PHYCOBILIPROTEIN COMPONENTS AND CHARACTERISTICS OF THE PHYCOBILISOME FROM A THERMOPHILIC CYANOBACTERIUM MYXOSARCINA CONCINNA1

A phycocyanin (PC) and three allophycocyanin (AP) components (designated PC, AP1, AP2, and AP3) were prepared from Myxosarcina concinna Printz phycobilisomes by the native gradient PAGE performed in a neutral buffer system combined with the ion exchange column chromatography on DEAE‐DE52 cellulose. PC contained one β subunit () and two α ones ( and ), and it carried two rod linkers ( and ) and one rod‐core linker (). AP1 and AP3 were characterized as peripheral core APs, whereas AP2 was an inner‐core one. AP2 and AP3 were demonstrated to function as the terminal emitters. Each of the three APs contained two β subunits ( and ), two α subunits ( and ) and an inner‐core linker (). AP2 and AP3 had another subunit of the allophycocyanin B (AP‐B) type () belonging to the β subunit group, and AP1 and AP3 carried their individual specific core linkers ( and ), respectively. No AP component was shown to associate with the core‐membrane linker L_CM. The functions of the linker polypeptides in the phycobilisome (PBS) construction are discussed.     

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.     

CHARACTERIZATION OF PHYCOCYANIN-DEFICIENT PHYCOBILISOMES FROM A PIGMENT MUTANT OF PORPHYRIDIUM SP. (RHODOPHYTA)

The structure and function of phycobilisomes in the rhodophyte Porphyridium sp. were investigated by comparing the properties of the wild type with a pigment mutant called C12. When grown under low light, cells of C12 were bright orange, while wild-type cells were deep red. The results obtained from a characterization of purified phycobilisomes of the mutant C12 led us to propose the existence in Porphyridium sp. phycobilisomes of two types of rods, some containing only phycoerythrin and others containing phycoerythrin bound to phycocyanin, which is in turn linked to the core by the linker L_RC. By studying the partitioning of phycobiliproteins between phycobilisomes and pools of free phycobiliproteins, we found that phycocyanin in the C12 mutant was only present in the pool of free proteins and that its specific linker, L_RC, was totally absent. Phycoerythrin was present in the free pool and in the purified phycobilisomes as well. One of the three specific phycoerythrin linkers γ was missing. In light of the fact that in the C12 mutant, the linker L_RC is absent and that there is no phycocyanin bound to the phycobilisomes, we propose that the rods in the mutant contain only phycoerythrin. These phycobilisomes are nevertheless functional and exhibit an efficient excitation transfer from phycoerythrin directly to allophycocyanin. Electron microscopy showed the purified phycobilisomes of C12 to be less dense than those of the wild type. This change was attributed to the disappearance of the rods containing the combination phycocyanin/phycoerythrin. Light still regulates phycobiliprotein synthesis in the mutant, as shown by the change in the color of the culture, which turned green-yellow when cells were shifted from low light to high light growth conditions. Light also regulates the structure of the phycobilisomes, which have fewer rods under high light growth conditions.     

CHLOROPLAST STRUCTURE AND PIGMENT COMPOSITION IN THE RED ALGA GRIFFITHSIA PACIFICA: REGULATION BY LIGHT INTENSITY1

The ratio of accessory phycobiliproteins to chlorophyll a is controlled by light intensity in the marine red alga Griffithsia pacifica. The greatest changes in pigment ratios are observed below 300 ft-c; above 300 ft-c the response approaches saturation. Ultrastructural examination of chloroplasts of plants grown at different intensities reveals that the number of phycobilisomes per unit of photosynthetic thylakoid changes in direct proportion to the pigment ratios and in inverse proportion to the light intensity.     

ULTRASTRUCTURE OF SELECTIVE CHLOROPLAST DESTRUCTION AFTER SOMATIC CELL FUSION IN GRIFFITHSIA PACIFICA KYLIN (RHODOPHYTA)1,2

Fusion of somatic cells from different geographic isolates of Griffithsia pacifica is followed by a cytoplasmic incompatibility reaction (CIR)⁵ in the cytoplasm donated by only one of the isolates. This CIR is characterized by the aggregation, fusion, and lysis of chloroplasts of the sensitive strain; the cholorplasts of the other strain are unaffected. Within a given cell, chloroplast fusions result in a 50% decrease in chloroplast number in the first 8 h; over the next 3–5 weeks, chloroplast lysis leads to the complete loss of photosynthetic pigments (18). Results of this ultrastructural study indicated that the primary effect of the cytoplasmic incompatibility reaction was a directed destruction of the sensitive strain chloroplasts. No aggregation or abnormalities were seen in organelles other than the chloroplasts. Chloroplast destruction was accompanied by extensive vesiculation of the cytoplasm and a definit change in the physiological state of the cell; the dictyosomes became inactive and there was a reduction in the amount of endoplasmic reticulum, cytoplasmic ribosomes and starch grains. This selective destruction of one chloroplast population in the presence of a second population of unaffected chloroplasts suggests that the incompatibility reaction observed in G. pacifica results from competitive interactions between the two chloroplast types.     

Characterization and transcript analysis of the major phycobiliprotein subunit genes from Aglaothamnion neglectum (Rhodophyta)

The genes encoding the and subunits of allophycocyanin, phycocyanin and phycoerythrin from the red alga Aglaothamnion neglectum were isolated and characterized. While the operons containing the different phycobiliprotein genes are dispersed on the plastid genome, the genes encoding the and subunits for each phycobiliprotein are contiguous. The subunit gene is 5 for both the phycocyanin and phycoerythrin operons, while the subunit gene is 5 for the allophycocyanin operon. The amino acid sequences of A. neglectum phycobiliproteins, as deduced from the nucleotide sequences of the genes, are 65–85% identical to analogous proteins from other red algae and cyanobacteria. The conserved nature of the plastid-encoded red algal and cyanobacterial phycobiliprotein genes supports the proposed origin of red algal plastids from cyanobacterial endosymbionts.Many environmental factors effect phycobilisome biosynthesis. The effect of both nutrient availability and light quantity on the level of A. neglectum phycobiliprotein subunits and the mRNA species encoding those subunits is described.     

REGENERATION AND SEXUAL DIFFERENTIATION OF GRIFFITHSIA JAPONICA (CERAMIACEAE,RHODOPHYTA) THROUGH SOMATIC CELL FUSION1

Hybrid cells were obtained from somatic cell fusion among male, female, and tetrasporangial plants in Griffithsia japonica Okamura by a wound-healing process. Isolated fusion cells regenerated new mature plants with mixed reproductive structures. The plants regenerated from hybrid cells between male and female plants developed into 1) spermatangiate, 2) carpogonial, 3) bisexual with spermatangia and carpogonial branches, 4) mixed-phase with spermatangia and tetrasporangia, or 5) bisexual/mixed-phase plants with spermatangia, carpogonial branches, and tetrasporangia. About 70% of the plants regenerated from hybrid cells between male and female plants produced tetrasporangia that were always formed with spermatangia on a single cell. Some of those tetrasporangia released tetraspores, six of which gave rise to mature plants. The plants regenerated from hybrid cells between male and tetrasporangial plants developed into spermatangiate, tetrasporangiate, or mixed-phase plants with spermatangia and tetrasporangia. The plants regenerated from hybrid cells between female and tetrasporangial plants developed into carpogonial, tetrasporangiate, or mixed-phase plants with carpogonial branches and tetrasporangia. All types of reproductive structures we re functional.     

GRACILARIOPSIS MCLACHLANII SP. NOV. AND GRACILARIOPSIS PERSICA SP. NOV. OF THE GRACILARIACEAE (GRACILARIALES,RHODOPHYCEAE) FROM THE INDIAN OCEAN1

Two new species of Gracilariopsis from the Indian Ocean are proposed—Gracilariopsis (Gp.) mclachlanii Buriyo, Bellorin et M. C. Oliveira sp. nov. from Tanzania and Gracilariopsis persica Bellorin, Sohrabipour et E. C. Oliveira sp. nov. from Iran—based on morphology and DNA sequence data (rbcL gene and SSU rDNA). Both species fit the typical features of Gracilariopsis: axes cylindrical throughout, freely and loosely ramified up to four orders, with an abrupt transition in cell size from medulla to cortex, cystocarps lacking tubular nutritive cells and superficial spermatangia. Nucleotide sequence comparisons of rbcL and SSU rDNA placed both species into the Gracilariopsis clade as distinct species from all the accepted species for this genus, forming a deeply divergent lineage together with some species from the Pacific. The new species are very difficult to distinguish on morphological grounds from other species of Gracilariopsis, stressing the importance of homologous molecular marker comparisons for the species recognition in this character‐poor genus.     

CHEMICAL COMPOSITION AND STRUCTURE OF THE CELL WALLS OF THE CONCHOCELIS AND THALLUS PHASES OF PORPHYRA TENERA (RHODOPHYCEAE)1,2

Purified cell walls were prepared from both the conchocelis and thallus phases of Porphyra tenera (Kjellm.). The nitrogen content of cell walls from the conchocelis was significantly greater than that for the thallus cell walls, being 3.35 ± 0.26% and 2.39± 0.03%, respectively. Amino acid analysis revealed important differences. The conchocelis cell wall hydrolyzates were richer in aspartic acid, glutamic acid, methionine, and basic amino acids. The thallus cell wall hydrolyzates, however, contained much more glycine and alanine than did those of the conchocelis. Hydroxyproline was not detected in cell walls of either phase. The neutral sugar content of cell wall hydrolyzates from the thallus was more than double that from the conchocelis being 83.6% and 34.5%, respectively. The former contained predominantly mannose which accounted for 72.2% of the neutral sugars while the latter was principally galactose (49.9%) and glucose (36.4%). Methylation analysis confirmed the presence of cellulose microfibrils in the conchocelis in contrast to xylan microfibrils in the thallus. The results establish that the conchocelis and thallus phases of P. tenera differ markedly in the structure and composition of the cell walls.     

CENTROCERAS INTERNITENS N. SP. (RHODOPHYCEAE,CERAMIACEAE) FROM THE WESTERN TROPICAL NORTH ATLANTIC OCEAN1

Centroceras internitens n. sp. (Rhodophyceae, Ceramiaceae) was found in the Florida Keys and Bermuda only as an epiphyte on Ceramium nitens (C. Agardh) J. Agardh. It is described on the basis of many collections, including an abundance of reproductive material representing all stages of the life history. Laboratory culture has not been successful. The type locality is a well-known limestone platform on the Atlantic Ocean side of West Summerland Key, Florida Keys. In addition to living material, C. internitens was found on eight herbarium specimens of Ceramium nitens at the United States National Museum, thereby establishing records of its occurrence throughout the Caribbean Sea and dating back to 1887.     

SPECIES OF ANTITHAMNION (RHODOPHYCEAE,CERAMIACEAE) OCCURRING ON THE SOUTHEAST AFRICAN COAST (NATAL)1

Three species of Antithamnion Naegeli were previously known from Natal, all reported since 1984. A more thorough collection of the flora in recent months has revealed that there are nine species in Natal, four of which are newly described. Vegetative characteristics are emphasized in this study of the taxonomy of Antithamnion, the most important being (1) size of axial cells, (2) form of whorl-branchlets, (3) size of whorl-branchlet basal cells, (4) size and shape of whorl-branchlet terminal cells, (5) size and position of gland cells, (6) primary or secondary derivation of indeterminate lateral branches, and (7) the relationship of primary indeterminate branches, if present, to normal arrangement of whorl-branchlets. Two closely related species, A. diminuatum Wollaston and A. eliseae sp. nov. have been grown in laboratory culture allowing for testing of stability and reliability of vegetative characteristics for use in taxonomic studies. Conclusions are that these criteria can be effectively used for these purposes. In addition to A. eliseae, A. nematocladellum, A. pterocladellum and A. adenocladellum also are described as new species. A tropical species previously known from the Caribbean and Red Seas, A. Iherminieri Nasr, a species previously recorded from Japan, A. secundum Itono, a species previously known from California and Japan, A hubbsii Dawson, and two species from Australia, A. diminuatum Wollaston and A. divergens (J. Agardh) J. Agardh, are also described from Natal.     

SPECIES OF ANTITHAMNION (RHODOPHYCEAE,CERAMIACEAE) OCCURRING ON THE SOUTHEAST AFRICAN COAST (NATAL)1

Three species of Antithamnion Naegeli were previously known from Natal, all reported since 1984. A more thorough collection of the flora in recent months has revealed that there are nine species in Natal, four of which are newly described. Vegetative characteristics are emphasized in this study of the taxonomy of Antithamnion, the most important being (1) size of axial cells, (2) form of whorl-branchlets, (3) size of whorl-branchlet basal cells, (4) size and shape of whorl-branchlet terminal cells, (5) size and position of gland cells, (6) primary or secondary derivation of indeterminate lateral branches, and (7) the relationship of primary indeterminate branches, if present, to normal arrangement of whorl-branchlets. Two closely related species, A. diminuatum Wollaston and A. eliseae sp. nov. have been grown in laboratory culture allowing for testing of stability and reliability of vegetative characteristics for use in taxonomic studies. Conclusions are that these criteria can be effectively used for these purposes. In addition to A. eliseae, A. nematocladellum, A. pterocladellum and A. adenocladellum also are described as new species. A tropical species previously known from the Caribbean and Red Seas, A. lherminieri Nasr, a species previously recorded from Japan, A. secundum Itono, a species previously known from California and Japan, A hubbsii Dawson, and two species from Australia, A. diminuatum Wollaston and A. divergens (J. Agardh) J. Agardh, are also described from Natal.     

SEASONAL VARIATION OF P CONTENT AND MAJOR N POOLS IN PALMARIA PALMATA (RHODOPHYTA)1

The annual variation of major nitrogen pools, phosphorus, carbon, ash, and thallus water content in relation to seasonal environmental changes was studied in two northern Spanish populations of the edible seaweed Palmaria palmata (Linnaeus) Kuntze. Observed patterns were investigated using Spearman rank order correlation coefficients. There were significant relationships between thallus nutrient content and nitrate and orthophosphate seawater concentration, irradiance, temperature, and wave force. The highest levels of total N and P and nitrogenous compounds were observed during autumn and winter because the thallus stored N‐ and P‐rich compounds in response to high nutrient seawater concentration when growth was limited by low light and temperature. Phycoerythrin and other proteins were the main N reserves. Thallus P content was higher in algae from the eutrophic site. During spring, reduced N and P thallus content and increased ash, water, and C content were observed in the growing fronds. N and P seawater concentrations were undetectable during summer when nutrient reserves were low and growth was reduced and eventually suppressed, suggesting nutrient limiting conditions. Palmaria palmata clearly could take advantage of elevated N and P concentrations to create storage reserves in winter to support early summer growth. This storage response reduced the dependence of algal nutrition on the external nutrient supply and supports the use of pulse fertilization to diminish summer nutrient limitation of cultured algae.     

PHYCOBILIN CONTENT OF THE CONCHOCELIS PHASE OF ALASKAN PORPHYRA (BANGIALES,RHODOPHYTA) SPECIES: RESPONSES TO ENVIRONMENTAL VARIABLES1

Variations of pigment content in the microscopic conchocelis stage of four Alaskan Porphyra species were investigated in response to environmental variables. Conchocelis filaments were cultured under varying conditions of irradiance and nutrient concentrations for up to 60 d at 11°C and 30 psu salinity. Results indicate that conchocelis filaments contain relatively high concentrations of phycobilins under optimal culture conditions. Phycobilin pigment production was significantly affected by irradiance, nutrient concentration, and culture duration. For Porphyra abbottiae V. Krishnam., Porphyra sp., and Porphyra torta V. Krishnam., maximal phycoerythrin (63.2–95.1 mg · g dwt^?1) and phycocyanin (28.8–64.8 mg · g dwt^?1) content generally occurred at 10 μmol photons · m^?2 · s^?1, f/4–f/2 nutrient concentration after 10–20 d of culture. Whereas for Porphyra hiberna S. C. Lindstrom et K. M. Cole, the highest phycoerythrin (73.3 mg · g dwt^?1) and phycocyanin (70.2 mg · g dwt^?1) content occurred at 10 μmol photons · m^?2 · s^?1, f nutrient concentration after 60 d in culture. Under similar conditions, the different species showed significant differences in pigment content. P. abbottiae had higher phycoerythrin content than the other three species, and P. hiberna had the highest phycocyanin content. P. torta had the lowest phycobilin content.     

ACCLIMATION OF SYNECHOCOCCUS STRAIN WH7803 TO AMBIENT CO2 CONCENTRATION AND TO ELEVATED LIGHT INTENSITY1

A CO₂ concentrating mechanism has been identified in the phycoerythrin-possessing Synechococcus sp. WH7803 and has been observed to be severely inhibited by short exposure to elevated light intensities. A light treatment of 300–2000 μmol quanta·m^?2·s^?1 resulted in a considerable decay in the variable fluorescence of PSII with time, suggesting decreased efficiency of energy transfer from the phycobilisomes, direct damage to the reaction center II, or both. Measurements of the activity of PSII and changes in fluorescence emission spectra during a light treatment of 1000 μmol quanta·m^?2·s^?1 indicated considerable reduction in the energy flow from the phycocyanin to the phycobilisome terminal acceptor and chlorophyll a. Consequently, whereas the maximal photosynthetic rate, at saturating light and Co₂ concentration, was hardly affected by a light treatment of 1000 μmol quanta·m^?2·s^?1 for 2 h, the light intensity required to reach that maximum increased with the duration of the light treatment.     

EFFECT OF SPECTRAL QUALITY ON GROWTH AND PIGMENTATION OF PICOCYANOBACTERIA1

The influence of spectral quality on growth and pigmentation was compared among five strains of marine and freshwater picocyanobacteria grown under the same photon flux density (28 μE · m^?2·s^?1). Growth and phycoerythrin (PE) concentration per unit carbon increased when marine Synechococcus WH7803 was grown under green light as compared to red light, but no change in phycocyanin concentration occurred. Marine Synechococcus strain 48B66 also showed greater levels of PE when grown under green light than under red light, but no concomitant growth increase occurred. Both strains thus exhibited Group II chromatic adaptation. Additionally, strain 48B66 increased the relative level of phycourobilin compared to phycoerythrobilin when grown under red light. In contrast, both marine and freshwater Synechococcus strains containing no PE showed decreased growth under green light. Chlorophyll a concentrations were greatest or among the greatest in all strains grown under green light. These results suggest that light quality, through its effects on growth rate, may be an important factor controlling the distribution and abundance of the various pigment types of Synechococcus.     

STRUCTURE AND COMPOSITION OF OOCYSTIS POLYMORPHA CELL WALLS ISOLATED FROM THE CULTURE MEDIUM1

Oocystis polymorpha and possibly other algae which shed their parent-cell walls during reproduction appear to be suitable organisms for cell wall research in that the cell walls can be readily collected from the culture medium. Electron micrographs of sectioned isolated cell walls, of the parent-cell walls, and of the daughter cells appear to be identical, suggesting that the isolated cell walls are representative of those of the intact cell. Chemically, the cell walls of Oocystis polymorpha resemble the cell walls of Cyanidium and the chemically resistant cuticle fraction of Cladophora and Porphyra in their high nitrogen and low sugar and lipid contents. Pink carotenoids are the major lipid components of the cell walls.     

THE EFFECTS OF LIGHT AND NITROGEN ON GROWTH,PIGMENT CONTENT,AND BIOCHEMICAL COMPOSITION OF GRACILARIA FOLIIFERA V. ANGUSTISSIMA (GIGARTINALES,RHODOPHYTA)1

The combined effects of light intensity and nitrogen (NO₃^?) on growth rate, pigment content, and biochemical composition of Gracilaria foliifera v. angustissima (Harvey) Taylor was investigated using outdoor continuous cultures. Growth of Gracilaria increased linearly with increasing light to 0.43 doublings d^?1 at high light levels (383 ly d^?1 of in situ light), suggesting that light may often limit growth of this plant in nature. Chlorophyll a and phycoerythrin contents were inversely proportional to light level and growth rate. However, pigment content did not affect the growth capacity of Gracilaria. There was no increase in growth or pigment content with increasing additions of nitrogen. The low nitrogen treatment was unenriched seawater that had higher NO₃^? levels than most coastal waters (influent = 8.61 μM; residual = 0.94 μM). When growing near its maximum rate under high light intensities, Gracilaria had a significantly (P < 0.001) lower phycoerythrin: chlorophyll a ratio (phyco: Chl a) than did Gracilaria growing more slowly under lower light (Phyco:Chl a of 2.8 ± 0.2 vs. 3.8 ± 0.3). Faster growing plants also had C:N ratios above 10, indicating N- limitation. In addition to harvesting light the phycobiliproteins of Gracilaria may store nitrogen. Growth rates of Gracilaria correlated negatively with ash (r =–0.85) and positively with the carbon: phycoerythrin ratio (r = 0.85), suggesting that these two indices can be used to estimate growth in the field.     

SEXUAL DIFFERENTIATION OF GRIFFITHSIA (CERAMIALES, RHODOPHYTA): NUCLEAR PLOIDY LEVEL OF MIXED-PHASE PLANTS IN G. JAPONICA1

Mixed-phase plants of Griffithsia japonica Okamura spontaneously occurred in a laboratory culture. Four female plants produced tetrasporangia and spermatangia in addition to their normal female reproductive structures (bisexual/mixed-phase plants), and four male plants produced tetrasporangia as well as spermatangia (male/mixed-phase plants). To determine the nuclear ploidy level of these mixed-phase plants, relative nuclear sizes of male, female, tetrasporangial, and mixed-phase plants were measured using a microscopic image analysis system. Haploid gametophytes could be distinguished from diploid tetrasporophytes by relative nuclear sizes, with the later having nuclei twice the size of the former. Relative nuclear sizes of the mixed-phase plants were similar to those of the haploid plants. Thus, the mixed-phase plants were determined to be haploid. Haploid mixed-phase plants of G. japonica have a potential to produce male, female and tetrasporangial reproductive structures. Sex determination models are discussed to explain "haploid" mixed-phase phenomena in red algae .