No abstract

Emiliania huxleyi (Lohmann) W. W. Hay et H. Mohler is a cosmopolitan coccolithophore occurring from tropical to subpolar waters and exhibiting variations in morphology of coccoliths possibly related to environmental conditions. We examined morphological characters of coccoliths and partial mitochondrial sequences of the cytochrome oxidase 1b (cox1b) through adenosine triphosphate synthase 4 (atp4) genes of 39 clonal E. huxleyi strains from the Atlantic and Pacific Oceans, Mediterranean Sea, and their adjacent seas. Based on the morphological study of culture strains by SEM, Type O, a new morphotype characterized by coccoliths with an open central area, was separated from existing morphotypes A, B, B/C, C, R, and var. corona, characterized by coccoliths with central area elements. Molecular phylogenetic studies revealed that E. huxleyi consists of at least two mitochondrial sequence groups with different temperature preferences/tolerances: a cool-water group occurring in subarctic North Atlantic and Pacific and a warm-water group occurring in the subtropical Atlantic and Pacific and in the Mediterranean Sea.

Morphological and culture studies were carried out on the genus Scytosiphon {Scytosiphonales, Phaeophyceae) growing in Japan. Morphologies of plurilocular zoidangia and sporophytic thalli were found to be useful as taxonomic characters. Two types of plurilocular zoidangial 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 gracilissp. 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 Oompsonemalike sporophytic thalli. Scytosiphon tenellus is characterized by a thin medullary layer, coherent plurilocular zoidangia, ascocysts among plurilocular zoidangial sori, and Stragutaria-llke sporophytic thalli. Scytosiphon tomentaria (Lyngbye) Link is characterized by a constricted gametophytic thallus, loose plurilocular zoidangia, the presence of ascocysts, and sporophytic thaili identified with Microspongium gelatlnosum Reinke.

The presence or absence of gamete fusion in the dioecious and isogamous brown alga Scytosiphon lomentaria (Scytosiphonaceae, Phaeophyceae) was investigated for thalli collected from three localities, Oshoro, Asari and Muroran in Hokkaido, northern Japan. Both sexual and asexual thalli were found in each locality. These sexual (35 samples) and asexual thalli (15 samples) were used to establish unialgal cultures. ITS2 sequences were determined in all cultures, the rbcL-spacer-S in 14, and cox3 in 25 cultures. Thirteen haplotypes (A1-4, B, C1-4, D, E1-3) were found in ITS2 sequences (241-252 bp). Ten haplotypes (A1-4, B, C1-4 and D) were found in sexual samples: haplotypes A and B were found in Oshoro, C and D in Asari, and C in Muroran. Haplotypes E1-3 were found in asexual samples. Sequence divergence values (including gap information) were less than 4.03% among sexual samples (A, B, C and D types), 0.38-0.77% among asexual samples (E types) and 4.39-5.70% between sexual and asexual samples. In the mitochondrial cox3 gene region (543 bp), six haplotypes (K1-3, L, M and N) were obtained. Cox3-K types were found in samples of ITS2-A types, cox3-L type in ITS2-B type, cox3-M type in ITS2-C and D types, and cox3-N type in ITS2-E types. For cox3 sequences, nucleotide differences were 0.18-4.42% among sexual samples (K, L and M types), but 8.66-10.31% between sexual and asexual samples (N type). In partial rbcL (174 bp)-spacer (188 bp)-partial rbcS (90 bp) sequences, six haplotypes (R-W) were found although there were 1-4 bp nucleotide differences among these haplotypes. R, U and W types were found in sexual samples, and S, T and V types in asexual samples. Results of the sequence analyses suggest that our presumed asexual thalli are genetically different from sexual thalli and may be derived from populations that lack sexual reproduction.

In isogamous brown algae, the sexuality of populations needs to be tested by laboratory crossing experiments, as the sexes of gametophytes are morphologically indistinguishable. In some cases, gamete fusion is not observed and the precise reproductive mode of the populations is unknown. In the isogamous brown alga Scytosiphon lomentaria in Japan, both asexual (gamete fusion is unobservable) and sexual populations (gamete fusion is observable) have been reported. In order to elucidate the reproductive mode of asexual populations in this species, we used PCR-based sex markers to investigate the sex ratio of three asexual and two sexual field populations. The markers indicated that the asexual populations consisted only of female individuals, whereas sexual populations are composed of both males and females. In culture, female gametes of most strains from asexual populations were able to fuse with male gametes; however, they had little to no detectable sexual pheromones, significantly larger cell sizes, and more rapid parthenogenetic development compared to female/male gametes from sexual populations. Investigations of sporophytic stages in the field indicated that alternation of gametophytic and parthenosporophytic stages occur in an asexual population. These results indicate that the S. lomentaria asexual populations are female populations that lack sexual reproduction and reproduce parthenogenetically. It is likely that females in the asexual populations have reduced a sexual trait (pheromone production) and have acquired asexual traits (larger gamete sizes and rapid parthenogenetic development).