Algicidal Sesquiterpene Hydroquinones from the Brown Alga Dictyopteris undulata

Flavoglaucin, its derivatives, and pyranonigrins, which are antioxidants produced by the molds used in fermented foods, were examined for their inhibition of tumor promotion by the Epstein-Barr virus early antigen activation test. Flavoglaucin and its derivatives exhibited high activity. Flavoglaucin and such a derivative as isodihydroauroglaucin inhibited mouse skin tumor promotion in a two-stage carcinogenesis test and appear to be antitumor promoters.     

Protoplast isolation in the marine brown alga Dictyopteris prolifera (Dictyotales)

Protoplasts were isolated from thalli of Dictyopteris prolifera using a mixture of crude enzymes from vicera of live oysters (Crassostrea gigas) and the following commercial enzymes: an abalone enzyme, cellulase, polygalacturonase and hemicellulase. The enzyme mixtures produced up to 3.3 × 10⁷ cells per l g of tissue fresh weight. The conversion to protoplasts of the cells was about 100% using the oyster enzyme or the abalone enzyme alone. The optimum pH for protoplast isolation was 6.0 and 20 hours were required for conversion to protoplasts.     

Photosynthetic performance of deep-water macroalgae (Phaeophyta,Dictyotales) off Bermuda

Photosynthetic performance and dark respiration rates were determined in situ for abundant macroalgae occurring between 27–49 m depths off Bermuda. Brown algae, particularly members of the order Dictyotales, predominated at all deep-water sites, and Stypopodium zonale was the most abundant species. Species showed net photosynthesis at very low ambient irradiances (<0.01 maximum I_o). Lobophora variegata, a species with a highly decumbent growth form, had low productivity across all irradiances. In contrast, Dictyota spp. (D. bartayresii, D. dichotoma, D. divaricata) and S. zonale had high photosynthetic capacity (ca. 400 µmol O₂ gdw^–1 h^–1), and light saturation was not evidenced at the highest ambient irradiance level (300 µE m^–1 s^–1) for species with thin, flat thalli. Light-harvesting pigment concentrations reflected tissue-nitrogen levels. C:N atom ratios, except for L. variegata and D. divaricata, were within the ratio for balanced growth. The repeated occurrence and photosynthetic efficiency of these Dictyotalean species in subtropical and tropical deep-water habitats emphasize their successful adaptation to low-light, nutrient-poor environments.     

Morphology and life history of Halopteris filicina (Sphacelariales, Phaeophyceae) from Korea

The vegetative morphology and life history of Halopteris filicina (Grateloup) Kutzing, collected from Korea, were examined in laboratory culture. Field plants attaining 3–5 cm in height were epilithic, tufted, yellowish-brown, and produced numerous erect axes with alternately distichous branches from compact basal discs. They were cultured under a 12:12 h LD photoperiod at 10°-C, 15°C and 20°C to observe the influence of temperature on reproduction. At 10°C plants grew only vegetatively, whereas at 15°C and 20°C they produced unilocular sporangia. Unispores released from sporangia developed into monoecious, anisogamous gametophytes that formed plurilocular female and male gametangia on the same lateral branches. The zygotes, by fusion of female macrogametes and male microgametes, developed into sporophytes bearing unilocular sporangia, whereas the unfused female gametes germinated parthenogenetically. This species was confirmed to have an isomorphic life history, basically similar to the other species of Sphacelariales.     

A virus infection inMyriotrichia clavaeformis (Dictyosiphonales,Phaeophyceae) from Argentina

Summary Virus-infected plants ofMyriotrichia clavaeformis from the coast of Argentina are sterile and produce viras particles in lateral vesicles homologous to plurilocular sporangia of normal plants. Virions are released into the surrounding sea water and can infect swimming spores of healthyMyriotrichia plants. Virus particles are hexagonal in cross section with a diameter of 170–180 nm and have an envelope with an electron dense core. Similar viruses are known for several genera in the order Ectocarpales. This is the first record of an infectious agent in the order Dictyosiphonales, which contains thalli with more complex tissues and life histories.Dedicated to Prof. Dr. Dr. h.c. Eberhard Schnepf on the occasion of his retirement     

Morphology and life history of Coelocladia arctica (Dictyosiphonales,Phaeophyceae), new to Japan*

The occurrence of Coelocladia arctica Rosenvinge (Dictyosiphonales, Phaeophyceae) is reported for the first time in the Pacific Ocean, from Oshoro, Hokkaido, Japan. The field material was approximately 30 mm in height, up to 300 M-μn in diameter, 2-3 times irregularly branched and provided with phaeophycean hairs and plurilocular sporangia. The thallus was composed of several (usually four) large, hyaline, rounded, isodiametric inner cells, with smaller sub-cortical and pigmented cortical cells. The plurilocular sporangia were 3-4 celled, often branched, and protruded from the cortical cells, arranged in a crown-shaped complex. In culture, the plurispores germinated unipolarly leaving an emptied original spore wall, and developed into a branched protonema. Cells of the protonema as well as the erect thallus contained several disc-shaped chloroplasts with pyrenoids. Uniseriate erect filaments arose from the protonema, then became polystichous and formed branches. Unilocular sporangia were never observed in the field or in cultured material. Erect thalli were formed under culture conditions of 5-15°C, and developed a thick parenchyma at 5-10°C, irrespective of the day length.     

Life history of Japanese Stypocaulon durum (Sphacelariales, Phaeophyceae)

Phenology, morphology, life history and responses to different temperature and photoperiod conditions were studied in Japanese Stypocaulon durum (Ruprecht) Okamura. Erect thalli of the species were collected year-round, but the mature thalli forming either uniloc-ular sporangia or two different types of plurilocular structures (evidently gametangia) on separate thalli were found only in winter. ln culture, an isomorphic life history is suggested for the species, alternating between a sporophyte forming unilocular sporangia and gam-etophytes forming plurilocular macro- (female) and micro- (male) gametangia. Contents of unilocular sporangia were not released, but germinated in situ, developing into erect thalli forming plurilocular gametangia. Macrogametangia released aplanogametes (oospores), but male gametangia appeared to be non-functional, although flagellated cells were once formed in the loc-uli. This is the first report of plurilocular gametangia in the species. Although the species grew well and matured under considerably lower temperature conditions than European Stypocaulon scoparium (L.) Sauvageau, its temperature requirements showed similarity to northwestern Atlantic Stypocaulon species. This supports the notion that northwestern Atlantic Stypocaulon is conspecific with S. durum.     

Intraspecific sterility barrier confirms that introduction of Sphaerotrichia divaricata (Phaeophyceae,Chordariales) into the Mediterranean was from Japan

Crossing studies revealed an intraspecific sterility barrier on the level of zygote formation between Japanese Sphaerotrichia divaricata and isolates of the same species from the Northeast Pacific and the North Atlantic. Because no consistent morphological differences exist between sporophytes from Japan and other areas, we propose not to distinguish the intersterile populations as different species. Japanese Sphaerotrichia and isolates from a recently detected population in the Étang de Thau, French Mediterranean coast, are interfertile. The crossing studies support the assumption that this Mediterranean population is a recent introduction from Japan.     

Ultrastructural study of plasmodesmata in the brown alga Dictyota dichotoma (Dictyotales, Phaeophyceae)

Plasmodesmata are intercellular bridges that directly connect the cytoplasm of neighboring cells and play a crucial role in cell-to-cell communication and cell development in multicellular plants. Although brown algae (Phaeophyceae, Heterokontophyta) are phylogenetically distant to land plants, they nevertheless possess a complex multicellular organization that includes plasmodesmata. In this study, the ultrastructure and formation of plasmodesmata in the brown alga Dictyota dichotoma were studied using transmission electron microscopy and electron tomography with rapid freezing and freeze substitution. D. dichotoma possesses plasma membrane-lined, simple plasmodesmata without internal endoplasmic reticulum (desmotubule). This structure differs from those in land plants. Plasmodesmata were clustered in regions with thin cell walls and formed pit fields. Fine proteinaceous "internal bridges" were observed in the cavity. Ultrastructural observations of cytokinesis in D. dichotoma showed that plasmodesmata formation began at an early stage of cell division with the formation of tubular pre-plasmodesmata within membranous sacs of the cytokinetic diaphragm. Clusters of pre-plasmodesmata formed the future pit field. As cytokinesis proceeded, electron-dense material extended from the outer surface of the mid region of the pre-plasmodesmata and finally formed the nascent cell wall. From these results, we suggest that pre-plasmodesmata are associated with cell wall development during cytokinesis in D. dichotoma.     

Accumulation,effects on growth,and localization of zinc in Padina gymnospora (Dictyotales,Phaeophyceae)

The ability of young plants of the brown alga Padina gymnospora to tolerate and accumulate zinc was determined for different zinc concentrations in seawater over a period of 21 days. This species, among 10 macroalgae species studied in a contaminated coastal area near Rio de Janeiro city, presented the highest zinc contents. The accumulation of zinc by this species determined from in vitro experiments increased approximately linearly with the zinc concentration in the water. A steady state was not reached even for high concentrations (1000 µg 1^–1). Analytical electron microscopy techniques used for subcellular localization of this metal showed that the zinc was present as dense granules along the cell walls, starting in a region 15 to 25 cells away from the thallus apex, but being absent in the cytoplasm of viable cells. It is suggested that the cell walls play the main role in the zinc accumulation in Padina gymnospora.This paper is dedicated in honor of Prof. Carlos Chagas Filho, founder of the Institute of Biophysics, on the occasion of its 50^th anniversary     

Life histories of Colpomenia sinuosa and Hydroclathrus clathratus (Scytosiphonaceae, Phaeophyceae) in culture

Life-history studies in culture were carried out on Colpomenia sinuosa (Mertens ex Roth) Derbès et Solier and Hydroclathrus clathratus (C. Agardh) Howe (Scytosiphonales, Phaeophyceae) from Japan. These species showed a heteromorphic life-history pattern with an alternation between erect thalli bearing plurilocular zoidangia and prostrate thalli bearing ectocarpoid plurilocular and unilocular zoidangia. Plurizoids released from erect and prostrate thalli developed into prostrate thalli. Unizoids, however, developed into erect thalli. Prostrate thalli produced plurilocular zoidangia in long-day conditions and unilocular zoidangia in short-day conditions at 10-20°C. Prostrate thalli of C. sinuosa formed ascocysts. Germlings of both species did not grow at 5°C.     

Glycolate metabolism and subcellular distribution of glycolate oxidase in Spatoglossum pacificum (Phaeophyceae, Chromophyta)

Seven enzymes participating in glycolate metabolism were demonstrated to be present in crude extract of the brown alga Spatoglossum pacificum Yendo. These were phosphoglycolate phosphatase, glycolate oxidase, glutamate-glyoxylate aminotransferase, serine hydroxymethyltransferase, amino acid-hydroxy-pyruvate aminotransferase, hydroxypyruvate reductase and catalase. Malate synthase, which is involved in glycolate metabolism in the xanthophycean alga, could not be detected. On demonstration of subcellular distribution of glycolate oxidizing enzymes by linear sucrose density gradient centrifugation, glycolate oxidase was detected in the same fraction at a density of 1.23 g cm^?3 with catalase: that is, the marker enzyme of peroxisome and serine hydroxymethyltransferase was found in the same fraction at a density of 1.21 g cm^?3 with isocitrate dehydrogenase, the marker of mitochondria. From the present data, it is proposed that the brown alga Spatoglossum possesses the ability to metabolize glycolate to glycerate via the pathway which may be similar to that of higher plants.     

Plastid Genome of <Emphasis Type="Italic">Dictyopteris divaricata</Emphasis> (Dictyotales,Phaeophyceae): Understanding the Evolution of Plastid Genomes in Brown Algae

Dictyotophycidae is a subclass of brown algae containing 395 species that are distributed worldwide. A complete plastid (chloroplast) genome (ptDNA or cpDNA) had not previously been sequenced from this group. In this study, the complete plastid genome of Dictyopteris divaricata (Okamura) Okamura (Dictyotales, Phaeophyceae) was characterized and compared to other brown algal ptDNAs. This plastid genome was 126,099 bp in size with two inverted repeats (IRs) of 6026 bp. The D. divaricata IRs contained rpl21, making its IRs larger than representatives from the orders Fucales and Laminariales, but was smaller than that from Ectocarpales. The G + C content of D. divaricata (31.19%) was the highest of the known ptDNAs of brown algae (28.94–31.05%). Two protein-coding genes, rbcR and rpl32, were present in ptDNAs of Laminariales, Ectocarpales (Ectocarpus siliculosus), and Fucales (LEF) but were absent in D. divaricata. Reduced intergenic space (13.11%) and eight pairs of overlapping genes in D. divaricata ptDNA made it the most compact plastid genome in brown algae so far. The architecture of D. divaricata ptDNA showed higher similarity to that of Laminariales compared with Fucales and Ectocarpales. The difference in general features, gene content, and architecture among the ptDNAs of D. divaricata and LEF clade revealed the diversity and evolutionary trends of plastid genomes in brown algae.     

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.     

The reproductive structures of the Mediterranean alga Laurencia pelagosae (Ceramiales,Rhodophyta)

Reproductive structures of Laurencia pelagosae are described for the first time. Tetrasporangia, in parallel arrangement, are cut off from the mother cells adaxially; antheridial branches, which are unramified, are inserted in depressions lacking a row of axial cells on their bases; cystocarps are sessile and subspherical. The occurrence of secondary pit-connections between epidermal cells and of lenticular thickenings in the walls of medullary cells is documented. The species is included in the new section Pelagosae within the subgenus Laurencia.     

A taxonomic study of Japanese Scytosiphon (Scytosiphonales, Phaeophyceae), including two new species

Morphological and culture studies were carried out on the genus Scytosiphon (Scytosiphonales, Phaeophyceae) growing in Japan. Morphologies of plurilocular zo-idangia and sporophytic thalli were found to be useful as taxonomic characters. Two types of plurilocular zo-idangial sori were recognized: (i) loosely coherent plurilocular zoidangia without a cuticular layer (loose type); (ii) tightly coherent plurilocular zoidangia with a cuticular layer (coherent type). The sporophytic thallus morphology was different among species in general appearance (tufted or discoid), degree of cohesion of erect filaments, and presence or absence of paraphyses. Two new species are described, Scytosiphon gracilis sp. nov. and Scytosiphon tenellus sp. nov, Scytosiphon gracilis is distinguished from other species by a thin medullary layer, coherent plurilocular zoidangia, no ascocysts associated with plurilocular zoidangia, and Compsonema-like sporophytic thalli. Scytosiphon tenellus is characterized by a thin medullary layer, coherent plurilocular zoidangia, ascocysts among plurilocular zoidangial sori, and Stragularia-like sporophytic thalli. Scytosiphon lo-mentaria (Lyngbye) Link is characterized by a constricted gametophytic thallus, loose plurilocular zoidangia, the presence of ascocysts, and sporophytic thaili identified with Microspongium gelatlnosum Reinke.