduction in experimental colonies of the Pacific tramp ant Cardiocondyla cf. kagutsuchi. Queens of Cardiocondyla are generally short-lived (0.5–2 years), which allows determining their lifetime reproductive success and establishing mortality tables. We show that the queen’s total number of sexual offspring is positively associated with its life-span. Regular counts of eggs, larvae, and pupae suggest that more fecund queens live longer than less fecund queens and that an early onset of sexual production does not negatively affect the queen’s life-span. This corroborates the view that the costs of reproduction are not born by the queen itself. The number of eggs present in colonies increased with queen’s age until shortly before death, indicating negligible reproductive senescence. Sex ratios were strongly affected by the origin of queens but neither by colony size nor total productivity. Several queens produced only males late in their lives, suggesting the occurrence of sperm depletion. Male life-span was not correlated with any other studied trait. Our study shows that the ant genus Cardiocondyla provides a suitable model system to investigate the evolution of life-span in social insect queens and males.     

Sexual reproduction in Acropora (Isopora) (Coelenterata: Scleractinia)

In contrast to the seasonal gamete and planula production of Acropora (Isopora) palifera on Heron Island reef (Lat. 23° S), populations on Lizard Island reef (Lat. 14° S), sampled in 1979, 1981 and 1983, and Salamaua and Busama reefs (Lat. 7° S), sampled from 1980 to 1983, planulated year-round. Intensive sampling of colonies at Salamaua and Busama showed that gametes ripened at two-month intervals and that up to six cycles of gametes and larvae could be produced by an individual colony. Gametes of only a portion of the population — usually close to 50% — ripened each month. The Salamaua population, on average, produced fewer and smaller planulae than the Heron Island population during each two-monthly reproductive cycle. Hypotheses correlating the annual periodicity of breeding in marine animals with latitudinal variation of temperature were tested. In general, the time of breeding in A. palifera at Heron Island reef is much more restricted than theories based on latitudinal variations of water temperature would predict. It is hypothesized that, in the near subtropical environment of Heron Island reef, this species has evolved a life-history strategy that limits the amount of energy allocated to reproduction and allocates more energy to growth.     

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