bluuebunny/crossref_metadata_2025 huggingface dataset:
A RIBOSOMAL DNA PHYLOGENY SUPPORTS THE CLOSE EVOLUTIONARY RELATIONSHIPS AMONG THE SPOROCHNALES, DESMARESTIALES, AND LAMINARIALES (PHAEOPHYCEAE)1
Authors: Tan Ian H. , Druehl Louis D.
Published: October 2004

We report six complete 18S ribosomal DNA (rDNA) sequences representing five brown algal orders: Sporochnus comosus C. A. Agardh (Sporochnales), Chorda tomentosa Lyngbye (Chordaceae, Laminariales), Saccorhiza polyschides (Lightfoot) Batters (Phyllariaceae, Laminariales), Desmarestia ligulata (Lightfoot) Lamouroux (Desmarestiales), Ectocarpus siliculosus (Dillwyn) Lyngbye (Ectocarpales), and Scytosiphon lomentaria (Lyngbye) J. G. Agardh (Scytosiphonales). These sequences were compared with published laminarialean (Alaria marginata Postel et Ruprecht [Alariaceae] and Macrocystis integrifolia Bory [Lessoniaceae]) and fucalean (Fucus gardneri Silva) rDNA sequences for phylogeny inference using both the distance‐matrix and parsimony methods. The inferred 18S phylogenies clustered Sporochnus, Desmarestia, Chorda, Saccorhiza, Alaria, and Macrocystis in an assemblage. This Sporochnales–Desmarestiales–Laminariales (S‐D‐L) complex was consistently separated from the Ectocarpales, Scytosiphonales, and Fucales by bootstrap analyses. The inferred phylogenies are consistent with several possible evolutionary processes leading to this S‐D‐L complex. Members in this assemblage lack eyespots in their sperm, and their sperm have the atypical brown algal flagellation: shorter anterior and longer posterior flagella. In addition, they are oogamous with a heteromorphic alternation of generations between a microscopic gametophyte and a macroscopic sporophyte. Members of the S‐D‐L complex can be separated into different phylogenetic lines based on the presence/absence of eyespots in their meiospores. Our findings support the contention that the Sporochnales, Desmarestiales, and Laminariales are closely related. In addition, our rDNA tree suggests that the Laminariales is paraphyletic.
EVOLUTIONARY CONSEQUENCES OF THE LOSS OF PHOTOSYNTHESIS IN CHLAMYDOMONADACEAE: PHYLOGENETIC ANALYSIS OFRrn18(18S rDNA) IN 13POLYTOMASTRAINS (CHLOROPHYTA)1
Authors: Rumpf Robert , Vernon Dawne , Schreiber David , Birky C. William
Published: October 2004

Complete sequences of theRrn 18genes were obtained from 13 strains of the nonphotosynthetic algal genusPolytoma.Phylogenetic analyses showed that these strains formed two clades. One clade shows only modest sequence diversity but is represented by strains collected at widely dispersed sites in Europe and America. The other clade consists of a single isolate from the Canary Islands. Both clades lie well within the extended clade that includes all species ofChlamydomonasfor which sequence data are available. The twoPolytomaclades are separated from each other by several green species, suggesting that the extant nonphotosynthetic Chlamydomonadaceae arose from photosynthetic ancestors at least twice. These results suggest that nonphotosynthetic mutants are capable of establishing lineages that can spread widely but have a higher probability of extinction than their photosynthetic congeners.
DICROGLOSSUM CRISPATULUM GEN. ET COMB. NOV. FROM WESTERN AUSTRALIA, REPRESENTING A NEW TRIBE WITHIN THE DELESSERIACEAE (RHODOPHYTA)1
Authors: Millar Alan J. K. , Huisman John M.
Published: October 2004

Dicroglossum gen. nov. (Delesseriaceae, Ceramiales) is a monotypic genus based on Delesseria crispatula, a species originally described by Harvey for plants collected from southwestern Western Australia. Distinctive features of the new genus include exogenous indeterminate branches; growth by means of a single transversely dividing, apical cell; absence of intercalary divisions in the primary, secondary, and tertiary cell rows; lateral pericentral cells not transversely divided; not all cells of the secondary cell rows producing tertiary cells rows; all tertiary initials reaching the thallus margin; midrib present but lateral nerves absent; determinate lateral bladelets arising endogenously; blades monostromatic, except, at the midrib; carpogonial branches restricted to primary cell rows, on both surfaces of unmodified blades; procarps produced on both blade surfaces, each procarp consisting of a supporting cell that bears two four‐celled carpogonial branches and one sterile‐cell group of three to four cells; and tetrasporangia borne in two layers, separated by a central row of sterile cells. The combination of exogenous indeterminate branching and bicarpogonial procarps is considered to warrant the recognition of a new tribe, the Dicroglosseae, within the subfamily Delesserioideae.
DASYA ROSLYNIAE SP. NOV. (DASYACEAE, RHODOPHYTA), WITH A DISCUSSION ON GENERIC DISTINCTIONS AMONG DASYA, EUPOGODON, RHODOPTILUM, AND POGONOPHORELLA1
Authors: Millar Alan J. K.
Published: October 2004

Dasya roslyniae sp. nov. (Dasyaceae, Rhodophyta) is described from subtidal habitats at Split Solitary Island (30°14′S, 153°11′E), New South Wales, Australia. The new species is distinct within the genus due to its strongly compressed and secondarily bilaterally branched axes, differing from the majority of Dasya species that are terete and secondarily radially organized. Pseudolaterals are quickly caducous on ventral and dorsal (transverse) surfaces but are persistent on lateral surfaces for short distances from the apex, leaving the bulk of the plants flattened and denuded. Its gross morphological characters are thus similar to those displayed by the genera Pogonophorella, Eupogodon (formerly known as Dasyopsis), and Rhodoptilum. Characters used for separating these genera and Dasya are, in some cases, overlapping and in need of critical evaluation. To the primarily radially organized taxa, determined by examination of divisions of the apical cell, are placed species of Dasya, six species now included in Eupogodon, and the type and only species of Pogonophorella californica. Examination of the activity of the apical cells of Eupogodon planum and Rhodoptilum plumosum, the type species of their genera, confirms the primary bilaterality of these two genera, and the traditional defining feature of Eupogodon (lack of discernible pericentral cells in cross‐section of indeterminate axes) is shown to be untenable. A secondary character that would separate Eupogodon and Rhodoptilum is the polysiphonous bases of otherwise monosiphonous laterals (pseudolaterals) in Eupogodon and the monosiphonous bases in Rhodoptilum.
THE GENUS PIKEA (DUMONTIACEAE, RHODOPHYTA) IN ENGLAND AND THE NORTH PACIFIC: COMPARATIVE MORPHOLOGICAL, LIFE HISTORY, AND MOLECULAR STUDIES1
Authors: Maggs Christine A. , Ward Barbara A.
Published: October 2004

A Pikea species attributed to Pikea californica Harvey has been established in England since at least 1967. Previously, this species was believed to occur only in Japan and Pacific North America. Comparative morphological studies on field‐collected material and cultured isolates from England, California, and Japan and analysis of organellar DNA restriction fragment length polymorphisms, detected using labeled organellar DNA as a non‐radioactive probe, showed that English Pikea is conspecific with P. californica from California. Both populations consist of dioecious gametophytes with heteromorphic life histories involving crustose tetrasporophytes; 96% of organellar DNA bands were shared between interoceanic samples. A second dioecious species of Pikea, P. pinnata Setchell in Collins, Holden et Setchell, grows sympatrically with P. californica near San Francisco but can be distinguished by softer texture, more regular branching pattern, and elongate cystocarpic axes. Pikea pinnata and P. californica samples shared 49–50% of organellar DNA bands, consistent with their being distinct species. Herbarium specimens of P. robusta Abbott resemble P. pinnata in some morphological features but axes are much wider; P. robusta may represent a further, strictly sub‐tidal species but fertile material is unknown. Pikea thalli from Japan, previously attributed to P. californica and described here as Pikea yoshizakii sp. nov., are monoecious and show a strikingly different type of life history. After fertilization, gonimoblast filaments grow outward through the cortex and form tetrasporangial nemathecia; released tetraspores develop directly into erect thalli. Tetrasporoblastic life histories are characteristic of certain members of the Phyllophoraceae but were previously unknown in the Dumontiaceae. Japanese P. yoshizakii shared 55 and 56% of organellar DNA bands with P. californica and P. pinnata, respectively; phylogenetic analysis indicated equally distant relationships to both species. Pikea yoshizakii or a closely similar species with the same life history occurs in southern California and Mexico.
GRADIENT ANALYSIS OF DIATOM ASSEMBLAGES IN WESTERN KENTUCKY WETLANDS1
Authors: Pan Yangdong , Stevenson R. Jan
Published: October 2004

Diatom and water chemistry data from 35 wetland sites in western Kentucky were used to assess diatoms as indicators of ecological conditions in wetlands. The wetlands were affected by different degrees of acid mine drainage and agriculture. Canonical correspondence analysis indicated that the distribution of diatoms was highly correlated with conductivity and total phosphorus (TP), two variables commonly associated with acidic mine drainage and agriculture, respectively.

Diatom‐based inference models were developed for use as quantitative indicators of two important environmental variables in wetlands: conductivity and TP. Diatom‐inferred conductivity and TP values were highly correlated with measured values. Cross‐validation with jackknifing procedures suggested that high apparent r2 between inferred and measured conductivity was overly optimistic and should be treated with caution. Jackknifing‐derived TP inference models performed poorly in predicting TP toward the ends of low and high TP concentrations. In general, the conductivity inference models based on plankton had better predictability than those based on epiphyton. Epiphyton‐based inference models can predict TP better than plankton. Therefore, diatoms in planktonic and epiphytic assemblages could provide complementary information on ecological conditions.

Our data suggest that diatoms can reflect major regional environmental gradients and therefore can be used as indicators of the ecological conditions in wetlands. Quantitative inference models with known predictive power can aid the development of realistic and ecologically sound biotic indices for this region.
CHANGES IN THE PLANKTONIC DIATOM FLORA OF A LARGE MOUNTAIN LAKE IN RESPONSE TO FERTILIZATION1
Authors: Yang Jing‐Rong , Pick Frances R. , Hamilton Paul B.
Published: October 2004

Fertilization (phosphorus and nitrogen) of Kootenay Lake, British Columbia, was begun in 1992 in an attempt to increase food resources for declining stocks of kokanee salmon. In 1992 and 1993, significant changes in diatom abundance, species composition, diversity, and morphology of individual species were observed in the fertilized North Arm in comparison to the unfertilized South Arm. Effects of fertilization were most obvious during the spring. During summer stratification, fertilization did not result in significant changes in total abundance, but changes in species composition were observed. In 1993, with continued fertilization of the North Arm, diatom abundance in both spring and summer was significantly higher than at the same sites in 1992. No significant differences were observed in the unfertilized South Arm. Diatom abundance was significantly correlated with dissolved inorganic nitrogen (positive) and dissolved reactive silica (negative). Diatom assemblages in the fertilized North Arm in both the spring and summer of 1993 were dominated by Asterionella formosa and Fragilaria crotonensis, taxa considered mesotrophic. Abundance of the oligotrophic taxa Rhizosolenia eriensis and Cyclotella michiganiana was higher in the South Arm than in the North Arm. Differences in diatom assemblages between the two arms in both years were further summarized by multivariate statistical analyses. Morphological variations in two small Cyclotella species (C. comensis and C. cf. pseudostelligera) were observed between the fertilized and “control” stations. Populations of both species from fertilized stations exhibited stronger silicification with coarser striations and a smaller mean size.
MAXIMUM AMMONIUM UPTAKE RATES OF SCENEDESMUS QUADRICAUDA (CHLOROPHYTA) AND MICROCYSTIS NOVACEKII (CYANOBACTERIA) GROWN UNDER NITROGEN LIMITATION AND IMPLICATIONS FOR COMPETITION1
Authors: Watanabe Taku , Miyazaki Tatsuo
Published: October 2004

We measured maximum ammonium uptake rates of the green alga Scenedesmus quadricauda (Turpin) Brébisson and the blue‐green alga Microcystis novacekii (Kom.) Comp. grown in nitrogen (ammonium)–limited chemostats. Maximum uptake rates per cellular carbon were larger in S. quadricauda than in M. novacekii. These rates increased with increased specific growth rates. Maximum uptake rates per cellular nitrogen were also larger in S. quadricauda than in M. novacekii. The maximum uptake rates per cellular nitrogen were nearly constant against increased cellular N:C ratios under nitrogen‐limited conditions. The higher maximum uptake rates indicate that S. quadricauda had higher uptake abilities for ammonium than M. novacekii when grown under nitrogen limitation. We examined the competition between both species under two distinct nutrient supply modes, using measured maximum uptake values and computer simulations. Microcystis novacekii prevailed in the small‐pulse, high‐frequency nutrient supply mode, whereas S. quadricauda became competitively superior in the large‐pulse, low‐frequency nutrient supply mode. These results indicate that we could control nuisance blooms of blue‐green algae in lakes and reservoirs by changing the nutrient supply modes.
EVOLUTIONARY AND ECOLOGICAL DIFFERENTIATION IN THE PANTROPICAL TO WARM‐TEMPERATE SEAWEED DIGENEA SIMPLEX (RHODOPHYTA)1
Authors: Pakker Hans , Klerk Hans , van Campen Jan Hein , Olsen Jeanine L. , Breeman Anneke M.
Published: October 2004

Genetic differentiation among geographic isolates of the pantropical to warm‐temperate red alga Digenea simplex (Wulfen) C. Agardh was investigated using random amplified polymorphic DNA (RAPD) markers, crossing studies, and temperature tolerances experiments. Eleven isolates representing populations from the Caribbean, eastern Atlantic, and Indo‐West Pacific were compared. RAPD analysis clearly revealed an Indo‐West Pacific group, a Caribbean/Cape Verde Islands group, and a Canary Islands group. Crossing studies showed different levels of inter fertility. In most crosses between Western Australian and Atlantic isolates, no hybrid tetrasporophytes were formed. In crosses between Caribbean and Cape Verde Islands isolates, tetrasporophytes developed, but the viability of tetraspores was reduced. Full sexual compatibility was observed among Cape Verde Islands isolates and among isolates from Bonaire. Temperature tolerance studies indicate that Pacific isolates have a broader temperature survival range than Atlantic isolates, which may be correlated to local temperature extremes. Despite the reduced level of sexual compatibility between Caribbean and Cape Verde Islands isolates, their shared position in the RAPD analysis and similar temperature responses suggest trans‐Atlantic dispersal in the near geological past. In addition to their discrete position in the RAPD distance analysis, the Canary Islands isolates were significantly more cold‐tolerant than the other Atlantic isolates. This finding is consistent with the hypothesis that the Canary Islands were recolonized from cold‐adapted eastern Mediterranean populations after the last Pleistocene glaciation.
THE EVOLUTION OF PARASITISM IN THE RED ALGAE: MOLECULAR COMPARISONS OF ADELPHOPARASITES AND THEIR HOSTS1
Authors: Goff Lynda J. , Moon Debra A. , Nyvall Pi , Stache Birgit , Mangin Katrina , Zuccarello Giuseppe
Published: October 2004

In several groups of parasites including insect, flowering plant, fungal, and red algal parasites, morphological similarities of the parasites and their specific hosts have led to hypotheses that these parasites evolved from their hosts. But these conclusions have been criticized because the morphological features shared by parasite and host may be the result of convergent evolution. In this study, we examine the hypothesis, originally put forth by Setchell, that adelphoparasitic red algae, that is, parasitic red algae that are morphologically very similar to their hosts, evolved from their specific red algal hosts. Rather than comparing morphological features of parasites and hosts, small‐subunit 18S nuclear ribosomal DNA and the internal transcribed spacer regions (ITSs) of the nuclear ribosomal repeat are compared for five parasites, their hosts, and related nonhosts from four red algal orders. These comparisons reveal that each of these adelphoparasites has evolved either directly from the host on which it is currently found, or it evolved from some other taxon that is closely related to the modern host. The parasites Gardneriella tuberifera, Rhodymeniocolax botryoides, and probably Gracilariophila oryzoides evolved from their respective hosts Sarcodiotheca gaudichaudii, Rhodymenia pacifica, and Gracilariopsis lemaneiformis, respectively. The parasite Faucheocolax attenuata evolved from either Fauchea laciniata or Fauchea fryeana and subsequently radiated onto the other host species. Presently this parasite is found on both hosts. Lastly, some parasitic genera such as Plocamiocolax are polyphyletic in their origins. A species of Plocamiocolax from an Antarctic Plocamium cartilagineum appears to have evolved from its host whereas the common Plocamiocolax pulvinata that occurs along the west coast of North America likely evolved from Plocamium violaceum and radiated secondarily onto its present day host, Plocamium cartilagineum.
PORPHYRA REDIVIVA SP. NOV. (RHODOPHYTA): A NEW SPECIES FROM NORTHEAST PACIFIC SALT MARSHES1
Authors: Stiller John W. , Waaland J. Robert
Published: October 2004

A new species, Porphyra rediviva (Bangiales, Rhodophyta), is described from the northeast Pacific based on morphological, cytological, reproductive, ecological, and molecular characters. This species occurs at high intertidal levels in salt marshes along the coasts of Washington, Oregon, and northern California and exhibits a growth optimum at reduced salinity. It is further distinguished by a distinct demarcation between male and female sectors of the gametophytic thalli of epilithic specimens. The species is found most commonly in the drift or trapped in Salicornia beds, but these detached blades never have been found with sporangia or gametangia. Molecular analyses using restriction fragment length polymorphism patterns of polymerase chain reaction–amplified ribosomal DNA (rDNA) show that this salt marsh Porphyra is conspecific throughout its range and is distinct from other Pacific Porphyra species with similar reproductive patterns. Based on molecular data, P. rediviva is related most closely to P. purpurea from the North Atlantic. Fixed rDNA polymorphisms between the two taxa, however, support ecological and cytological evidence that they should be considered different species.
FIRST INSIGHTS INTO IMPROVEMENT OF EICOSAPENTAENOIC ACID CONTENT IN PHAEODACTYLUM TRICORNUTUM (BACILLARIOPHYCEAE) BY INDUCED MUTAGENESIS1
Authors: Alonso Diego López , Segura del Castillo Clara I. , Grima Emilia Molina , Cohen Zvi
Published: October 2004

A strain improvement program was initiated based on mutagenesis with the goal of commercial production of eicosapentaenoic acid (EPA)from EPA‐overproducing microalgal strains. Two rounds of mutation and selection were conducted using Phaeodactylum tricornutum Bohlin UTEX #640 as the parent strain. After the first round of mutagenesis, a putative mutant (provisionally labeled 114) was obtained. The EPA content (% of dry weight) of this mutant strain was 37% higher than that of the wild type. 114 was further mutated and another putative mutant (provisionally called II242) was isolated, the EPA content of which was 44% higher than that of the wild type. When cultured with aeration in 1‐L flasks, EPA content of the wild type and putative mutants 114 and II242 was, 17.3 mg · g−1, 31.5mg · g−1, and 38.6 mg · g−1 dry biomass, respectively. EPA productivity was 3.48 mg · L−1· d−1 4.01 mg · L−1· d−1, and 4.98 mg · L−1· d−1 respectively. These figures compare favorably with many other promising EPA‐producing microorganisms and suggest that the use of a single methodology such as mutation and selection is a way to improve the polyunsaturated fatty acid content of microalgae and other microorganisms.
A METHOD FOR THE CRYOCONSERVATION OF DUNALIELLA SALINA (CHLOROPHYCEAE): EFFECT OF GLYCEROL AND COLD ADAPTATION1
Authors: Mortain‐Bertrand Anne , Etchart Freddy , Boucaud Marie‐Thérèse de
Published: October 2004

The unicellular green alga Dunaliella salina Teod. was frozen according to the following procedure: 3 days cold adaptation at 4°C, addition of 3.5 M glycerol as a cryoprotectant, slow cooling to –40°C, immersion in liquid nitrogen, and rapid thawing. The survival rate was higher when cells were grown, before freezing, in the presence of 2 M NaCl instead of 1 M NaCl (78 and 48% survival, respectively). This difference is probably due to the intracellular amount of glycerol, which increases with external NaCl concentration and, therefore, may enhance cell protection. Although cells grown in 4 M NaCl accumulated a large amount of glycerol in response to osmotic stress, they did not withstand freezing. The use of cryoprotectant was absolutely necessary for the cells to recover from storage at –196°C. Glycerol was used because it is naturally produced by Dunaliella salina and therefore is not toxic. Provided it was added slowly to avoid osmotic shock, 3.5 M glycerol gave better results than 1M glycerol (48 and 18% survival, respectively).

Cold adaptation in the dark increased postthaw viability. Cells grown in 1 M or 2 M NaCl had a survival rate of 48 and 78%, respectively, when cold‐adapted, against 10 and 42% when not cold‐adapted. This adaptation could be due to the synthesis, at low temperature, of specific proteins because two bands (28–29 kDa) appeared when electrophoretically separated proteins from cold‐adapted cells and control cells were compared. Also, it could be due to the degradation of starch that occurs in the dark and leads to glycerol accumulation. Our procedure has never been used to cryopreserve microalgae and could enhance reported survival rates.
THE STORAGE GLUCAN OFPHAEOCYSTIS GLOBOSA(PRYMNESIOPHYCEAE) CELLS1
Authors: Janse Ingmar , van Rijssel Marion , van Hall Pieter‐Jan , Gerwig Gerrit J. , Gottschal Jan C. , Prins Rudolf A.
Published: October 2004

A non‐colony‐forming axenic strain ofPhaeocystis globosa(Harlot) Lagerheim was shown to produce a water‐soluble β‐d‐glucan. This glucan consisted of about 20 glucose units, mainly (l→3)‐linked, with branching at position 6. Therefore, it can be classified as a chrysolaminaran. Glucan production occurred mainly during the stationary growth phase and resulted in concentrations as high as 76 pg glucose per cell. When cultures were deprived of light the glucans were consumed, which supports their possible role as compounds used for temporary storage of energy.
CHANGES IN CELL COMPOSITION AND LIPID METABOLISM MEDIATED BY SODIUM AND NITROGEN AVAILABILITY IN THE MARINE DIATOM PHAEODACTYLUM TRICORNUTUM (BACILLARIOPHYCEAE)1
Authors: Larson Tony R. , Rees T. Alwyn V.
Published: October 2004

The effects of nitrogen starvation in the presence or absence of sodium in the culture medium were monitored in batch cultures of the marine diatom Phaeodactylum tricornutum Bohlin. During nitrogen starvation in the presence of sodium, cell nitrogen and chlorophyll a decreased, mainly as a consequence of continued cell division. These decreases were accompanied by decreases in the rates of photosynthesis and respiration. There was no change in either cell volume or carbohydrate, but both carbon and lipid increased. During nitrogen starvation in the absence of sodium, cell division ceased. Cell nitrogen and chlorophyll a remained constant, and respiration did not decrease, but the changes in the photosynthetic rate and the lipid content per cell were similar to cultures that were nitrogen‐starved in the presence of sodium. The carbon‐to‐nitrogen ratio increased in both cultures. Nitrogen, in the form of nitrate, and sodium were resupplied to cultures that had been preconditioned in nitrogen‐ and sodium‐deficient medium for 5 d. Control cultures to which neither nitrate or sodium were added remained in a static state with respect to cell number, volume, and carbohydrate but showed slight increases in lipid. Cells in cultures to which 10 mM nitrate alone was added showed a similar response to cultures where no additions were made. Cells in cultures to which 50 mM sodium alone was added divided for 2 d, with concomitant small decreases in all measured constituents. Cell division resumed in cultures to which both sodium and nitrate were added. The lipid content fell dramatically in these cells and was correlated to metabolic oxidation via measured increases in the activity of the glyoxylate cycle enzyme, isocitrate lyase. We conclude that lipids are stored as a function of decreased growth rate and are metabolized to a small extent when cell division resumes. However, much higher rates of metabolism occur if cell division resumes in the presence of a nitrogen source.
CHANGES IN INTRACELLULAR NITROGEN POOLS AND FEEDBACK CONTROLS ON NITROGEN UPTAKE IN CHAETOMORPHA LINUM (CHLOROPHYTA)1
Authors: McGlathery Karen J. , Pedersen Morten Foldager , Borum Jens
Published: October 2004

Changes in the size of intracellular nitrogen pools and the potential feedback by these pools on maximum N uptake (NH4+ and NO3−) rates were determined for Chaetomorpha linum (Müller) Kützing grown sequentially under nutrient‐saturating and nutrient‐limiting conditions. The size of individual pools in N‐sufficient algae could be ranked as residual organic N (RON) comprised mainly of amino acids and amino compounds > protein N > NO3− > NH4+ > chlorophyll N. When the external N supply was removed, growth rates remained high and individual N pools were depleted at exponential rates that reflected both dilution of existing pools by the addition of new biomass from growth and movement between the pools. Calculated fluxes between the tissue N pools showed that the protein pool increased throughout the N depletion period and thus did not serve a storage function. RON was the largest storage reserve; nitrate was the second largest, but more temporary, storage pool that was depleted within 10 days. Upon N resupply, the RON pool increased 3 × faster than either the inorganic or protein pools, suggesting that protein synthesis was the rate‐limiting step in N assimilation and caused a buildup of intermediate storage compounds. Maximum uptake rates for both NH4+ and NO3− varied inversely with macroalgal N status and appeared to be controlled by changes in small intracellular N pools. Uptake of NO3− showed an initial lag phase, but the initial uptake of NH4+ was enhanced and was present only when the intracellular NH4+ pool was depleted in the absence of an external N supply. A strong negative correlation between the RON pool size and maximum assimilation uptake rates for both NH4+ and NO3− suggested a feedback control on assimilation uptake by the buildup and depletion of organic compounds. Enhanced uptake and the accumulation of N as simple organic compounds or nitrate both provide a temporary mechanism to buffer against the asynchrony of N supply and demand in C. linum.
MODULATION OF THE HEAT SHOCK UBIQUITIN POOL IN SKELETONEMA COSTATUM (BACILLARIOPHYCEAE)1
Authors: Penna Antonella , Crinelli Rita , Magnani Mauro
Published: October 2004

Immunoblotting experiments performed with an anti‐ubiquitin antibody revealed that Skeletonema costatum (Grev.) Cleve cells contained free ubiquitin as well as ubiquitin conjugated to various endogenous proteins. A temperature shift from 18° to 30°C greatly increased the total amount of ubiquitin and particularly the ubiquitin fraction in high molecular mass conjugates. A solid‐phase immunoassay indicated values of 0.031 ± 0.004 pmol·10−6 cells for free ubiquitin and 0.046 ± 0.004 pmol·10−6 cells for conjugated ubiquitin for cells grown at 18°C, and 0.056 ± 0.008pmol·10−6cells and 0.21 ± 0.03 pmol·10−6cells, respectively, after a temperature increase from 18° to 30°C. Cell‐free extracts of S. costatum were equally able to form thiol ester linkages with 125I‐ubiquitin in an adenosine triphosphate–dependent manner at 18° C and at 30°C. Cell‐free extracts were also able to conjugate 125I‐ubiquitin to endogenous proteins, but the ubiquitin conjugation rate at 30°C was lower than at 18°C. Incubation of S. costatum for 3 h at 30°C and then for 3 h at 18°C resulted in the formation of high amounts of ubiquitin conjugates, suggesting that partially inactive or denaturated proteins accumulate during heat stress. These denaturated proteins are then conjugated to ubiquitin very efficiently when the physiological temperature is restored. Thus, S. costatum cells contain ubiquitin and an active ubiquitin conjugation system responding to stress conditions (temperature stress). The intracellular concentration of ubiquitin conjugates is most likely limited by the availability of protein substrates to be conjugated rather than by ubiquitin‐conjugating activity.
ANALYSIS OF ALEXANDRIUM (DINOPHYCEAE) SPECIES USING SEQUENCES OF THE 5.8S RIBOSOMAL DNA AND INTERNAL TRANSCRIBED SPACER REGIONS1
Authors: Adachi Masao , Sake Yoshihiko , Ishida Yuzaburo
Published: October 2004

The 5.8S ribosomal RNA gene (rDNA) and flanking internal transcribed spacers 1 and 2 (ITS1 and ITS2) from 7 isolates of Alexandrium catenella (Wedon et Kofoid) Taylor, 13 isolates of A. tamarense (Lebour) Balech, 2 isolates of A. affine (Fukuyo et Inoue) Balech, and single isolates of A. fundyense Balech, A. insuetum Balech, and A. pseudogonyaulax (Biecheler) Horiguchi ex Yuki et Fukuyo comb. nov. from Japan, Thailand, and the United States were amplified using the polymerase chain reaction (PCR), sequenced, and subjected to phylogenetic analysis. The sequences ranged from 518 to 535 base pairs (bp) exclusive of the 18S and 28S rDNA coding regions. Sequence comparisons revealed seven divergent “ITS types” designated as follows: 1) catenella type, 2) tamarense type, 3) WKS‐1 type, 4) Thai type, 5) affine type, 6) insuetum type, and 7) pseudogonyaulax type. Isolates of the tamarense type from various locations in Japan and the United States and of A. fundyense from the United States were closely related to each other and were clearly divergent from isolates of A. tamarense WKS‐1 (WKS‐I type) or A. tamarense CU‐15 (Thai type). These latter two strains carried unique ITS types, although they were not distinguishable from isolates of the tamarense type by morphological criteria. Distance values between isolates of the tamarense type and the WKS‐1 or Thai type were quite high (about 0.21 and 0.39, respectively). Seven isolates of A. catenella from Japan (catenella type) clearly diverged from the other ITS types already mentioned. Distance values between isolates of the catenella type were extremely low (<0.01), whereas distance values of ITS between the catenella type and the tamarense, WKS‐1, or Thai type were 0.17, 0.18, and 0.40, respectively. Isolates of A. affine, A. insuetum, and A. pseudogonyaulax all carried unique ITS types. The ITSs of the tamarense type exhibited two distinct ITS sets, the “A gene” and the “B gene.” The two sequences occurred in a 1:1 ratio in PCR products. In contrast, the ITSs of all other isolates appeared homogeneous. Sequence comparisons also showed that the variations in the 3′ end of ITS1 (150‐177 bp) were low within each ITS type but extremely high between ITS types. The number of different nucleotides among the seven Alexandrium types in this 28‐bp region is more than 10. High diversity of this region may facilitate the design of DNA probes specific for each ITS type/species of Alexandrium.
GENETIC CHARACTERIZATION OF STRAINS OF CYANOBACTERIA USING PCR‐RFLP OF THE cpcBA INTERGENIC SPACER AND FLANKING REGIONS1
Authors: Bolch Christopher J. S. , Blackburn Susan I. , Neilan Brett A. , Grewe Peter M.
Published: October 2004

Oligonucleotide primers, specific for conserved regions of the genes encoding the β‐ and α‐phycocyanin subunits of phycobilisomes (cpcB and cpcA) of cyanobacteria, were used to amplify a DNA fragment containing the intervening intergenic spacer region (cpcBA‐IGS) of 19 strains of three morphospecies of cyanobacteria. Six Australian strains were identified as Anabaena circinalis Rabenhorst, six strains were identified as Microcystis aeruginosa Kützing, and seven strains were identified as Nodularia spumigena Mertens. Restriction enzyme digestion of the amplification products from the strains revealed restriction fragment length polymorphism (RFLP) within all three morphospecies. Strains corresponding to M. aeruginosa were highly polymorphic: 11 of the 14 restriction enzymes used displayed RFLPs. The A. circinalis and N. spumigena strains were less variable: three of 14 enzymes and seven of 14 enzymes, respectively, showed RFLPs. The presence of genetic variation between strains within these three divergent morphospecies, which span two orders of cyanobacteria (Chroococcales Wettstein and Nostocales (Borzi) Geitler), show that the cpcBA‐ IGS fragment has broad application as a molecular marker for intrageneric studies of cyanobacteria systematics and genetics.
UTILITY OF THE MITOCHONDRIAL nad4L GENE FOR ALGAL AND PROTISTAN PHYLOGENETIC ANALYSIS1
Authors: Chesnick Joby M. , Tuxbury Kathyrn , Coleman Annette , Burger Gertraud , Lang B. Franz
Published: October 2004

Mitochondrial gene sequences, in contrast to their plastid counterparts, have been rarely exploited for use in determining algal phylogenetic relationships. Their utilization would prove advantageous particularly for the assessment of evolutionary relationships among protistan lineages that include photo synthetic species and their colorless heterotrophic dependents that contain only a reduced plastid genome as well as ancestral nonphotosynthetic relatives. Recent studies that have examined mitochondrial protein (coxl, cox2, cob, and nad5) rather than ribosomal RNA genes confirm that these algal sequences are sufficiently conserved to be used routinely in algal systematics. In this investigation, we sequenced the mitochondrial nad4L gene, encoding a small (ca. 99‐amino acid) polypeptide subunit of the NADH dehydrogenase complex, from two chromophytic algae and an oomycete to determine its suitability for use in determining algal and protistan evolutionary relationships. Our analysis demonstrated that nad4L could clearly resolve these three organisms as a distinct clade, the stramenopiles, to the exclusion of terrestrial plants/chlorophytes and eufungi. However, the position of deeply rooted species within the stramenopiles proved tenuous and would best be examined in conjunction with the analysis of longer mitochondrial protein gene sequences.
TWO DISTINCT COLONIAL MORPHOTYPES OF AMPHORA COFFEAEFORMIS (BACILLARIOPHYCEAE) CULTURED ON SOLID MEDIA1
Authors: Garduño Rafael A. , Hall Beverley D. , Brown Leslie , Robinson Maurice G.
Published: October 2004

When cultured on different types of solid media, the marine‐fouling diatom Amphora coffeaeformis (Ag.) Kütz. consistently formed two distinct colonial morphotypes named tight and fuzzy. Tight colonies were comprised mainly of small, morphologically distorted, nonmotile cells, whereas morphologically normal and highly motile cells formed the fuzzy colonies. Cells from tight colonies were less adherent to glass, grew more slowly in liquid media, and had a slightly decreased viability on plates with copper than cells from fuzzy colonies. Whereas the protein profiles of the two types of cells were nearly identical in polyacrylamide gels stained with Coomassie blue, cells from tight colonies produced a significantly lower amount of a protease‐resistant, low Mr polysaccharide or glycoconjugate as detected in silver‐stained gels. The frequency of appearance of the fuzzy and tight morphotypes was not influenced by the mode of nutrition or the type of substratum to which the algal cells adhered. However, certain formulations of solid medium and the presence of growth‐inhibitory concentrations of copper in agar plates favored the formation of tight colonies. Due to their frequencies and patterns of appearance, it was clear that the two naturally formed morphotypes were not the consequence of spontaneous mutations, genetic rearrangement, or selection of stable natural variants, and we have hypothesized that they were linked to a normal physiological behavior. The tight colonial morphotype was used as a valuable marker to screen for true motility/adhesion mutants within an ultraviolet‐mutagenized population of A. coffeaeformis. Seven mutants were isolated that were non‐motile on agar plates, poorly adherent to glass, and distinguished from naturally formed cells from tight colonies by their inability to form fuzzy colonies upon subculture on solid media.
TEMPERATURE REQUIREMENTS FOR GROWTH AND SURVIVAL OF ANTARCTIC RHODOPHYTA1
Authors: Bischoff‐Bäsmann Bettina , Wiencke Christian
Published: October 2004

The temperature requirement for growth and the upper survival temperatures (USTs) of 15 Antarctic red algal species collected on King George Island (South Shetland Islands) and Signy Island (South Orkney Islands) were determined. Two groups with different temperature requirements were identified. 1) A “eurythermal” group includes Rhodymenia subantarctica, Phyllophora ahnfeltioides, Gymnogongrus antarcticus, and Rhodochorton purpureum, growing between 0° and 10°C with optimum values at (0°) 5°(l0°)C. The USTs of these species and of Porphyra endiviifolium, Delesseria lancifolia, and Bangia atropurpurea were between 22° and 16°C. These species survived temperatures in a similar range as most endemic Arctic or Arctic/cold‐temperate species but exhibited a lower temperature demand for growth, suggesting an earlier contact with low temperatures than Arctic species. 2) A stenothermal group includes Pantoneura plocamioides, Myriogramme mangini, Ballia callitricha, Phyllophora antarctica, Gigartina skottsbergii, Georgiella confluens, and Plocamium cartilagineum growing at 0° or ≤5°C with optimum values at 0° or 5°C. The USTs of these species and of Phycodrys austrogeorgica were between 14° and 7°C. The species of this group must have had an even earlier contact with the Antarctic cold‐water environment than species of the “eurythermal” group. Gigartina skottsbergii, Georgiella confluens, Plocamium cartilagineum, and Pantoneura plocamioides were probably exposed longer to low temperatures than the other species of this group or Antarctic green and brown algae because they show the lowest temperature requirements so far determined in seaweeds. The results are discussed in the context of present local temperature regimes at the localities where the isolates were collected. Moreover, an attempt was made to explain the geographic distribution of individual species by the temperature requirements determined in this study. Only a few of the distribution limits are determined by temperature growth and/or survival characteristics. In many species (Rhodymenia subantarctica, Ballia callitricha, Gigartina skottsbergii, Bangia atropurpurea, Rhodochorton purpureum, and Plocamium cartilagineum), the development of temperature ecotypes is evident.
CELL‐ASSOCIATED PROTEOLYTIC ENZYMES FROM MARINE PHYTOPLANKTON1
Authors: Berges John A. , Falkowski Paul G.
Published: October 2004

Despite their central importance in cell metabolism, little is known about proteases in marine phytoplankton. We surveyed caseinolytic and leucine aminopeptidase (LAP) activities in log‐phase cultures of the chlorophyte Dunaliella tertiolecta Butcher, the diatom Thalassiosira weissflogii (Gru.) Fryxell et Hasle, the chrysophyte Isochrysis galbana Parke, the coccolithophorid Emiliania huxleyi (Lohm.) Hay et Mohler, and the cyanobacterium Synechococcus sp. (WH 5701). LAP activity was very low at pH < 6 and peaked between pH 7.5 and 8.5 in all species, whereas caseinolytic activity in most species showed only minor peaks in the pH 4–5 range and broad maxima above pH 8. Thus, acidic vacuolar proteases apparently represented only a small fraction of total protease activity. Attempts to classify proteases using selective inhibitors were inconclusive. Neither the serine/cysteine protease inhibitor leupeptin nor the aspartic protease inhibitor pepstatin. A inhibited caseinolytic or LAP activity in any species. The metalloprotease inhibitor EDTA was only effective against LAP activity in some species, causing average decreases of 30–50%, whereas the cysteine/serine protease inhibitor phenyl methyl sulfonylfluoride achieved at best a 30–60% decrease in caseinolytic activity. Caseinolytic activities were remarkably stable. At pH 7.5 and 25°C, extracts of D. tertiolecta, E. huxleyi, and Synechococcus showed no changes in activity after 24 h, whereas activity declined by less than 50% in the other species. Incubation of cell extracts for 1 h at 25°C in pH 7.5 buffer did not alter patterns of cell proteins, suggesting that endogenous proteases did not effectively degrade endogenous proteins. Casein zymograms were used to identify >200‐and <20‐kDa proteases in homogenates of log‐phase T. weissflogii; only the smaller protease was found in D. tertiolecta. Antibodies to the ATPase subunit (C) of the conserved, chloroplastic Clp protease from Pisum cross‐reacted with proteins in Synechococcus, D. tertiolecta, and I. galbana, but no cross‐reactions were found for any species with antibodies against the ClpP subunit from either E. coli or Nicotiana. Our results show that phytoplankton contain a diverse complement of proteases with novel characteristics.
CHANGES IN PHOTOSYNTHETIC YIELD, AMINO ACIDS, AND ORGANIC ACIDS ARE INDUCED BY AMMONIUM ADDITION TO CELLS OF PHORMIDIUM LAMINOSUM (CYANOPHYCEAE)1
Authors: Ochoa de Alda Jesús A. G. , Tapia María I. , Llama María J. , Serra Juan L.
Published: October 2004

The effect of NH4+ addition to NO3−‐growing cells of the non‐N2‐fixing cyanobacterium Phormidium laminosum (Agardh) Gomont (strain OH‐1‐pCl1) on photo‐synthetic and respiratory electron transport as well as on the intracellular levels of amino acids and some organic acids was studied. Addition of ammonium to nitrate‐growing cells resulted in substantial increases in the pool size of most amino acids and a transient decrease in the pool size of organic acids. The high demand for organic acids was partially overcome by degradation of stored carbohydrates, more than by newly fixed carbon, as indicated by the large stimulation of the respiration rate upon ammonium addition. Following ammonium addition, the photosynthetic yield of the in vivo noncyclic electron transport decreased, and the sensitivity of photosystem II to photodamage increased. Results indicate that cells balance their photosynthetic and respiratory activities depending on nitrogen availability and point to an important involvement of respiration in providing energy for ammonium assimilation until adaptation of bioenergetic processes to the new nitrogen source is complete.
SODIUM DEPRIVATION UNDER ALKALINE CONDITIONS CAUSES RAPID DEATH OF THE FILAMENTOUS CYANOBACTERIUM SPIRULINA PLATENSIS1
Authors: Schlesinger Penina , Belkin Shimshon , Boussiba Sammy
Published: October 2004

Like other alkaliphiles, the cyanobacterium Spirulina platensis (Norst.) Geitler requires sodium to function properly at elevated pH values. At pH 10.0, 150–250 mM Na+ were required for optimal growth, whereas 2.5 mM were sufficient for short‐term photosynthetic oxygen evolution. The complete absence of sodium, however, caused S. platensis to deteriorate. O2 evolution stopped, the absorbance at 620 nm corresponding to phycocyanin decreased, and the cells lysed within 1 h, a process accelerated by light. The activity of photosystem II, but not that of photosystem I, was affected in the process, which was irreversible unless sodium was readded within 15 minfrom the onset of the deprivation. The effect was mimicked, even in the presence of sodium, by the ionophore nigericin. We suggest that the cascade of events leading to cell lysis is primarily due to the inability of S. platensis to maintain a proton gradient (acid inside), possibly due to inactivity of a sodium/proton antiporter, as demonstrated for other alkaliphiles.
PHOTOSYNTHETIC RESPONSE OF THE GIANT KELP MACROCYSTIS PYRIFERA (PHAEOPHYCEAE) TO ULTRAVIOLET RADIATION1
Authors: Clendennen Stephanie K. , Zimmerman Richard C. , Powers Dennis A. , Alberte Randall S.
Published: October 2004

Solar ultraviolet radiation (UVA + UVB) impairs photosynthesis in marine algae. Canopy blades of the giant kelp Macrocystis pyrifera (L.) C. Agardh are exposed to high levels of solar UV in the field. To determine the effects of UV radiation on photosynthesis in the giant kelp and to identify sites of UV damage, O2 evolution, reaction center organization, light harvesting, and energy transfer efficiency were measured in canopy blades that had been exposed to elevated levels of UV in the laboratory. UV treatment reduced both the light‐saturated rate and the light‐limited rate of photosynthesis by 50% but produced no significant change in the rate of dark respiration. A significant impairment of photosystem II (PSII) reaction center function was observed, suggesting that PSII is a major site of damage in chromophytes. Reduced quantum efficiency of photosynthesis and loss of energy transfer from light‐harvesting pigments (fucoxanthin, chlorophyll a, and chlorophyll c) to PSII indicate that the major light‐harvesting complex of M. pyrifera, the fucoxanthin‐chlorophyll protein complex (FCPC), was another site of UV damage. These measures provide the first evidence of a direct effect of UV radiation on specific sites in the photosynthetic apparatus of chromophytes and indicate that in situ fluorescence excitation analysis may be a simple means to detect UV stress in algae.
EFFECT OF IRRADIANCE ON GROWTH AND REPRODUCTIVE PROCESSES DURING THE CELL CYCLE IN SCENEDESMUS ARMATUS (CHLOROPHYTA)1
Authors: Tukaj Zbigniew , Kubínová Anna , Zachleder Vilém
Published: October 2004

Synchronized populations of the chlorococcal alga Scenedesmus armatus (Chod.) Chod. were grown under five irradiance levels. During the cell cycles of these populations, reproductive processes such as DNA replication, nuclear division, protoplast fission, and daughter cell release and growth processes such as RNA and protein accumulation were followed. The amount of RNA and proteins increased stepwise with a short time interval between individual steps during which the rate of RNA and protein accumulation decreased. At each of the steps, the amount of RNA and protein approximately doubled and the number of steps increased with irradiance. At the end of each of the growth steps, a commitment to trigger the sequence of reproductive events (DNA replication, nuclear division, protoplast fission) was attained. After attaining the commitment point, the cells were able to trigger and terminate the whole reproductive sequence without any further growth, that is, even in the dark when the external supply of energy was cut off. With increasing irradiance, the number of commitment points attained during one cell cycle increased from one to four. Consequently, one to four sequences of the reproductive steps were triggered, and each of them ended by doubling the reproductive structures, which resulted in the formation of 2, 4, 8, or 16 daughter cells. The length of the precommitment periods shortened with increasing irradiance as the result of an increasing rate in growth. The length of postcommitment periods showed light independence and remained constant at the range of irradiances at which the number of growth steps and, consequently, the number of sequences of reproductive events did not change. At higher irradiances, the number of sequences of reproductive events increased, which caused a prolongation of postcommitment periods. The length of the cell cycle varied as a result of this distinct effect of irradiance on pre‐ and postcommitment periods.