PHYCOBILIPROTEIN COMPOSITION AND CHLOROPLAST STRUCTURE IN THE FRESHWATER RED ALGA COMPSOPOGON COERULEUS (RHODOPHYTA)1

Serial sections of uncorticated axial cells of Compsopogon coeruleus revealed a single interconnected parietal chloroplast. Phycobilisomes in such chloroplasts were hemidiscoidal in shape with a broad-face diameter of ca. 25–30 nm. The molar ratio of phycobiliproteins in whole cell extracts was IPE:3PC:1APC, similar to isolated phycobilisomes. Two spectrally distinct C-phycocyanin forms (A618 nm, F648 nm and A630 nm, F652 nm) were resolved in dissociated phycobilisomes along with B-phycoerythrin and allophycocyanin.     

Phycobilisomes from blue-green and red algae: isolation criteria and dissociation characteristics

A general procedure for the isolation of functionally intact phycobilisomes was devised, based on modifications of previously used procedures. It has been successful with numerous species of red and blue-green algae (Anabaena variabilis, Anacystis nidulans, Agmenellum quadruplicatum, Fremyella diplosiphon, Glaucosphaera vacuolata, Griffithsia pacifica, Nemalion multifidum, Nostoc sp., Phormidium persicinum, Porphyridium cruentum, P. sordidum, P. aerugineum, Rhodosorus marinus). Isolation was carried out in 0.75 molar K-phosphate (pH 6.8 to 7.0) at 20 to 23 C on sucrose step gradients. Lower temperature (4 to 10 C) was usually unfavorable resulting in uncoupling of energy transfer and partial dissociation of the phycobilisomes, sometimes with complete loss of allophycocyanin. Intact phycobilisomes were characterized by fluorescence emission peaks of 670 to 675 nanometers at room temperature, and 678 to 685 nanometers at liquid nitrogen temperature. Uncoupling and subsequent dissociation of phycobilisomes, in lowered ionic conditions, varied with the species and the degree of dissociation but occurred preferentially between phycocyanin and allophycocyanin, or between phycocyanin and phycoerythrin.     

Investigations on the arrangement and fine structure ofPorphyridium cruentum phycobilisomes

Summary Phycobilisomes ofPorphyridium cruentum were investigated in ultrathin sectionsin situ and after isolation and positive or negative staining. The phycobilisomes have the approximate shape of one half of a compressed rotation ellipsoid with the following dimensions: length 2 a=40±4 nm, width 2 b=19±2 nm, height c=28±3 nm. The phycobilisomes form rows and pillars on the thylakoid membranes. A plug-like structure (foot) which apparently fixes the phycobilisomes to the thylakoid membrane is described.     

Observations on gymnoascaceae. IX. A new species of pseudoarachniotus from soil-burial beds

Summary A new species ofPseudoarachniotus P. marginosporus, is described and illustrated. The developmental morphology of this new ascomycete is discussed. On a standard medium such as YpSs,P. marginosporus can be distinguished from other species of the genus by colonial coloration which varies from ochre-rust to very pale yellow at different stages of maturation. The colony ofP. roseus is red, red-orange or orange-yellow;P. citrinus is lemon yellow to brownish yellow in age;P. reticulatus is red orange andP. hyalinosporus is yellowish and usually with a great deal of green pigmentation in the mycelium and in the medium. Furthermore ascospores ofP. marginosporus are oblate, small and thin-walled, measuring 2.2–3.0×4.0–4.4 µ, with a minute elevated rim around the long axis, while ascospores ofP. reticulatus are globose and reticulate, and those ofP. roseus andP. citrinus are large and thick-walled, measuring about 4.4×7.7 µ. Ascospores ofP. hyalinosporus are similar in size and shape to those ofP. marginosporus, but lack a rim or other markings.     

The Supramolecular Structure of Photosystem II — Phycobilisome-Complexes of Porphyridium cruentum

The structure and arrangement of phycobilisomes of the unicellular red alga Porphyridium cruentum is compared with the organization of the thylakoid freeze-fracture particles in order to determine the relationship between phycobilisomes and photosystem II. The hemi-ellipsoidal phycobilisomes, 20 nm thick, are predominantly organized into rows; their centre to centre periodicity is 30–40 nm, so that they are well separated by a gap of 10–20 nm. The phycobilisomes are cleaved by a central faint furrow, parallel to the long axis from top to base. The organization of the exoplasmic particles in rows is similar to the arrangement of the phycobilisomes so that a structural relationship between both systems, previously demonstrated in cyanobacteria, is evident. Within the rows, the 10 nm EF-particles are grouped in tetrameric complexes separated by distances similar to those observed for phycobilisomes. We propose that the tetrameric EF-particle complexes correspond to tetrameric photosystem II complexes which bind one hemi-ellipsoidal phycobilisome on the stroma exposed surface of the thylakoid. A hypothetical model of this photosystem II-phycobilisome complex is presented.     

STRUCTURE AND PHYCOBILIPROTEIN COMPOSITION OF PHYCOBILISOMES FROM GRIFFITHSIA PACIFICA (RHODOPHYCEAE)1

Phycobilisomes in Griffithsia pacifica are closely spaced on the thylakoid membrane. By negative staining, attached and isolated phycobilisomes have been shown to have a block shaped appearance. They are 63 nm long, 38 nm high, and 38 nm wide, making them the largest thus far reported. Isolated phycobilisomes, shown to be functionally intact by their 675 nm fluorescence emission (excitation 545 nm) were stable for more than a day. Phycobiliproteins from dissociated phycobilisomes, separated on sucrose gradients and by polyacrylamide electrophoresis, yielded large (R-) and small (r-) molecular weight species of phycoerythrin (ca. 4:1 respectively) constituting 89% of the phycobiliprotein content, with R-phycocyanin 8%, and allophycocyanin 3% accounting for the rest. Phycobilisomes of Griffithsia pacifica and Porphyridium purpureum (Bory) Drew and Ross (P. cruentum) are structurally very similar with phycoerythrin being on the outside and surrounding a core of R-phycocyanin and allophycocyanin.     

Cyanobacterial Picoplankton from Lake Constance*

Four types of unicellular cyanobacteria were classified by pigment composition and cell size. These originated from the picoplankton fraction of Lake Constance, Germany. β-Carotene and zeaxanthin were found to be the main carotenoids of three Synechococcus isolates. In this group, coccoid forms with phycocyanin-rich phycobilisomes also produce caloxanthin and nostoxanthin, which are xanthophylls with 3 and 4 hydroxy groups, respectively. In addition, these rare carotenoids are observed in rod-forming Synechococcus isolates which contain phycoerythrin-rich phycobilisomes, but they are very low or absent in the coccoid phycoerythrin-rich isolates. Due to size and pigment content the coccoid forms are similar to Synechococcus leopoliensis (SAUG B 1402-1, formerly Anacystis nidulans) and S. rubescens (SAUG B 3.81) while the rod-forming isolates differ from S. elongatus (PCC 6716) in phycobilisome composition. The isolate BO 8402 was tentatively assigned to Synechocystis but differs in pigment composition from all strains described as yet. The green cultures exhibited a faint red glow due to an unusual high in vivo autofluorescence from phycocyanin. Neither were phycobilisomes found in a standard preparation nor was allophycocyanin present. The most abundant carotenoids are β-carotene and caloxanthin, while zeaxanthin, with 12 % of all colored carotenoids, is low. All forms described in this paper lack complementary chromatic adaptation, indicating that the pigment composition is a reliable parameter to identify these freshwater isolates.     

PHOTOREGULATION OF PHYCOBILISOME STRUCTURE DURING COMPLEMENTARY CHROMATIC ADAPTATION IN THE MARINE CYANOPHYTE PHORMIDIUM SP. C861

Changes in the molecular structure of phycobilisomes during complementary chromatic adaptation were studied in the marine cyanophyte Phormidium sp. C86. This strain forms phycoerythrin (PE)-less phycobilisomes under red light but synthesizes PE-rich phycobilisomes under green light. Analysis of phycobiliprotein composition and electron microscopic examination of phycobilisomes in ultra-thin sections of cells and of isolated phycobilisomes were performed for cells acclimated to red and green light, respectively. The structure of phycobilisomes formed under red light conditions was typically hemidiscoidal. Phycobilisomes in cells acclimated to green light were twice as large in size as those in cells acclimated to red light. This increase in phycobilisome size was a result of the increase in the molar ratio of antenna pigment (PE and phycocyanin) to allophycocyanin, from 3.5 to 11.3. Pigment composition and fine structure of phycobilisomes formed under green light were similar to those of “nonhemidiscoidal” phycobilisomes reported in Phormidium persicinum. These results suggest that changes occur not only in the molecular species of peripheral rods but also in the structure of rods and probably of cores in relation to their connection with rods during chromatic adaptation of Phormidium sp. C86.     

Light and electron microscopical studies onHafniomonas gen. nov. (Chlorophyceae,Volvocales), a genus resemblingPyramimonas (Prasinophyceae)

Based on light and electron microscopical studies ofPyramimonas reticulata the genusPyramimonas is shown to contain a number of unrelated flagellates.P. reticulata andP. montana are transferred to the new genusHafniomonas, cells of which differ fromPyramimonas in shape, in the absence of scales and hairs on the body and flagellar surfaces, in details of the chloroplast, the position of the nucleus, the Golgi apparatus, the internal structure of the flagellar apparatus, and in cell division. The prasinophytePyramimonas contains a characteristic association of a large microbody and a rhizoplast, situated on the nuclear surface. A similar association is being found in an increasing number of prasinophycean flagellates, but is absent inHafniomonas, which is considered related to chlorophycean rather than prasinophycean flagellates. The phylogenetic position ofHafniomonas is discussed, based in particular on details of the unique flagellar apparatus.     

Cell shape in the algaPediastrum (Hydrodictyaceae; Chlorophyta)

Summary Species ofPediastrum, a genus in which the colonies assemble from aggregating zoospores, differ in the number and form of prongs on peripheral cells and the amount of space between cells of the colony; cell shape appears to be genetically based. Peripheral cells of theP. boryanum colony, for example, have two prongs per cell;P. simplex has one prong per cell. Prong extension is suppressed in the interior cells ofP. boryanum, but prong sites have been reported in scanning electron micrographs of the cell walls. A mutant unicellular strain in which cells of the colony separate after attaining typical form reveals several prong sites (6 or more) in each cell. Multiple suppressed prong sites are evident inP. simplex cells as well. Polyeders, 4- and 5-pronged unicells, occur in the life cycle ofP. simplex. Based on these observations and a recent report byMarchant (1979) of a microtubule organizing center associated with the prongs, it is suggested that several microtubule organizing centers are to be found in zoospores ofPediastrum species and may be related to species differences in cell shape.Research supported in part by Argonne Center for Educational Affairs, U.S. Department of Energy, under contract No. W-31-109-ENG-38.     

Laboratory culture and taxonomy of two species ofPedobesia (Bryopsidales,Chlorophyceae) in Japan

The morphology ofPedobesia lamourouxii andDerbesia ryukyuensis, both collected in Shimoda and the adjacent areas in central Japan, was studied from field specimens and laboratory cultures. Specimens which had the same morphology as EuropeanP. lamourouxii produced stephanokont zoospores which developed into either prostrate filaments or expanded discoidal thalli similar to those described by Feldmann and Codomier (1974) and Feldmannet al. (1975). Erect filament identical with the thallus found in nature developed directly from prostrate filaments. The specimens which had morphology similar to that ofDerbesia ryukyuensis described by Yamada and Tanaka (1938) also produced stephanokont zoospores which developed similarly to those ofP. lamourouxii. This species is, therefore, a member ofPedobesia, and it is made a new combinationP. ryukyuensis (Yamada et Tanaka) Kobara et Chihara, comb. nov.     

A new type of complementary chromatic adaptation exemplified byPhormidium sp. C86: Changes in the number of peripheral rods and in the stoichiometry of core complexes in phycobilisomes

The marine cyanobacterium Phormidium sp. strain C86 changes the phycobilisome type depending on light quality. Red-light-adapted cells contained hemidiscoidal phycobilisomes with a photosystem II:phycobilisome ratio of 2.2, while green-light-adapted cells exhibited hemiellipsoidal phycobilisomes with a photosystem II:phycobilisome ratio of 4.4, as determined by a combined analysis of freeze-fractured thylakoid membranes and ultrathin sections and by photochemical determinations of photosystems and phycobilisomes. Core complexes of phycobilisomes of red- and green-light-adapted cells were isolated by affinity chromatography and were subsequently separated into two allophycocyanin-containing fractions. The high-molecular-weight fraction, with a sedimentation coefficient of 24 S and a calculated mol. wt. of 860,000, contained complexes of the quaternary structure (α^AP ₉β^AP ₈β^19.5AP)₂· (L_CM)₂ and tricylindrical shape, previously designated AP_CM. This fraction was similar in size in red- and green-light-adapted cells; however, differences were detected in the low-molecular-weight allophycocyanin fraction containing the "trimeric" complexes with a sedimentation coefficient of 6 S. As shown by comparison of spectral and stoichiometric data of intact phycobilisomes and isolated core complexes, the amount of the α^APB-containing core complex (α^AP ₂α^APBβ^AP ₃) · L_C ¹⁰ was greater in core fractions of green-light phycobilisomes, whereas the amount of the core complexes (α^AP ₃β^AP ₃) · L_C ¹⁰, designated AP · L_C ¹⁰, was higher in cores of red-light phycobilisomes. Phormidium sp. is the first organism examined that exhibits a new type of complementary chromatic adaptation by altering the composition of the phycobilisome core and the number and composition of peripheral rods and by changing the ratio of photosystem II to phycobilisomes. A model summarizing the structural consequences of the results is presented. Received: 5 December 1995 / Accepted: 10 April 1995     

Subparticles of Anabaena Phycobilisomes I. Reconstitution of Allophycocyanin Cores and Entire Phycobilisomes

From the dissociation of Anabaena variabilis phycobilisomesfour phycocyanin (PC) and four allophycocyanin (APC) subparticlesspecies were isolated. In high phosphate a 14.5 S APC fractionwas capable of associating with an APC trimer (6.1 S) fractionat the ratio of 2 : 4 to yield 23 S APC particles. The 23 SAPC particles by electron microscopy were square in shape withthe dimension of 16 x 23 nm, strongly resembling the core ofnative phycobilisomes. When the 23 S APC particles were mixedwith PC subparticles, phycobilisomes resulted which were indistinguishablefrom native phycobilisomes in shape as well as in the capacityof transferring excitation energy. It appears that phycobilisomesconsist of two far-emitting APC moieties, each of which hascontact with one PC rod. ¹ This paper is dedicated to late Prof. Dr. Joji Ashida.     

Co‐suppression in Pyropia yezoensis (Rhodophyta) Reveals the Role of PyLHCI in Light Harvesting and Generation Switch

The red macroalga Pyropia yezoensis is an economically important seaweed widely cultured in Asian countries and is a model organism for molecular biological and commercial research. This species is unique in that it utilizes both phycobilisomes and transmembrane light‐harvesting proteins as its antenna system. Here, one of the genes of P. yezoensis (PyLHCI) was selected for introduction into its genome to overexpress PyLHCI. However, the co‐suppression phenomenon occurred. This is the first documentation of co‐suppression in algae, in which it exhibits a different mechanism from that in higher plants. The transformant (T1) was demonstrated to have higher phycobilisomes and lower LHC binding pigments, resulting in a redder color, higher sensitivity to salt stress, smaller in size, and slower growth rate than the wildtype (WT). The photosynthetic performances of T1 and WT showed similar characteristics; however, P700 reduction was slower in T1. Most importantly, T1 could release a high percentage of carpospores in young blades to switch generation during its life cycle, which was rarely seen in WT. The co‐suppression of PyLHCI revealed its key roles in light harvesting, stress resistance, and generation alternation (generation switch from gametophytes to sporophytes, and reproduction from asexual to sexual).     

Ultrastructure of the motile cells of the prostrate filamentous green algaeProtoderma sarcinoidea andChamaetrichon capsulatum

The biflagellate zoospores ofProtoderma sarcinoidea and the quadriflagellate zoospores ofChamaetrichon capsulatum are each covered by an amorphous, mucous material and a single layer of square scales, and the pyrenoid matrix is traversed by one or more thylakoid membranes. In the flagellar apparatus the basal bodies ofP. sarcinoidea and the upper basal bodies ofC. capsulatum are displaced in the counterclockwise absolute orientation, while the lower basal bodies ofC. capsulatum are directly opposed. Other components of the flagellar apparatus observed in each alga include: cruciately arranged d and s rootlets, each associated with an electron-dense component; simple terminal caps comprised of large and small subunits; a terminal electron-dense mass located near the proximal end of each basal body inP. sarcinoidea and near the upper basal bodies inC. capsulatum; and two rhizoplasts. Components specific to one or the other species include a single accessory basal body inP. sarcinoidea and a fibrous, electron-opaque band that links the upper and the lower basal bodies inC. capsulatum. The flagellar apparatus architecture ofP. sarcinoidea resemblesGayralia oxysperma, while that ofC. capsulatum is similar toTrichosarcina polymorphum andUlothrix species, all of which are included in theUlothrix-group,Ulotrichales, Ulvophyceae.     

Reproduction and larval development of the gastropodAlaba vladivostokensis in Vostok Bay, the Sea of Japan

The reproduction and development of the prosobranch gastropodAlaba vladivostokensis Bartsch were studied. In Vostok Bay,A. vladivostokensis reproduces in the warmest season at water temperatures of 19–23°C. The females lay egg masses in the shape of flat, coiled bands on eelgrass leaves and on sargosso thalluses. The total period of development from egg laying to larval settling takes 25–27 days. The reproduction and development patterns and larval morphology ofA. vladivostokensis are similar to that ofAustralaba picta (A. Adams), which inhabits the coast of Japan. It is suggested that the population ofA. vladivostokensis in Peter the Great Bay should considered as part of the wide geographical range ofA. picta.     

A new species ofLentinula (Agaricales) from Central America

Lentinula aciculospora sp. nov. is described from a single Province in Costa Rica. Stature and habit of basidiomata of this species are similar to some otherLentinula taxa (i.e.L. edodes), but the elongate-cylindrical shape of basidiospores is a distinctive character ofL. aciculospora. The mating system ofL. aciculospora is tetrapolar as also found in other species of the genus, but this species is genetically isolated fromL. boryana, and all morphological variants ofL. edodes.     

The structure of Gloeobacter violaceus and its phycobilisomes

The fine structure of the atypical cyanobacterium Gloeobacter violaceus has been studied on frozen-etched replicas and compared to that of a typical unicellular strain: Synechocystis 6701. The complementary fracture faces of G. violaceus cytoplasmic membrane contain particles less numerous and more heterogenous in size than either the cytoplasmic membrane or the thylakoid membranes of Synechocystis. The most frequently observed particles of the exoplasmic fracture (EF) face of the G. violaceus cytoplasmic membrane are 11 nm in diameter and occasionally form short alignments. This particle class is similar in appearance to the numerous, aligned EF particles of Synechocystis thylakoid membranes. In replicas of cross-fractured G. violaceus, a layer 50–70 nm thick, composed of rod-like elements, underlies the inner surface of the cytoplasmic membrane. The rods, 12–14 nm in diameter, are oriented perpendicularly to the cytoplasmic membrane and show a 6 nm repeat along their length.Isolated phycobilisomes of G. violaceus appear, after fixation and negative staining, as bundles of 6 parallel rodshaped elements connected to an ill-defined basal structure. The bundles are 40–45 nm wide and 75–90 nm long. The rods are 10–12 nm in width; their length varies between 50 and 70 nm. These rods are morphologically similar to those observed at the periphery of hemidiscoidal phycobilisomes of other cyanobacteria, with a strong repeat at 6 nm intervals and a weaker one at 3 nm intervals along their length.The calculated molar ratio of phycobiliproteins in isolated G. violaceus phycobilisomes corresponds to 1:3.9:2.9 for allophycocyanin, phycocyanin and phycoerythrin respectively. When excited at 500 nm, isolated phycobilisomes exhibit a major fluorescence emission band centered at 663 nm.Abbreviations PBS phycobilisome(s) - PBP phycobiliprotein(s) - AP allophycocyanin - PC phycocyanin - PE phycoerythrin - K–PO₄ buffer KH₂PO₄ titrated with KOH to a given pH     

Structure and organization of phycobilisomes on membranes of the red alga Porphyridium cruentum

In the present work, electron microscopy and single particle averaging was performed to investigate the supramolecular architecture of hemiellipsoidal phycobilisomes from the unicellular red alga Porphyridium cruentum. The dimensions were measured as 60 × 41 × 34 nm (length × width × height) for randomly ordered phycobilisomes, seen under high-light conditions. The hemiellipsoidal phycobilisomes were found to have a relatively flexible conformation. In closely packed semi-crystalline arrays, observed under low-light conditions, the width is reduced to 31 or 35 nm, about twice the width of the phycobilisome of the cyanobacterium Synechocystis sp. PCC 6803. Since the latter size matches the width of dimeric PSII, we suggest that one PBS lines up with one PSII dimer in cyanobacteria. In red algae, a similar 1:1 ratio under low-light conditions may indicate that the red algal phycobilisome is enlarged by a membrane-bound peripheral antenna which is absent in cyanobacteria. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users. Ana A. Arteni and Lu-Ning Liu equally contributed to the work.     

Post-larval and larval shells ofJuranomia Fürsich & Werner 1989, andAnomia Linnaeus 1758 (Anomüdae, Bivalvia)

Post-larval and larval shells ofJuranomia calcibyssata from the Bathonian to Callovian of Poland and RecentAnomia membranacea from the Mediterranean are described and compared to other fossil and Recent members of the family Anomüdae. The stratigraphic range of the monospecific genusJuranomia, which, up to now, was only known from the Kimmeridgian, can be extended to include the Lower Bathonian. The state of preservation of the fossil species allows recognition of an internal aragonitic, branching and complex cross-lamellar shell layer in the post-larval left valve, which previous studies could only assume to be present.Anomia membranacea is a member of theA. ephippium lineage as proved, among other characters, by the presence of an outer calcitic prismatic layer in its right valve. It possesses an anterior pedal retractor in the left valve, which, in the original discussion of the phylogenetic affinities ofJuranomia, was thought to be lacking in species ofAnomia. Consequently, the generaJuranomia andAnomia only differ in two important shell characters: closeness or distance between the three central muscles and thickness of the inner aragonitic shell layer of the left valve. Larval shells ofJuranomia are similar to those of Recent anomiids in shape, size, the presence of a byssal notch in the right valve, and an external sinus and internal shelly process in the left valve. The last three features are parts of a single character which is considered as an autapomorphy of the stem species of the Anomiiudae. The small P I size ofJuranomia calcibyssata suggests a purely planktic-planktotrophic development and thus, high potential of dispersal, just as its modern counterparts. Irrespective of the general similarity in shell size, the mean dimensions of the P II are likely species-specific.