Phycological Research

Symbiotic algae of the ciliate Paramecium bursaria (Ehrenberg) Focker are key species in the fields of virology and molecular evolutionary biology as well as in the biology of symbiotic relationships. These symbiotic algae were once identified as Zoochlorella conductrix Brandt by the Dutch microbiologist, Beijerinck 120 years ago. However, after many twists and turns, the algae are today treated as nameless organisms. Recent molecular analyses have revealed several different algal partners depending on P. bursaria strains, but nearly all P. bursaria contains a symbiont belonging to either the so-called ‘American’ or ‘European’ group. The absence of proper names for these algae is beginning to provoke ill effects in the above-mentioned study areas. In the present study, we confirmed the genetic autonomy of the ‘American’ and ‘European’ groups and described the symbionts as Chlorella variabilis Shihira et Krauss and Micractinium reisseri Hoshina, Iwataki et Imamura sp. nov., respectively (Chlorellaceae, Trebouxiophyceae).

Field and culture samples of the red algal genera Bostrychia and Stictosiphonia from all around the world were analyzed for the polyols D-sorbitol and dulcitol, that function as osmolytes, as well as for the heteroside digeneaside by using 13C-NMR spectroscopy and HPLC. While all plants exhibited D-sorbitol, the occurrence of dulcitol and digeneaside was highly variable. Therefore, different types of low molecular weight carbohydrate distribution patterns were found in Bostrychia and Stictosiphonia. The presence of D-sorbitol seems to be a reliable chemosystematic character for both genera, because no other red alga is known to contain this compound. The lack of dulcitol may be correlated with the geographical origin of the Bostrychia and Stictosiphonia samples: while all tropical isolates exhibited both sugar alcohols, in cold-temperate plants only D-sorbitol was determined. In warm-temperate species, however, both polyol distribution types may occur. These data are discussed in terms of possible temperature sensitivity of the dulcitol pathway. However, the biological function of digeneaside (the main photo-assimilated compound in members of the order Ceramiales) is still obscure.

Two Synechococcus strains from the Culture Collection of the Institute for Marine Sciences of Andalusia (Cádiz, Spain), namely Syn01 and Syn02, were found to be closely related to the model strain Synechococcus sp. PCC7002 according to 16S rDNA (99% identity). Pigment and lipid profiles and crtR genes of these strains were ascertained and compared. The sequences of the crtR genes of these strains were constituted by 888 bp, and showed 99% identity between Syn01 and Syn02, and 94% identity of Syn01 and Syn02 to Synechococcus sp. PCC7002. There was coincidence in photosynthetic pigments between the three strains apart from the pigment synechoxanthin, which could be only observed in Synechococcus sp. PCC7002. Species of sulfoquinovosyl-diacyl-glycerol (SQDG), phosphatidyl-glycerol (PG), mono- and di-galactosyl-diacyl-glycerol (MGDG and DGDG) were detected by high performance liquid chromatography-mass spectrometry analysis of lipid extracts. The most abundant species within each lipid class were those containing C18:3 together with C16:0 fatty acyl substituents in the glycerol backbone of the same molecule. From these results it is concluded that these cyanobacterial strains belong to group 2 of the lipid classification of cyanobacteria.

Dr Shigeo Yamanouchi was born in Yamagata Prefecture and completed his secondary education at Tokyo Higher Normal School (THNS) where he was also a professor until 1904. In 1905, he went to the University of Chicago in the USA and earned a PhD in Botany in 1907. He is best noted for his excellent research on the cytology and life histories of the marine algae Polysiphonia, Fucus, Cutleria, Aglaozonia and Zanardinia, published between 1906 and 1921 while he was associated with the University of Chicago. He also described the freshwater green alga Hydrodictyon africanum. In 1910, he returned to THNS as a Professor and wrote several botanical textbooks, receiving his DSc degree in 1911 and traveling in England and the USA as an advisor for the Japanese Ministry of Education during 1911–1913. For much of the time between 1920 and 1942 he remained in the USA, returning to Japan following the advent of World War II, During his later life, he was in obscurity, and sadly there is very little recorded of his activities in the post-war years. He died in Tokyo on 2 February 1973 at the age of 96.

SUMMARYA marine araphid pennate diatom Plagiostriata goreensis is described from the sand grains of Goree Island, Dakar, Republic of Senegal, based on observations of fine structure of its frustule. The most striking feature of the species is its striation, which is angled at approximately 60° across the robust sternum. The other defining features of the species are its one highly reduced rimoportula and apical pores located at both ends of the valve margin. In the 18S rDNA phylogeny, the species appears as a member of a ‘small-celled clade’ of araphid pennate diatoms that consist of Nanofrustulum, Opephora and Staurosira. The results of the phylogenetic analyses suggest that the distinct characters of the diatom; namely, oblique striae and apical pores, may have been acquired independently. However, it remains unclear whether the rimoportula of P. goreensis is a reduced state or P. goreensis acquired its morphologically curious rimoportula independently after the loss of an ancient rimoportula at the root of the small-celled clade.

Chattonella verruculosa Y. Hara et Chihara was re-examined by molecular methods and microscopic examination. The 18S rDNA phylogenetic analysis clearly indicated that C. verruculosa is a member of the Dictyochophyceae, with a specific affinity to Florenciella parvula. The morphological features in C. verruculosa– namely the proximal helix with two gyres and many scattered DNA-containing areas in the chloroplasts – display the evolutionary link to the Dictyochophyceae, instead of the Raphidophyceae. Similarly, unique pyrenoid morphologies are shared between C. verruculosa and the dictyochophycean algae. Combining the molecular data and morphological characteristics, C. verruculosa is transferred to Pseudochattonella gen. nov. of the class Dictyochophyceae as Pseudochattonella verruculosa (Y. Hara et Chihara) Hosoi-Tanabe, Honda, Fukaya, Inagaki et Sako comb. nov.

To elucidate the phylogeny of the Dunaliellales sensu Ettl. the taxon often thought to be intermediate between primitive green flagellates such as the prasinophytes and the advanced chlorophycean algae, the sequences of the nuclear-encoded small subunit ribosomal RNA gene (18SrDNA) were determined and analyzed for five green algae, including three dunaliellalean algae. Phylogenetic trees based on 18SrDNA suggest that Oltmannsiellopsis viridis (Margraves et Steele) Chihara et Inouye represents an early divergence in the Ulvophyceae/Trebouxio-phyceae/Chlorophyceae clade and has no close relationship to any other green algae, as also suggested from ultrastructural characters. We propose Oltmannsiellopsidates ord. nov. for this genus. Hafniomonas and Polytomella are included in the clade which is characterized by clockwise basal bodies (CW group). The 18SrDNA trees suggest that multiple losses of the cell wall of the flagellate cell occurred in the CW group, and that the Dunaliellales sensu Ettl has a polyphyletic nature. This study also suggests that Planophita terrestns Groover et Hof-stetter (Chaetopeltidales) and Chaetophora incras-sata (Hudson) Hazen (Chaetophorales) are distinct lineages in the Chlorophyceae.

Nuclear-encoded small subunit ribosomal RNA gene (18SrDNA) sequences were determined for Chlamydomonas moewusii Gerloff and five chlorococcalean algae (Chlorococcum hypnosporum Starr; Chlorococcum oleofaciens Trainor et Bold; Chlorococcum sp.; Tetracystis aeria Brown et Bold; Protosiphon botryoides (Kützingl Klebs). All these algae are characterized by a clockwise CCW) flagellar apparatus. Phylogenetic trees were constructed from sequences from these algae together with 20 green algae. All algae with a CW flagellar apparatus form a monophyletic clade (CW group). Three principal clades can be recognized in the CW group, although no morphological character supports monophyly of any of these three clades. The 18SrDNA trees clearly demonstrate the non-monophyly of the Chlamydomonadales and Chlorococcales, suggesting that vegetative morphology does not reflect phylogenetic relationships in the CW group. The paraphyly or polyphyly of the genus Chlamydomonas and Chlorococcum are also revealed. Present analysis suggests that the presence or absence of a zoospore's cell wall and the multinucleate condition have limited taxonomic values at higher taxonomic ranks.

The use of nitrogen-to-protein conversion factors (N-Prot factors) is the most practical way of determining protein content. The accuracy of protein determination by this method depends on the establishment of N-Prot factors specific to individual species. Experimental data are needed to allow the use of this methodology with seaweeds. The present study was designed to characterize the amino acid composition and to establish specific N-Prot factors for six green, four brown and nine red marine algae. Mean values for individual amino acids tended to be similar among the three groups, but some differences were found. Green algae tended to show lower percentages of both aspartic acid and glutamic acid than the other two groups of algae. The percentages of both lysine and arginine were higher in red algae, while brown algae tended to show more methionine than green and red algae. The actual protein content of the species, based on the sum of amino acid residues, varied from 10.8% (Chnoospora minima, brown algae) to 23.1% (Aglaothamnion uru-guayense, red algae) of the dry weight. Nitrogen-to-protein conversion factors were established for the species studied, based on the ratio of amino acid residues to total nitrogen, with values ranging from 3.75 (Cryptonemia seminervis, red algae) to 5.72 (Padina gymnospora, brown algae). The relative importance of non-protein nitrogen is greater in red algae, and consequently lower N-Prot factors were calculated for these species (average value 4.59). Conversely, protein nitrogen content in both green and brown algae tends to be higher, and average N-Prot factors were 5.13 and 5.38, respectively. An overall average N-Prot factor for all species studied of 4.92 ± 0.59 (n = 57) was established. This study confirms that the use of the traditional factor 6.25 is unsuitable for seaweeds, and the use of the N-Prot factors proposed here is recommended.

Based on examination of a clay sample comprising part of the type material of diatoms described by Skvortzow in 1936 from Lake Biwa, lectotypes of the following 14 pennate diatom taxa were designated: Caloneis nipponica Skvortzow; Cocconeis disculus var. nipponica Skvortzow; Cymbella nipponica Skvortzow; Cymbella turgidula var. nipponica Skvortzow; Gomphonema lingulatum var. elongatum Skvortzow; Navicula costu-lata var. tenuirostris Skvortzow; Navicula gastrum fo. nipponica Skvortzow; Navicula hasta var. gracilis Skvortzow; Navicula lambda var. nipponica Skvortzow; Navicula lanceolata var. nipponica Skvortzow; Navicula radiosa fo. nipponica Skvortzow; Navicula rostellata var. biwaensis Skvortzow; Navicula similis var. nipponica Skvortzow; and Navicula undulata Skvortzow. Navicula lanceolata var. nipponica, N. costulata var. tenuirostris, N. undulata, G. lingulatum var. elongatum and N. lambda var. nipponica are probably later synonyms of other taxa. Navicula lambda var. nipponica and N. similis var. nipponica are given the new combinations Sellaphora lambda var. nipponica (Skvortzow) Ohtsuka and Placoneis clementis var. nipponica (Skvortzow) Ohtsuka, respectively. Cymbella turgidula var. nipponica, N. hasta var. gracilis, and N. rostellata var. biwaensis are each elevated to the independent species under the names Cymbella rhe-ophila Ohtsuka, Navicula subhasta Ohtsuka and Navicula biwaensis (Skvortzow) Ohtsuka, respectively.

The Culture Collection of Algae and Protozoa (CCAP) and the National Institute for Environmental Studies (NIES) act as service collections in the UK and Japan, respectively. Since 1991, the collections have collaborated on a wide range of topics of mutual interest. These include: algal taxonomy, bio-informatics, collection management, conservation of microbial biodiversity, cryopreservation, and cyanobacterial toxin research. The main activities have focused on the re-examination of cyanobacterial isolates housed in both collections and cryopreservation research. The preliminary data generated in the survey of cyanobacterial strains were used in the production of the latest CCAP catalogue and database. This project is ongoing at NIES and forms a key component of a major project on cyanobacterial taxonomy. To date, many protists are apparently freeze-recalci-trant. A primary mode of lethal cell injury is the formation of intracellular ice. In this study, intracellular ice formation was observed in Tetraselmis suecica (Ky-lin) Butcher under suboptimal cooling conditions, Tetraselmis chui Butcher under both fast and slow cooling and in both pigmented and achlorophylous strains of Euglena gracilis Klebs. These findings have significant implications for the development of successful cryopreservation strategies. In addition to the above activities, a variety of algal strains have been examined in taxonomic and ultra-structure research projects and cyanobacterial cultures have been screened for novel toxins.

We report the isolation, characterization and expression of a cDNA encoding a polypeptide elongation factor-1 (EF-1) from the marine red alga, Porphyra yezoensis. A cDNA clone was isolated from a leafy gametophyte cDNA library and the sequence was analyzed. The clone contained an open reading frame for a protein of 449 amino acids which exhibits sequence similarity to the known EF-1. Comparison of the deduced amino acid sequence showed higher similarity to the Porphyra pur-purea EF-1tef-c (97%) than to the P. purpurea EF-1tef-s (61%). The mRNA was detected both in the leafy gametophyte and the filamentous sporophyte.

In the Yellow Sea of China, large-scale green tides have broken out for three consecutive years from 2007 to 2009. As part of the efforts to localize the algal source, two cruises were conducted in the early stage and the outbreak stage of the bloom in 2009. We analyzed the morphological and genetic diversity of drifting Ulva specimens and culture-derived isolates from seawater sampled in different localities. For phylogenetic analyses, the nuclear encoded ribosomal DNA internal transcribed spacer region (ITS nrDNA) and the plastid encoded large subunit of ribulose-1, 5-bisphosphate carboxylase/oxgenase gene (rbcL) were used. Our molecular and morphological data indicate that the dominant free-floating Ulva species in 2008 and 2009 possibly belonged to a single strain of the U. linza-procera-prolifera (LPP) clade. The ITS sequences from bloom-forming algal samples with dense branches were identical to those from U. linza-like specimens without branches derived from the Yellow Sea. Microscopic individuals of the dominant Ulva strain were detected in eight stations, revealing that spore dispersal in the water helped to enlarge biomass in the water during the outbreak stage of green tide in the Yellow Sea.

Changes in photosynthetic activity and trehalose levels in field-isolated, natural colonies of the terrestrial cyanobacterium Nostoc commune responding to desiccation and salt stress were investigated. As the water content decreased in N. commune colonies during desiccation, photosynthetic O2-evolving activity decreased and no activity was detected in desiccated colonies. A high level of O2 evolution was restored in the colonies as they absorbed atmospheric moisture, indicating that only a small amount of water is required for reactivation of photosynthesis. No detectable trehalose was found in fully hydrated N. commune colonies; however, trehalose accumulation occurred in response to water loss during desiccation and high levels of trehalose were detected in the air-dried colonies. Moreover, a 0.2 M NaCl treatment also induced trehalose accumulation to a level equivalent to that by desiccation. Photosynthetic O2 evolution was inhibited by 0.2 M NaCl, indicating that N. commune can tolerate only low levels of salt. These results suggest that cessation of photosynthesis and trehalose accumulation occur in response to both matric water stress (desiccation) and osmotic water stress (high salt concentration), and that while trehalose may be a less effective osmoprotective compound than others, it is important for the extreme tolerance to desiccation observed in terrestrial cyanobacterium.

Tandem repeats of the 5S ribosomal RNA gene (rDNA) were confirmed for almost all laminarian, cymathaerean and kjellmaniellan species distributed in northern Japan. The nucleotide sequence of the spacer region between tandemly repeated 5S rDNA was investigated for 79 samples from 31 sites. Phylogenetic analysis of the 29 different sequences detected revealed two lineages: (1) Laminaria coriacea group, including Laminaria coriacea Miyabe, Laminaria cichorioides Miyabe, Laminaria sachalinensis (Miyabe) Miyabe, Laminaria yendoana Miyabe, Cymathaere japonica Miyabe et Nagai, Kjellmaniella gyrata (Kjellman) Miyabe and Kjellmaniella crassifolia Miyabe; (2) Laminaria japonica group including Laminaria japonica Areschoug, Laminaria religiosa Miyabe, Laminaria ochotensis Miyabe, Laminaria diabolica Miyabe, Laminaria longipedalis Okamura, Laminaria angustata Kjellman and Laminaria longissima Miyabe. In addition, the latter group was divided into two: subgroup (2a) including L. angustata and L. longissima and subgroup (2b) including L. japonica, L. religiosa, L. ochotensis, L. diabolica and L. longipedalis. Members of the three groups differ from each other in the appearance of ornaments (bullation, gyration and folds) on the surface of the blade. These are absent in group (2a), only present in the early stages of the lifespan of group (2b), and present for the duration of the lifespan in group (1). Genetic distances among samples were extremely small within group (2a). Together with previous crossing studies and data on ocean currents and distribution, these findings suggest that gene flow occurs within group (2b).

The salinity tolerance of 62 strains of Pfiesteria and Pfiesteria-like heterotrophic dinoflagellates was measured. All strains were acclimated at 12 psu for at least 1 year before experimentation. Strains isolated from the Chesapeake Bay and Neuse River systems tolerated lower salinities than strains isolated from the Wilmington River system (P< 0.005). Swimming cells were still observed after 5 days at 0.5 psu for one strain, and at 1 psu for most other Chesapeake Bay and Neuse River strains. Swimming cells for the Wilmington River were still observed after 5 days at 3–5 psu, but no swimming cells were observed at ≤ 2 psu. With regard to the upper salinity tolerance, the Wilmington River strains tolerated higher salinities than the Chesapeake Bay and Neuse River systems (P< 0.005). Most Wilmington River strains were swimming after 5 days at salinities ≥ 50 psu, whereas the Chesapeake Bay and Neuse River system strains rarely had swimming cells at salinities exceeding 35–45 psu. For all three water systems and for both lower and higher salinities, cells apparently encysted in many instances. However, when salinities were returned to 12 psu, swimming cells often re-appeared. Statistically significant geographic differences in salinity tolerance suggest a geographic adaptation has occurred and that salinity tolerance is under genetic control. The results also suggest there is diversity among the strains.

The relationship between the abundance of nitrogenase and its activity was studied in the marine unicellular cyanobacterium Gloeothece sp. 68DGA cultured under different light/dark regimens. The Fe- and MoFe-protein of nitrogenase and nitrogen (N2)-fixing (acetylene reduction) activity were detected only during the dark phase when the cells were grown under a 12 h light/12 h dark cycle (12L/12D). Nitrogenase activity appeared about 4 h after entering the dark phase. Maximum nitrogenase activity occurred at around the middle of the dark phase, and the activity rapidly decreased to zero before the start of the light phase. The rapid decrease of nitrogenase activity and the Fe-protein of nitrogenase near the end of the dark phase in 12L/12D were partly recovered by the addition of l-methionine-sulfoximine, an inhibitor of glutamine synthetase. Diurnal oscillation of the abundance of nitrogenase was maintained in the first subjective dark phase (i.e. the period corresponding to the dark phase) after the cells were transferred from 12L/12D to continuous illumination. However, enzyme activity was detected only when photosynthetic oxygen (O2) evolution was completely suppressed by reducing the light intensity or by the addition of 3-(3,4-dichlorophenyl)-1,1-dimethylurea. Nitrogenase always appeared in the cells about 16 h after starting the light phase, even when the 12L/12D cycle was modified by the addition or subtraction of a single 6 h period of light or dark. These results suggest the following: (i) N2-fixation by Gloeothece sp. 68DGA is primarily regulated by an endogenous circadian oscillator at the level of nitrogenase synthesis. (ii) The endogenous circadian rhythm resets on a shift of the timing of the light phase. (iii) Nitrogenase activity is not always reflected in the presence of nitrogenase. (iv) The activity of nitrogenase is negatively regulated by fixed nitrogen and the concentration of ambient O2.

The ntcA and ntcB genes, encoding cAMP-receptor-protein (CRP) type and LysR type transcriptional regulators, respectively, were isolated from the transformable marine cyanobacterium Synechococcus sp. strain PCC 7002 to study the inactivation of these genes by interposon mutagenesis. The ntcA gene could not be disrupted in this cyanobacterial strain, indicating that the ntcA gene is essential for cell growth on either urea or nitrate as the sole nitrogen source. The ntcB mutant cells grew very slowly on nitrate or nitrite as the sole nitrogen source but could grow on urea at the same rate as wild-type cells. The consumption of nitrate and nitrite was barely detectable in the ntcB mutant cells, indicating that nitrate and nitrite use was impaired in the ntcB mutant. In wild-type cells, mRNA levels for the three genes required for nitrate assimilation, nrtP (nitrate permease), narB (nitrate reductase) and nirA (nitrite reductase), increased when cells were shifted to nitrate growth conditions, but no increase in transcription of these genes occurred in the ntcB mutant. The mRNA level of ntcA was nearly identical in the wild-type and ntcB mutant cells. These results demonstrate that NtcB specifically controls the transcription of the three genes for nitrate assimilation in this marine cyanobacterium.

Partial rbcL sequences of type specimens, historically significant specimens and recently collected comparative specimens are used to demonstrate that Prionitis decipiens (Montagne) J. Agardh (type locality: Paita, Peru), Prionitis angusta (Harvey) Okamura (type locality: Shimoda, Japan, currently known as Grateloupia angusta (Harvey) Kawaguchi et Wang) and Prionitis cornea (Okamura) E. Y. Dawson (type locality: Shizuoka, Japan, currently known as Grateloupia cornea Okamura), all previously reported from the northeast (NE) Pacific (California, USA and Baja California, Mexico), were not, and currently are not, present. Historically significant specimens from the NE Pacific specifically annotated by E. Y. Dawson as belonging to P. cornea or specifically annotated by I. A. Abbott as belonging to P. angusta were sequenced and shown to belong to either Prionitis linearis Kylin (type locality: La Jolla, California, USA) or to Prionitis filiformis Kylin (type locality: San Francisco, California, USA). The lectotype of Gelidium decipiens Montagne (basionym of Prionitis decipiens) is narrowed to a single specimen illustrated by Montagne, and a lectotype is designated for P. linearis. The utility of partial rbcL sequences (less than 200 base pairs) for determining the identity of specimens is discussed, as is the importance of sequencing type and historically significant specimens.

Sulcochrysis biplastida gen. et sp. nov., a golden, marine, mixotrophic flagellate is described. Cells resemble Ochromonas in light microscopic features, but they are distinct at the electron microscopic level from Ochromonas or any other typical chrysophyte. Ultrastructural features that discriminate Sulcochrysis from the Chrysophyceae are: (i) a proximal helix in the flagellar transition region; (ii) basal bodies situated in the anterior depression of the nucleus; (iii) the lack of the rhizoplast; and (iv) simple flagellar hairs lacking lateral filaments. These features suggest that Sulcochrysis is a relative of the Pedinellophyceae, Dictyochophyceae and Pelagophyceae. However, Sulcochrysis has a flagellar root system similar to that of the Ochromonas-type cell and it may use the R3 root for prey capture, as do ochromonadalean algae. The R3 root and the phagotrophic mechanism using the R3 root are interpreted as a plesiomorphy, because these are also distributed in primitive heterokonts such as the bicosoecids. Sulcochrysis has one microtubule probably homologous with the x-fibre of Bicosoeca maris Picken. Based on these features it is suggested that Sulcochrysis is an organism that links bicosoecids (Bicosoeco-phyceae), Pedinellophyceae, Dictyochophyceae and Pelagophyceae.

SUMMARYUV-absorbing mycosporine-like amino acid compounds (MAA) were identified and quantified in 13 macroalgal Chlorophyceae, six Phaeophyceae and 28 Rhodophy-ceae collected in the intertidal zone from the tropical island Hainan, People's Republic of China, as well as from tropical mangrove locations in America, Africa, Australia and Japan. All of these habitats receive naturally high solar ultraviolet (UV) irradiances. The study revealed that all Rhodophyceae contained several MAA, which are assumed to function as natural UV sunscreens. Within all species investigated eight distinct compounds were found, seven of which were identified as mycosporine-glycine, shinorine, porphyra-334, pal-ythine, asterina-330, palythinol and palythene. The unknown substance had an absorption spectrum with a maximum at 357 nm. This compound was restricted to two red algal species from Hainan. In contrast, the Chlorophyceae and Phaeophyceae did not contain MAA or exhibited only trace concentrations. Compared with data from the literature, the amount of all MAA in the tropical Rhodophyceae seemed to be higher than in temperate organisms, probably reflecting acclimation to the stronger solar radiation which is typical for lower latitudes. The data suggest that accumulation of MAA may represent a natural defence system against exposure to biologically harmful UV radiation.

Filament density of Aphanizomenon flos-aquae (Lemmerm.) Ralfs, water temperature and soluble reactive phosphorus (SRP) were measured from April to August in 1993–1996 in Lake Barato, Hokkaido, Japan. In addition, growth characteristics and internal phosphorus (P) utilization of Aph. flos-aquae were evaluated under P limitation at three temperatures (15, 20 and 25†C) to clarify the role of internal accumulated P for its growth in the incubation experiment. The filament density was highest in early July 1994, when SRP concentration had not yet decreased and the water temperature was high. These are important factors favoring an increase in abundance of this species in L. Barato. During batch culture, the time course of the stationary phase was shortest at 25†C and longest at 15†C; the cellular C:P molar ratio was 111 under P sufficiency and increased eight- to 12-fold under P limitation. As the C:P ratio was significantly higher in the decreasing phase at 15†C, Aph. flos-aquae may be more adaptable to Plimitation at 15†C than at 20†C and 25†C. However, the low temperatures did not favor the abundance of Aph. flos-aquae in 1996. This indicates that the filament density of Aph. flos-aquae decreases before it reaches the maximum value for some reason under P limitation in L. Barato.

To investigate which genes may be important for growth under extreme conditions such as very low or high salinities, a survey of the Dunaliella sp. transcriptome was performed with a cDNA microarray which had been generated previously representing 778 expressed sequence tags. The comparative microarray analysis indicated that 142 genes differed in expression levels by more than twofold in cells grown at extreme salinities (0.08 M and 4.5 M NaCl) when compared with cells grown at intermediate salinity (1.5 M NaCl). Of these genes, 28 had increased expression and 57 were suppressed in cells grown at low salinity. In cells grown at high salinity, 43 genes showed increased expression and 69 genes showed suppressed expression. However, we did observe a large overlap in the expression of extreme salinity-responsive genes based on Venn diagram analysis, which found 55 genes that responded to both of the two extreme salinity conditions. Further, we found that several genes had similar expression levels under low and high salinities, including some general stress response genes that were upregulated in both extreme salinity conditions. For confirmation of the validity of the cDNA microarray analysis, expression of several genes was independently confirmed by the use of gene-specific primers and real-time polymerase chain reaction. The present study is the first large-scale comparative survey of the transcriptome from the microalga Dunaliella sp. acclimated to extreme salinities, thus providing a platform for further functional investigation of differentially expressed genes in Dunaliella.

Using nuclear magnetic resonance spectroscopy, we identified and characterized accumulated compatible solutes in cells of the haptophyte alga Pavlova sp. strain CCMP504. The predominant organic solutes were d-1,4/2,5-cyclohexanetetrol (CHT), 1,3,5/2,4-cyclohexanepentol (CHP) and scyllo-inositol. We then profiled the intracellular organic solutes present in Pavlova sp. grown in medium with salinity ranging from 23 practical salinity units (PSU) (hyposaline) to 35 PSU (optimum salinity for growth), to 47 PSU (hypersaline). The results of these analyses reveal progressive accumulation of CHT and CHP in response to increasing growth medium salinity. We also observed altered accumulation of CHT and CHP in samples subjected to salinity shock. To further characterize the CHT and CHP biosynthesis in Pavlova sp., we carried out stable isotope feeding experiments. Specific labeling of CHT and CHP with d-13C-glucose suggested that d-glucose is a biosynthetic precursor of these cyclitols. The salinity-induced accumulation of CHT and CHP suggests that these cyclitols act as compatible solutes. Our results therefore provide new evidence supporting classification of CHP as a compatible solute.

Concerning the accumulation of S042- in cells, three types of species are known in the Dictyotales: (i) acidic type (high H2S04 accumulation); (ii) high MgS04 accumulation type; and (iii) nonacidic type (low S042- accumulation). Seasonal changes of intracellular pH and concentrations of inorganic ions were examined in six dictyotalean species. In acidic species (Dictyopteris prolifera (Okamura) Okamura and Spatoglossum eras-sum J. Tanaka), intracellular concentrations of S042- and H+ estimated by pH were high through all seasons. In high MgS04 accumulating species (Dictyotasp. and Padina arborescens Holmes), intracellular concentrations of S042- and Mg2+ were high through all seasons. In nonacidic species (Dictyota dichotoma (Hudson) Lamouroux and Dictyopteris undulata Holmes), intracellular concentration of sulfuric acid ion was low all year round.

Fragments of Ecklonia cava Kjellman were cultured under controlled laboratory conditions of light irradiance, water temperature, and photoperiod. To clarify the relationship between the maturation of E. cava and the photosynthetic products, laminaran, the content in the fragments was measured with the progress of maturation. The culture conditions ranged from 12.5 to 100 µmol m−2 s−1, 10–25°C, and 14 : 10 h LD (light : dark) to 10 : 14 h LD. In the case of low light conditions, despite an optimum temperature for maturation, the fragments did not form sori and laminaran was not accumulated during the culture period. In the case of sufficient light and non-optimum temperature conditions, the fragments did not form sori, but laminaran was accumulated. When the fragments were cultured under optimum light and temperature conditions for maturation, laminaran was accumulated in the early stage of maturation, just before or after cortex of the bladelets thickened, and decreased with the progress of maturation, and all fragments matured regardless of the length of the photoperiod. So, these results support the idea that laminaran is used as the main respiratory substrate in the maturation of E. cava.

Cellular pH estimated from cell extract pH, and the ion compositions of major inorganic ions (Na+, NH4+, K+, Mg2+, Ca2+, Cl−, Br−, NO3−, S042-) were studied by ion chromatography in 15 species of 4 orders (Cladophorales, Codiales, Siphonocladales and Ulvales) of Ulvophyceae and 49 species of 8 orders (Bangiales, Ceramiales, Corallinales, Cryptonemiales, Gelidiales, Gigartinales, Nemaliales and Rhodymeniales) of Rhodophyceae. Among the Rhodophyceae, relatively low intracellular pH (approximately 2.0 within cells) and high concentration of S042- was demonstrated in Plocamium telfairiae(Harvey) Harvey. Furthermore, five species of Hypnea Lamouroux were shown to contain high concentrations of S042- balanced by relatively high concentrations of Na+. Among the Ulvophyceae, Codium cylindricum Holmes and Ulva pertusa Kjellman contained high concentrations of S042- balanced by relatively high concentrations of Mg2+.

The Gulf of Mannar on the southeast coast of India, which has recently been declared as biosphere reserve, is the main harvesting place of Gelidiella acerosa for the Indian agar industry. The biomass harvested from natural stocks was estimated to be over 10 000 tonnes dry weight (DW) in the last two decades and is a serious cause of concern on long term sustainability of this valuable resource from this region. The present study reports the seasonal variation in the biomass and agar yield and quality from G. acerosa in four different habitats such as Rameswaram (rocky intertidal), Ervadi (lower intertidal), Sethukarai (subtidal) and Krusadai Island (lagoon reef area). A maximum biomass of 260 ± 26 g DW m−2 was recorded from Ervadi, whereas Sethukarai showed the highest percentage cover (69.83 ± 4.83%) and density (208.20 ± 30.16 plants m−2). Biomass, percentage cover and density were lowest in Rameswaram and significantly lower (P < 0.001) than the other three stations. A single peak in biomass was observed with autumn maxima in Ervadi and Rameswaram and southwest monsoon maxima in Sethukarai and Krusaidai Island. Length of G. acerosa was maximum (9.65 ± 0.25 cm) in Ervadi and the branch index value was maximum (24.70 ± 4.01 br−1 g−1 DW) for Rameswaram. Agar yield (37.24 ± 7.59%) and gel strength (448.66 ± 6.50 g/cm2) were higher in Sethukarai plants. The agar yields attained a peak in the northeast monsoon and decreased in summer, whereas a reverse trend was observed for gel strength. Evidently, agar yield showed a strong negative correlation with gel strength (P < 0.0001) and gelling (P < 0.0001) and melting temperatures (P < 0.01). The single annual peak in biomass observed in the present study, as compared with two peaks reported earlier, are attributed to the continuous overharvesting that would have had an effect on the overall annual biomass production. The findings of this study reveal that a single harvest during the autumn months (January to March) could yield optimum biomass with moderate agar and that would be the best for sustainable usage and conservation of this resource from this region.

Advances have been made in cell and tissue culture of seaweeds to define a unique branch of in vitro techniques; however, they are lagging far behind those of land plants and have limited applications. Explants can be cultivated axenically in enriched or artificial seawater culture media, and regeneration and even callus formation are achieved. In this state of the art technique, seaweed tissue culture may be already useful for certain biotechnological applications, such as clonal propagation of seed material for mariculture. Nevertheless, the absolute control of growth and development as it is exerted in higher plant tissue culture is lacking, and it is required for more complex biotechnological applications in seaweeds. Definitively, we need appropriate cells (competent cells) to induce growth with the most effective chemical regulators in culture medium adjusted towards the addition of carbon sources. Still, free cells and protoplast isolation and regeneration in marine seaweeds constitute the most developed topic in seaweed tissue culture. The regulation of growth and development of seaweed free cell and protoplast cultures may sustain a purposeful use of techniques in the era of genomic applications.

The present study clarifies the fine structure of the vegetative frustules, initial valves and perizonium of Achnanthes crenulata Grunow. The valves of the vegetative cell are distinctly linear-lanceolate with an undulate margin. The valve face is quite flat and in girdle view is smoothly curved as in species of Gephyria (Bacillariophyceae). However, the valve face of the initial cells is slightly rounded and does not have an undulate margin. Furthermore, the rapheless sternum is centrally positioned along the apical axis of the araphid initial valve. As this taxon develops from auxospore to initial valve, it forms only longitudinal perizonial bands; no transverse bands arise. The perizonium consists of three silicified bands: one large, central longitudinal plate and two bands that underlie this plate; these two bands are either open or closed. This taxon has several conspicuous structures compared to other marine species of Achnanthes, but the structure of the perizonium supports the position of A. crenulata within Achnanthes sensu stricto.

Composition of fatty acids in Boekelovia hooglandii Nicolai et Baas Becking (Chrysophyceae) was investigated as a function of salinity. It was confirmed by gas chromatography that the composition of fatty acids in cells cultured in a 50 mmol L−1 NaCl medium consisted of C14:0, C15:0, C16:0, C16:1, C18:0, C18:1, C18:2, C18:3, C18:4, C20:0, C20:4, C20:5, C22:5 and C22:6, in which C14:0, C16:0, C16:1, C18:4, C20:0, C20:5, C22:5 and C22:6 were main constituents. When the cells were cultured in a medium with different concentrations of NaCl ranging from 50 to 800 mmol L−1, the mole percentage of fatty acids such as C14:0, C16:0 and C16:1 decreased with increases in the salinity, while the mole percentage of highly polyunsaturated fatty acids such as C18:4, C20:5, C22:5 and C22:6 increased. When the cells were transferred from a 200 mmol L−1 NaCl medium to a 600 mmol L−1 NaCl medium, a decrease in mole percentage of C14:0, C16:0 and C16:1, and an increase in C18:4, C20:5, C22:5 and C22:6 were observed within 4 h. However, no change in the compositions of fatty acids was observed within 4 h when the cells were transferred from a 600 mmol L−1 NaCl medium to a 200 mmol L−1 NaCl one. The increase in the content of highly polyunsaturated fatty acids was considered to reflect the rapid response to upshock and to be the characteristic of salt tolerance in B. hooglandii.

The Pinguiophyceae class. nov., a new class of photo-synthetic stramenopiles (chromophytes), is described. The class includes five monotypic genera, Glossomastix, Phaeomonas, Pinguiochrysis (type genus), Pinguio-coccus and Polypodochrysis. These algae have an unusually high percentage of polyunsaturated fatty acids, especially 20:5 (n-3)(EPA, eicosapentaenoic acid). These fatty acids are the basis for choosing the Latin noun ‘Pingue’ (= fat, grease) as the root for the class name. Analyses of nuclear-encoded 18S rRNA and chloroplast-encoded rbcL gene sequence data showed that these algae formed a monophyletic group that could not be placed in any other class. Morphologically, the species are all single-celled microalgae from picoplanktonic size to over 40 urn in length. Each cell has one (or two) typical chloroplast(s) with a girdle lamella and a surrounding chloroplast endoplasmic reticulum. Pyrenoids occur within the chloroplast, varying from embedded to stalked, and membranes penetrate into the pyrenoid in all five genera. Phaeomonas has motile cells with two flagella, and the forward-directed flagellum bears mastigonemes (tripartite flagellar hairs). Two other genera (Glossomastix, Polypodochrysis) produce zoospores that possess only one smooth flagellum (no mastigonemes), and this flagellum apparently is the mature flagellum, a feature previously unknown in the photosynthetic stramenopiles. The major carotenoid pigments in the pinguiophytes are fucoxanthin, violaxanthin, zeaxanthin and P-carotene, as well as chlorophyll a and chlorophyll c-related pigment(s). These features support recognition of the Pinguiophyceae class. nov. as a unique group of algae.

The patterns of occurrence of photosynthetic pigments and fatty acids among seven available species (11 strains) of marine raphidophytes were determined and used as chemotaxonomic markers. All currently recognized genera of marine raphidophytes were included for analysis: that is, Chattonella, Fibrocapsa, Heterosigma, Olisthodiscus and Haramonas. The characteristic pigment composition was shown to be chlorophyll a, chlorophylls c1 and/or c2, fucoxanthin as the major carot-enoid, β,β-carotene and any or all of zeaxanthin, violaxanthin and an auroxanthin-like pigment as the minor carotenoids. The carotenoid composition of all marine raphidophyte genera investigated was virtually the same, except in Fibrocapsa and Haramonas, which differed due to the occurrence of fucoxanthinol and 19′-butanoyloxyfucoxanthin, respectively. These fucoxanthin derivatives, in addition to fucoxanthin, have potential chemotaxonomic use for differentiating the two species. In all 11 strains, 15 fatty acids (saturated, mono-unsaturated and polyunsaturated) were determined. Significant taxonomic distinctions between genera were reflected by their fatty acid profiles. A rapid key for the differentiation of genera, in addition to morphological features, may be the absence of the 18:4 fatty acid in Olisthodiscus; presence of 18:5 in Heterosigma; the presence of fucoxanthinol in Fibrocapsa and presence of 19′-butanoyloxyfucoxanthin in Haramonas.

Physiological properties of the temperate hermatypic coral Acropora pruinosa Brook with symbiotic algae (zooxanthellae) on the southern coast of the Izu Peninsula, Shizuoka Prefecture, central Japan, were compared between summer and winter. Photosynthesis and respiration rates of the coral with symbiotic zooxanthellae were measured in summer and winter under controlled temperatures and irradiances with a differential gasvolumeter (Productmeter). Net photosynthetic rate under all irradiances was higher in winter than in summer at the lower range of temperature (12–20°C), while lower than in summer at the higher range of temperature (20–30°C). The optimum temperature for net photosynthesis was apt to fall with the decrease of irradiance both in summer and winter, whereas it was higher in summer than in winter under each irradiance. At 25/ 50/100 μmol photons nr2 s−1, it was nearly the sea-water temperature in each season. Dark respiration rate was higher in winter than in summer, especially in the range from 20–30°C. In both seasons the optimum temperature for gross photosynthesis was 28°C under 400 μmol photons nr2 s−1 and lowered with decreasing irradiance up to 22°C under 25 μmol photons nr2 s−1 in summer, while 20°C under the same irradiance in winter. The optimum temperature for production/respiration (P/R) ratio was higher in summer than in winter under each irradiance. Results indicated that metabolism of coral and zooxanthellae is adapted to ambient temperature condition under nearly natural irradiance in each season.

The behaviors of nuclei and microtubules (MT) in Acrosiphonia duriuscula (Ruprecht) Collins were observed in detail using fluorescence and electron microscopy. Numerous nuclei exist in cells of A. duriuscula (multinucleate cells). Cortical MT radiate from the apex of the tip cell and run parallel to its long axis. Between 30 and 40% of nuclei in the upper part of cytoplasm migrate downward to the region where cytokinesis will take place, and these numerous nuclei form a ‘nuclear ring’ before mitosis. The parallel array of the cortical MT changes to a transverse orientation at the region where cytokinesis will take place, and finally forms a characteristic circumferential band. Mitosis starts from the nuclei in the ring. Cortical MT disappear in the region of the nuclear ring and many mitotic spindles form. The band-shaped array of MT remains. Mitosis spreads in an apparent wave to the other nuclei. After mitosis, daughter nuclei that formed a nuclear ring migrate apically and repopulate the apical daughter cell. When the numerous daughter nuclei have relocated, a rearrangement of the cortical MT occurs. They are randomly arranged at first, but finally become parallel to the long axis of the cell. Cytokinesis occurs by furrowing of the cell, and the band-shaped array of MT could be detected at the leading edge of the furrow.

Cyclin-dependent kinases, the most notable of which is cdc2, are key regulators of the cell cycle and are highly conserved in evolution. We have cloned and analyzed one cDNA containing an open reading frame of 337 amino acids from a multinucleate, multicellular green alga, Acrosiphonia duriuscula (Ruprecht) Collins. The deduced protein, named Adcdc2-1, showed 51% and 54% amino acid sequence identity to yeast cdc2/cdc28 and human cdc2, respectively. Several domains that characterize the cdc2-related kinases were identified from this sequence, although the PSTAIRE motif was replaced with PPTTIRE. Furthermore, this protein has conserved Tyr and Thr residues that are sites of phosphorylation in cdc2-related kinases and are important for regulating kinase activity. The present results suggest that the universal cdc2 is conserved in algae with unique structural characteristics.

The morphology, life history, and the geographical distribution of Spongomorpha spiralis Sakai in Hokkaido, Japan, was studied. The thallus is characterized by its digitate tufts (bundles) of slender filaments with hooked branchlets. The diameter of the filaments is usually 20-50 μm in the basal portion, 50-130 (70-110) μm in the middle portion of main axes, and 30-150 μm in the upper portion. The diameter of the middle portion of the main axes varies from locality to locality. At Muroran, for example, the range is 70-130 μm (average approximately 100 μm), while at other localities it is 50–100 μm (average approximately 70 μm). The diameter of filaments in the upper portion decreases toward the end of the growing season. The cells are multinucleate with four chromosomes in each nucleus of the haploid plant. The manner of cell division is identical to the process reported previously for the Spongomorpha-Acrosiphonia complex. Sexual reproduction is isogamous with anteriorly biflagellate gametes. Plants are unisexual. The life history involves an alternation of heteromorphic generations: the gametophytic phase is a macroscopic, filamentous thallus and the sporophytic phase is a microscopic, ellipsoidal or club-shaped cell. Optimal culture condition for growth and reproduction of both stages was 5°C and long daylength. The gametophyte developed abnormally at 15°C. The sporophyte developed normally at 15°C, but did not produce zoospores.

Chloroplast banding occurs in Griffithsia pacifies Kylin in a daily rhythm. At the start of the light period, chloroplasts have a uniform distribution. During the light period chloroplasts move away from the ends of the cell leaving two chloroplast-free regions by mid-afternoon. Later in the light period these bands disperse as chloroplasts return to the ends of cell where they remain throughout the dark period. After enzyme treatment with β-glucuronidase to permeabilize cell walls, non-banded plants did not form bands after the addition of 5 μg ml−1 cytochalasin B in dimethyl sulfoxide (DMSO). When enzyme-grown plants with chloroplast bands were treated with cytochalasin B, bands were retained for at least three days. Plants grown in media supplemented with β-glucuronidase or DMSO alone gave banding consistent with untreated controls. Staining of microfilaments with rhodamine-phalloidin before and after treatment with cytochalasin B gave results consistent with chloroplast movement studies. This is the first report of actin-mediated organelle movement in red algae.

Catalase (CAT; EC 1.11.1.6) and ascorbate peroxidase (APX; EC 1.11.1.11) activities, as well as malondialdehyde (MDA) and reduced glutathione (GSH) and oxidized glutathione (GSSG) contents, were determined during the growth of the unicellular marine alga Lingulodinium polyedrum (Stein) Dodge in batch-cultures. CAT and APX activity peaks were detected at the beginning of algal exponential growth, although declining trends were subsequently identified in both enzymes, with a slight increase in CAT activity at the end of the experimental period. MDA content attained maximum values from day 0–3 and at the end of the experimental period (day 21), declining halfway from day 10–14. GSH and GSSG contents presented the highest values at the beginning of the growth curve, decreasing from day 3 onwards. Despite the depletion of the GSH pool, an upward trend was observed in the (GSH) (0.5 GSSG + GSH)−1 ratio, indicating that the L. polyedrum cells were able to maintain an increasing redox potential along exponential and linear growth phases in their efforts to prevent oxidative stress.

Under limited nutrient availability (i.e. unenriched sea-water) and under 75 mol photons m–2 s–1 irradiance 12:12 LD, thalli of Solieria chordalis J. Agardh accumulated floridean starch and floridoside. When they were transferred into nutrient-enriched seawater (150 umol L−1 NO31- and 7 umol L−1 P043i at 35 umol photons m−2 s−1 in irradiance 12:12 LD, starch and floridoside levels decreased. The main nucleotide diphosphate (NDP) sugars (i.e. UDP-glucose, UDP-galactose and ADP-glucose) and the activities of UDP-glucose pyrophosphorylase [Enzyme Code (EC) 2.7.7.9] and UDP-glucose-4-epimerase (EC 5.1.3.2) were measured under these controlled culture conditions. Both UDP-glucose and UDP-galactose in the thal l i increased under conditions known to favor the accumulation of floridean starch and floridoside, whereas they decreased under conditions leading to floridean starch and floridoside breakdown. On the other hand, ADP-glucose level only varied slightly. Although UDP-glucose pyrophosphorylase activity rose under conditions of floridean starch synthesis, little variation was observed in UDP-glucose-4-epimerase activity. These results suggest a possible enzymatic regulation of the NDP-sugar and carbohydrate pool in which UDP-glucose pyrophosphorylase would play a major role.

In the search for antioxidant substances/radical scavengers and cytotoxic substances, extracts from four species (10 strains) of epiphytic dinoflagellates in laboratory cultures were subjected to screening for production of bioactive metabolites. Assays for antioxi-dants were performed using microsomal lipids prepared from rat livers, which were oxidized with Fe3+-nitrilotriacetic acid complex (Fe3+-NTA). Generated lipid peroxides were determined using the thiobarbituric acid (TBA) colorimetric method. Screening tests for cytotoxic activity were carried out using P388 leukemic cells of which the survival ratio was assessed using the tetrazolium salt (MTT) method. Extracts from two strains (Gymnodinium sp. and Gambierdiscus toxicus) and those from five strains (two of G. toxicus, two of Coolia monotis, and one of Prorocentrum sp.) were found to contain active constituents for antioxidant activity and for cytotoxic activity, respectively.

The seasonal growth and reproductive phenology of Neorhodomela aculeata (Perestenko) Masuda and Ceramium kondoi Yendo, and the food preferences of herbivorous snails were examined to elucidate (i) why snails select the fronds of N. aculeata for their habitat; and (ii) the survival strategies of the two red algae under grazing pressures. The maximal lengths and weights of both algal species were recorded for each season over a 12-month period beginning with the spring of 2003. C. kondoi grew in length at a faster rate than N. aculeate, whereas the turf alga N. aculeata produced new branches from the tips of broken branches. The reproductive period of C. kondoi was between the spring and summer but the reproductive organs of N. aculeata were observed throughout the year. The algal loss rate of fresh N. aculeata to snails was low but snails had a food preference for N. aculeata when compared to C. kondoi in an artificial food experiment. These results indicate that snails may adapt to chemical compounds characteristic of N. aculeata and that the alga further reduces predation damage by its structural resistance. In conclusion, the survival strategies of C. kondoi appear to be rapid growth, seasonal sexual reproduction, and a delicately branched frond morphology that reduces stable feeding patterns of its predators plus high tissue nitrogen content, whereas the survival strategy of N. aculeata includes regenerative growth responses, structural toughness and chemical defenses while under the grazing pressure of herbivorous snails.

Four species of eukaryotic algae proliferate in the sulfureous, acidic (pH 3.1) water of the largest geothermal pond on Vulcano Island (southern Italy). Consequently, this pond constitutes a natural laboratory for analysis of adaptation by phytoplankters to extremely stressful conditions. To distinguish between the pre-selective or post-selective origin of adaptation processes allowing the existence of phytoplankters in the pond, a Luria-Delbrück fluctuation test was carried out with the chlorophycean Dictyosphaerium chlorelloides and the cyanobacterium Microcystis aeruginosa, both isolated from non-extreme waters; natural water from the Vulcano Island pond was used as selective factor. Preselective, resistant D. chlorelloides cells appeared with a frequency of 4.7 × 10−7 per cell per generation. We propose that the micro-algae inhabiting this stressful pond could be the descendents of chance mutants that arrived in the past or are even arriving at the present. The genetic adaptation of D. chlorelloides to Vulcano waters could help to explain the survival of photosynthesizers in very stressful geothermal waters during the Neoproterozoic ‘snowball Earth’, a period when primary production collapsed in the biosphere. On the other hand, adaptation to these conditions was not observed in M. aeruginosa, suggesting that cyanobacteria may not be able to develop any kind of adaptation to Vulcano pond water.

Gametophyte cultures of seven genetically distinct, filamentous members of the Bangiales were grown under a range of temperatures (10°C, 12°C, 15°C and 20°C) and photoperiods, as well as lowered salinity (5 psu), in order to compare their responses to differing environmental regimes, and to evaluate the utility of a range of morphological, physiological and life history characters for distinguishing these entities. Significant differences in morphological characters such as filament width and cell size were found between some lineages. While these may have some regional diagnostic value, it was concluded that the practical use of these morphometric analyses is limited given the experimental growth period and replication required. The most useful characters for delineating filamentous Bangiales taxa were found to be the germination rates of spores, growth rates of filaments, timing and amount of spore release, and mortality of filaments. Different lineages appear to be adapted to particular environmental conditions, here represented by temperature/photoperiod, and salinity. The results of these comparative culture experiments provide some explanation for the seasonal occurrence and biogeographic distribution of the tested lineages around New Zealand.

The halogenated secondary metabolite constitution of four species of the red algal genus Laurencia (Rhodomelaceae) from southern Japan is reported. Laurencia composita Yamada from Tanegashima Island (Kagoshima Prefecture) bears five sesquiterpenoids (2,10-dibromo-3-chloro-α-chamigrene and 2,10-dibromo-3-chloro-9-hydroxy-α-chamigrene, in addition to pre-pacifenol epoxide, johnstonol and pacifenol, which are known in other populations of this species). Laurencia intricata Lamouroux from Chinzei (Saga Prefecture) and Oomura Bay (Nagasaki Prefecture) bear a C15 aceto-genin, okamurallene. Laurencia majuscula (Harvey) Lucas from Tanegashima Island produces three sesquiterpenoids, (Z)-10,15-dibromo-9-hydroxy-chamigra-1, 3(15),7(14)-triene, 10-bromo-7-hydroxylaurene and 10,11-dibromo-7-hydroxylaurene, corresponding to those of one of its chemical races. Laurencia venusta Yamada from Tanegashima Island produces two sesquiterpenoids, cupalaurenol and cyclolaurenol, which were known only from a sea hare, Aplysia dactylomela Rang. This strongly suggests that Aplysia consumes L. venusta and concentrates these halogenated compounds.

The red alga Laurencia nipponica Yamada (Rhodomelaceae, Ceramiales) is known to contain several chemical races, each of which is characterized by a particular, major halogenated secondary metabolite. Both field-collected and cultured plants of a population of this species found recently at Shikanoshima Island, Fukuoka Prefecture, southern Japan, produced C15 bromoethers, (3E)-laureatin and (3E)-isolaureatin, and sesquiterpenoids, 2,10-dibromo-3-chloro-9-hydroxy-α-chamigrene and 2,10-dibromo-3-chloro-α-chamigrene. Laurencia nipponica can be referred to as a further chemical race that is characterized by the production of two C15 bromoethers, (3E)-laureatin and (3E)-isolaureatin, and a sesquiterpenoid, 2,10-dibromo-3-chloro-9-hydroxy-α-chamigrene as major compounds.

The aim of the present paper is to identify the possible existence of groups of species in the genera Audouinella (Rhodophyta), Cystoseira (Phaeophyceae) and Cladophora (Chloropyta) with significantly similar distribution patterns (chorotypes), in the western Mediterranean Sea and the Adriatic Sea. Of the 98 species studied, 59 were grouped into 11 chorotypes, whereas 39 species remained ungrouped. Thirty-eight species were included in a generalist chorotype, whereas 6 chorotypes were monospecific. The relationships with the environmental factors that could explain the chorotypes are discussed.

An isolate of the very small marine dinoflagellate Biecheleriopsis adriatica gen. et sp. nov. (12–15 µm long) has been examined by light, scanning and transmission electron microscopy, combined with partial sequencing of nuclear-encoded large subunit rRNA. Biecheleriopsis is a genus of thin-walled dinoflagellates, related to Biecheleria and the taxonomic group of Polarella, Protodinium and Symbiodinium, the latter comprising mainly symbionts of marine invertebrates. The mixotrophic Biecheleriopsis adriatica is characterized by: (i) a special type of apical furrow apparatus; (ii) an eyespot of Type E sensu Moestrup and Daugbjerg; (iii) an unusual type of pyrenoid; and (iv) a spiny resting cyst. Thin sections showed the presence a fibrous connection between the flagellar apparatus and a finger-like extension of the nucleus (‘rhizoplast’). It forms a physical connection between the flagella and the nucleus. This unusual structure has previously been considered to characterize the ‘true’ gymnodinioids, represented by Gymnodinium sensu Daugbjerg et al. and related forms. However, the apical furrow apparatus and the nuclear envelope of Biecheleriopsis are woloszynskioid rather than gymnodinioid. The related genus Biecheleria lacks a rhizoplast, and it also lacks a 51-base pair fragment of domain D2 of the large subunit rRNA, which is present in other woloszynskioids. A physical connection between the flagellar apparatus and the nucleus mediated by a fibrous structure is known in other groups of protists, for example, the ‘rhizoplast’ of many heterokont flagellates, some green algal flagellates, etc. The phylogenetic significance of a rhizoplast in two groups of dinoflagellates that are only distantly related is presently difficult to assess.

Cells of the unicellular green alga Scenedesmus obtusiusculus Chod. were cultivated for 2–24 h in nutrient media with low (60 ?mol/L) or high (1000 ?mol/L) phosphorus (P) concentration, and in the presence or absence of 222 ?mol/L aluminum chloride (Al). Cell aggregation was studied by using light microscope, sedimentation and centrifugation. After 2 h, Al was adsorbed to the cell surface and cell aggregates were formed by the attraction of the cells to each other. Aluminum is bound by the negative charges of the cell walls, and studies at different pH showed that a high proportion of positively charged Al forms promote cell aggregation. This effect was most pronounced in low phosphorus cultures as phosphate reduces the effect of Al on cell aggregation by forming aluminum-phosphate. Algae cultivated in the absence of Al did not show any cell aggregation tendencies.

Spontaneous formation and development of adventive embryos were observed in cauline leaves of Sargassum macrocarpum in laboratory culture. Semi-spherical swellings, which were 200–250 μm in diameter, arose from the surface of cauline leaves of thalli cultured for 4 months from zygotic embryos. Swellings became cylindrical protuberances and grew into ‘daughter’ thalli with one or two small cauline leaves. These thalli detached from ‘mother’ thalli and attached to the surface of culture vessels by rhizoids produced within 1 week after detachment. Each daughter thallus developed into an individual thallus exhibiting the same morphological processes as zygotic embryos.