HYBRIDIZATION OF MACROCYSTIS (PHAEOPHYTA) WITH OTHER FLOAT-BEARING KELPS1,2

Sporophytes were produced in 22 out of 23 attempts to cross Macrocystia angustifolia Bory with Pelagophycus porra (Lemon) Setchell. Representative samples of these plants were raised in a surge-tank under greenhouse conditions to several meters in length. The plants resulting from this intergeneric cross were intermediate in morphology between the parental genera. The intermediate plants were similar to those found fry one of the authors (MN) in 1957, and by others since then, in California kelp beds where Macrocystis and Pelagophycus co-occur, A cross between M. angustifolia and Nereocystis luetkeana (Mertens) P. & R., also produced normal sporophytes. The taxonomic, evolutionary, morphogenetic and genetic implications of these hybridizations are discussed.     

Low Divergence in rDNA ITS Sequences Among Five Species of Fucus (Phaeophyceae) Suggests a Very Recent Radiation

Sequences from the two ribosomal DNA internal transcribed spacers (ITS1 and ITS2) were compared among five species of Fucus. Based on the present taxon sampling, parsimony analysis showed that Fucus serratus is the sister-group of the remaining Fucus species when Ascophyllum nodosum was used as an outgroup. The topology of the tree was (Fucus serratus (F. lutarius (F. vesiculosus (F. spiralis+F. ceranoides)))). The extremely low variation observed suggests a very recent radiation of the genus which supports the view widely accepted that the Fucales are among the most evolutionarily advanced of the brown algae. We further note that sequence differences between Fucus and Ascophyllum were 28%: this does not rule out the utility of ITS sequences within the Fucaceae. The very low number of informative positions allows to demonstrate empirically that distance matrix methods group on the basis of symplesiomorphies. Received: 17 February 1997 / Accepted: 17 April 1997     

Centrosome and microtubule dynamics in apical cells ofSphacelaria rigidula (Phaeophyceae) treated with nocodazole

Summary Treatment of young thalli ofSphacelaria rigidula with 0.04 g of nocodazole (Nz) per ml for up to 36 h affects microtubules (Mts) only slightly, but blocks a large number of mitotic cells in metaphase, without disruption of the metaphase plate. Higher concentrations of Nz (0.1 g/ml) depolymerize interphase Mts. Only a few perinuclear and some short Mts resist and remain associated with the centrosomes. Fragmented Mts or groups of Mts sometimes remain in the apical dome. After treatment with 0.1 g of Nz per ml, prometaphase cells are blocked at metaphase, while post-metaphase cells become binuclear, due to the failure of cytokinesis. With anticentrin immunofluorescence, a positive centrin signal is always observed in the centrosome area. Centrosome duplication is not affected by Nz, but separation is disturbed. After recovering for 2–4 h, most of the blocked metaphases proceed normally. In such cells duplicated centrosomes are seen in different stages of separation. In some cells independent aster-like microtubule configurations appear in the apical dome, occasionally displaying centrin at their centre. During recovery various configurations of bimitosis or multipolar mitosis were found. The multipolar spindles may share common centrosomes. Up to four centrosomes may accompany each nucleus. In some 24 h treated cells, as well as in cells recovering for 2 h, the centrin-positive structure is rod-like, extending in opposite directions from the usual position to the poles. Electron microscopical examination of thin sections revealed that the growth pattern of the apical cells is disrupted after Nz treatment. The observations show that: (a) the Mt cytoskeleton is involved in maintaining the polarity and growth pattern of apical cells, (b) mitosis is blocked by low concentrations of Nz without significant depolymerization of Mts, (c) the centrosome cycle is independent of the nuclear cycle, (d) centrosome separation and differentiation are disturbed by Nz treatment, (e) during recovery from Nz treatment, centrosomal material that may have separated from the centrosomes, as well as Mt fragments that resisted depolymerization, may operate as Mt nucleation centres.Abbreviations DIC differential interference contrast - EM electron microscope - Mt microtubule - MTOC microtubule-organizing center - Nz nocodazole - NBBC nucleus-basal body connector     

Carbohydrates of Padina tetrastromatica

Extraction with hydrochloric acid (pH 2.5) of the brown alga Padina tetrastromatica afforded water-soluble and water-insoluble polysaccharides. The water-soluble polysaccharide was fractionated using cetyltritmethyl ammonium bromide and chromatography on DEAE-cellulose and Sephadex G-100. A neutral laminaran like glucan and two new sulphated heteropolysaccharides comprising d-glucuronic acid, l-fucose, l-rhamnose, d-xylose, d-arabinose, d-galactose, d-glucose and half-ester sulphate were obtained. The alginic acid isolated from this brown seaweed was found to be predominantly of poly 1 → 4β-d-mannuronic acid type. The water-soluble sulphated polymer showed high anticoagulant activity.     

Brominated phlorethols and nonhalogenated phlorotannins from the brown alga Cystophora congesta

The following phlorotannins were isolated from the brown alga Cystophora congesta and characterized as their acetates: phloroglucinol triacetate, bromodiphlorethol pentaacetate, diphlorethol pentaacetate, bromotriphlorethol-A₁-heptaacetate, bromotriphlorethol-A₂-heptaacetate, tetraphlorethol-C-nonaacetate and fucodiphlorethol-D-decaacetate. The substances bromodiphlorethol pentaacetate, bromotriphlorethol-A₁-heptaacetate and bromotriphlorethol-A₂-heptaacetate are the first brominated members of this series to be described. Triphlorethol-A- heptaacetate was isolated previously from C. congesta.     

Laminarionema elsbetiae gen. et sp. nov. (Ectocarpales, Phaeophyceae), a new endophyte in Laminaria sporophytes

Laminarionema elsbetiae gen. et sp, nov, (Ectocarpaceae, Ectocarpales. Phaeophyceae), a new endophyte of Laminaria japonica Areschoug (Laminariales, Phaeophyceae), is described from Muroran. Hokkaido, Japan. Laminarionema elsbetiae grows in the host tissues forming networks in the epidermal and subcortical layers as well as penetrating into the cortical and medullary layers. Only phaeophycean hairs emerge from the surface of the host tissue. No reproductive cells were found in field material. However, under host-free culture the species formed three morphologically different reproductive structures. Macrosporangia containing a single large motile spore were formed under long and short day conditions below 20°C, transformed from vegetative cells, conical to elongated in shape, 50–75 μm in length and ca. 10 μm in diameter. Microsporangia were linear to lanceolate, sometimes branched, formed under long and short day conditions below 15°C. Unilocular sporangia were more or less irregular in shape, formed under short day conditions of 5–15°C, 60–75 μm in length and 40–45 μm in diameter. Sexual fusion between macro- and microspores was not seen. In mixed cultures of L. elsbetiae with young sporophytes of L Japonica Areschoug as well as Saccorhiza dermatodea (de la Pylaie) J. Agardh, L elsbetiae infected both hosts, grew in the same manner as in natural hosts, and formed macrosporangia between host epidermal cells.     

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.     

SECRETION OF PHENOLIC SUBSTANCES INTO THE ZYGOTE WALL AND CELL PLATE IN EMBRYOS OF HORMOSIRA AND ACROCARPIA (FUCALES, PHAEOPHYCEAE)

We have used light and electron microscopy to study the secretion of phenolic substances into the primary walls and cross walls of zygotes and embryos of the brown algae Hormosira banksii (Turner) Decaisne (Hormosiraceae) and Acrocarpia paniculata (Turner) Areschoug (Cystoseiraceae). Phenolic compounds packaged within physodes are a major cytoplasmic component of brown algae. Physodes accumulate at the zygote periphery early in development and are secreted into the primary zygote wall. At germination, physodes accumulate at the rhizoid tip. The first sign of impending cytokinesis is the lining up of these phenolic vesicles in the plane of cell division. This precedes the arrival of other wall constituents. Physodes, together with other wall components, contribute to the development of the cell plate. We conclude that phenolic compounds play an important role in cell-wall construction in these species.     

A MOLECULAR ANALYSIS OF ANALIPUS AND RALFSIA (PHAEOPHYCEAE) SUGGESTS THE ORDER ECTOCARPALES IS POLYPHYLETIC1

We report partial 18S ribosomal DNA sequences of Analipus japonicus (Harvey) Wynne and Ralfsia fungiformis (Grunnerus) Setchell et Gardner. These sequences were compared with the corresponding sequences of 13 brown algae representing six phaeophycean orders: Dictyotales, Ectocarpales, Fucales, Laminariales, Sphacelariales, and Syringodermatales. These 15 brown algae included 10 ectocarpoids representing nine ectocarpacean (sensu Gabrielson et al. 1989) families: Chordariaceae, Dictyosiphonaceae, Ectocarpaceae, Elachistaceae, Heterochordariaceae, Leathesiaceae, Punctariaceae, Ralfsiaceae, and Scytosiphonaceae. We addressed the controversial taxonomic placement of A. japonicus and R. fungiformis in the Ectocarpales by analysis of DNA sequences. Neighbor-joining and maximum parsimony-inferred phylogenies provided evidence that A. japonicus and R. fungiformis are not closely associated with the other representatives of the Ectocarpales. Bootstrap analyses suggest polyphyly of the order Ectocarpales (sensu Gabrielson et al.). However, our analysis failed to resolve the phylogenetic relationship between A. japonicus and R. fungiformis. Our results suggest that the ectocarpoids are just as distantly related to A. japonicus and R. fungiformis as they are to members of the advanced orders Desmarestiales, Dictyotales, Fucales, Laminariales, Sphacelariales, and Syringodermatales.     

ISOLATION AND CHARACTERIZATION OF A DNA VIRUS INFECTING FELDMANNIA SIMPLEX (PHAEOPHYCEAE)1

We report on the isolation and characterization of a virus that is formed in modified zoidangia of the marine brown alga Feldmannia simplex (Crouan) Hamel (Ectocarpales, Phaeophyceae). Isolated virus particles had a buoyant density of about 1.35 g·mL^?1 in CsCl equilibrium gradients. They contained one major polypeptide (MW = 55,000) and at least six additional polypeptides (MW = 15,000–120,000). Four of these proteins were glycosylated. The viral genome consisted of double-stranded DNA and formed two freely migrating fractions in pulsed-field-gel electrophoresis, namely linear DNA with a size of 220 kilobase pairs, and fragments of 10–60 kilobase pairs. However, electron microscopic examination revealed that a substantial fraction of the viral DNA occurred as closed circles. We suggest that the viral DNA in native particles is circular but tends to break at random sites during the preparation.