Brown algae (Phaeophyceae) are complex photosynthetic organisms with a very different evolutionary history to green plants, to which they are only distantly related(1). These seaweeds are the dominant species in rocky coastal ecosystems and they exhibit many interesting adaptations to these, often harsh, environments. Brown algae are also one of only a small number of eukaryotic lineages that have evolved complex multicellularity (Fig. 1). We report the 214 million base pair (Mbp) genome sequence of the filamentous seaweed Ectocarpus siliculosus (Dillwyn) Lyngbye, a model organism for brown algae(2-5), closely related to the kelps(6,7) (Fig. 1). Genome features such as the presence of an extended set of light-harvesting and pigment biosynthesis genes and new metabolic processes such as halide metabolism help explain the ability of this organism to cope with the highly variable tidal environment. The evolution of multicellularity in this lineage is correlated with the presence of a rich array of signal transduction genes. Of particular interest is the presence of a family of receptor kinases, as the independent evolution of related molecules has been linked with the emergence of multicellularity in both the animal and green plant lineages. The Ectocarpus genome sequence represents an important step towards developing this organism as a model species, providing the possibility to combine genomic and genetic(2) approaches to explore these and other(4,5) aspects of brown algal biology further

The algal pyrenoid is a large plastid body, where the majority of the CO 2 -fixing enzyme, ribulose-1,5-bisphosphate carboxylase/oxygenase (RubisCO) resides, and it is proposed to be the hub of the algal CO 2 -concentrating mechanism (CCM) and CO 2 fixation. The thylakoid membrane is often in close proximity to or penetrates the pyrenoid itself, implying there is a functional cooperation between the pyrenoid and thylakoid. Here, GFP tagging and immunolocalization analyses revealed that a previously unidentified protein, Pt43233, is targeted to the lumen of the pyrenoid-penetrating thylakoid in the marine diatom Phaeodactylum tricornutum. The recombinant Pt43233 produced in Escherichia coli cells had both carbonic anhydrase (CA) and esterase activities. Furthermore, a Pt43233:GFP-fusion protein immunoprecipitated from P. tricornutum cells displayed a greater specific CA activity than detected for the purified recombinant protein. In an RNAi-generated Pt43233 knockdown mutant grown in atmospheric CO 2 levels, photosynthetic dissolved inorganic carbon (DIC) affinity was decreased and growth was constantly retarded; in contrast, overexpression of Pt43233:GFP yielded a slightly greater photosynthetic DIC affinity. The discovery of a θ-type CA localized to the thylakoid lumen, with an essential role in photosynthetic efficiency and growth, strongly suggests the existence of a common role for the thylakoid-luminal CA with respect to the function of diverse algal pyrenoids. marine diatom | CGHR domain | luminal carbonic anhydrase | CO 2 -concentrating mechanism | pyrenoid M arine diatoms are major primary producers, which are responsible for up to 20% of annual global carbon fixation (1, 2). To overcome the difficulties of CO 2 limitation in alkaline and high-salinity seawater, diatoms use a CO 2 -concentrating mechanism (CCM) for the intracellular accumulation of dissolved inorganic carbon (DIC) (3). It is known that the marine pennate diatom, Phaeodactylum tricornutum, uses solute carrier 4 (SLC4) family transporters to take up HCO 3 − actively from the surrounding seawater (4). Based upon physiological measurements of cellular DIC flux, it has been hypothesized that accumulated HCO 3 − is further concentrated in the chloroplast and that an ample flux of CO 2 to ribulose-1,5-bisphosphate carboxylase/ oxygenase (RubisCO) is facilitated by the pyrenoidal β-carbonic anhydrases (CAs), PtCA1 and PtCA2 (5, 6). In this process, α-type CAs present in the matrices of the four-layered chloroplast membranes are thought to prevent leakage of CO 2 from the chloroplast in P. tricornutum (7,8).Algal CCMs are distinct from their carboxysomal counterparts in cyanobacteria, and were most likely acquired by an extensive convergent evolution process (9). It is postulated that the algal CCM is composed of active DIC transport systems at the plasma membrane and the chloroplast envelope, as well as a highly localized CO 2 formation system within close proximity to RubisCO. The possibility remains that the latter process occurs within the py...

Brown algae are one of the most developmentally complex groups within the eukaryotes. As in many land plants and animals, their main body axis is established early in development, when the initial cell gives rise to two daughter cells that have apical and basal identities, equivalent to shoot and root identities in land plants, respectively. We show here that mutations in the Ectocarpus DISTAG (DIS) gene lead to loss of basal structures during both the gametophyte and the sporophyte generations. Several abnormalities were observed in the germinating initial cell in dis mutants, including increased cell size, disorganization of the Golgi apparatus, disruption of the microtubule network, and aberrant positioning of the nucleus. DIS encodes a TBCCd1 protein, which has a role in internal cell organization in animals, Chlamydomonas reinhardtii, and trypanosomes. Our study highlights the key role of subcellular events within the germinating initial cell in the determination of apical/basal cell identities in a brown alga and emphasizes the remarkable functional conservation of TBCCd1 in regulating internal cell organization across extremely distant eukaryotic groups.

The new type blue light (BL) receptor aureochrome (AUREO) was recently discovered in a stramenopile alga, Vaucheria (Takahashi et al. Proc Natl Acad Sci USA 104(49):19625-19630, 2007). AUREO has a bZIP (basic region/leucine zipper) and BL-sensing light-oxygen-voltage (LOV) domain and functions as a BL-activated transcription factor. It mediates BL-induced branching and regulates the development of the sex organ in V. frigida. Although AUREO sequences have previously been found in Fucus and some diatoms, here we report that AUREO orthologs are commonly conserved in photosynthetic stramenopiles. Five AUREO orthologs were isolated from three stramenopile genera (Fucus, Ochromonas, and Chattonella). By BLAST search, several AUREO sequences were also detected in genomes in Aureococcus anophagefferens (Pelagophyceae). However, AUREO was not found in heterotrophic stramenopiles or in closely related phyla, such as haptophytes and cryptophytes, or in green plants. Stramenopiles do not possess phototropin, the well-known BL receptor for phototropism of green plants. From comparative analysis of LOV domains, together with kinship analysis of AUREO bZIP domains, AUREO can be regarded as the BL receptor specific to phototrophic stramenopiles. The evolution of AUREO and the phylogeny of LOV domains in stramenopiles and green plants are discussed.

The ultrastructure of mitosis and cytokinesis in Scytosiphon lomentaria (Lyngbye) Link zygotes was studied by freeze fixation and substitution. During mitosis, the nuclear envelope remained mostly intact. Spindle microtubules (MTs) from the centrosome passed through the gaps of the nuclear envelope and entered the nucleoplasm. In anaphase and telophase, two daughter chromosome masses were partially surrounded with endoplasmic reticulum. After telophase, the nuclear envelope was reconstructed and two daughter nuclei formed. Then, several large vacuoles occupied the space between the daughter nuclei. MTs from the centrosomes extended toward the mid-plane between two daughter nuclei, among the vacuoles. At that time, Golgi bodies near the centrosome actively produced many vesicles. Midway between the daughter nuclei, small globular vesicles and tubular cisternae accumulated. These vesicles derived from Golgi bodies were transported from the centrosome to the future division plane. Cytokinesis then proceeded by fusion of these vesicles, but not by a furrowing of the plasma membrane. After completion of the continuity with the plasma membrane, cell wall material was deposited between the plasma membranes. The tubular cisternae were still observed at the periphery of the newly formed septum. Microfilaments could not be observed by this procedure. We conclude that cytokinesis in the brown algae proceeds by fusion of Golgi vesicles and tubular cisternae, not by a furrowing of the plasma membrane.

From these results, it is suggested that PPD would be associated with the cell wall development during cytokinesis of D. dichotroma.

During cytokinesis in brown algal cells, Golgi derived vesicles (GVs) and flat cisternae (FCs) are involved in building the new cell partition membrane. In this study, we followed the membrane fusion process in Silvetia babingtonii zygotes using electron microscopy together with rapid freezing and freeze substitution. After mitosis, many FCs were formed around endoplasmic reticulum clusters and these then spread toward the future cytokinetic plane. Actin depolymerisation using latrunculin B prevented the appearance of the FCs.Fusion of GVs to FCs resulted in structures that were thicker and more elongated (EFCs; expanded flat cisternae). Some complicated membranous structures (MN; membranous network) were formed by interconnection of EFCs and following the arrival of additional GVs. The MN grew into membranous sacs (MSs) as gaps between the MNs disappeared. The MSs were observed in patches along the cytokinetic plane. Neighboring MSs were united to form the new cell partition membrane. An immunocytochemical analysis indicated that fucoidan was synthesized in Golgi bodies and transported by vesicles to the future cytokinetic plane, where the vesicles fused with the FCs. Alginate was not detected until the MS phase. Incubation of sections with cellulase-gold showed that the cellulose content of the new cross wall was not comparable to that of the parent cell wall.

A small prostrate filamentous brown alga, isolated from the warm temperate Pacific coast of Japan, was cultured to investigate its life history, morphology, ultrastructure and molecular phylogenetic position. The isolate had an isomorphic, diphasic life history with anisogamous sexual reproduction. Gametogenesis was regulated by temperature and occurred only at temperatures 14 C. At higher temperatures, gametophytes reproduced by means of asexual zoids. All swarmers from plurilocular and unilocular zoidangia lacked an eyespot. Both sporophytes and gametophytes had oligostichous filamentous axes, lateral hairs with a basal sheath and no erect thalli. Three to eight oblong chloroplasts, each with a protruding pyrenoid, aggregated to form a stellate chloroplast configuration. All these morphological features suggested that this alga belongs to the genus Asterocladon. Plurilocular gametangia produced in an intercalary or terminal position and zoids lacking eyespots, are, however, unlike those of Asterocladon lobatum, formerly the only member of the genus. Therefore, we described the alga as a new taxon, Asterocladon interjectum sp. nov. Molecular phylogenetic analyses based on the large RUBISCO subunit (rbcL) and partial SSU nuclear ribosomal RNA gene sequences showed that A. interjectum, A. lobatum and Asteronema rhodochortonoides together formed a monophyletic clade, sister to the Ectocarpales sensu lato. Because the molecular phylogenetic analyses showed Asteronema to be paraphyletic, but monophyly of Asterocladon spp. and Asteronema rhodochortonoides was supported by the absence of cytoplasmic invaginations in pyrenoids, Asteronema rhodochortonoides was transferred to Asterocladon and an emended generic circumscription of Asterocladon was included.