Do reproductive tactics vary with habitat heterogeneity in the intertidal sea anemone Actinia tenebrosa?

Cnidarians display a diverse range of reproductive tactics including sexual and asexual modes of reproduction. Although few studies have looked for intraspecific variation in reproductive tactics, flexible expression of such life-history traits may be favoured in species that occupy a range of habitats. We tested this in the sea anemone Actinia tenebrosa by comparing cycles of reproductive activity and the mode of production of brooded larvae in local populations occupying boulder fields and stable rock platforms. We determined the mode of production of broods from eight rock platforms (separated by up to 1600 km) and two boulder shores on the south east coast of Australia using a combination of allozyme data and four newly characterised microsatellite markers.

We determined seasonal patterns of brooding and gonad development by monthly dissection of 15–30 adults from each of two boulder fields and two stable platforms. Previous genetic studies have shown that populations of A. tenebrosa on rock platforms can be highly clonal, whereas anemones on more heterogeneous boulder habitats display levels of genotypic diversity similar to that expected for sexual reproduction. We genotyped a total of 221 juveniles from 37 brooding adults including 11 broods and 80 juveniles from boulder shores. We did not detect any evidence of sexual production of broods. All brooded juveniles displayed identical multi-locus genotypes to their brood parent irrespective of habitat of origin or location, including 28 broods (200 juveniles) that were heterozygous at one or more locus. Similarly, we found that temporal patterns of gonad formation and brooding were consistent across habitats and locations. We detected 346 mature males, 234 non-reproductive or immature individuals, and no mature females within a total of 580 dissected individuals. These data suggest that the reproductive tactics of A. tenebrosa are essentially fixed and that variation in the genotypic diversity of populations may reflect variation in factors such as the input of sexually derived planktonically dispersed recruits or post-settlement processes. However, the apparent lack of females paradoxically implies that sexual reproduction, and hence recruitment, must be rare or no longer possible within some populations, and highlights the need for long-term studies of these populations.

Relações evolutivas entre Passiflora actinia Hooker e Passiflora elegans masters (Passifloraceae)

Em estudos prévios sobre a filogenia de Passiflora, as espécies P. actinia e P. elegans destacaram-se pela sua grande similaridade genética, apesar de sua classificação em séries taxonômicas distintas. As duas espécies apresentam distribuição geográfica muito diferente. Enquanto P. actinia é encontrada em áreas de Mata Atlântica desde o estado do Espírito Santo até o Rio Grande do Sul (RS), P. elegans está restrita ao RS e a poucas regiões limítrofes. Para melhor avaliar as relações evolutivas entre estas duas espécies foram realizadas coletas intensivas em todo o estado e desenvolvidos testes quanto às seqüências dos espaçadores intergênicos cloroplasmáticos trnL-trnF e psbA-trnH, e dos espaçadores transcritos dos genes ribossomais nucleares ITS de plantas de diferentes localidades. As análises revelaram uma baixa variabilidade intraespecífica, e evidenciaram um perfil genético próprio a cada espécie. Nas comparações interespecíficas, foram utilizadas seqüências de espécies do subgênero (Passiflora) estudadas previamente, pertencentes às séries Simplicifoliae e Lobatae, as mesmas de P. actinia e P. elegans, respectivamente. Nos três marcadores as menores distâncias genéticas encontradas foram entre estas duas espécies, sugerindo o pouco tempo de divergência entre elas. Estas comparações não mostraram diferenças marcantes nas diversidades dentro e entre as duas séries, indicando similaridade genética entre elas Apesar da intensa amostragem realizada na área limítrofe das distribuições de P. actinia e P. elegans, somente foi encontrado um híbrido entre as duas. Além do fenótipo morfológico intermediário, o híbrido pôde ser reconhecido através das suas características genéticas, o espaçador nuclear ITS apresentando padrão aditivo nos sítios variáveis destas duas espécies; as seqüências dos marcadores cloroplasmáticos foram iguais às de P. actinia, indicando que esta é a espécie doadora deste genoma. Os padrões genéticos e geográficos destas duas espécies sugerem que o processo de especiação que se desenvolveu entre as duas seja recente e tenha ocorrido em alopatria, estando provavelmente ligado aos eventos geológicos do Holoceno que influenciaram a migração da Mata Atlântica no RS. A investigação das características abióticas das regiões de ocorrência das espécies não apresentou grandes dissimilaridades, podendo indicar que a atual segregação espacial deva-se à fragmentação florestal ou que haja exclusão competitiva entre elas, pois apresentam nichos ecológicos muito semelhantes.

Absence of consistent genetic differentiation among several morphs of Actinia (Actiniaria: Actiniidae) occurring in the portuguese coast

Actinia equina, the beadlet sea anemone, is a very labile species, displaying variable colour patterns, broad habitat choice and diverse modes of reproduction. Historically, studies using genetic markers such as allozymes and differences in habitat choice lead several authors to propose that different colour morphs could represent different species. One of the species defined was A. fragacea. In this paper, the relationships between brown, red and green colour morphs of A. equina and A. fragacea were studied, using two DNA fragments (one mitochondrial and one nuclear). Individuals were sampled from three different areas in Portugal separated by a maximum distance of 500 km. This is the first study applying direct sequencing of selected gene fragments to approach the validity of Actinia morphs as different genetic entities. The results show that, at least in the Portuguese coast, these colour morphs do not correspond to the two valid species recognized in the literature. The existence of cryptic species is discussed.

Asexual origin of brooding in the sea anemones Actinia equina and A. schmidti: Molecular evidence from the portuguese shore

Reproduction by sexual or asexual viviparity is a common phenomenon in some anemone species. In this short communication, the origin of the brooded young of Actinia equina and A. schmidti from the Portuguese shore was investigated. DNA was extracted from 56 brooding adult Actinia sp. and the nuclear gene that codes for the 28S ribosomal subunit was sequenced. Species identity was then assessed using GenBank. In total, 50 individuals were A. schmidti, five were A. equina and one had a hybrid origin. Three adult anemones (the hybrid, one A. equina and one A. schmidti) possessed two different 28S sequences and so their offspring was selected for further analysis using the same molecular procedure. Each brooded polyp was found to possess the exact same sequence as its parent, strongly suggesting the asexual origin of broods in A. equina and A. schmidti.

Proteomics of the neurotoxic fraction from the sea anemone Bunodosoma cangicum venom: Novel peptides belonging to new classes of toxins

In contrast to the many studies on the venoms of scorpions, spiders, snakes and cone snails, tip to now there has been no report of the proteomic analysis of sea anemones venoms. In this work we report for the first time the peptide mass fingerprint and some novel peptides in the neurotoxic fraction (Fr III) of the sea anemone Bunodosoma cangicum venom. Fr III is neurotoxic to crabs and was purified by rp-HPLC in a C-18 column, yielding 41 fractions. By checking their molecular masses by ESI-Q-Tof and MALDI-Tof MS we found 81 components ranging from near 250 amu to approximately 6000 amu. Some of the peptidic molecules were partially sequenced through the automated Edman technique. Three of them are peptides with near 4500 amu belonging to the class of the BcIV, BDS-I, BDS-II, APETx1, APETx2 and Am-II toxins. Another three peptides represent a novel group of toxins (similar to 3200 amu). A further three molecules (similar to similar to 4900 amu) belong to the group of type 1 sodium channel neurotoxins. When assayed over the crab leg nerve compound action potentials, one of the BcIV- and APETx-like peptides exhibits an action similar to the type 1 sodium channel toxins in this preparation, suggesting the same target in this assay. On the other hand one of the novel peptides, with 3176 amu, displayed an action similar to potassium channel blockage in this experiment. In summary, the proteomic analysis and mass fingerprint of fractions from sea anemone venoms through MS are valuable tools, allowing us to rapidly predict the occurrence of different groups of toxins and facilitating the search and characterization of novel molecules without the need of full characterization of individual components by broader assays and bioassay-guided purifications. It also shows that sea anemones employ dozens of components for prey capture and defense. (C) 2008 Elsevier Inc. All rights reserved.

Fine-scale adaptation in a clonal sea anemone

Local adaptation in response to fine-scale spatial heterogeneity is well documented in terrestrial ecosystems. In contrast, in marine environments local adaptation has rarely been documented or rigorously explored. This may reflect real or anticipated effects of genetic homogenization, resulting from widespread dispersal in the sea. However, evolutionary theory predicts that for the many benthic species with complex life histories that include both sexual and asexual phases, each parental habitat patch should become dominated by the fittest and most competitive clones. In this study we used genotypic mapping to show that within headlands, clones of the sea anemone Actinia tenebrosa show restricted distributions to specific habitats despite the potential for more widespread dispersal. On these same shores we used reciprocal transplant experiments that revealed strikingly better performance of clones within their natal rather than foreign habitats as judged by survivorship, asexual fecundity, and growth. These findings highlight the importance of selection for fine-scale environmental adaptation in marine taxa and imply that the genotypic structure of populations reflects extensive periods of interclonal competition and site-specific selection.

Ecologia de actiniários no entremarés rochoso: mensuração do tamanho, estrutura da comunidade e avaliação do deslocamento

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)