Cell-cell-recognition during fertilization in Ectocarpus siliculosus (Phaeophyceae)

Two cellular communication systems are involved in sexual recognition of complementary gametes of Ectocarpus siliculosus. The first is mediated via the pheromone ectocarpene and is not species-specific. A second system has been identified which is based on the interaction of carbohydrate and carbohydrate receptor. Cell fusion is inhibited selectively by wheat germ agglutinin (WGA) applied to female gametes or with the complementary N-acetylglucosamine (GlcNAc) applied to male gametes. Both, WGA and GlcNAc showed gametic specificity in their effects. Fertilization was not affected by the lectins GNA, LCA and SBA. The maximum fertilization success was 50.2% and followed Michaelis-Mentenkinetics (I₅₀ = 6 minutes).     

Verosphacela silvae sp. nov. (Onslowiaceae, Phaeophyceae) from the Mediterranean Sea

We describe Verosphacela silvae sp. nov., from the Mediterranean Sea. It consists of horizontal filaments living on the lower face of the red alga Peyssonnelia rubra (Greville) J. Agardh, from which erect filaments up to 1.5 mm high rise and grow upright after passing through the thallus of the supporting species. There are both horizontal and erect filaments growing by apical cells. In the subapical cells, 1–2 longitudinal divisions occur (more frequently in the erect filaments) but no secondary transverse divisions occur. Erect filaments bear lateral propagules on a stalk of one to three (rarely more) cells. Propagules, with neither apical cells nor arms, consist of seven cells. Zoidangia are borne at the apex of erect laterals. The new species differs from V. ebrachia Henry mainly in habit, propagules and zoidangia. In addition, distinct from V. ebrachia, filaments of V. silvae never penetrate between the cuticle and the cell wall of the supporting alga. Moreover, propagules of V. silvae consist of seven cells, whereas those of V. ebrachia consist of 9–13 cells, and zoidangia are terminal on laterals in V. silvae, whereas in V. ebrachia they are sessile on both axes and laterals.     

A Virus Infection in the Marine Brown Alga Ectocarpus siliculosus (Phaeophyceae)

Laboratory cultures of Ectocarpus siliculosus originating from New Zealand showed a defect in gametangium formation. Nuclear divisions in gametangium initials are not followed by cell wall formation. In the resulting multinucleate cells nuclear DNA increases dramatically, and nuclear membranes disintegrate. Eventually, the entire structure is filled with hexagonal particles of approximately 130 nm diameter. These were isolated and shown by EM to consist of a dense core surrounded by a 3-layered shell. They are released into the culture medium when the host cells burst. Ectocarpus gametes from healthy cultures could be infected by these particles. The resulting partheno-sporophytes developed pathological symptoms, suggesting that the particles are viruses. The expression of the defect is temperature dependent. At 10°C all gametangia are abnormal, while between 15 and 20 °C defective and normal gametangia and gametes are formed on the same plant. Partheno-sporophytes developing from such gametes carry the viral particles expressed in deformed unilocular and plurilocular sporangia.     

Chorotypes of seaweeds from the western Mediterranean Sea and the Adriatic Sea: An analysis based on the genera Audouinella (Rhodophyta), Cystoseira (Phaeophyceae) and Cladophora (Chlorophyta)

The aim of the present paper is to identify the possible existence of groups of species in the genera Audouinella (Rhodophyta), Cystoseira (Phaeophyceae) and Cladophora (Chloropyta) with significantly similar distribution patterns (chorotypes), in the western Mediterranean Sea and the Adriatic Sea. Of the 98 species studied, 59 were grouped into 11 chorotypes, whereas 39 species remained ungrouped. Thirty‐eight species were included in a generalist chorotype, whereas 6 chorotypes were monospecific. The relationships with the environmental factors that could explain the chorotypes are discussed.     

Ultrastructural analysis of flagellar development in plurilocular sporangia of Ectocarpus siliculosus (Phaeophyceae)

Flagellar development in the plurilocular zoidangia of sporophytes of the brown alga Ectocarpus siliculosus was analyzed in detail using transmission electron microscopy and electron tomography. A series of cell divisions in the plurilocular zoidangia produced the spore-mother cells. In these cells, the centrioles differentiated into flagellar basal bodies with basal plates at their distal ends and attached to the plasma membrane. The plasma membrane formed a depression (flagellar pocket) into where the flagella elongated and in which variously sized vesicles and cytoplasmic fragments accumulated. The anterior and posterior flagella started elongating simultaneously, and the vesicles and cytoplasmic fragments in the flagellar pocket fused to the flagellar membranes. The two flagella (anterior and posterior) could be clearly distinguished from each other at the initial stage of their development by differences in length, diameter and the appendage flagellar rootlets. Flagella continued to elongate in the flagellar pocket and maintained their mutually parallel arrangement as the flagellar pocket gradually changed position. In mature zoids, the basal part of the posterior flagellum (paraflagellar body) characteristically became swollen and faced the eyespot region. Electron dense materials accumulated between the axoneme and the flagellar membrane, and crystallized materials could also be observed in the swollen region. Before liberation of the zoospores from the plurilocular zoidangia, mastigoneme attachment was restricted to the distal region of the anterior flagellum. Structures just below the flagellar membrane that connected to the mastigonemes were clearly visible by electron tomography.     

Qualitative and quantitative determination of pheromone secretion in female gametes of Ectocarpus siliculosus (Phaeophyceae)

Pheromone secretion in living female gametes of Ectocarpus siliculosus was examined by closed-loop extraction and gas chromatography. The pheromone (ectocarpene) recovery efficiency was calibrated with synthetic ectocarpene and was found to depend strongly on the volume of the closed-loop system. Maximum extraction efficiency of ectocarpene (17.4%) was obtained with a small-volume system. In addition to the main compound ectocarpene, the pheromone bouquet of E. siliculosus contained minor amounts of hormosirene, multifidene and dictyotene, which until now are known only as sperm-attractants in other brown algae. The pheromone bouquet was identical in 4 clones of different geographic origins. Settled female gametes of Ectocarpus siliculosus continued to secrete pheromone and to attract male gametes for up to 7 h after their release with an ectocarpene secretion rate of 1 x 10(5) molecules s-1 cell-1.     

Organotin Compounds (OTCs) in Saccharina latissima (Phaeophyceae) from the Barents Sea

Doklady Biological Sciences - The degree of contamination of Saccharina latissima, the dominant species among macrophyte algae in the sublittoral zones of the Kola Bay and Eastern Murman of the...     

An immunofluorescence study on lectin binding sites in gametes of Ectocarpus siliculosus (Ectocarpales, Phaeophyceae)

The distribution of binding sites for the lectins WGA, DSA. RCA I, PNA, AAA, MAA. SNA, GNA. and Con A in gametes of both sexes of the brown alga, Ectocarpus siliculosus (Dillw.) Lyngbye, was investigated by fluorescence microscopy. Digoxigenin-conjugated lectins and an FITC-anti-digoxigenin antibody were used as a high sensitivity detection system. Organelles and other distinct cellular domains could be distinguished by their binding specificities. Glycoconjugates associated with one flagellum were found to be associated with the axoneme by lectin binding to isolated flagellar apparatuses. In addition, changes in the distribution of carbohydrate epitopes during the attachment of gametes to the substratum were revealed by differential lectin binding.     

Effects of temperature on total phenolic compounds in Cystoseira amentacea (C. Agardh) Bory (Fucales,Phaeophyceae) from southern Mediterranean Sea

The aim of this study was to test the effects of temperature on phenolic content of the brown seaweed Cystoseira amentacea. Phenolic compounds are secondary metabolites involved in different protection mechanisms as, for example, against grazers, epiphytes and UV radiation. Seasonal variations of phenolic content in C. amentacea were analysed and laboratory experiments, in which C. amentacea was exposed to an increase of temperature (25°C and 30°C), were performed. Total phenolic content (TPC) was determined colorimetrically with the Folin–Ciocalteu reagent. In C. amentacea, a seasonal pattern in TPC was observed, with a maximum value in winter-spring. C. amentacea responded significantly to the temperature treatments with a consistent decrease in TPC after 48 h.     

On the sexual reproduction ofDictyosiphon foeniculaceus (Phaeophyceae,Dictyosiphonales)

Dictyosiphon foeniculaceus from Sweden and Newfoundland was studied in laboratory culture. Zoids from unilocular sporangia developed into dioecious microscopic filamentous gametophytes which produced uniseriate plurilocular gametangia in low temperatures (0 to 8 °C). Zygotes and unfused isogametes gave rise to filamentous protonemata on which parenchymatous macroscopic sporophytes were formed. Isolates from Sweden and Newfoundland were interfertile. Although formed in culture, genetically unisexual sporophytes were not detected in nature. Female gametes ofD. foeniculaceus produced a sexual pheromone. It was identified as finavarrene, which is also known as the sperm attractant inAscophyllum nodosum.     

Experimental evidence against sexual fusions of spores from unilocular sporangia of Ectocarpus siliculosus (Phaeophyta)

Evidence that sexual fusion does not occur in zoids from unilocular sporangia of Ectocarpus siliculosus is presented as follows: (1) Zoids from various types of unilocular sporangia do not interact with genuine gametes; (2) Aggregates of two zoids, previously claimed to be zygotes, were frequently seen in cultured material but could be demonstrated to be the result of incomplete cell separation during sporogenesis; (3) Culture studies showed that there is no fusion of male and female nuclei in such aggregates.     

Parthenogenesis in the brown alga Lessonia nigrescens (Laminariales,Phaeophyceae) from central Chile1*

Parthenogenesis, the development of female gametes without fertilization, is relatively common in brown algae, although limited quantitative information on the phenomenon is available. Its occurrence is reported for the first time in Lessonia nigrescens Bory, a member of the Laminariales and a key ecological component of the benthic algal communities along the Chilean coast. Isolated female gametophytes developed into parthenosporophytes throughout the year, with a maximum in spring to early summer. Isolated male gametophytes, on the other hand, never developed fronds. Parthenosporophytes obtained in the laboratory developed normally when cultivated under greenhouse conditions, and the resulting individuals were indistinguishable in size, shape, texture, and color from heterozygous sporophytes. Quantification of DNA of various tissues demonstrated that early during their development, parthenosporophytes duplicated their DNA content, displaying levels similar to heterozygous sporophytes and almost twice the level found in gametophytes. One out of 45 individuals from a field population yielded only female gametophytes, strongly suggesting that parthenogenesis does occur in wild stands of L. nigrescens.     

Biochemical composition of the Atlantic bonito Sarda sarda from the Aegean Sea (eastern Mediterranean Sea) in different stages of sexual maturity

The content (% wet mass) in water, ash, lipid, crude protein, DNA and RNA of different tissues was determined during sexual maturation of bonitos Sarda sarda from the Aegean Sea. A total of 220 specimens were collected in the following stages of sexual maturity: immature, resting, developing, mature, spawning and spent. Highest lipid levels in the white muscle, red muscle and liver were measured in immature specimens, while lowest levels were found in spawning bonitos. The gradual percentage of lipid reduction from immature to spawning bonitos was relatively higher in the liver (females 71·2% and males 64·4%) than in the white (females 59·2% and males 53·5%) and red (females 62·1% and males 51·7%) muscle. Lipid levels in the gonads increased gradually from the immature to spawning stage. The decrease of lipid in the somatic tissues was more intense in females than in males, and gonadal lipid content was higher in females than in males. There was a strong reverse correlation between water and lipid percentage in all tissues. Protein content decreased significantly only in spawning bonitos. The percentage of protein reduction from immature to spawning stage was relatively higher in males than in females in both white (females 3·4% and males 4·6%) and red (females 4·6% and males 5·1%) muscles. Protein content in the liver was significantly lower than in the other tissues, being highest in mature females. Gonadal protein content in females increased with maturation and decreased after spawning. The content in ash exhibited considerable stability. The RNA:DNA ratio exhibited a similar pattern of variation in both muscles. The RNA:DNA ratio increased during gonadal development gradually from the developing to spent stage. It was concluded that in S. sarda during gonadal development, there was an increase in gonadal lipid accompanied by a decrease in somatic tissue lipid reserves. Thus, reproductive inactive bonitos have more lipid in their edible part and a higher nutritional value than active ones.     

Size and age at sexual maturity of female bluefin tuna (Thunnus thynnus L. 1758) from the Mediterranean Sea

The ovaries of 501 female eastern Atlantic bluefin tuna (Thunnus thynnus Linnaeus, 1758) captured in the Mediterranean Sea from May to September between 1998 and 2004 were analysed histologically. Body size at median sexual maturity (L₅₀) was 103.6 cm fork length (FL), while 100% maturity was reached above 135 cm FL. The age analysis, based on the count of the translucent zones of the first spiniform ray of the first dorsal fin, showed that most of the specimens with FL = L₅₀ were 3 years old while 100% maturity was reached between 4 to 5 years. The reported evidence indicates that for the eastern Atlantic bluefin tuna stock, the size and age of first sexual maturity of females was lower than in the western Atlantic stock.     

Hormonal interaction in sexual reproduction of Desmarestia aculeata (Phaeophyceae)

Male and female gametophytes of Desmarestia aculeata have been grown as clonal cultures. Red light permits vegetative growth only. Differentiation of oogonia and antheridia takes place under illumination with blue light as has been previously reported for several species of Laminaria. Morphological details for differentiation of oogonia and antheridia are presented, stressing the strong resemblance with the respective processes in Laminaria. Experimental evidence indicates the presence of a hormonal factor secreted by the eggs, which is responsible for discharge and subsequent attraction of spermatozoids to the egg.     

Repopulation techniques for Macrocystis integrifolia (Phaeophyceae: Laminariales) in Atacama,Chile

The giant kelp Macrocystis (integrifolia) has been intensely harvested in northern Chile for several years. In order to prevent a future disaster, we developed two different techniques for restoration of damaged Macrocystis integrifolia beds in the Atacama region of Chile. (1) Explantation: Laboratory-grown juvenile sporophytes were fixed to different substrata (plastic grids, ceramic plates, or boulders) by elastic bands or fast-drying glue (cyanoacrylate). Explants reached 150–200 cm in length within 5 months (relative growth rate?≈?1.3–1.7 % day^?1), and reproductive maturity in 5–7 months. (2) Seeding of spores: Mature sporophylls were placed at 8 m depth on the sea bottom, supported by cotton gauze sleeves attached to boulders of different origin. Sixty percent of clean boulders collected on the beach produced up to seven recruits per boulder. In contrast, 20 % of the boulders from the sea bottom, colonized by epibionts, showed up to two recruits. Relative growth rates, however, were similar (≈2.4–2.6 % day^?1). Practical applications of our findings are: laboratory-produced juvenile sporophytes fixed to various substrata by elastic bands or cyanoacrylate glue can be used to colonize rocks or artificial reefs. In cases, where laboratory-grown seedlings are unavailable, mature sporophylls from nearby Macrocystis beds can be used to establish new recruits on rocky substrata.     

Feeding ecology of Mediterranean razorfish Xyrichthys novacula in the Tyrrhenian Sea (Central Mediterranean Sea)

The stomach contents of 196 razorfish ( Xyrichthys novacula ), collected in the Tyrrhenian Sea (Ponza Island, Italy) between July and December 1994, were examined in order to assess the diet and to analyse size-related dietary variation of the species. The diet of X. novacula showed a strict relationship with well-sorted fine sand (SFBC) benthic species, mostly Mollusca and Echinodermata, as pointed out by the analysis at the specific level of the stomach contents. Mollusca Pelecipoda, essentially Acanthocardium tuberculatum and Echinodermata, with Echinocardium cordatum , made up 90% of the volume of prey of X. novacula. Mysidacea and Gastropoda were frequent but volumetrically less important. The study showed that male and female X. novacula were not effectively segregated by trophic dimension in the study area. The niche overlap between males and females in prey composition was pronounced, as evidenced by the significantly high values of Schoener's index with larger males that showed a greater tendency towards predation of large prey. The increment in length (L.I.) in males, despite a decrease in the percentage body weight increment (B.W.I.), could represent the result of a male reproductive feature: the lack of sperm competition allows the males much more energy to devote to growth than do that of the females of this species.     

Effect of Extracellular Ca2+ and Ca2+-Antagonists on the Movement and Chemoorientation of Male Gametes of Ectocarpus siliculosus (Phaeophyceae)

Chemoorientation in male gametes of Ectocarpus siliculosus in response to sexual pheromones is effected by two distinct mechanisms, chemokinesis and chemoklinotaxis. These are characterized by a strongly asymmetric bending pattern of the anteriorly-directed flagellum and transient unilateral bending of the hind flagellum, respectively. Removal of extracellular Ca²⁺ showed that normal flagellar movement and chemokinesis require millimolar concentrations of Ca²⁺ in the medium. The response to pheromones is strongly inhibited by La³⁺, whereas the Ca²⁺-channel drugs, verapamil and nifedipine, have only little effect. Nifedipine nethertheless effectively inhibited accumulation at pheromone sources. These results are interpreted as an indication for the involvement of two pharmacologically distinct Ca²⁺-channels in chemokinesis and chemoklinotaxis. The calmodulin-antagonist, trifluoperazine, induces, at low concentrations, the same flagellar response in chemokinesis as the pheromone, the mechanism of action remaining unknown.     

Six new Richtersia species (Nematoda, Selachinematidae) from the Mediterranean Sea

Six new species of the genus Richtersia Steiner, 1916 are described from the Mediterranean Sea: R. bathyalis sp.n., R. spinosa sp.n., R. heipi sp.n., R. mediterranea sp.n., R. staresensis sp.n. and R. coomansi sp.n. Some morphological adaptations of R. bathyalis, R. spinosa and R. heipi are discussed in relation to their small body size. The structure of the stomatal region, labial region and spicules is described in detail and probably holds for the entire genus.