Isolation and Characterization of a Sex-Specific Lectin in a Marine Red Alga,Aglaothamnion oosumiense Itono

In red algae, spermatial binding to female trichogynes is mediated by a lectin-carbohydrate complementary system. Aglaothamnion oosumiense is a microscopic filamentous red alga. The gamete recognition and binding occur at the surface of the hairlike trichogyne on the female carpogonium. Male spermatia are nonmotile. Previous studies suggested the presence of a lectin responsible for gamete recognition on the surface of female trychogynes. A novel N-acetyl-d-galactosamine-specific protein was isolated from female plants of A. oosumiense by affinity chromatography and named AOL1. The lectin was monomeric and did not agglutinate horse blood or human erythrocytes. The N-terminal amino acid sequence of the protein was analyzed, and degenerate primers were designed. A full-length cDNA encoding the lectin was obtained using rapid amplification of cDNA ends-PCR (RACE-PCR). The cDNA was 1,095 bp in length and coded for a protein of 259 amino acids with a deduced molecular mass of 21.4 kDa, which agreed well with the protein data. PCR analysis using genomic DNA showed that both male and female plants have this gene. However, Northern blotting and two-dimensional electrophoresis showed that this protein was expressed 12 to 15 times more in female plants. The lectin inhibited spermatial binding to the trichogynes when preincubated with spermatia, suggesting its involvement in gamete binding.     

Vegetative and reproductive morphology of Herpochondria corallinae (Martens) Falkenberg and H. elegans (Okamura) Itono (Ceramiaceae,Rhodophyta) from Korea

The vegetative and reproductive morphology of two epiphytic Herpochondria species, H. corallinae (Martens) Falkenberg (the type species) and H. elegans (Okamura) Itono from Korea were investigated. They are bilaterally compressed and alternate-distichously branched, and lateral branches are produced by oblique divisions of the apical cell. Procarps are restricted to the first periaxial cell of the branches. Spermatangial mother cells produce one or two spermatangia. H. corallinae forms tetrasporangia on six periaxial cells, whereas H. elegans has them on only the two lateral periaxial cells. The prostrate habit, the production of six periaxial cells in opposite pairs, and tetrasporangial stichidia are confirmed as diagnostic characters of the genus Herpochondria.     

The species of Ceramium (Ceramiaceae,Rhodophyta) around Taiwan

A taxonomic study was recently carried out on the species of Ceramium occurring around Taiwan. One hundred and fifty-one collections were made from January 1993 through spring 1994 at 75 sites distributed around the island. A total of thirteen species were found, including three for which names cannot currently be assigned: Ceramium aduncum, C. ciliatum, C. cimbricum, C. cliftonianum, C. flaccidum, C. gracillimum var. byssoideum, C. mazatlanense, C. nakamurai, C. paniculatum, C. tenerrimum and unidentified species 1, 2 and 3. Among these are three new records for Taiwan, C. cimbricum, C. cliftonianum, and C. mazatlanense. This paper summarizes the taxonomic characters of these species and presents a key for their identification. Distribution records of all 42 species reported from the tropical western Pacific are compared. This report is the first detailed record of the genus for Taiwan.     

Daylength and development in four species of Ceramiaceae (Rhodophyta)

Developmental patterns in four species of Ceramiaceae were determined using excised thallus apices grown under a range of light periods. Models of thallus development and organization based on these patterns are presented. Increased rates of apical cell division, greater growth of apical fragments and increased average cell size were found with increasing number of hours light per day between 8–16 and 16–8 h. No aspect of growth investigated was associated with photoperiodic phenomena, and growth occuring during the light break (8-7.5-1-7.5 h) was intermediate between that in 8–16 and 12–12 h. Three patterns of cell elongation were found in the four species in which (1) cell age, (2) cell age and position and (3) cell age, cell position and light period determined cell length at different axial cell positions. Elongation was followed within cells, along axes ofAntithamnion spirographidis for plants grown under different day lengths. Three regions of development were found along main axes: (1) an apical region in which basipetal expansion was greater than acropetal expansion. (2) a zone of stability with equal elongation in apical and basal growth region of cells, and (3) a basal region with greater acropetal expansion. With increasing daylength, the zone of stability was extended to greater ranges of cell length.     

Isolation of Functionally Intact Rhodoplasts from Griffithsia monilis (Ceramiaceae, Rhodophyta)

A procedure is described for isolating photosynthetically active rhodoplasts (“red algal chloroplasts”) from the marine alga Griffithsia monilis. The rhodoplasts exhibited rates of CO₂ fixation and CO₂-dependent O₂ evolution in the order of 200 micromoles per milligram chlorophyll a per hour when illuminated with red or green light and were approximately 80% intact. The response of the rate of photosynthesis to the inorganic phosphate and pyrophosphate concentrations in the medium was qualitatively similar to that previously reported for spinach chloroplasts. Osmotically shocked rhodoplasts evolved O₂ from ferricyanide in red, but not in green, light and were completely uncoupled. Rhodoplast envelope rupture appeared to be accompanied by phycobilisome loss from the thylakoids.     

Female reproductive organization and systematic position ofPlumariella yoshikawae Okamura (Ceramiaceae,Rhodophyta)

A detailed account of female reproductive features ofPlumariella yoshikawae is presented.Plumariella yoshikawae is recognized as morphologically similar toDelesseriopsis elegans in several features, particularly in the sequence of branch initiation, the presence of gland cells on the abaxial sides of the basal segments of the lateral branches, having carpogonial branches on the basal segments of unmodified lateral branches, and the maturation of the carposporophyte at well below the thallus apices. These features indicate thatPlumariella yoshikawae is best removed from the Ptiloteae and is correctly placed in the tribe Delesseriopsideae. The generaPlumariella andBalliella are retained along withDelesseriopsis within the tribe Delesseriopsideae.     

Cell-Cell Recognition during Fertilization in a Red Alga, Antithamnion sparsum (Ceramiaceae, Rhodophyta)

A gamete recognition mechanism in Antithamnion sparsum Tokidais proposed based on experiments using various lectins and carbohydrates.Spermatial binding to trichogynes is inhibited by pre-incubationof spermatia with concanavalin A (ConA) and/or L-fucose, whiletrichogyne receptors are blocked by the complementary carbohydrate-methyl D-mannose and/or the lectin Ulex europaeus agglutinin(UeA1). Binding inhibition (40–50%) was observed with10–50 mM carbohydrates and 25–50 µg ml^-1 lectins.The inhibitory effects of ConA and UeA1 is partially reversed(to 80–90% of controls) by addition of -methyl D-mannoseand L-fucose, respectively. Lectin binding to spermatial surfaceswas visualized by Fluorescein isothiocyanate (FITC) conjugatedConA, whereas carbohydrate receptors along the trichogyne andspermatium were localized with -mannosylated-FITC-albumin andL-fucosylated-FITC-albumin, respectively. These results suggestthat gamete recognition in Antithamnion sparsum is mediatedby a double-docking recognition system consisting of spermatiapossessing surface L-fucose receptors and -methyl D-mannosemoiety, and trichogynes possessing the complementary receptors. (Received December 5, 1995; Accepted April 22, 1996)     

Genes encoding phycobilisome linker polypeptides on the plastid genome of Aglaothamnion neglectum (Rhodophyta)

The genes encoding the phycobilisome anchor protein (apcE) and rod-core linker (cpcG) are on the plastid genome of the red alga Aglaothamnion neglectum. The apcE gene product is 5 to and in the same operon as the and subunit genes of allophycocyanin. This arrangement is identical to the arrangement observed in many cyanobacteria. The cpcG gene product is 5 to the operon encoding the and subunits of phycoerythrin, but is transcribed from the opposite DNA strand. This gene arrangement is different from that observed in cyanobacteria.The amino acid sequences of the A. neglectum anchor protein and rod-core linker polypeptide, as deduced from the nucleotide sequences of the genes, are approximately 50% identical to analogous polypeptides from cyanobacteria and another eukaryotic alga Cyanophora paradoxa. The conserved nature of these proteins suggests that the structure of the core and the rod-core interface are very similar in phycobilisomes of cyanobacteria and eukaryotic red algae.     

Characterization and transcript analysis of the major phycobiliprotein subunit genes from Aglaothamnion neglectum (Rhodophyta)

The genes encoding the and subunits of allophycocyanin, phycocyanin and phycoerythrin from the red alga Aglaothamnion neglectum were isolated and characterized. While the operons containing the different phycobiliprotein genes are dispersed on the plastid genome, the genes encoding the and subunits for each phycobiliprotein are contiguous. The subunit gene is 5 for both the phycocyanin and phycoerythrin operons, while the subunit gene is 5 for the allophycocyanin operon. The amino acid sequences of A. neglectum phycobiliproteins, as deduced from the nucleotide sequences of the genes, are 65–85% identical to analogous proteins from other red algae and cyanobacteria. The conserved nature of the plastid-encoded red algal and cyanobacterial phycobiliprotein genes supports the proposed origin of red algal plastids from cyanobacterial endosymbionts.Many environmental factors effect phycobilisome biosynthesis. The effect of both nutrient availability and light quantity on the level of A. neglectum phycobiliprotein subunits and the mRNA species encoding those subunits is described.     

Karyology and cytophotometric estimation of nuclear DNA content variation in Gracilaria,Gracilariopsis and Hydropuntia (Gracilariales,Rhodophyta)

Chromosome numbers of 1 N=24 were determined for three species of Gracilaria (G. flabelliforme P. Crouan et H. Grouan ex Schramm et Maze, G. mammillaris Montagne and G. tikvahiae McLachlan) and 1 N=32 for two species of Gracilariopsis (G. lemaneiformis (Bory) Dawson, Acleto et Folvik and G. tenuifrons (Bird et Oliveira) Fredericq et Hommersand). Karyotypes for these species exhibit a characteristic size difference between largest and smallest chromosomes. Polyvalents were a common feature of meiotic nuclei. Microspectrophotometry with the DNA-localising fluorochrome DAPI was used to quantify nuclear genome sizes. A 2 C genome size of 0·37–0·40 pg was determined for five species of Gracilaria (G. chilensis Bird, McLachlan et Oliveira, G. flabelliforme, G. mammillaris, G. pacifica Abbott, G. tikvahiae) and 0·33 pg for an isolate of G. verrucosa (Hudson) Papenfuss from Pas de Calais, France. Species of Hydropuntia (H. cornea (J. Agardh) Wynne and H. dentata (J. Agardh) Wynne) and Gracilariopsis (G. lemaneiformis and G. tenuifrons) were found to have slightly larger 2 C genome contents of 0·42–0·47 pg. No intraspecific variation in 2 C genome sizes was found in regional populations of Gracilaria tikvahiae and Gracilariopsis tenuifrons.     

181 Two Lineages Within Griffithsia (Ceramiaceae, Rhodophyta) Based on Plastid Protein-Coding psaA, psbA and rbcL Genes

"Green tides" or blooms of ulvoid green algae are frequent in Yaquina Bay estuary on the central Oregon coast, USA. Measurements of their biomass were made from late spring to early winter in 1999 at six intertidal sites in the estuary, and were continued through the winter of 2002 at two sites that showed the greatest accumulation. The dominant blooming species were the Enteromorpha linza complex, Ulva fenestrata, E. flexuosa , and E. intestinalis . Red and brown algal abundance was negligable. Nutrients in the central channel were monitored, along with incident light and water column absorbence, and sediment temperature. The abundance of benthic amphipods was negatively correlated with the concentration of dissolved sulfides in porewater. However, dissolved sulfide levels were not correlated with green macroalgae biomass. Additional correlations were sought between the variables measured in an effort to suggest or negate major cause and effect relationships. This research is relevant to discerning the effects of eutrophication as a stressor on estuarine processes and could contribute toward an understanding of anthropogenic impacts on biological communities in coastal ecosystems.     

Microcladia exserta sp. nov. (Ceramiaceae,Rhodophyta) from the East coast of South Africa

Microcladia exserta, a new species of the red algal genus Microcladia Greville (Ceramiaceae, Ceramiales), is described from the Natal coast of South Africa. This small, creeping alga, an epiphyte on the coralline alga Amphiroa anceps (Lamarck) Decaisne, is distinguished from other species in the genus by the following combination of characteristics: the prostrate habit, the exerted position of the tetrasporangia, and the presence in the cortex of numerous and conspicuous vesicle cells. Evidence is presented to demonstrate that the criterion of an erect habit in Microcladia vs. a prostrate habit in Herpochondria used to separate these genera is not sound.     

Evidence for two independent lineages of Griffithsia (Ceramiaceae, Rhodophyta) based on plastid protein-coding psaA, psbA, and rbcL gene sequences

The ceramiaceous red algal genus Griffithsia has characteristic large vegetative cells visible to the unaided eye and thousands of nuclei in a single cell at maturity. Its members often occur intertidally along temperate to tropical coasts. Although previous morphological studies indicated that Griffithsia is subdivided into four groups, there is no molecular phylogeny for the genus. We present the multigene phylogeny of the genus based on plastid protein-coding psaA, psbA, and rbcL genes from ten samples of eight Griffithsia species, eight samples of five putative relatives, such as Anotrichium and Halurus, and three outgroup taxa. Saturation plots for each of the three datasets showed no evidence of saturation at any codon position. The partition homogeneity test indicated that none of the individual datasets resulted in significantly incongruent trees. All the analyses of individual and concatenated datasets separated Griffithsia into two well-defined lineages: Lineage 1 was composed of Griffithsia corallinoides, Griffithsia pacifica, and Griffithsia tomo-yamadae, while lineage 2 encompassed Griffithsia antarctica, Griffithsia japonica, Griffithsia teges, Griffithsia traversii, and Griffithsia sp. Our results support the monophyly of the four Anotrichium species and cast a question on the autonomy of Halurus. The monophyly of the tribe Griffithsieae is well resolved, although interrelationships among Griffithsia, Anotrichium, and Halurus were unclear. Our study indicates that the psaA and psbA genes are powerful new tools for the genus-level phylogeny of red algal groups, such as Griffithsia. This is the first report on the multigene phylogeny of the Ceramiales algae based on three protein-coding plastid genes.     

Karyology and evolution inSesamoides (Resedaceae)

The meiotic behaviour abnormalities, fertility and size of pollen of 6 taxa ofSesamoides have been analysed. Besides diploids (2x), polyploids (4x, 6x, 8x) have been found. The chromosome base number is x = 10, but an origin from x = 5 is suggested.     

Systematic revision of the genus Reinboldiella (Ceramiaceae,Rhodophyta) from Taiwan based on comparative morphology and rbcL sequence analyses,including two new species of Reinboldiella

The genus Reinboldiella essentially consists of tiny, membranous blades, epiphytic on benthic red algae, with a wide distribution in the north-western Pacific Ocean (Taiwan, Korea and Japan). In this study, we re-examined the generitype R. schmitziana from Taiwan and neighbouring islands. Two new species of Reinboldiella, R. orientalis sp. nov. and R. taiwanensis sp. nov. are recognized from Taiwan based on comparative morphological studies and molecular analyses of RuBisCO large subunit (rbcL) gene sequences. Reinboldiella orientalis is characterized by its rosette-like appearance, with both erect and prostrate, membranous flat blades whereas R. taiwanensis is distinguishable by mainly consisting of erect, membranous blades with few, short prostrate blades in basal parts of the thalli. Thalli of R. schmitziana also have a rosette-like appearance, but their erect flat blades have cylindrical bases or stipes. The rbcL phylogenetic analyses also support the separation of R.‘schmitziana’ from Taiwan and R. schmitziana from Japan and Korea.     

The morphology and reproduction of Sphondylothamnion multifidum (Hudson) Naegeli (Ceramiaceae)

Sphondylothamnion multifidum (Huds.) Naeg., while a typical member of the Ceramiaceae in its structure and reproduction, differs in post-fertilisation details from other known genera. The sterile cells associated with the procarp system, i.e. the apical cell, the sterile cell on the supporting cell, and the sterile pericentral cell, all divide actively after fertilisation to produce branched involucral systems which surround the gonimoblast.     

New Systematic Insights in the Ceramium Sinicolacomplex: Resurrection of C. Interruptum S. & G. (Ceramiaceae,Rhodophyta)

Setchell & Gardner (1924) provided a taxonomic treatment for the genus Ceramium from lower California and the Gulf of California, Mexico, in which they described several new species, including C. sinicola S. & G. described from Ensenada, BCN and C. interruptum S. & G. described from the vicinity of La Paz, BCS. The latter was later reduced to variety rank in C. sinicolaby Dawson (1950), a taxonomic decision that has subsequently been widely adopted. Phylogenetic analyses inferred from three molecular markers (chloroplast‐encoded rbcL, the RUBISCO spacer, and nuclear encoded SSU rDNA) from recent collections from the Pacific North‐west (California, Oregon) and the Gulf of California reveal a well supported assemblage of three corticated taxa: C. codicolaJ. Ag. 1894, C. sinicola and C. sinicola var. interruptum. Sequence divergence values among the three taxa are sufficient to warrant separate species ranking; hence, we reinstate C. interruptum as a widely distributed epiphyte for the region. C. sinicolais more closely related to C. codicolathan to C. interruptum, with the former two restricted to the host Codium. The molecule‐based relationships are congruent with evolutionary trends in cortication pattern and attachment mode.     

Incubation temperatures, sex ratios and sex determination in a population of Nile crocodiles (Crocodylus niloticus)

A demographic study of the Nile crocodile Crocodylus niloticus at Lake Ngezi, Zimbabwe, revealed that females predominated in all size classes and among embryos. The sex of C. niloticus was shown to be determined by the temperature of egg incubation in constant temperature laboratory experiments. At 31 °C and below only females were produced. The threshold temperature for maleness was between 31 ° and 34 °C, but appeared to vary between clutches. The duration of the incubation period varied with temperature and was 110 days at 28 °C, falling to 85 days at 34 °C. Incubation temperature affected hatchling length, but not mass. Hatchlings from incubation at 34 °C were shorter on average than those from incubation at 28 °C and 31 °C, but by three months had outgrown them. There was no sex-related difference in length in a random sample of 200 two-year-old C. niloticus on a crocodile farm. Mean temperatures in wild nests were consistently lower than 31 °C and therefore the male threshold as determined in the laboratory. Embryonic development was slow and hatching success poor. The shallowest eggs in a nest had higher mean temperatures and more advanced embryos than the deepest eggs. They also experienced daily temperature fluctuations of up to 10 °C during which the maximum occasionally rose to 35 °C. Constant temperature incubation was not a good model of field conditions, but the correlation between nest temperatures and embryonic sex is consistent with temperature-dependent sex determination in the wild.     

Erythropeltidaceen (Bangiophyceae,Rhodophyta) von Helgoland

Ontogenesis and reproduction of the Helgolandian taxa of the Erythropeltidaceae have been studied. In all species monospores are only produced from differentiated sporangia. FilamentousConchocelis-like stages have not been observed. Sexual reproduction was formerly demonstrated in the heteromorphous genusErythrotrichopeltis (Kornmann, 1984). Based on these features a revised classification for the family is presented.Porphyropsis imperfecta, a new species, is a widespread epiphyte in sublittoral habitats.