Sexual and asexual reproduction in geographically separated populations of the fissiparous asteroid Coscinasterias calamaria (Gray)

Sexual reproduction and asexual reproduction by fission were studied in four populations of Coscinasterias calamaria (Gray), two in Otago Harbour in the South Island of New Zealand and two in the North Island near Auckland. The annual reproductive cycle in both islands of New Zealand is clearly defined with a spawning season between November and January. In both sites the pyloric caeca index was approximately inverse to the gonad index cycle as found in other forcipulate asteroids. There are substantial differences in the sex ratios of mature starfish at each site studied, with 1:1 ratios in two populations, one population heavily biased towards females and the fourth consisting almost entirely of males. Morphometric variation in arm number due to splitting was studied and the frequency of splitting varied considerably between the four populations. Generally speaking sublittoral C. calamaria divide less frequently than intertidal starfish and populations in which food is less abundant or of poorer quality are more fissiparous and put less energy into sexual reproduction, than populations with plentiful readily available food in the form of mussels. The significance of the different reproductive patterns in C. calamaria is discussed.     

Reproductive biology of Dugesia sanchezi,a fresh-water planarian from Chile

Dugesia sanchezi reproduces in nature both sexually and by fission. Laboratory cultures have shown that fissioning is controlled by genetic factors. From sexual specimens randomly mated, 105 sexual and five fissiparous offspring were obtained. The former produced almost exclusively sexual descendents while the latter gave rise to fissiparous individuals, although successively many of them became sexual. These ex-fissiparous specimens show a high fertility and their offspring were almost all fissiparous, with the same characteristics displayed by the preceeding fissiparous generation. Some hypotheses are advanced in order to attempt a genetical interpretation of these results.     

Clonal structure through space and time: High stability in the holothurian Stichopus chloronotus (Echinodermata)

Sea cucumbers are increasingly exploited for human consumption and for their curative properties, and many wild populations are now depleted or in danger of extinction. While aquaculture is seen as an alternative to fisheries and as a mean to restore wild populations, more knowledge is needed on their reproductive strategies to render this practice efficient, notably for fissiparous holothurians, which are some of the mobile animals able of asexual reproduction by transverse fission. Little information is available on their population genetic diversity and structure. Here, the clonal structure of populations of the fissiparous sea cucumber Stichopus chloronotus has been investigated using nine microsatellite loci and a random sampling, at different spatial (intra‐reef and inter‐reef) and temporal (inter‐season and inter‐year) scales. Our findings highlight the importance of asexual reproduction in maintaining these populations, and the prevalence of the “initial seedling recruitment” strategy (ISR), leading to a high stability of clonal composition over seasons and years. It also seemed that clonal propagation was limited to the reef scale (<10 km) while reefs were connected by sexual dispersal. This is the first time that clonal structure in sea cucumbers has been studied at such a fine scale, with a specific sampling strategy. It provides key findings on the genetic diversity and structure of fissiparous sea cucumbers, which will be useful for the management of wild populations and aquaculture.     

Studies of the fissiparous holothurian Holothuria parvula (Selenka) (Echinodermata: Holothuroidea)

A Bermudan population of the fissiparous holothurian Holothuria parvula (Selenka) was sampled over a 13-month period (1984–1985). Fission was most frequent in the summer when water temperatures were > 25 °C. During fission, the holothurian split into roughly equal parts, and there was little difference in survival of the oral and anal ends. Regeneration of a new gut is a priority and feeding was possible within 2 months of fission. The majority of growth following fission occurred between April and July, just prior to the peak occurrence of fission. Many individuals were fully regenerated within a year, so fission is possibly an annual event. Individuals showing evidence of multiple fission were found. The capacity for sexual reproduction was limited and it appeared to occur mainly during the summer, which was also the peak period for asexual reproduction. No small (< 18 mm) individuals were ever found suggesting that larval recruitment to this population had not recently been successful. The population has probably been maintained recently by fission.     

Individual recognition and phenotypic variability in Acanthaster planci (Echinodermata: Asteroidea)

Summary A coding method for identifying individuals by utilizing natural attributes (numbers and disposition of arms, madrporites, anuses, spines and deformities) permits individual recognition of crown-of-thorns sea stars Acanthaster planci. Because natural injuries altering parts of the code are frequent in A. planci (present in 20% to 40% of the individuals in a population), the physiognomy of the centrally located periproct, which is much less frequently injured than peripherally located areas, was used in combination with the arrangement of arms and madreporites. Identical madreporite/arm codes occurred in fewer sea stars in Guam (0%) than in Panamá (14%). The greater variability among sea stars at the western Pacific locality could be due in part to the higher incidence of gross deformities (presumed injuries) in Guam (43%) compared with Panamá (17%).     

Regenerative capacity and biochemical composition of the sea star Luidia clathrata (Say) (Echinodermata: Asteroidea) under conditions of near-future ocean acidification

The present study examines sublethal effects of near-future (year 2100) ocean acidification (OA) on regenerative capacity, biochemical composition, and behavior of the sea star Luidia clathrata, a predominant predator in sub-tropical soft-bottom habitats. Two groups of sea stars, each with two arms excised, were maintained on a formulated diet in seawater bubbled with air alone (pH 8.2, approximating a pCO₂ of 380 μatm) or with a controlled mixture of air/C0₂ (pH 7.8, approximating a pCO₂ of 780 μatm). Arm length, total body wet weight, and righting responses were measured weekly. After 97 days, a period of time sufficient for 80% arm regeneration, pyloric caecal indices, and protein, carbohydrate, lipid, and ash levels were determined for body wall and pyloric caecal tissues of intact and regenerating arms of individuals held in both seawater pH treatments. The present study indicates that predicted near-term levels of ocean acidification (seawater pH 7.8) do not significantly impact whole animal growth, arm regeneration rates, biochemical composition, or righting behavior in this common soft bottom sea star.     

Asexual Reproduction and Segmental Regeneration, but not Morphallaxis, are Inhibited by Boric Acid in Lumbriculus variegatus (Annelida: Clitellata: Lumbriculidae)

Body fragmentation, in some animal groups, is a mechanism for survival and asexual reproduction. Lumbriculus variegatus (Müller, 1774), an aquatic oligochaete worm, is capable of regenerating into complete individuals from small body fragments following injury and reproduces primarily by asexual reproduction. Few studies have determined the cellular mechanisms that underlie fragmentation, either regenerative or asexual. We utilized boric acid treatment, which blocks regeneration of segments in amputated fragments and blocks architomic fission during asexual reproduction, to investigate mechanistic relationships and differences between these two modes of development. Neural morphallaxis, involving changes in sensory fields and giant fiber conduction, was detected in amputated fragments in the absence of segmental regeneration. Furthermore, neural morphallactic changes occurred as a result of developmental mechanisms of asexual reproduction, even when architomy was prevented. These results show that fragmentation in L. variegatus, during injury or asexual reproduction, employs developmental and morphallactic processes that can be mechanistically dissociated by boric acid exposure. In regeneration following injury, compensatory morphallaxis occurred in response to fragmentation. In contrast, anticipatory morphallaxis was induced in preparation for fragmentation during asexual reproduction. Thus, various forms of regeneration in this lumbriculid worm can be activated independently and in different developmental contexts.     

Adaptive advantages of patterns of growth and asexual reproduction of the sea anemone Metridium senile (L.) in intertidal and submerged populations

Individual size, rate of growth, and mode and frequency of asexual reproduction are life-history traits of primary importance for sea anemones. These traits determine sexual reproductive output, affect an individual's probability of survival, and are crucial in adapting an individual to its environmental surroundings. The sea anemone Metridium senile (L.) is highly variable in ecological distribution and life history, including rate of growth, individual size, and rate of asexual reproduction. Gonad size (measured as cross-sectional area of gonadal tissue) increases with body weight, so individuals should grow as large and as rapidly as possible to maximize individual sexual reproductive output. Cessation of growth and small body size in intertidal populations suggest that growth is constrained by genetic or environmental conditions. The growth of intertidal individuals transplanted to harbor-float panels demonstrated that growth limits are imposed by environmental factors, most probably limited food and feeding time and damage from wave exposure (which stimulates fragmentation). Individuals in harbor-float populations, which are continuously immersed, grow much larger, and large individuals comprise a greater proportion of the population than in the intertidal zone. The highest rate of fragmentation observed was on harbor-float panels. Patterns of growth and asexual reproduction provide adaptive advantages for M. senile. For harborfloat individuals, large individual size increases gamete production and may increase feeding efficiency. For intertidal individuals, asexual reproduction allows growth despite individual size constraints and rapid population growth, with specific advantages resulting from clone formation.     

A non‐lethal method for estimation of gonad and pyloric caecum indices in sea stars

Abstract. Gonad and pyloric caecum indices are widely used indicators of reproductive effort and nutritional condition in asteroids. Current methods of quantification generally require sacrificing multiple animals and the resulting reduction in local sea star density could have an unintended impact on benthic communities. Using the intertidal sea star Pisaster ochraceus, we developed and tested a method for estimating organ indices through the non‐lethal sampling of single arms. Indices estimated via dissections of single arms accurately predicted the values obtained by sacrificing whole animals. In laboratory and field trials, we compared two methods of sampling single arms: (1) arm removal, and (2) organ extraction through an incision (without arm removal). Two years after these treatments, organs had regenerated in the affected arms of most sea stars, but were small relative to those in unmanipulated arms. The extent of organ recovery did not differ between the two treatments, but sea stars in the arm removal group were relocated in the field more frequently than those in the arm incision group. Our results suggest that sampling via the removal of single arms can be an effective, non‐lethal method for estimating gonad and pyloric caecum indices in asteroids.     

Patterns of asexual reproduction in the fissiparous seastar Coscinasterias acutispina (Asteroidea: Echinodermata) in Japan

The morphological and anatomical characteristics of the fissiparous seastar Coscinasterias acutispina were investigated at 23 sites around Japan in order to clarify the environmental factors determining asexual reproduction. The frequency of recent fission among the sites differed, although no significant differences were evident among the three major sea areas around Japan (the Sea of Japan, Seto Inland Sea, and Pacific Ocean), indicating that the spatial variability in incidence of fission might be determined on a local scale. Morphological observations revealed that the seastars endoparasitized by Dendrogaster okadai had not undergone fission recently, suggesting that fission was suppressed by parasitism. Stepwise multiple regression analysis of the frequency of seastars that had recently undergone fission revealed that nutrient conditions affected the incidence of fission. The relationships between the pyloric caeca index and arm length ratio at the individual level suggested that good nutritional conditions were required before the occurrence of fission. Additionally, regression analysis showed that high temperature was one of the triggers of fission in C. acutispina. Thus, the process of asexual reproduction in C. acutispina appears to be governed by intricate interactions among several factors, including parasitism by D. okadai, nutrient conditions, and high temperature.     

Effect of the degree of autotomy on feeding, growth, and reproductive capacity in the multi-armed sea star Heliaster helianthus

The term autotomy refers to the process by which some species lose limbs or parts of limbs in response to adverse biotic or abiotic conditions, as for example, predation or abnormally high temperatures. The multi-armed sea star Heliaster helianthus is a key predator of the intertidal and the shallow rocky subtidal communities of north-central Chile. Natural populations of this sea star have been found with up to 60% of the individuals showing some degree of autotomy. The present study evaluated the effects of autotomy on feeding rate and growth of juvenile and adult H. helianthus after experimentally induced autotomy of 17% and 33% of their arms, as well as on the energy content of the pyloric caeca and gonads of adults during the reproductive period. Experimental juvenile sea stars were maintained and fed in the laboratory over a period of five months and adult sea stars for one month, Intact individuals were maintained as parallel controls. The results showed that juveniles undergoing 33% autotomy decreased their feeding rates, and as a consequence showed lowered net individual growth. In contrast, adults with 17% and 33% autotomy showed marked reductions in feeding. The results showed that autotomized adults had between five and seven times lower contents of carbohydrates, lipids, and proteins (and thus energetic content) in their pyloric caeca and gonads. The loss of the arms not only decreased the capacity for feeding in sea stars, but also allocated energy away from growth and reproduction into the process of regeneration of arms. This suggests that autotomy reduces the fitness of H. helianthus. Growth was reduced in the juveniles, while adults became limited in their ability to store energy which then limited their reproductive potential. Finally, based on the important effect of autotomy on reducing the feeding capacity of H. helianthus, the role of this sea star as a predator in the environment may be strongly affected.     

Effects of environmental stress on intertidal mussels and their sea star predators

Consumer stress models of ecological theory predict that predators are more susceptible to stress than their prey. Intertidal mussels, Mytilus californianus, span a vertical stress gradient from the low zone (lower stress) to the high zone (higher thermal and desiccation stress), while their sea star predators, Pisaster ochraceus, range from the low zone only into the lower edge of the mussel zone. In summer 2003, we tested the responses of sea stars and mussels to environmental stress in an experiment conducted on the Oregon coast. Mussels were transplanted from the middle of the mussel bed to cages in the low and high edges of the mussel bed. Sea star predators were added to half of the mussel cages. Mussels and sea stars were sampled between June and August for indicators of sublethal stress. Mussel growth was measured, and tissues were collected for heat shock protein (Hsp70) analyses and histological analyses of reproduction. Sea stars were weighed, and tissues were sampled for Hsp70 analyses. Mussels in high-edge cages had higher levels of total Hsp70 and exhibited spawning activity earlier in the summer than mussels in the low-edge cages. Sea stars suffered high mortality in the high edge, and low-edge sea stars lost weight but showed no differences in Hsp70 production. These results suggest that stress in the intertidal zone affected the mobile predator more than its sessile prey, which is consistent with predictions of consumer stress models. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Autotomy reduces feeding, energy storage and growth of the sea star Stichaster striatus

We evaluated the effect of autotomy on feeding, energy storage and growth of juvenile Stichaster striatus kept in the laboratory for five months with a limited supply of the mussel Semimytilus algosus. Autotomy strongly decreased feeding, energy storage and growth. Intact juveniles showed a ∼ 3 fold higher feeding rate than autotomized individuals throughout the experiment. Intact juveniles also had a higher (∼ 5 fold) energy content per pyloric caeca in each arm. This was mainly due to higher lipid content, the main proximate constituent of pyloric caeca. Intact juveniles showed a greater growth rate and reached a greater size than autotomized individuals, more evident for underwater mass than radius length. The reduced capacity to feed reduced energy intake in autotomized individuals. However, low energy reserves along with low growth in autotomized sea stars, support the hypothesis that juveniles of this species allocate energy to regeneration to the detriment of growth. This was also supported by the ∼ 25% of arm length regeneration after 5 mo. Remaining small could increase risk of lethal predation, however, S. striatus may reduce predation risk by using crevices and kelp holdfasts as refuges from predators. Given the strong impact of autotomy on feeding, regeneration of arms to recover full capacity to forage and grow seems a better strategy for juvenile S. striatus, than merely growing.     

Switching from asexual to sexual reproduction in the planarian Dugesia ryukyuensis: change of the fissiparous capacity along with the sexualizing process

Asexual worms of fissiparous strain of the planarian Dugesia ryukyuensis switch from asexual to sexual reproduction, if they are fed with sexually mature worms of Bdellocephala brunnea. This suggests that the sexually mature worms have a sexualizing substance(s) that induces the sexuality in the asexual worms. Here, we found by analysis of the sexualization that the cessation of the fission, namely their asexual reproduction, occurs immediately after the acquisition of sexuality. This result suggests that the downstream mechanisms induced by the putative sexualizing substance in B. brunnea become responsible for the cessation of fission. We also found that the decapitation triggers fission in the worms even after the acquisition of sexuality if they are not sexually mature, while the fully sexualized worms never fission even though they are decapitated. This result suggests that the cessation of fission takes place via at least two steps: (1) the mechanisms associated with the cephalic system; (2) other mechanisms independent of cephalic control.     

Sexual and asexual reproduction of the holothurian Stichopus chloronotus (Echinodermata): a comparison between La Réunion (Indian Ocean) and east Australia (Pacific Ocean)

Summary

Stichopus chloronotus (Brandt, 1835) is one among nine aspidochirotide holothurian species known to reproduce both sexually by broadcast spawning and asexually by transverse fission. New data on the sexual cycle of this species in La Réunion are presented here and information available on sexual and asexual reproduction in this species is summarised. Sexual reproduction on La Réunion shows a distinct seasonality with a main spawning period in the warm season (November-February). The spawning period the Great Barrier Reef appears to be at the same time. Some intriguing deviations from unity in sex-ratio, usually biased towards more male individuals, have been observed in both geographic regions (sex ratio at La Reunion 31:1). New data on the asexual reproduction of this species in La Réunion confirm the high rates of fission. The peak of asexual reproduction in both the Indian and Pacific Ocean was observed in winter (June-July). Thus, asexual reproduction in this species occurs outside the season for sexual reproduction. The rate of asexual reproduction appears to vary between sample locations. However, results of population genetic studies on S. chloronotus (Uthicke et al., 1999; Uthicke et al., 2001) indicated that in most populations investigated a maximum of about 60% of all individuals may be derived from sexual recruitment. Cluster analyses on genetic distances between populations grouped populations within Oceans together, with the exception of one sample from a nearshore reef of the GBR. Although genetic differences between the two regions exist, these are relatively small regarding the large geographic distance. We conclude that asexual reproduction in S. chloronotus is important to maintain local population sizes, but that larval exchange between populations mediated by sexual reproduction is important for colonisation of new areas and to provide connectivity between populations. Here, we present the first synthesis of these phenomena for a holothurian species.     

<Emphasis Type="Italic">Afuni</Emphasis>, a novel transforming growth factor-β gene is involved in arm regeneration by the brittle star <Emphasis Type="Italic">Amphiura filiformis</Emphasis>

The bone morphogenetic proteins (BMPs) are a family of the transforming growth factor- (TGF-) superfamily that perform multiple roles during vertebrate and invertebrate development. Here, we report the molecular cloning of a novel BMP from regenerating arms of the ophiuroid Amphiura filiformis. The theoretically translated amino acid sequence of this novel BMP has high similarity to that of the sea urchin BMP univin. This novel BMP has been named afuni. Whole-mount in situ hybridisation implicates afuni in arm regeneration. Expression occurs in distinct proximal and distal regions of late regenerates (3- and 5-week postablation). These sites are at different stages of regeneration, suggesting multiple roles for this gene in adult arm development. Cellular expression of this gene occurs in migratory cells within the radial water canal (RWC) of regenerating and nonregenerating arms. These migrating coelomocytes suggest a key role for the coelomic RWC as a source of the cellular material for use in arm regeneration by A. filiformis.     

Growth and asexual reproduction of the sea anemone Metridium: comparative laboratory studies of three species

The rate of growth, individual size attained, and mode and frequency of asexual reproduction were investigated in three species of the sea anemone Metridium. The species differ in ecological distribution, characteristic habitat, and morphology. M. exilis Hand is characteristically intertidal and occurs in aggregations of numerous small (0.2 cm² pedal disc area, pda) individuals. Growth in laboratory culture ceased at 1.2 cm² pda despite copious feeding. Feeding accelerated the rate of asexual reproduction by binary fission but did not result in larger individuals. Individuals of M. senile (L.) are larger and reproduce both sexually and asexually by fragmentation. The species occurs from the mid-intertidal to the shallow subtidal in central California. Individuals from intertidal populations grew rapidly in laboratory culture when fed daily (mean pda after 5 months in culture was 45 cm²). The regenerative ability of M. senile is excellent: 100% of excised fragments (including column and pedal disc tissue) had completely regenerated, including tentacle formation, after 3 wk with no mortality. An undescribed species of Metridium is much larger (individuals grow >/ l m in height) and apparently is exclusively subtidal in central California. Asexual reproduction is rare or absent and no pedal lacerates were produced by individuals during the period of study. The regenerative ability of Metridium sp. is poor: 53% of the excised fragments had regenerated after 19 wk with 25% mortality. The three species thus show distinct patterns of growth and asexual reproduction, paralleling their morphological, ecological, and genetic distinctness.     

Observations on Fission, Maturation and Reproduction in Dugesia biblica (Turbellaria, Tricladida) from Israel

Laboratory studies on the normally fissiparous Dugesia biblica from Israel (3 n =27+ x supernumeraries) have been made in order to investigate fission rates and spontaneous maturation of four fissioning populations and one clone. Laboratory induced maturation in a sixth population of D. biblica resulted in the laying of fertile cocoons, the offspring of which were either 3 n =27+ x , 2 n = 18+ x or 2 n = 18+ x It is suggested that triploidy is primarily responsible for the lack of sexual reproduction in the field, and that unequal inheritance of supernumerary chromosomes during mitosis may explain the differing tendencies to undergo spontaneous maturation by individuals of a clone of D. biblica. The karyotype, habitat and reproduction of D. biblica is compared with a closely related but diploid, normally sexually reproducing Dugesia form and possible relationships between these two are discussed.     

Early segregation of germ and somatic lineages during gonadal regeneration in the annelid Enchytraeus japonensis

Although regeneration studies are useful for understanding how organs renew, little information is available about regeneration of reproductive organs and germ cells. We here describe the behavior of germ-cell precursors during regeneration of the oligochaete annelid worm Enchytraeus japonensis, which has the remarkable feature of undergoing asexual (by fission) and sexual reproduction . We first found that the gonad can regenerate from any body fragment yielded by fission during asexual reproduction. We then examined behavior of germ-cell lineage during this regenerative process, by using a homolog of the Piwi gene (Ej-piwi) as a marker. We found that in asexually growing animals, specialized cells expressing Ej-piwi are distributed widely in the body as single cells. These cells seem to serve as a reservoir of germ-cell precursors because during asexual propagation these cells migrate into the regenerating tissue, where they ultimately settle in the prospective gonads, and give rise to germ cells upon sexualization. These cells are distinct from the neoblasts, thought to be stem cells in other animals. This is the first report to directly show that the germ and somatic lineages are segregated in asexually growing animals and behave differently during regeneration.     

Population size structure,growth, arm number and damage in the sea star Archaster angulatus Müller and Troschel, 1842 (Echinodermata: Asteroidea)

Monthly sampling for the sea star Archaster angulatus revealed at least four cohorts with linear growth in the 0+ and 1+ year groups to about 26 months after which growth slows and conforms to the von Bertalanffy model of growth. The appearance of small individuals (arm radius (R) < 20 mm) from March to May is consistent with the discrete December/January spawning period. Sea stars attain R of 38.7 mm at one year, 73.7 mm at two years and maturity occurs in the second year. The 3+ year and 4+ year cohorts were difficult to detect reliably, but sample number peaks at 90–94 mm and 105–107 mm in January of both 2010 and 2011 are indicative of size at three and four years. Two methods of estimating von Bertalanffy growth parameters both yielded estimates of 127 mm for L_inf and k values of 0.41 and 0.49. Comparison of size at age estimates from growth models and cohort analysis suggested the actual value of k after maturity lies between 0.35 and 0.45. Four-armed (1.48%), six-armed (1.26%) and seven-armed (0.11%) individuals were present in the population and arm damage, thought to have resulted from sublethal predation was observed in 13.8% of sea stars.