Anticoagulant activity of a proteoglycan in extracts ofCodium fragile ssp.atlanticum

Aqueous extracts ofCodium fragile ssp.atlanticum possess marked anticoagulant activity. The compounds responsible appear to be proteoglycans with molecular weights, determined by a low angle, laser light-scattering method, of 1.8 × 10⁶ daltons, with a polydispersity value of 1.2.     

Isolation of organellar DNA from Codium fragile (Codiaceae, Codiales, Ulvophyceae)

Organellar DNA was isolated from Codium fragile (Suringar) Hariot (Codiaceae, Codiales, Ulvophyceae) by CsCI-buoyant density centrifugation in the presence of Hoechst dye 33258. Three bands were formed by ultracentrifugation and each fraction of DNA was identified by Southern hybridization. The uppermost fraction was identified as chloroplast DNA, the middle fraction was nuclear DNA and the bottom fraction was mitochondrial DNA. Nuclear rDNA was isolated in the same fraction as mitochondrial DNA. The estimated genome size of mitochondrial DNA by analysis with restriction endonucleases was more than 141.6 kb, which was larger than that of microalgae but smaller than land plants. Restriction endonuclease analysis of the chloroplast DNA showed no difference with that known of C. fragile in New York.     

The Colonisation of Human-made Structures by the Invasive Alga Codium fragile ssp. tomentosoides in the North Adriatic Sea (NE Mediterranean)

Human-made structures, such as groynes, breakwaters, seawalls, pier pilings and floating pontoons, are becoming common features of the landscape in urbanised coastal and estuarine areas. Despite this tendency few studies have focused on their ecology or on their potential impacts on natural assemblages of organisms. When artificial structures are introduced in areas with little or no hard substrata, they not only provide novel habitats, which enables the colonisation of sandy areas by hard-bottom dwelling species, but they can also provide suitable habitats for exotic species. Along the north-east coast of Italy, sandy shores are protected from erosion by a line of breakwaters, which runs almost uninterrupted for about 300 km. These structures provide habitat for a variety of macroalgae and invertebrates and also for the invasive green alga Codium fragile ssp. tomentosoides. The aim of this study was, therefore, to investigate patterns of distribution of this alga on breakwaters in Cesenatico. In particular, we compared the density of thalli, biomass, length and degree of branching of C. fragile ssp. tomentosoides between the landward and the seaward sides of breakwaters, to test the hypothesis that sheltered habitats (landward) represent more suitable habitats than exposed habitats (seaward). In general, the landward side of breakwaters supported greater numbers of thalli of C. fragile ssp. tomentosoides than seaward sides. Thalli grew longer and more branched in sheltered habitats, leading to an overall larger biomass of the alga on the landward side of breakwaters. The presence of sheltered human-made hard substrata in the vicinity of major trading ports and sources of eutrophication could enhance the dispersal of invasive species across regional and geographic scales. Thus, the effects of artificial structures and introduced species on coastal assemblages cannot be evaluated separately, but their synergistic nature should be considered in planning strategies for conservation of biodiversity in coastal habitats.     

Anticoagulant properties of sulphated polysaccharides and a proteoglycan from Codium fragile ssp. atlanticum

A high molecular weight sulphated (18.4%) proteoglycan was isolated from extracts of Codium fragile ssp. atlanticum by molecular exclusion chromatography on Sepharose 2B. Ion exchange chromatography, using Sepharose CL-6B, of lower molecular weight components eluted from the Sepharose 2B column gave two major products with sulphate contents of 10.2% and 7.5%, respectively. Anticoagulant activities of each of the three products were assessed using coagulation techniques and chromogenic substrate assays. An increase in anticoagulant effect was demonstrated by increasing concentration and sulphate content of each algal component. The mechanism of anticoagulant action was shown to be, principally, anti-thrombin in character due to potentiation of heparin cofactor II and antithrombin III activity. Although the anticoagulant substances described are unlikely to be used as antithrombotic therapeutic agents, they have uses as biomedical reagents for investigation of the processes of thrombin inhibition.     

A chitin-like glycan in the cell wall of a Chlorella sp. (Chlorococcales,Chlorophyceae)

The stable amino-sugar fraction of the cell wall of the symbiotic Chlorella strain Pbi (Chlorophyceae) was isolated and investigated by sugar analysis, infra-red spectroscopy, lectin binding, enzymatic degradation, X-ray diffraction and electron microscopy. The results indicate the existence of a glycosaminoglycan which can be regarded as a chitin-like glycan. This carbohydrate structure is unusual for algae and reported here for the first time in unicellular chlorophycean algae.Abbreviations FITC fluorescein isothiocyanate - IR infra-red - NMR nuclear magnetic resonance - TFA trifluoroacetic acid - WGA wheat germ agglutininWe thank Peter Zugenmaier, Institut für Physikalische Chemie der Technischen Universität Clausthal-Zellerfeld, Germany, for valuable advice on X-ray diffraction techniques and for taking the Debye-Scherrer images. Wilfried Diekmann and David G. Robinson, Pflanzenphysiologisches Institut der Universität Göttingen, Germany, kindly carried out the freeze-etching. This work was supported by a fellowship from the Friedrich-Ebert-Stiftung to the first author and a grant from the Deutsche Forschungsgemeinschaft to the second author.     

Ultrastructure of Draparnaldia glomerata (Chaetophorales, Chlorophyceae) II. Mitosis and cytokinesis

The mitosis and cytokinesis of Draparnaldia glomerata as examined here by transmission electron microscopy are in many aspects similar to those described earlier for other chaetophoralean algae. The standard chaetophoralean model of the mechanism of mitosis/cytokinesis is described in detail. Characteristic in this pattern is the movement of sets of centrioles towards the nuclear poles followed by a proliferation of extranuclear microtubules at prophase, the (partial) fusion of centrioles with the spindle poles at metaphase and anaphase, the simultaneous separation of chromosomes apparently caused by both spindle elongation and shortening of the chromosomal microtubules at anaphase, the expulsion of the centrioles by daughter nuclei and finally the non–persistent spindle at telophase. Cytokinesis takes place by formation of a cell plate associated with phycoplast microtubules. The possible function of the phycoplast in cytokinesis in Draparnaldia is discussed.     

Light and electron microscopical studies onHafniomonas gen. nov. (Chlorophyceae,Volvocales), a genus resemblingPyramimonas (Prasinophyceae)

Based on light and electron microscopical studies ofPyramimonas reticulata the genusPyramimonas is shown to contain a number of unrelated flagellates.P. reticulata andP. montana are transferred to the new genusHafniomonas, cells of which differ fromPyramimonas in shape, in the absence of scales and hairs on the body and flagellar surfaces, in details of the chloroplast, the position of the nucleus, the Golgi apparatus, the internal structure of the flagellar apparatus, and in cell division. The prasinophytePyramimonas contains a characteristic association of a large microbody and a rhizoplast, situated on the nuclear surface. A similar association is being found in an increasing number of prasinophycean flagellates, but is absent inHafniomonas, which is considered related to chlorophycean rather than prasinophycean flagellates. The phylogenetic position ofHafniomonas is discussed, based in particular on details of the unique flagellar apparatus.     

Limnothrix redekei (Van Goor) Meffert (Cyanobacteria) Strains from Lake Kastoria, Greece Form a Separate Phylogenetic Group

Three strains of Limnothrix (Cyanobacteria) isolated from Lake Kastoria, Greece, were characterized based on their morphological features and 16S rRNA gene sequences. The Limnothrix isolates 007a, 165a, and 165c can morphologically be assigned to Limnothrix redekei (Van Goor) Meffert. The 16S rRNA gene of the Limnothrix strains showed a 99% similarity to the 16S rRNA gene of Planktothrix sp. FP1. Limnothrix redekei strains 165a, 165c, 007a and Planktothrix sp. FP1 formed a separate cluster in the cyanobacterial 16S rRNA gene tree. It was distinct from the Pseudanabaena cluster, which included the other Limnothrix strains isolated from northern temperate lakes. This is the first report on the phylogeny of L. redekei strains originating from a Mediterranean lake (southern Europe) and provides new data about the genus Limnothrix.     

Growth pattern of isolated zoospores in Hydrodictyon reticulatum (Chlorococcales, Chlorophyceae)

Zoospores at various developmental stages in Hydrodictyon reticulatum were isolated from parent cells and cultured in Waris medium. Isolated zoospores grew to mature vegetative cells, and were able to reproduce zoo-spores that formed daughter hexagonal nets. Three types of shape appeared in cells 24 h after isolation: cylindrical, Y-shaped and 4-armed type. Protrusions of Y-shaped or 4-armed cells were formed at an angle of about 120° to the long axis of the cell. When cells were isolated at later stages, more cells became cylindrical in shape and fewer ceils became Y-shaped or 4-armed, Direction of cell growth also seemed to depend largely on the developmental stages of the zoospores. The later the isolated stages were, the more the cells elongated along the long axis of the zoospores.     

New species of the genus Zygnemopsis (Zygnematales,Chlorophyceae) from India

Three new species of the genus Zygnemopsis (Skuja) Trans. 1934 (Z. benghalensis sp. nov.; Z. pseudolahaulensis sp. nov., and Z. scrobiculata sp. nov.) are described from West Bengal, India.     

Upper limits of temperature for growth inChlorella (Chlorophyceae)

The upper limit of temperature for growth is a species-specific character in the genusChlorella. The limits of 14Chlorella species range from 26–30°C (C. saccharophila) to 38–42°C (C. sorokiniana), withC. fusca var.vacuolata (34°C) andC. kessleri (34–36°C) assuming an intermediate position. Thus, there is no wide gap in the temperature limits between the normal (“low-temperature”) species ofChlorella and the “high-temperature” species,C. sorokiniana.     

Photoacclimation of Dunaliella tertiolecta (Chlorophyceae) Under Fluctuating Irradiance

We investigated photoacclimation of Dunaliella tertiolecta (Butcher) in irradiance (I) regimes simulating mixed layer conditions of turbid estuarine waters or lakes. D. tertiolecta was exposed to a range of fixed I regimes to establish baseline physiology-I relationships that were compared with subsequent photoacclimation to a simulated mixed layer. Measured indices of photoacclimation included cellular pigmentation, chlorophyll variable fluorescence, and effective photosystem 2 antenna size. While D. tertiolecta grown under fluctuating I maintained division rates comparable to cells grown at high I, the cells exhibited characteristics of photoacclimation consistent with cells grown under a stable regimes at irradiances considerably lower than the average I of the simulated mixed layer.     

Silene rothmaleri P. Silva (Caryophyllaceae), a rare, fragmented but genetically diverse species

Silene rothmaleri is an endemic Portuguese species considered extinct until 1992, when it was rediscovered in the wild with a highly fragmented distribution. These rare plants occur along the southwestern Portuguese coast in small populations, which in addition to phenological differences that occur along the north–south gradient could create a pattern of genetic isolation. To evaluate the degree of genetic diversity and estimate the relationship between population fragmentation and genetic variability, we analysed the five known populations of S. rothmaleri using random amplified polymorphic DNA. Degree of polymorphism and Shannon Index of phenotypic diversity revealed high levels of diversity, found mainly within populations. PCo and cluster analysis revealed a distinct north–south cline, which was confirmed by spatial autocorrelation (Mantel) analysis. This indicates the existence of gene flow between small nearby populations and its insufficiency between widely separated populations. Levels of gene flow (Nm) estimated from the Shannon Index reveal a pattern consistent with a larger past distribution that went through a period of contraction and lack of gene flow followed by population differentiation. The central and largest population probably acts as a core of genetic variability inherited as a relict from a larger and more diverse ancestral population.     

Typification of Cerastium fontanum ssp. vulgare (Caryophyllaceae)

The basionym of Cerustium fontanum ssp. vulgare (Hartman) Greuter and Burdet, C. vulgare Hartman, is neotypified, based on a specimen in UPS. Flora Nordica notes No. 14.     

Characterization of microsatellite loci in polyploid Lavatera assurgentiflora ssp. assurgentiflora and ssp. glabra (Malvaceae)

Lavatera assurgentiflora (Malvaceae) is one of four species of the genus Lavatera native to California and Baja California. Two geographically defined subspecies are recognized: L. a. assurgentiflora on the northern islands and L. a. glabra on the southern islands. We isolated nine polymorphic microsatellite loci that amplify in both subspecies of L. assurgentiflora. Substantial levels of polymorphism were observed at many of the loci. Four loci exhibited more than 10 alleles, polymorphism information content ranged from 0.4 to 0.8, and up to six alleles were found in some individuals, supporting reports that these taxa are hexaploid. All loci also amplified in Lavatera lindsayi from Guadalupe Island, and we anticipate that they will cross‐amplify in other California Lavatera species as well.     

Culture studies of Acrochaete leptochaete comb. nov. and A. wittrockii comb. nov. (Chaetophoraceae, Chlorophyceae)

A culture study of the plants known as Phaeophila leptochaete (Huber) Nielsen and P. wittrockii (Wille) Nielsen proved that these are distinct species. Both have the Acrochaete type of hairs, and therefore, the new combinations Acrochaete leptochaete and A. wittrockii are introduced. Several isolates of both species were started from plants collected at different places in Europe.
A. leptochaete in culture was mainly characterised by having two or more (up to six) pyrenoids in many of its cells, while in A. wittrockii there invariably was only one. The swarmers formed by A. leptochaete had two or four flagella. Several hairs from one basal swelling were sometimes observed in one of the isolates. Another isolate often formed a hair on the germinated zoospore. The isolates of A. wittrockii had a variable morphology, from unicellular plants to large pseudoparenchymatous cell masses. Different kinds of swarmers were observed; zoospores with three flagella seemed typical, but ones with two or four flagella were also observed. Small, pale biflagellate swarmers were assumed to be gametes. One of the isolates of A. wittrockii differed from the rest as the only swarmer type observed was zoospores with two flagella. Chlorophilum ephemerum is considered identical to A. wittrockii. Observations on hair formation in A. repens suggested that the hair structure represents a separate cell.     

Production of extracellular carbohydrate in cultures of Ankistrodesmus densus Kors. (Chlorophyceae)

An extracellular polysacchande, released into the culture mediumby Ankistrodesmus densus is shown to be a product of activephotosynthesis rather than diagenesis. Although produced inthe exponential phase, its accumulation is most pronounced inthe stationary phase under conditions of nitrogen depletion.It is a glycan, containing residues of galactose, mannose, glucose,xylose, arabinose and a trace of rhamnose.