Purification of α-1,4-glucan lyase from red algae and fungi has previously been reported. The α-glucan lyase converts α-glucans to 1,5-anhydro-D-fructose. In this study, an endophyte was detected between the algal cells of Gracilariopsis sp., but not penetrating the cell walls. Histological staining was consistent with the endophyte being fungal and immunohistochemistry revealed that it possessed an α-1,4-glucan lyase differing from that of the host. The endophyte α-glucan lyase was recognized by antisera against both fungal and red algal α-glucan lyases, whereas the α-glucan lyase found in the algal tissue was recognized only by the antisera against algal α-glucan lyase. The lyases occur independently in the host and in the endophyte, and were sometimes detected simultaneously. The endophyte was only observed in Gracilariopsis sp. collected in China; no endophyte could be detected in Gracilariopsis sp. from California or in Gracilaria chilensis.

The Japanese endemic charalean species Nitella gracilens Morioka (=N. furcata subsp. furcata var. sieberi f. gracilens) was thought to be extinct until it was rediscovered at the type locality, Lake Ashinoko, Kanagawa Prefecture. Uni-charalean cultures of N. gracilens were established and sexual organs developed and matured. The morphology of this alga, examined by light microscopy, corresponds closely with the original description by Morioka, particularly with regard to the oospores. The mature oospore walls of N. gracilens are orange or yellowish-brown and the fossa wall of the oospore is smooth or has a finely granulate ornamentation. In addition, scanning electronic microscopic examination demonstrated that the fossa walls have minute granulate structures that are compactly arranged. These observations contrast with a previous report that the oospore wall ornamentation of this taxon is reticulate. Since oospore wall ornamentation and the form of the dactyls in N. gracilens are essentially different from those of other infraspecific taxa of N. furcata sensu Wood (1965), N. gracilens should be maintained as a distinct morphological species within the genus Nitella.

The zygote wall in developing Durvillaea potatorum embryos is secreted soon after fertilization. Mucilage covers the surface of the zygote wall ensuring rapid adhesion of the newly fertilized egg. The zygote wall is replaced by new cell walls secreted by growing, dividing embryonic cells. Separation of the zygote wall from the embryo is evident 4 days after fertilization and is visible at the edge of the cap flanking the side of the developing embryo and/or at the curved part of the cap where it lifts away from the top of the embryo. At this stage the embryo has undergone an average of 8 cell divisions, increased 3 times in length, and started to increase in width. The zygote wall is eventually shed as it cannot accommodate embryo expansion. The growing embryo flicks out of the rigid zygote wall, which usually remains connected to the developing rhizoid by a mucilaginous strand. By day 5 the zygote wall has detached from the embryo in 22% of individuals and this increases to nearly 70% of individuals by day 10. Removal of alginate-gelling cations from the artificial sea medium indicates that the zygote wall has different properties from the underlying embryonic cell walls. Shedding of the zygote wall allows continued embryo growth, frees the embryonic thallus from the substratum and cleans the embryo surface of phenolic bodies and mucilage that are secreted soon after fertilization; it also removes micro-organisms colonizing the mucilage.

The red algae Chondrophycus patentirameus (Montagne) Nam (‘patentiramea’) and L. paniculata (C. Agardh) J. Agardh were investigated on the basis of type material and recent collections. Both species show the following features: (i) production of two vegetative pericentral cells from each axial segment; (ii) absence of secondary pit connections between cortical cells; (iii) lack of projecting cortical cells near the apex; (iv) absence of lenticular thickenings in the walls of medullary cells; and (v) perpendicular arrangement of tetrasporangia, each of which is produced from the second pericentral cell in each fertile segment with no additional tetrasporangial pericentral cells. However, C. patentirameus has a stoloniferous holdfast, divaricate branching (with branches arising at angles up to 90°) and non-palisade-like superficial cortical cells, whereas L. paniculata has a discoid holdfast, non-divaricate branching and a palisade-like cortical layer. From this study it is concluded that C. patentirameus (here lectotypified) is distinct from L. paniculata. The latter species is here transferred to the genus Chondrophycus (Tokida & Saito) Garbary & T. Harper as follows: Chondrophycus paniculatus (C. Agardh) G. Furnari.

A new member of Delesseriaceae (Ceramiales, Rhodophyta) is described from Southern Taiwan and the Philippines. On the basis of comparative vegetative and reproductive morphology, and phylogenetic analysis inferred from nuclear-encoded large-subunit ribosomal DNA sequences (LSU rDNA), we conclude that it belongs in the genus Drachiella, tribe Schizoserideae, subfamily Phycodryoideae. The new taxon shares with other Drachiella species the absence of macro- and microscopic veins; diffuse growth by marginal and intercalary meristematic cells; a polystromatic, lobed thallus; abundance of rhizoidal marginal proliferations used for attachment; convoluted plastids in surface cells; abundant secondary pit connections among adjacent vegetative cells; large intercellular spaces between surface cells; procarps confined to the upper side of the thallus, circular in outline, consisting of a supporting cell bearing a strongly curved carpogonial branch and two sterile groups that remain undivided; vertical division of gonimoblast initial from auxiliary cell, and unilateral, monopodial branching of gonimoblasts; and mature cystocarps with a massive candelabrum-like fusion cell of fused gonimoblasts bearing carposporangia in branched chains. It is distinguished from the other members of the genus by thalli that consist of extensive tangled mats of prostrate and overlapping decumbent blades, procarps confined to the upper side of the thallus, and the lack of basal stalks or stipes. Whereas the Schizoserideae is predominantly a Southern Ocean tribe, one of the tribe's four genera, Drachiella, was known only from the eastern Atlantic and Mediterranean. We herein report the first record of the genus for the Indo-Pacific Ocean, and describe Drachiella liaoii, sp. nov., as a fourth species in the genus.

The bchI gene (synonym: chlI), whose product is involved in magnesium chelatase activity, is located in the small single-copy region of the plastid genome of Heterosigma carterae (heterokont alga, Raphidophyceae; formerly named Olisthodiscus luteus). As a unique feature, bchI in H. carterae overlaps an upstream open reading frame (ORF97) of unknown function by 23 base-pairs. Cells contain mRNAs for the full (1.6 kb) cotranscript as well as transcripts for bchI (1.2 kb) and ORF97 (0.4 kb). Transcription initiation in a plastid run on assay of the ORF97/bchI gene cluster is approximately 5-fold higher in cultures sampled in the light versus the dark. In contrast, Northern analysis shows that all three transcripts are in equal abundance at both light and dark sampling times. An antibody raised against a BchI fragment expressed in Escherichia coli recognized a protein of the expected size in the plastid fraction of H. carterae on a Western blot. Analysis of BchI amino acid sequences suggests that the protein may be membrane translocated and could bind ATP. Phylogenetic analysis of bchI sequences documents a deep evolutionary branching between chlorophyll a/b and non-chlorophyll b plastids.

In a recently published examination of the newly re-discovered holotype of Leptophytum (Lithophyllum) laeve, the characters of the holotype specimen were judged to be taxonomically inadequate. Consequently, an epitype was selected which was determined to belong to Phymatolithon lenormandii, rendering Leptophytum and its type L. laeve junior synonyms of Phymatolithon and P. lenormandii, respectively. We have re-studied the holotype specimen in Stockholm, and in this paper we demonstrate that it is consistent with the protologue and exhibits features that are fully consistent with the modern circumscription of Leptophytum laeve, based on population studies. Our results demonstrate that the holotype specimen is taxonomically adequate and that the selection of an epitype was not necessary. Accordingly, we propose the re-instatement of Leptophytum Adey 1966, based on its type L. laeve (Foslie) Adey (basionym: Lithophyllum lenormandii f. laeve Foslie 1891). The genera Leptophytum and Phymatolithon Foslie are similar in some anatomical respects (such as in having diminutive sub-epithallial initials, i.e. meristem cells, a polystromatic non-coaxial hypothallium, and roundish to flattened epithallial cells in transverse section). The genera differ somewhat in morphology and markedly in most reproductive features (such as spermatangial systems, origin of gonimoblasts, position of conceptacle primordia, and the presence or absence of distinct asexual conceptacle pore cells). The taxonomic, ecological and biogeographic characteristics of the two genera lead us to conclude that the similarities between Phymatolithon and Leptophytum have very likely resulted from parallel evolution.

Glutamate-l-semialdehyde (GSA) aminotransferase catalyses the final step in the C5 pathway converting glutamate to the tetrapyrrole precursor δ-aminolaevulinic acid. This enzyme is sensitive to gabaculine (2,3-dihydro-3-amino benzoic acid) and to 4-amino-5-fluoropentanoic acid (AFPA), which are irreversible, mechanism-based inhibitors of pyridoxal phosphate-dependent enzymes. Spontaneous mutants of Synechococcus PCC6301 resistant to these inhibitors contain altered enzyme that displays corresponding resistance to high concentrations of the inhibitor. The enzyme from strain GR6, resistant to both inhibitors, contains a three-amino-acid deletion at positions 5–7 and a Met248→Ile substitution. The enzyme from strain K40 resistant to AFPA but not to gabaculine, contains a Ser163→Thr substitution. GSA aminotransferases containing either the deletion or the substitution that are characteristic of the GR6 mutant were produced in Escherichia coli using the expression vector pMalc2. These engineered mutant enzymes were characterized in terms of their catalytic parameters and sensitivities to gabaculine and AFPA. Furthermore, maltose binding protein/aminotransferase fusion proteins were characterized spectrophotometrically to monitor the interaction of bound cofactor with diamino- and dioxo-compounds related to the substrate and both inhibitors. Results were compared with those for similarly produced recombinant wild-type, K40 and GR6 GSA aminotransferases. The engineered products with either the N-terminal deletion or the Met248→Ile substitution displayed catalytic efficiencies that were intermediate between the wild-type and GR6 or K40 enzymes. However, with respect to their absorption spectra, sensitivity to inhibitors and the reactivity of bound cofactor, they were essentially wild-type. These in vitro studies demonstrate that both changes in enzyme structure are necessary to obtain the distinctive properties of the GR6 aminotransferase, including resistance to high concentrations of gabaculine and AFPA.

Field emission scanning electron microscopy (FESEM) preparation techniques have been successfully adapted for visualization of the internal and external ultrastructure of Mougeotia filaments and protoplasts. FESEM of the innermost layer of cell wall in Mougeotia filaments revealed that microfibrils are deposited parallel to each other in an interconnected mesh and are oriented perpendicular to the direction of elongation. For the first time, the surface of protoplasts at different stages of regeneration has been observed using FESEM. Nascent microfibril deposition occurs between 1 and 2 h after isolation and arrangement of these microfibrils is random for at least 8 h. Observation of the inner surface of the plasma membrane in burst protoplasts showed that microtubules are not strongly attached for at least 3 h after protoplast isolation.

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.

The red alga Caulacanthus okamurae Yamada occurs commonly on the coasts of South Korea, but reproductive plants have rarely been found in the field. Seasonality of field populations was examined and the temperature responses of growth and reproduction were determined in laboratory cultures. In the field, monthly average temperatures of surface seawater ranged from 12·7 °C±0·73 (SD) to 21 °C±1·19. Monthly mean biomass in 25×25 cm quadrats (n = 3) ranged from 11·2±9·6 g m−2 (SD) to 148·8±19·2 g m−2 in wet weight with minimal biomass in February 1995 and maximal values in August. In culture, the growth of vegetative plants and germination of tetraspores of C. okamurae occurred at temperatures from 13 to 27 °C. Tetrasporangia, gametangia and cystocarps were formed at 19 °C and above, but not at 13 or 16 °C, indicating that reproduction is directly affected by seawater temperature. On the basis of these data, seawater temperature was above the critical temperature for reproduction from August to October (20–21 °C) yet only three plants with tetrasporangial branches were found in the field. In September the biomass of C. okamurae declined sharply following severe storm damage. Thus, the absence of reproductive organs in field populations of C. okamurae may occur because they inhabit a seasonally stressful habitat, in which typhoon damage removes branches before the water temperature becomes favourable for reproduction.

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 marine red alga Ahnfeltiopsis triquetrifolia sp. nov. (Phyllophoraceae, Gigartinales) is described from Japan. Foliaceous unisexual gametophytes are characterized by having three-edged axes, and crustose tetrasporophytes are distinguished by the following combination of features: tightly coalescent perithallial filaments, nemathecial sori, small numbers of both tetrasporangia (3 or 4) and sterile cap cells (1 or 2) in each fertile filament, and the absence of hypobasal tissue. Gonimoblast filaments develop from the auxiliary cell after it has fused with a carpogonium. Carposporangia develop from gonimoblast cells, from medullary cells contacted by the gonimoblast filaments, and from the cells produced by such medullary cells. Mature, compact cystocarps are immersed within the medullary layer and discharge carpospores through carpostomes formed in the thickened cortex. Cultured tetrasporophytes were induced to sporulate by transfer from a long-day to a short-day regime at 20°C, and one single short-day cycle was effective for tetrasporangial induction. Tetraspore germlings gave rise to basal discs from which upright axes developed. The upright axes showed a morphology similar to that of field-collected gametophytes, although they remained vegetative during 18 months in culture.

A mechanism of bicarbonate uptake with a high sensitivity to the putative anion-exchange inhibitor 4,4′-diisothiocyanostilbene-2,2′-disulphonic acid (DIDS) has been previously reported in green algae. In this study, DIDS inhibited net oxygen evolution by Dunaliella tertiolecta by up to 22 %, but internal pH regulation, intracellular CO2 accumulation, carbon fixation and affinity for dissolved inorganic carbon (DIC) in Dunaliella tertiolecta showed low or insignificant sensitivity to DIDS. However, in cells grown and tested at pH 9·5, treatment with DIDS elevated the k0·5(HCO3-), suggesting there may be a minor role for a DIDS-sensitive anion-exchange-type HCO3- transporter in DIC acquisition by D. tertiolecta at high pH. In contrast, significant external carbonic anhydrase (CAext) activity and up to 70 % inhibition of DIC-dependent O2 evolution by acetozolamide (AZ) suggest that CAext has an important role in DIC acquisition in D. tertiolecta, in normal seawater conditions and at elevated pH. Furthermore, the rate of DIC-dependent photosynthesis at high pH, in the presence of AZ, was 12 times higher than the calculated uncatalysed rate of CO2 supply from HCO3-. This requires some system for direct HCO3- uptake by D. tertiolecta, which may include a DIDS-insensitive mechanism. The effects of DIDS upon indirect measures of DIC acquisition should be interpreted cautiously as DIDS may have non-specific effects upon whole cell function, and affect ion transport processes not directly related to HCO3- uptake.

Sexually reproducing populations of the marine red alga Peyssonnelia rosenvingii Schmitz (Peyssonneliaceae, Gigartinales), for which only spermatangia and monosporangia, borne on the same thalli, have previously been known, are reported from the cold-temperate region of Japan. Plants are reproductive in late autumn and early winter. Each carpogonial branch consists of four or five cells, and the suprabasal cell of three- to five-celled auxiliary cell branches functions as an auxiliary cell. The absence of spermatangia and the presence of large, non-reproductive plants during the fertile season suggest that the spermatangial sori are ephemeral. After presumed fertilization a connecting filament develops from the hypogynous cell and runs in the horizontal plane. It branches irregularly and fuses with several auxiliary cells, forming several gonimoblasts along its length. Each gonimoblast produces five to eight carposporangial chains in conspicuously elevated nemathecia up to 350 μm high. Tetrasporangia are also formed in similar nemathecia up to 300 μm high and have three- to eight-celled pedicels.

Polypeptides of 21, 36 and 37 kDa are induced in the unicellular green alga Chlamydomonas reinhardtii when cells are transferred from high (5%) to low (0·03%) CO2 concentrations. The synthesis of these polypeptides is correlated with the induction of the CO2-concentrating mechanism. Interaction between the induction of low-CO2-inducible polypeptides, the CO2-concentrating mechanism and photorespiration has been studied in wild-type C. reinhardtii with the aim of clarifying whether the glycolate pathway is involved in algal acclimation to limiting CO2 conditions. Our results showed that the induction of the 37 kDa periplasmic carbonic anhydrase and 21 kDa polypeptide under low-CO2 conditions was not observed in the presence of aminooxyacetate, an inhibitor which completely blocks glycolate metabolism. However, the induction of the 36 kDa polypeptide was not affected by this inhibitor. The presence of aminooxyacetate during the acclimation to low CO2 conditions also inhibited the increase in the photosynthetic affinity for inorganic carbon shown by low-CO2-acclimated Chlamydomonas cells without the inhibitor. Our results suggest that the induction of the CO2-concentrating mechanism may require the function of the glycolate pathway. In addition, our results also indicate that there is differential regulation of the induction of these three low-CO2-inducible polypeptides in Chlamydomonas reinhardtii.

We recently reported (Gueneau et al., 1998) the existence of a GC-rich region downstream of the trnR gene which was conserved in the plastidial genome of three algae: Thalassiosira weissflogii, Odontella sinensis (Bacillariophyceae) and Porphyra purpurea (Rhodophyceae). We identified this GC-rich region as a possible vestigial ORF and noted additional high similarity to an open reading frame (ORF) in a plastid from a fourth alga, Pavlova lutherii (Haptophyceae); sequence data from P. lutherii did not extend to the TΨC tRNA arm. We now report that this region instead encodes a recently recognized type of RNA known as tmRNA (Fig. 1A).

An elachistacean epiphyte, Neoleptonema yongpilii E.-Y. Lee & I.K. Lee, gen. et sp. nov., is reported from Korean coasts. The plants are distinguished by having unbranched assimilatory filaments with intercalary plurilocular sporangia as well as lateral plurilocular sporangia from the cortex. The new genus differs from the genera Leptonematella P. Silva and Halothrix Reinke by having pod-shaped plurilocular sporangia on the medulla, and from Elachista Duby and Proselachista Y.P. Lee & Garbary by having intercalary plurilocular sporangia and a poorly developed medulla. The phylogenetic relationships of Neoleptonema yongpilii were inferred from the spacer sequences between the genes coding for the large and small subunits of the RuBisCO gene. The new genus is a member of a poorly resolved clade consisting of several genera within the Elachistaceae, within which it is more closely related to Halothrix and Elachista nipponica than to Leptonematella.

Detailed studies on the sporophytes and gametophytes of Dictyopteris australis and Dictyopteris muelleri from Australia have demonstrated the usefulness of several reproductive characters in species discrimination. In D. australis, spherical sporangia that project above the thallus surface and are clustered around reflexed bundles of paraphyses contrast markedly with the angular sporangia of D. muelleri, which are embedded in the thallus and are scattered in broad fertile zones on the upper branches. In D. muelleri, tetrads of released spores enveloped by the inner sporangial wall are temporarily retained above the thallus surface on the ends of mucilaginous stalks. Sporangial stalks have not been reported in any other species of the brown algae, although egg stalks are produced by several fucalean species. Differences in oogonial structure can also be used to distinguish D. australis and D. muelleri. The spherical oogonia of D. australis, which project above the thallus surface, are conspicuously different from the angular embedded oogonia of D. muelleri. Detailed comparative studies on other species of Dictyopteris are now required to ensure that reproductive characters are used not only to define species of Dictyopteris but also to effectively circumscribe the genus.