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.

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.

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.

Distribuição geográfica da fauna e flora da Baía de Guanabara

The author studied, the horizontal and vertical distribution of most common part of the flora and fauna of the bay of Guanabara at Rio de Janeiro. In this paper the eulittoral, poly, meso and oligohaline regions were localised and studied; and the first chart of its distribution was presented (fig. 2). The salinity of superficial waters was established through determinations based on 30 trips inside the buy for collecting biological materials. Some often 409 determinations which were previous reported together with the present ones served for the eleboration of a salinity map of the bay of Guanabara (fig. 1). This map of fig. 2 shows the geographic locations of the water regions. EULITTORAL WATER REGIME — Fig. 3 shows the diagram scheme of fauna and flora of this regime. Sea water salinity 34/1.000, density mean 1.027, transparent greenish waters, sea coast with moderate bursting waves. Limpid sea shore with white sand, gneiss with the big barnacle Tetraclita squamosa var. stalactifera (Lam. Pilsbry. Vertical distributions: barna¬cles layers with a green region in which are present the oyster Ostrea pa-rasitica L., the barnacles Tetraclita, Chthamalus, Balanus tintinnabulum var. tintinnabulum (L.) e var. antillensis Pilsbry in connection with several mollusca and the sea beatle Isopoda Lygia sp. Covered by water and exposed to air by the tidal ritms, there is a stratum of brown animals that is the layer of mussels Mytilus perna L., with others brown and chestnut animals : the Crustacea Pachygrapsus, the little crab Porcellana sp., the stone crab Me-nippe nodifrons Stimpson, the sea stars Echinaster brasiliensis (Mull. & Tr.), Astropecten sp. and the sea anemones Actinia sp. Underneath and never visible there is a subtidal region with green tubular algae of genus Codium and amidst its bunches the sea urchin Lycthchinus variegatus (Agass.) walks and more deeply there are numerous sand-dollars Encope emarginata (Leske). The microplancton of this regime is Ceratiumplancton. POLYHALINE WATER REGIMB — Water almost sea water, but directly influenced by continental lands, with rock salts dissolved and in suspension. Salinity: 33 to 32/1.000. This waters endure the actions of the popular nicknamed «water of the hill» (as the waters of mesohaline and oligohaline regimes), becoming suddenly reddish during several hours. That pheno¬menon returns several times in the year and come with great mortality of fishes. In these waters, according to Dr. J. G. FARIA there are species of Protozoa : Peridinea, the Glenoidinium trochoideum St., followed by its satellites which he thinks that they are able to secret toxical substances which can slaughter some species of fishes. In these «waters of the hill» was found a species of Copepoda the Charlesia darwini. In August 1946 the west shore of the Guanabara was plenty of killed fishes occupying a area of 8 feet large by 3 nautical miles of lenght. The enclosure for catching fishes in the rivers mouthes presents in these periods mass dead fishes. The phenomenon of «waters of the hill» appears with the first rains after a period of long dryness. MESOHALINE WATER REGIME — Fig. 4 shows the the diagramm scheme. Salt or brackish water from 30 to 17/1.000 salinity, sometimes until 10/1.000. Turbid waters with mud in suspension, chestnut, claveyous waters; shore dirty black mud without waving bursting; the waters are warmer and shorner than those of the polihaline regime. Mangrove shore with the mangrove trees : Rhizophora mangle L., Avicennia sp., Laguncularia sp., and the »cotton tree of sea» Hibiscus sp. Fauna: the great land crab «guaimú» Cardisoma guanhumi Latr., ashore in dry firm land. There is the real land crab Ucides cordatus (L.) in wetting mud and in neigh¬ bourhood of the burrows of the fiddler-crabs of genus Uca. On stones and in the roots of the Rhizophora inhabits the brightly colored mangrove-tree-crab («aratu» Portuguese nickname) Goniopsis cruentata (Latreille) and the sparingly the big oyster Ostrea rhizophorae Guild. Lower is the region of barnacles Balanus amphitrite var. communis Darwin and var. niveus Darwin; Balanus tintinnabulum var. tintinnabulum (L.) doesn't grow in this brackish water; lower is the region of Pelecipoda with prepollency of Venus and Cytherea shell-fishes and the Panopeus mud crab; there are the sea lettuce Ulva and the Gastreropod Cerithium. The Paguridae Clibanarius which lives in the empty shells of Gasteropod molluscs, and the sessile ascidians Tethium plicatum (Lesuer) appears in some seasons. In the bottom there is a black argillous mud where the «one landed shrimps» Alpheus sp. is hidden. OLIGOHALINE WATER REGIME — The salinity is lower than 10/1.000. average 8/1.000. There are no barnacles and no sea-beetles Isopods of genus Lygia; on the hay of the shore there are several graminea. This brackish water pervades by mouthes of rivers and penetrates until about 3 kilometers river above. While there is some salt dissolved in water, there are some mud crabs of the genus Uca, Sesarma, Metasesarma and Chasmagnatus. The presence of floating green plants coming from the rivers in the waters of a region indicated the oligohaline waters, with low salt content because when the average of NaCl increases above 8/1.000 these plants die and become rusty colored.

Enxertia de maracujazeiro-azedo em estacas herbáceas enraizadas de espécies de passifloras nativas

As doenças provocadas por patógenos do solo em maracujazeiro constituem-se em um dos principais problemas para essa cultura no Brasil. Uma das alternativas de controle dessas doenças seria a utilização de porta-enxertos resistentes. Várias espécies de passifloras nativas vêm apresentando resistência a essas doenças, mas a utilização destas como porta-enxertos oriundos de sementes tem sido dificultada pelas diferenças de diâmetro entre o porta-enxerto e o enxerto da espécie comercial, o que não aconteceria caso fossem utilizadas as estacas herbáceas como porta-enxerto. No presente experimento, utilizaram-se estacas herbáceas retiradas da parte mediana de ramos de plantas de Passiflora setacea (acesso EC-PS 1), P. nitida (acesso EC-PN 1), P. caerulea (acesso EC-PC 1), P. actinia (acesso EC-PA 1) e de um híbrido F1 entre P. setacea x P. edulis f. flavicarpa comercial e tratadas com ácido naftaleno acético (ANA) a 500 mg/L e mantidas em câmaras de nebulização. As enxertias do tipo "garfagem lateral no topo" foram efetuadas aos 40; 55 e 70 dias após a coleta e plantio das estacas, utilizando garfos de uma única planta de maracujazeiro-azedo. As avaliações foram efetuadas aos 145 e 150 dias após o plantio das estacas, determinando-se a porcentagem de pegamento da enxertia e de enxertos brotados e o comprimento do broto do enxerto. A produção de mudas por enxertia em estacas herbáceas enraizadas de Passiflora nitida e do híbrido F1 (P. setacea x P. edulis f. flavicarpa) foi tecnicamente viável.

Enraizamento de estacas de três espécies silvestres de Passiflora

Em ambiente com nebulização controlada, estacas herbáceas com um par de folhas, contendo 2 ou 3 nós, foram testadas quanto ao enraizamento, utilizando-se de bandeja de poliestireno com célula de 95cm³ e saco plástico de 15x25x0,02cm com 1.730 cm³. Foram testadas estacas de Passiflora actinia, P. serrato-digitata e P. setacea. Observou-se que P. serrato-digitata apresentou 94,3% de estacas enraizadas com brotos e 2,4% de mortalidade; enquanto P. actinia e P. setacea apresentaram, respetivamente, 30,5% e 28,6% de estacas enraizadas com brotos e 56,8% e 60,7% de mortalidade. A alta mortalidade das estacas foi atribuída ao estado fenológico das matrizes de P. actinia e P. setacea e ao ataque de larvas de bradisia (Bradysia spp.) Estacas com dois e três nós não apresentaram diferenças significativas, e o recipiente saco plástico de 1.730 cm³ proporcionou melhor desenvolvimento das mudas.

Confirmação de híbridos interespecíficos artificiais no gênero passiflora por meio de marcadores RAPD

A obtenção de híbridos interespecíficos de maracujazeiro é um processo de grande valia para proporcionar ganhos agronômicos à espécie comercial Passiflora edulis em programas de melhoramento genético, obter novos materiais genéticos com potencial para uso como porta-enxertos e também como alternativas para o mercado de plantas ornamentais. Neste trabalho, marcadores moleculares RAPD foram utilizados visando à confirmação do sucesso de 17 hibridações interespecíficas. Amostras de DNA genômico do suposto híbrido e de seus prováveis genitores foram extraídas, e 12 primers decâmeros foram utilizados para a obtenção de marcadores RAPD. Os marcadores gerados foram analisados quanto à presença ou não de bandas informativas para a confirmação da fecundação cruzada. Foram confirmados os cruzamentos P. laurifolia x P. nitida; P. edulis f. flavicarpa GA2 x RC1 (GA2 x P. coccinea); P. caerulea x P. amethystina; P. glandulosa x P. galbana; P. coccinea x P. actinia; P. glandulosa x P. edulis f. flavicarpa GA2; P. sidaefolia x P. actinia; P. galbana x P. actinea; F1 (P. coccinea x P. setacea) x P. coccinea; F1 (P. coccinea x P. setacea) x P. mucronata; P. eichleriana x P. gibertii; P. galbana x P. edulis f. flavicarpa GA2; P. glandulosa x P. edulis edulis cinza TO; P. glandulosa x P. sidaefolia; P. coccinea x P. setacea. Constatou-se a existência de compatibilidade genética entre essas espécies, sendo possível a sua utilização em programas de melhoramento. Os marcadores RAPD mostraram-se excelentes ferramentas para verificar a ocorrência da fecundação cruzada no gênero Passiflora.

A wolf in sheep"s clothing: carnivory in dominant sea urchins in the Mediterranean

Arbacia lixula and Paracentrotus lividus are the dominant sea urchins in the Mediterranean sublittoral, where they are key structuring species due to their grazing activity. It has been commonly accepted that competition between both species is minimized by specializing in different algal foods. A. lixula is considered to feed mainly on encrusting coralline algae, while P. lividus prefers fleshy macroalgae. We used stable isotope analysis to test if these species occupy different trophic positions at 3 locations in the western Mediterranean and one in Macaronesia. Our results show unambiguously that A. lixula always occupies a higher trophic level than P. lividus, with a δ15N comparable in some locations to strict carnivores such as Actinia schmidti or Marthasterias glacialis. A temporal monitoring at one locality showed that this signature of a higher trophic level is consistent throughout the year. These results are incompatible with the current belief of an herbivorous diet for A. lixula and suggest that it must be considered an omnivore tending to carnivory in Mediterranean ecosystems, feeding at least partially on sessile animals such as Cirripedia, Hydrozoa or Bryozoa. A parallel analysis of gut contents showed a predominance of vegetal items in both species, although A. lixula consistently had a higher abundance of animal components than P. lividus. Our results challenge the validity of using gut content observations alone for characterizing the trophic behaviour of omnivorous marine invertebrates that feed on a variety of food sources with different digestibility.

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.

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)