The taxonomic status of three closely related species of the brown algal genus Elachista, E. tenuis Yamada, E. sargassicola Noda and E. nipponica Umezaki (Elachistaceae, Ectocarpales sensu lato) was reassessed on the basis of morphological observations including laboratory culture experiments and molecular phylogenetic analyses. These species share characteristic assimilatory filaments of constant width throughout their length except for slightly constricted bases and slightly tapering apices. Although E. sargassicola has been characterized by the presence of phaeophycean hairs, our culture experiments revealed that the presence or absence of such hairs is not a stable taxonomic feature in E. tenuis and E. sargassicola. Furthermore, molecular analyses showed that the rRNA ITS1 and ITS2 regions of E. tenuis and E. sargassicola were nearly identical and grouped them in a strongly supported monophyletic clade. Thus, E. sargassicola should be reduced to synonymy with E. tenuis. On the other hand, E. nipponica, which has been characterized by the production of cylindrical cells in a basal hemispherical cushion, formed cylindrical cells under experimental culture conditions, indicating that the production of such cells is a specific feature of E. nipponica. However, another reported feature of E. nipponica, the production of intercalary plurizoidangia in assimilatory filaments, was rejected, because reexamination of type material revealed that filaments bearing such reproductive structures are referable to Leptonematella fasciculata (Reinke) Silva. Molecular phylogenetic analyses support the distinction of E. tenuis and E. nipponica.

A continuous culture experiment was conducted with the marine diatom Ditylum brightwellii to assess the photo-oxidative effects of transition from dim light to high light at three successive levels. The focus was on temporary stress by ambient UVA at the water surface, via a simulation of the periodic ascent of diatoms caused by vertical mixing. Within a 14 h dim-light period, cells were exposed daily to 4 h of: (A) moderate PAR (100 μmol photon m-2 s-1), (B) high PAR (400 μmol photon m-2 s-1) and (C) high PAR (400–700 nm) plus 6·7 μmol photon m-2 s-1 (2·2 W m-2) UVA. Cell shape, life cycle and sinking rates were not affected by high PAR per se (mode B). The daily 4 h high-irradiance mode C, with additional UVA (UVA: PAR = 0·016), caused an increase in cell diameter, a decrease in length axes, and a production of vegetative resting stages. Superoxide dismutase (SOD) activities, oxidized glutathione (GSSG) pools and malondialdehyde (MDA) contents increased as well, which indicates that this biologically effective UVA dose (0·78 W m-2) promoted active oxygen production, oxidative stress and lipid peroxidation. Sinking rates in UVA-exposed D. brightwellii were higher than in those grown without UVA. A close correlation between sinking rates and MDA contents suggests that UVA-induced lipid peroxidation has altered the membrane functions which regulate buoyancy in D. brightwellii. Although UVA-induced formation of resting cells will contribute to accelerated sinking, it is not evident that this can be considered as a stress avoidance mechanism under adverse light (UVA) conditions. D. brightwellii proved to be sensitive to transitions from shade to natural surface irradiance conditions. In coastal waters, such large diatom species may experience oxidative stress after a few hours' residence near the water surface at ambient UVA levels.

The sublittoral red alga Delesseria sanguinea was pre-cultivated in the laboratory in a predominantly green light field at 10 μmol m−2 s−1 (10 °C), with an approximately 0·5% daily increase in blade area. Such pre-cultivated blades were exposed horizontally in flow-through chambers for 5–6 days in April and May to solar radiation reduced to 19% and 11%, respectively, by neutral density screens. In all three experiments, the full solar spectrum (UVB+UVA+PAR) reduced growth rate significantly to 0–50% of values obtained in PAR alone, while this occurred with UVA+PAR in only one experiment. The growth rate of a Delesseria blade may thus be used as a sensitive and reliable biological indicator of UVB in natural solar radiation at a pre-chosen, neutral reduction level. Rapid measurements of growth rate of apical parts of Delesseria blades during and after UVB+UVA pulses of 2, 3 or 6 h duration were performed in the laboratory by measuring thallus area every 2 min by means of a CCD camera coupled to an on-line, computer-aided image analysis system. A single pulse of 2 or 3 h duration administered during the light phase caused a temporary drop in growth rate during and after the pulse, with recovery starting 2·5 h after the end of the pulse and completed by the end of the light phase. A single 6 h pulse at a biologically effective UVB dose (BEDDNA300 nm) of only 0·5 kJ m−2 reduced growth rate by 55% if administered around noon, but halted growth almost completely if supplied at night, when no photoreactivation was possible. The UVB-sensitive behaviour of Delesseria compares well with the highly sensitive phytoplankton alga Emiliania huxleyi whose growth was reported to be halted at a daily, weighted BEDDNA300 nm of 0·4 kJ m−2 administered during 4 days for 3 h in the middle of the light period.

Laurencia Lamouroux (Rhodophyta) was recently separated into three genera – Laurencia, Chondrophycus (Tokida et Saito) Garbary et Harper and Osmundea Stackhouse – each of which was newly defined based on vegetative and reproductive structures. In this study, the previously unknown vegetative and reproductive morphology of two Australian endemic species of Laurencia, L. clavata Sonder and L. elata (C. Agardh) Harvey, was studied, particularly in the context of the revised generic delineation. These species exhibit vegetative axes with four pericentral cells and trichoblast-type spermatangial development. Tetrasporangia are abaxially produced from the existing third and fourth pericentral cells. L. clavata has terete thalli with distinctive verticillate branching and is similar to Chondria C. Agardh, rather than to Laurencia, in having an unusually marked constriction at the base of the branches and starch accumulation in subcortical and medullary cells. Compared to Laurencia, apical cells of this species exhibit a less oblique division; the resulting recognizable axial cell rows extend somewhat below the branches, and particularly at a young stage they are also clear throughout branchlets. However, other vegetative and spermatangial structures show that L. clavata is more closely allied to Laurencia than to Chondria, and it is placed in Laurencia. By contrast, L. elata exhibits morphology typical of Laurencia and is characterized by large, robust, compressed thalli with fastigiately distichous branching and an extensive secondary cortex. Furthermore, it appears to be distinct from similar species in sometimes having a parasitic species of Janczewskia Solms-Laubach (Rhodophyta). Taxonomy of Laurencia is discussed on the basis of these and previous studies.

The taxonomy of Vitreochlamys species (Volvocales, Chlorophyta) was studied with light and electron microscopy; DNA sequence data were obtained for nine strains from various localities in Japan, the Czech Republic, Slovak Republic and Germany. The strains could be delineated clearly into six species – V. fluviatilis (Stein) Batko, V. gloeocystiformis (Dill) Nakazawa, comb. nov., V. ordinata (Skuja) Nakazawa, comb. nov., V. nekrassovii (Korshikov) Nakazawa, comb. nov., V. aulata (Pascher) Batko, and V. pinguis Nakazawa, sp. nov. – on the basis of differences in cell and protoplast shape, numbers of contractile vacuoles and pyrenoids, form and position of the stigma, location of the nucleus, degree of longitudinal striations on the chloroplast surface, and ultrastructure of the pyrenoid and stigma. Although V. pinguis and V. aulata share common chloroplast morphology, including stigma and pyrenoid ultrastructure, these two species can be distinguished clearly by cell shape, the number of contractile vacuoles and location of the nucleus. In addition, phylogenetic sequence data of the rbcL (large subunit of Rubisco, ribulose-1,5-bisphosphate carboxylase/oxygenase) gene from all of the Vitreochlamys strains strongly suggest that V. pinguis from Japan is robustly separated from the clade composed of four strains of V. aulata from the Czech Republic, Slovak Republic and Japan. The sequence data also showed that Vitreochlamys is composed of three clades, which are largely consistent with the ultrastructural features of the stigma and pyrenoids.

Eighteen collections of red-coloured Audouinella from Central Mexico and southeastern Brazil detected three species. The most common species, A. eugenea, is characterized by macroscopic thalli, the erect system consisting of filaments with cylindrical cells, undifferentiated into proximal and distal parts, and relatively large monosporangia ([ges ] 12·0 μm long). Spermatangia and possible propagules were observed in some Mexican populations. This is the third Audouinella species observed to have gametangia and the first member of the Acrochaetiales with putative propagules. The second species, from Central Mexico, was characterized by the following features: macroscopic thalli, the erect system differentiated into proximal parts with cylindrical cells, unbranched or rarely branched, and distal parts with barrel-shaped cells, abundantly branched to form dense fascicles, with alternate or dichotomous branching, some at right-angles to the axis, and relatively large monosporangia ([ges ] 12·0 μm long). The morphologically distinct proximal and distal portions of the erect system, the latter forming dense fascicles, was a consistent character so far unknown in Audouinella; thus, we propose a new species, A. huastecana sp. nov. The third species is a microscopic epiphyte, A. meiospora, with a well-developed prostrate system composed of creeping and loosely aggregated filaments, and a short homogeneous erect system ([les ] 15 cells) of filaments with cylindrical or barrel-shaped cells and small monosporangia ([les ] 13·0 μm long). A. eugenea and A. meiospora are characterized for the first time from the Southern and Northern Hemispheres, respectively, both occurring mostly in areas of tropical or subtropical rainforests. A. meiospora is reported from new macroalgal hosts. A. eugenea and A. huastecana tended to occur in warm, alkaline waters with a high ion content that were moderate to fast flowing, whereas A. meiospora was not associated with particular habitats.

The red algae generally are considered to have no motile stages. We have confirmed sporadic reports of motile spores in a few red algae by recording freshly released live spores with time-lapse videomicroscopy. Of the 25+ species investigated, only a few had spores that appeared to be immobile. Cells of unicellular species often displayed active movement although were otherwise indistinguishable from nearby immobile cells, and these may be equivalent to the motile spores released after differentiation in more complex multicellular species. There was considerable variation among members of the Bangiophycidae. Cells of Porphyridium often moved at around 0·66 μm s-1 and had conspicuous mucilage tails. Flintiella and Rhodospora showed no cellular movement. Rhodosorus sometimes moved a little and all four strains displayed continuous cytoplasmic rotation within the wall. Glaucosphaera was inert but displayed formation and transport of vesicles from the perinuclear region to the surface where they were discharged. Spores (whether monospores, tetraspores or carpospores) of almost all other species or strains available displayed various forms of motility. Some (e.g. Batrachospermum-Chantransia monospores) showed smooth, directional and continuous gliding (c. 2·2 μm s-1) for shorter or longer distances, clearly detectable to the naked eye through the microscope. In others, movement could be almost as fast or slower, non-continuous and undirectional. Certain spores (e.g. monospores of the filamentous Audouinella sp.) were amoeboid, and in some cases (e.g. Sahlingia) cells actively squeezed through the irregular interstices of a crustose colony. The mechanism of movement is unknown; polysaccharide secretion may be involved in some cases. These forms of motility may be significant as dispersal mechanisms, by moving spores out of the stagnant boundary layer, and in settling processes, by allowing spores limited ability to optimize their site of germination.

Three Cocconeis Ehrenberg species, epiphytic on the leaves of Posidonia oceanica (L.) Delile, were studied by light and electron microscopy. A new species, C. pseudonotata, is described which is closely related to C. notata Petit and C. diruptoides Hustedt. Gross morphology and the presence of a sigmoid raphe and sternum are the characters shared by the three species, whereas the central areas of sternum and raphe-sternum valves, and the stria and areola densities are the main distinctive characters for the species when observed by light microscopy. Scanning and transmission electron microscopic observations revealed that the fine structure of areolae must be also considered in order to identify the species. Comparison among the three species has been carried out by observations on material from our samples, as well as on the type material of C. diruptoides and cleaned samples from the Hustedt collection. The study also provides new information on the geographical distribution of these species.

The development of auxospores and initial cells of the freshwater chain-forming diatom Ellerbeckia arenaria forma arenaria was studied using light and electron microscopy. Whether the auxospore mother cell is formed vegetatively (parthenogenesis) or through autogamy is still a matter for speculation. Because no signs of spermiogenesis could be detected in our material, we exclude oogamy. Further development involves retraction of the protoplast from the epivalve and the formation of a featureless siliceous cap with a central opening on its free surface. The homology and function of this cap remain uncertain. At the other end of the cell the protoplast remains tightly attached to the hypovalve throughout its further development. This hypovalve always has a concave valve face. An organic wall bearing many layers of overlapping, small silica scales begins to be formed while the cell is still enclosed within the mother cell wall, and allows the auxospore to swell, thus breaking the siliceous cap and girdle bands. External and internal surfaces of scales can be distinguished morphologically. Initial valves are formed internal to the scales at an oblique angle to the parent chain. The initial epivalve always faces the hypotheca of the auxospore mother cell, and the initial valves already show elements of the heterovalvy manifest in the vegetative cells. The significance of the observations in terms of ecology and phylogeny is discussed. The combinations Ellerbeckia arenaria (Moore ex Ralfs) R. M. Crawford, and E. arenaria forma teres (Brun ex Schmidt) R. M. Crawford are made.

A new benthic marine dinoflagellate, Bysmatrum granulosum Ten-Hage, Turquet, Quod & Couté, sp. nov., was obtained from sediment and coral samples from sites of La Réunion Island (SW Indian Ocean). This new species is described and illustrated by light and scanning electron micrographs. Cells are 40–50 μm long and 40–46 μm wide. The epitheca is conical and smaller than the hypotheca, which is trapezoidal with convex sides. Plate tabulation is typical for the genus (P0, X, 4′, 3a, 7′′, 6c, 5s, 5′′′, 2′′′′); epithecal plates 3′ and 4′′ separate the intercalary plates 2a and 3a. Thecal plates are perforated by pores and covered by both small and minute wart-like projections, linearly arranged, radiating from the apical pore. This new species differs from the three others of the genus Bysmatrum in the following characters: cell shape and size, size of the apical pore complex and features of thecal plates (plate shapes and ornamentations).

An image analysis system was used to count and measure acetoorcein-stained mitotic chromosomes of the acidothermophilic unicellular red algae Galdieria maxima, G. partita, G. sulphuraria and Cyanidium caldarium (Cyanidiophyceae). Chromosome numbers fell into two groups: n = 2 for all three species of Galdieria and n = 5(7) for C. caldarium. It seems that a separation of C. caldarium from Galdieria is karyologically justified. A chromosome number of 2 appears to be indicative of the genus Galdieria and could be used as a marker to distinguish this taxon from Cyanidium. The two smaller chromosomes in the karyotype of C. caldarium were about 0·4 μm long whereas the other three were 0·5–0·7 μm long. In karyotypes of Galdieria species, the two chromosomes differed in length, the smaller chromosome ranging from 0·8 to 1·8 μm and the larger one from 1·2 to 2·3 μm. The visualization of these extremely small chromosomes was possible due to pretreatment of the cells with the DNA intercalator 9-aminoacridine. The mean absolute length of each chromosome of the three members of Galdieria had statistically significant interspecies differences. Nuclear 1C DNA contents were estimated in the algal cells by the Feulgen technique. All species investigated had genome sizes of 1·50–2·25×10−2 pg. Thus it seems that the members of Cyanidiophyceae have the smallest known genomes of all photosynthetic eukaryotes.

Morphology, sexual reproduction and phylogeny of a colonial green alga collected from Germany were studied in culture. Light and electron microscopy of the gelatinous (extracellular) matrix of vegetative colonies, the absence of obligately somatic cells, and the anisogamous sexual reproduction with sperm packets in this alga indicated that it is assignable to the genus Eudorina. This German alga was similar to E. elegans Ehrenberg in its multiple pyrenoids of nearly identical size and almost identical sized vegetative cells in the colony, but differed from it in having a prominent tubular structure (flagellar sheath) surrounding each flagellum in the gelatinous matrix of the vegetative colonies. Sexual reproduction was homothallic and dioecious. On the other hand, the vegetative morphology agreed well with that of Pandorina minodii Chodat with regard to multiple pyrenoids and the prominent flagellar sheaths. Thus, a new combination, Eudorina minodii (Chodat) Nozaki et Krienitz, is proposed. Molecular phylogenetic analyses of the rbcL-atpB gene sequences from the colonial Volvocales resolved that E. minodii and several heterothallic strains of E. elegans constituted a robust clade. Therefore, prominent flagellar sheaths and homothallic sexual reproduction in E. minodii may be derived characters that evolved recently within the clade.

Thermal acclimation following changes from 6 °C to 15 °C and from 15 °C to 25 °C was investigated in the diatom Chaetoceros calcitrans. Cell division rate increased with temperature. Cellular carbon showed a slight decrease with an increase in temperature from 6 °C to 15 °C but showed no further change with an increase to 25 °C. Cells grown at 6 °C had low levels of light-harvesting components and a high carotenoid to chlorophyll a (Chla) ratio due to an increase in abundance of photoprotective pigments. The abundance of light-harvesting pigments increased with temperature and this was mirrored by a decrease in carotenoid:Chla ratio. All pigments showed an immediate change in abundance following each temperature shift. However, changes in photoprotective pigments reached a plateau after 12 h, in contrast to the light-harvesting pigments that took 4 days to become acclimated. Light-saturated rates of photosynthesis were at a minimum at 6 °C and increased twofold with temperature between 6 °C and 25 °C. Light-limited photosynthesis showed little change with temperature. It is suggested that, at low temperatures, the rate of carbon fixation is impaired due to a decrease in the activity of photosynthetic enzymes. This imparts an increase in excitation pressure due to an imbalance between light absorption and utilization. Cells respond to this by decreasing their light-harvesting capacity and increasing their ability to dissipate the excess energy. This fairly rapid regulation may be ecologically advantageous in areas where there are transient temperature changes.

Bitectatodinium tepikiense Wilson cysts, first described from Pleistocene sediments and generally associated with sediments in temperate to cold-temperate waters, were germinated to establish cultures. The motile thecate cells derived were referable to Gonyaulax digitale (Pouchet) Kofoid. For the first time detailed morphological information is given for the motile stage of this cyst–theca relationship. Cultures were re-encysted; the cultured cysts showed a range of morphology from spherical cysts to those with an apical boss. The taxonomy of Gonyaulax digitale and cyst–theca relationships within the Spiniferites plexus are discussed but, given the debate surrounding the status of the genera Gonyaulax and Spiniferites, no formal taxonomic reassignments are proposed. These observations confirm that establishing cultures of cyst taxa in problematic dinoflagellate groupings and meticulous high-resolution scanning electron microscopic observation of the specimens have great potential for elucidating cyst–theca relationships and clarifying dinoflagellate taxonomy.

Examination of sediment samples from oligo-and dystrophic ponds, lakes and streams in Papua New Guinea, Australia and New Zealand revealed a hitherto unknown diversity of the diatom genus Actinella Lewis. Six new species are proposed, viz. Actinella aotearoaia sp. nov., A. giluwensis sp. nov., A indistincta sp. nov., A. muylaertii sp. nov., A. parva sp. nov. and A. pulchella sp. nov. All species are heteropolar, both in girdle and valve view. Novel information on the genus Actinella includes the observations of two ribbon-shaped, valve-appressed plastids in A. aotearoaia and A. pulchella, and the presence of long mucilage stalks in A. aotearoaia. It is argued that, despite recent proposals to reduce the genera Actinella and Desmogonium Ehrenberg to the rank of subgenera of Eunotia, they should be kept separate until the taxonomic significance of their distinctive morphological features (such as heteropolarity) is fully assessed. The new species appear to be endemic to Australasia and have distinct biogeographies within this region. Except for A. aotearoaia, all species are present in Tasmania; A. indistincta and A. pulchella have also been found in New Zealand (Stewart Island). A. aotearoaia is common in several localities in New Zealand but was also found near Sydney on the Australian mainland. Actinella giluwensis has only been observed in material from Papua New Guinea. The record of A. punctata for the latter country constitutes the first confirmed record for this species outside North America and Europe. The discovery of the new Actinella species again confirms the importance of the Australasian region as a major centre of microalgal biodiversity and endemism.

Comparison of karyology, restriction patterns and DNA methylation levels (as gender separation could be related to some epigenetic mechanism, e.g. DNA methylation) has been performed between male and female individuals of the dioecious species, Chara tomentosa. In contrast to angiosperms, sex determination systems in Charophyceae have not been studied. High-performance liquid chromatography (HPLC) showed that the cytosine methylation level in female plants of Chara tomentosa reached 5·9 %, while in male plants it reached 9·1 %. Karyological analysis did not show any differences in metaphase chromosome number or morphology between sexes. Cytophotometric measurements of DNA content revealed differences in 1C DNA content: 7·0 pg in females and 7·4 pg in males (p < 0·001). Computer image analysis showed that in the interphase nuclei condensed chromatin consists of 34·3 % of the total chromatin in female plants, while in males it was 29·6 % of total chromatin (p < 0·001). Digestion of isolated DNAs with 12 enzymes showed poor enzymatic cleavage (for example with SacI) or restriction patterns which differed between the sexes (TaqI, Bst0I). The restriction endonucleases RsaI and BamHI revealed repetitive DNA families in males. BamHI generated bands of over 3000 bp while RsaI revealed populations of fragments 650–2000 bp long. DNA isolated from male individuals of C. tomentosa was less susceptible than DNA isolated from female plants to digestion with enzymes that are inhibited by methylation of dC residues (HpaII, HhaI).

Photosynthetic responses of rewetted Nostoc flagelliforme to CO2, desiccation, light and temperature were investigated under emersed conditions in order to characterize its ecophysiological behaviour in nature. Net photosynthesis increased to a maximum rate at about 30% water loss, then decreased, while dark respiration always decreased with the progress of desiccation. Light-saturated photosynthesis and dark respiration were significantly reduced at 8 °C, but remained little affected by changes of temperature within the range of 15–35 °C. Photosynthetic efficiency (α) was maximal at the beginning of desiccation and then reduced with increased water loss. Saturating irradiance for photosynthesis was about 194–439 μmol quanta m−2 s−1, being maximal at about 30% water loss. No photoinhibition was observed at irradiances up to 1140 μmol m−2 s−1. Light compensation points were about 41–93 μmol m−2 s−1. Photosynthesis of N. flagelliforme was CO2-limited at the present atmospheric CO2 level. The CO2-saturated photosynthesis increased with increase of irradiance (190–1140 μmol m−2 s−1) and temperature (8–25 °C) and decreased significantly with water loss (0–75%). Photosynthetic affinity for CO2 was sensitive to temperature and irradiance. The CO2 compensation point (Γ) increased significantly with increased temperature and was insensitive to irradiance. Desiccation did not affect Γ values before water loss exceeded 70%. Photorespiratory CO2 release did not occur in N. flagelliforme at the current atmospheric CO2 level.

There is increasing evidence that bacterial–algal interactions play a role in Harmful Algal Bloom (HAB) ecology. Bacteria that are associated with bloom-forming algal species, specifically toxic dinoflagellate algae, have been implicated in the production and biotransformation of paralytic shellfish toxins (PSTs). To clarify the role that these bacteria may play in the production of PSTs, it is desirable to identify and localize the bacteria associated with the dinoflagellates and enumerate them during the course of the algal blooms that the toxic dinoflagellates produce. Because 16S rRNA-targeted probes offer the possibility of both, we previously made and tested probes for some putatively toxigenic bacteria isolated from cultures of the PSP-related dinoflagellates Alexandrium tamarense, A. affine and A. lusitanicum. The bacteria isolated from the dinoflagellates belong primarily to the alpha-proteobacterial group of Roseobacter and the gamma-proteobacterial group of Alteromonas. Here, we report the successful application of these probes to Lugol's-fixed seawater samples. We detected these bacteria in high numbers in the water column when Alexandrium spp. were both present and absent, and during periods when mussels contained PSTs.

Alaria (Alariaceae, Phaeophyceae) is a common genus of kelps in the northern hemisphere. Fourteen species are currently recognized, of which three, Alaria esculenta (L.) Greville, A. pylaii (Bory de Saint-Vincent) Greville and A. grandifolia J. Agardh, are reported for the cold-temperate North Atlantic Ocean. Alaria esculenta, the type species described originally from the North Atlantic, exhibits a range of biogeographically correlated morphotypes suggesting the possibility of multiple species, subspecies or hybrids. In Ireland we discovered an A. esculenta population with unusually long stipes resembling the type specimen of A. grandifolia described from Spitsbergen by J. Agardh in 1872. These and other plants of A. esculenta from Ireland were compared with plants from Spitsbergen fitting the description of A. grandifolia, using sexual hybridization, relative growth rate measurements and DNA sequence comparisons. Complete interfertility was observed between the different isolates. Three nucleotide substitutions (0·37 %) were found in the rbcL and RuBisCo spacer of A. grandifolia, and two in the partial 18S rRNA gene and ITS1 sequences. The relative growth rate at 10 °C of an Irish self-cross was significantly lower than those of all the other crosses. Comparison of RuBisCo spacer sequences of the Spitsbergen A. grandifolia and six A. esculenta isolates showed that A. grandifolia was identical to A. esculenta from Halifax, Canada. The partial 18S rRNA gene and ITS1 sequence of A. grandifolia was identical to that of A. praelonga from Japan and differed by a single substitution from A. esculenta from Scotland and by two nucleotide substitutions from the isolate from Ireland. The intraspecific differences in A. esculenta, together with the hybridization and morphometric results, suggest that A. grandifolia is to be considered conspecific with A. esculenta, and that A. grandifolia is most probably a large deep-water morphological variant, subspecies or ecotype of A. esculenta.

Complete nuclear 18S rRNA and chloroplast 16S rRNA coding sequences of the fusiform green alga Closteriopsis acicularis (G. M. Smith) Belcher et Swale (Chlorophyta) were used to determine its phylogenetic position within the Chlorophyta. Phylogenetic trees inferred from both genes show that C. acicularis is most closely related to Chlorella kessleri Fott et Nováková (Trebouxiophyceae) and does not belong to the family Selenastraceae (Chlorophyceae) as proposed previously. Despite its morphological distinctness, C. acicularis has to be placed close to the genus Chlorella Beijerinck, and especially to C. kessleri. The chloroplast 16S rRNAs of C. acicularis and C. kessleri possess a unique insertion resulting in an additional AT-rich helix in a length-conserved region of the gene. A close relationship to the genus Chlorella is further confirmed by the pyrenoid ultrastructure. The 16S rRNA gene of C. acicularis contains a group I subgroup IA3 intron which is most similar to the first intron in the corresponding gene of Chlamydomonas pallidostigmatica King (Chlorophyta). In helix P6, the intron carries an additional extension related to intron-encoded endonucleases of the LAGLIDADG family. It is the first group I intron reported in chloroplast 16S rDNA of a species other than Chlamydomonas.