Showing papers in "Journal of Phycology in 1996"

Stress tolerance in intertidal seaweeds

Abstract: Intertidal seaweeds are periodically exposed to air where they experience a variety of potentially stressful environmental conditions, including nutrient limitation, high light, high and low temperature, desiccation, and osmotic stress. This paper considers the current understanding of stress tolerance in intertidal seaweeds and discusses ways in which future research could increase our understanding of the role of environmental factors in the ecology and physiology of these algae. We believe research is required in at least three areas. 1) Laboratory physiological studies have established that correlations exist between stress tolerance and the vertical distribution of species. However, little information is available on the importance of stress in determining community structure in nature. Field experiments are essential to relate the impact of single or multiple stresses on the survival, growth, and reproductive output of macroalgae. In paticular, it is necessary to clarify the role of sublethal stress in determining the outcome of competitive interactions. 2) With the exception of obvious lethal effects or damage associated with extreme environmental conditions, such as unusually hot and dry weather, it is difficult to evaluate the occurrence and severity of stress in natural populations of seaweeds. There is a need to develop molecular and biochemical markers specific for individual stresses or groups of stresses to allow the unambiguous and direct determination of stress in situ. 3) Despite the apparent importance of stress in intertidal seaweeds, we are largely ignorant of the mechanistic basis of tolerance. The application of currently available tools of molecular and cell biology to the investigation of stress-induced transcriptional and translational changes could enormously increase our understanding of both the sites of, and pathways involved in, stress tolerance. In summary, there are numerous unanswered fundamental questions about the stress tolerance of intertidal seaweeds, providing opportunities for research ranging from field ecology to molecular biology and biochemistry.

Microenvironments and microscale productivity of cyanobacterial desert crusts

Abstract: We used microsensors to characterize physicochemical microenvironments and photosynthesis occurring immediately after water saturation in two desert soil crusts from southeastern Utah, which were formed by the cyanobacteria Microcoleus vaginatus Gomont, Nostoc spp., and Scytonema sp. The light fields within the crusts presented steep vertical gradients in magnitude and spectral composition. Near-surface light-trapping zones were formed due to the scattering nature of the sand particles, but strong light attenuation resulted in euphotic zones only ca. 1 mm deep, which were progressively enriched in longer wavelengths with depth. Rates of gross photosynthesis (3.4–9.4 mmol O2·m−2·h−1) and dark respiration (0.81–3.1 mmol O−2·m−2·h−1) occurring within 1 to several mm from the surface were high enough to drive the formation of marked oxygen microenvironments that ranged from oxygen supersaturation to anoxia. The photosynthetic activity also resulted in localized pH values in excess of 10, 2–3 units above the soil pH. Differences in metabolic parameters and community structure between two types of crusts were consistent with a successional pattern, which could be partially explained on the basis of the microenvironments. We discuss the significance of high metabolic rates and the formation of microenvironments for the ecology of desert crusts, as well as the advantages and limitations of microsensor-based methods for crust investigation.

Diatom resting stages

Abstract: Resting cells and spores are common to many groups of algae (J. S. Davis 1972) and, particularly in the diatoms, they appear to have many roles. This review discusses the morphology and physiology of diatom resting stages as well as the classical and more recent hypotheses about the functions of these cells. Most of the literature suggests that resting stages are a means of longor short-term survival. However, the timing of formation and germination of these cells may also be important for species succession, dispersal, and cycling of nutrients through the water column.

Growth and nitrogen fixation of the diazotrophic filamentous nonheterocystous cyanobacterium trichodesmium sp. ims 101 in defined media: evidence for a circadian rhythm

Abstract: Trichodesmium sp. IMS 101, originally isolated from coastal western Atlantic waters by Prufert-Bebout and colleagues and maintained in seawater-based media, was successfully cultivated in two artificial media. Its characteristics of growth, nitrogen fixation, and regulation of nitrogen fixation were compared to those of natural populations and Trichodesmium sp. NIBB 1067. Results indicate that the culture grown in artificial media had nitrogen fixation characteristics similar to those when the culture is grown in seawater-based medium and to those of Trichodesmium sp. in the natural habitat. The study provides practical artificial media to facilitate the physiological studies of these important diazotrophic cyanobacteria, as well as the cultivation of other Trichodesmium species in future studies. Manipulations of the light/dark cycle were performed to determine whether or not the daily cycle of nitrogen fixation is a circadian rhythm. Cultures grown under continuous light maintained the cycle for up to 6 days. We demonstrated that the daily cycle of nitrogen fixation in Trichodesmium sp. IMS 101 was at least partially under the control of a circardian rhythm.

Effects of harvest stage and light on the biochemical composition of the diatom thalassiosira pseudonana1

Abstract: The marine diatom Thalassiosira pseudonana (Hustedt, clone 3H) Hasle and Heimdal was cultured under three different light regimes: 100 μmol photon · m−2· s−1 on 12:12 h light : dark (L:D) cycles; 50 μmol photon · m−2· s−2 on 24:0 h L:D; and 100 μmol photon · m−2· s−1 on 24:0 h L:D. It was harvested during logarithmic and stationary phases for analysis of biochemical composition. Across the different light regimes, protein (as % of organic weight) was highest in cells during logarithmic phase, whereas carbohydrate and lipid were highest during stationary phase. Carbohydrate concentrations were most affected by the different light regimes; cells grown under 12:12 h L:D contained 37–44% of the carbohydrate of cells grown under 24:0 h L:D. Cells in logarithmic phase had high proportions of polar lipids (79 to 89% of total lipid) and low triacylglycerol (≤10% of total lipid). Cells in stationary phase contained less polar lipid (48 to 57% of total lipid) and more triacylglycerol (22 to 45% of total lipid). The fatty acid composition of logarithmic phase cells grown under 24:0 h L:D were similar, but the 100 μmol photon · m−2· s−1 (12:12 h L:D) cells at the same stage contained a higher proportion of polyunsaturated fatty acids (PUFAs) and a lower proportion of saturated and monounsaturated fatty acids due to different levels of 16:0, 16:1(n-7), 16:4(n-1), 18:4(n-3), and 20:5(n-3). With the onset of stationary phase, cells grown at 100 μmol photon · m−2· s−1 (both 12:12 and 24:0 h L:D) increased in proportions of saturated and monounsaturated fatty adds and decreased in PUFAs. Concentrations (% organic or dry weight) of 14:0, 16:0, 16:1(n-7), 20:5(n-3), and 22:6(n-3) increased in cells of all cultures during stationary phase. The amino acid compositions of cells were similar irrespective of harvest stage and light regime. For mariculture, the recommended light regime for culturing T. pseudonana will depend on the nutritional requirements of the animal to which the alga is fed. For rapidly growing bivalve mollusc larvae, stationary-phase cultures grown under a 24:0 h L:D regime may provide more energy by virtue of their higher percentage of carbohydrate and high proportions and concentrations of energy-rich saturated fatty acids.

The effect of phosphate status on the kinetics of cyanophage infection in the oceanic cyanobacterium synechococcus sp. wh78031

Abstract: Phycoerythrin-containing Synechococcus species are considered to be major primary producers in nutrient-limited gyres of subtropical and tropical oceanic provinces, and the cyanophages that infect them are thought to influence marine biogeochemical cycles. This study begins an examination of the effects of nutrient limitation on the dynamics of cyanophage/Synechococcus interactions in oligotrophic environments by analyzing the infection kinetics of cyanophage strain S-PM2 (Cyanomyoviridae isolated from coastal water off Plymouth, UK) propagated on Synechococcus sp. WH7803 grown in either phosphate-deplete or phosphate-replete conditions. When the growth of Synechococcus sp. WH7803 in phosphate-deplete medium was followed after infection with cyanophage, an 18-h delay in cell lysis was observed when compared to a phosphate-replete control. Synechococcus sp. WH7803 cultures grown at two different rates (in the same nutritional conditions) both lysed 24 h postinfection, ruling out growth rate itself as a factor in the delay of cell lysis. One-step growth kinetics of S-PM2 propagated on host Synechococcus sp. WH7803, grown in phosphate-deplete and-replete media, revealed an apparent 80% decrease in burst size in phosphate-deplete growth conditions, but phage adsorption kinetics ofS-PM2 under these conditions showed no differences. These results suggested that the cyanophages established lysogeny in response to phosphate-deplete growth of host cells. This suggestion was supported by comparison of the proportion of infected cells that lysed under phosphate-replete and-deplete conditions, which revealed that only 9.3% of phosphate-deplete infected cells lysed in contrast to 100% of infected phosphate-replete cells. Further studies with two independent cyanophage strains also revealed that only approximately 10% of infected phosphate-deplete host cells released progeny cyanophages. These data strongly support the concept that the phosphate status of the Synechococcus cell will have a profound effect on the eventual outcome of phage-host interactions and will therefore exert a similarly extensive effect on the dynamics of carbon flow in the marine environment.

Microenvironmental control of photosynthesis and photosynthesis‐coupled respiration in an epilithic cyanobacterial biofilm1

Abstract: The photosynthetic performance of an epilithic cyano-bacterial biofilm was studied in relation to the in situ light field by the use of combined microsensor measurements of O2, photosynthesis, and spectral scalar irradiance. The high density of the dominant filamentous cyanobacteria (Oscillatoria sp.) embedded in a matrix of exopolymers and bacteria resulted in a photic zone of < 0.7 mm. At the biofilm surface, the prevailing irradiance and spectral composition were significantly different from the incident light. Multiple scattering led to an intensity maximum for photic light (400–700 nm) of ca. 120% of incident quantum irradiance at the biofilm surface. At the bottom of the euphotic zone in the biofilm, light was attenuated strongly to < 5–10% of the incident surface irradiance. Strong spectral signals from chlorophyll a (440 and 675 nm) and phycobilins (phycoerythrin 540–570 nm, phycocyanin 615–625 nm) were observed as distinct maxima in the scalar irradiance attenuation spectra in the upper 0.0–0.5 mm of the biofilm. The action spectrum for photosynthesis in the cyanobacterial layer revealed peak photosynthetic activity at absorption wavelengths of phycobilins, whereas only low photosynthesis rates were induced by light absorption of carotenoids (450–550 nm). Respiration rates in light- and dark-incubated biofilms were determined using simple flux calculations on measured O2 concentration profiles and photosynthetic rates. A significantly higher areal O2 consumption was found in illuminated biofilms than in dark-incubated biofilms. Although photorespiration accounted for part of the increase, the enhanced areal O2 consumption of illuminated biofilms could also be ascribed to a deeper oxygen penetration in light as well as an enhanced volumetric O2 respiration in and below the photic zone. Gross photosynthesis was largely unaffected by increasing flow velocities, whereas the O2 flux out of the photic zone, that is, net photosynthesis, increased with flow velocity. Consequently, the amount of produced O2 consumed within the biofilm decreased with increasing flow velocity. Our data indicated a close coupling of photosynthesis and respiration in biofilms, where the dissolved inorganic carbon requirement of the photo-synthetic population may largely be covered by the respiration of closely associated populations of heterotrophic bacteria consuming a significant part of the photosynthetically produced oxygen and organic carbon.

Widespread phagocytosis of ciliates and other protists by marine mixotrophic and heterotrophic thecate dinoflagellates1

Abstract: An electron microscopic examination of large amorphous inclusions located in a variety of photosynthetic thecate dinoflagellates (Alexandrium ostenfeldii (Paulsen) Balech et Tangen, Gonyaulax diegensis Kofoid, Scrippsiella sp., Ceratium longipes (Bailey) Gran, and Prorocentrum micans Ehrenberg) and a nonphotosynthetic thecate species (Amylax sp.) revealed each inclusion to be a food vacuole, the majority of which were ingested ciliate prey. Recognizable features of these ciliates included linear arrays of basal bodies and cilia consistent with oligotrich polykinetid structure, characteristic macronuclei, chloroplasts (evidently kleptoplastids), cup-shaped starch plates, and cylindrical extrusomes. Three species contained (apparent) nonciliate prey: Scrippsiella sp., whose food vacuoles consistently contained unusual and complex extrusome-like cylindrical bodies having a distinctive six-lobed, multilayered structure; P. micans, which contained an unidentified encysted cell; and a single A. ostenfeldii cell, containing a Dinophysis sp. dinoflagellate cell. Several food vacuoles of ciliate origin had a red hue. This, together with the resemblance of A. ostenfeldii cells to planozygotes, suggests that similar structures previously identified as accumulation bodies may in fact be food vacuoles and that feeding may in some cases be associated with sexual processes.

Analysis of alexandrium (dinophyceae) species using sequences of the 5.8s ribosomal dna and internal transcribed spacer regions

Abstract: 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.

Pfiesteria piscicida gen. et sp. nov. (pfiesteriaceae fam. nov.), a new toxic dinoflagellate with a complex life cycle and behavior

Abstract: The newly described toxic dinoflagellate Pfiesteria piscicida is a polymorphic and multiphasic species with flagellated, amoeboid, and cyst stages. The species is structurally a heterotroph; however, the flagellated stages can have cleptochloroplasts in large food vacuoles and can temporarily function as mixotrophs. The flagellated stage has a typical mesokaryotic nucleus, and the theca is composed of four membranes, two of which are vesicular and contain thin plates arranged in a Kofoidian series of Po, cp, X, 4′, 1a, 5″, 6c, 4s, 5″′, and 2″″. The plate tabulation is unlike that of any other armored dinoflagellate. Nodules often demark the suture lines underneath the outer membrane, but fixation protocols can influence the detection of plates. Amoeboid benthic stages can be filose to lobose, are thecate, and have a reticulate or spiculate appearance. Amoeboid stages have a eukaryotic nuclear profile and are phagocytic. Cyst stages include a small spherical stage with a honeycomb, reticulate surface and possibly another stage that is elongate and oval to spherical with chrysophyte-like scales that can have long bracts. The species is placed in a new family, Pfiesteriaceae, and the order Dinamoebales is emended.

Isolation and characterization of a virus infecting phaeocystis pouchetii (prymnesiophyceae)1

Abstract: A virus infecting the haptophyte Phaeocystis pouchetii (Hariot) Lagerheim was isolated from Norwegian coastal waters in May 1995 at the end of a bloom of this phytoplankter. The virus was specific for P. pouchetii because it did not lyse 10 strains of P. globosa Scherffel, Phaeocystis sp., and P. antarctica Karsten. It was a double-stranded DNA virus, and the viral particle was a polyhedron with a diameter of 130–160 nm. The virus had a main polypeptide of about 59 kDa and at least five minor polypeptides between 30 and 50 kDa. The latent period of the virus when propagated in cultures of P. pouchetii was 12–18 h, and the time required for complete lysis of the cultures was about 48 h. The burst size was estimated to be 350–600 viral particles per lysed cell.

Changes in intracellular nitrogen pools and feedback controls on nitrogen uptake in chaetomorpha linum (chlorophyta)1

Abstract: 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 (Muller) Kutzing 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.

IDENTIFICATION OF CULTURED PSEUDO‐NITZSCHIA (BACILLARIOPHYCEAE) USING SPECIES‐SPECIFIC LSU rRNA‐TARGETED FLUORESCENT PROBES

Abstract: Some, but not all, marine pennate diatoms of the genus Pseudo-nitzschia H. Peragallo are associated with the production of domoic acid, a naturally occurring amino acid responsible for amnesic shellfish poisoning. Distinguishing between potentially toxic and nontoxic representatives of this genus is time-consuming and difficult because it demands scanning electron microscopy of cleaned frustules. The objective of this work is to speed and ease identification of these organisms by using whole-cell (in situ) hybridization and species-specific large-subunit ribosomal RNA (LSU rRNA)-targeted oligonucleotide probes. Toward that end, cultures of P. australis Frenguelli, P. pungens (Grunow) Hasle, P. multiseries (Hasle) Hasle, P. fraudulenta (P. T. Cleve) Heiden, P. heimii Manguin, P. delicatissima (P. T. Cleve) Heiden, P. pseudo-delicatissima (Hasle) Hasle, and P. americana (Hasle) Fryxell were screened with a suite of 15 putative species-specific probes. Of those, a subset of eight probes was found that distinguished each species tested. In addition, Pseudo-nitzschia chloroplasts were labeled with a probe directed against a eubacterial-conserved sequence. Identification of new cultures based on their reactivity toward a set of probes agreed with species designations as defined by morphological criteria. Whole-cell hybridization is a rapid, simple, and cost-effective technique for discriminating among cultured Pseudo-nitzschia species.

Changes in domoic acid production and cellular chemical composition of the toxigenic diatom pseudo-nitzschia multiseries under phosphate limitation1

Abstract: Production of domoic acid (DA), a neurotoxin, by the diatom Pseudo-nitzschia multiseries (previously Nitzschia pungens f. multiseries) Hasle and its cellular chemical composition were studied in phosphate-limited chemostat continuous cultures and in subsequent batch cultures. Under steady-state chemostat conditions, DA production increased from 0.01 to 0.26 pg DA · cell−1· d−1 as the growth rate decreased. When the nutrient supply was discontinued (to produce a batch culture), DA production was enhanced by a factor of ca. 3. DA production was temporarily suspended upon addition of phosphate to the batch cultures but resumed 1 d later at a higher rate coincident with the decline of phosphate uptake. In both steady-state continuous culture and batch culture, more DA was produced when alkaline phosphatase activity (APA) was high. The association of high DA production with high levels of APA and high cellular N:P ratios strongly suggests that phosphate limitation enhances DA production. Also, DA production was high when other primary metabolism (e.g. uptake of carbon, nitrogen, phosphorus and silicon, and cell division) was low, but chlorophyll a and adenosine triphosphate were generally high. This suggests that the synthesis of DA requires a substantial amount of biogenic energy.

Quantitative method for determining a representative algal sample count1

Abstract: A method for determining a representative count of a sample dependent on number of species is presented for application to various algal communities. Constant species curves are calculated as efficiency = (number of individuals–number of species)/number of individuals and diagrammed on a plot of efficiency versus number of individuals counted. Efficiency is defined as the probability that a new species encountered is minimal. That is, as the ratio of number of species to number of individuals approaches 1, more individuals will need to be counted in order to achieve a representative count. Data and calculations of efficiency from two algal communities are presented for illustration.

Further SEM study of marine dinoflagellates : the genus Ostreopsis (Dinophyceae)

Abstract: This paper presents a comprehensive examination of the taxonomy of the genus Ostreopsis Schmidt. The morphology of six species of marine dinoflagellates, Ostreopsis siamensis Schmidt 1902. Ostreopsis lenticularis Fukuyo 1981, Ostreopsis ovata Fukuyo 1981, Ostreopsis heptagona Norris, Bomber, et Balech 1985, Ostreopsis mascarenensis Quod 1994, and Ostreopsis labens Faust et Morton 1995 from three geographical regions (Japan, Southwest Indian Ocean, and the Caribbean) and three marine habitats (sand, water column, and macroalgal surfaces) are described from scanning electron micrographs. Differences in the following morphological characteristics differentiated the species: cell shape and size, and ornamentation of the epitheca, cingulum, and hypotheca. The thecal plate formula of the six Ostreopsis species is Po, 3′, 7″, 6C, 6S?, Vp, Rp, 5′″, 1p, 2″″, with differences in thecal plate size and shape. The cingulum in ventral view has two prominent structures: a ventral plate (Vp) with a ventral pore (Vo) and a ridged plate (Rp) that distinguishes Ostreopsis species from any other dinoflagellate taxa. This paper also includes ecological and toxicity information regarding the six Ostreopsis species.

Bacteria that induce morphogenesis in ulva pertusa (chlorophyta) grown under axenic conditions1

Abstract: Marine foliaceous green macroalgae such as Ulva lose their typical morphology when cultured aseptically in defined synthetic media. However, after reinfection by certain marine bacteria (isolated from unialgal cultures of Ulva pertusa Kjellman), the organisms regain their typical foliaceous or tubular morphology. To investigate the morphogenesis (MG) induced in U. pertusa by bacteria, we isolated and identified bacteria with MG activity on U. pertusa and studied the distribution of such bacteria in seawater and on various marine macroalgae. We isolated 1555 bacterial strains from 18 species of marine macroalgae (six Chlorophyta, five Phaeophyta, and seven Rhodophyta), from seawater and from sediment collected at the beach at Omaezaki, Shizuoka Prefecture; Japan. Of these, 676 bacterial strains (43.5%) showed MG activity. They were classified into six bacterial groups, Flavobacterium, Vibrio, Pseudomonas, Deleya, Escherichia, and gram-positive cocci. These bacteria were ubiquitous among the samples and were not specific to U. pertusa. Several plant growth regulators had no MG activity. Filter-sterilized supernatants of culture media of MG-active bacteria strains did not induce MG. Cocultivation of Ulva with active bacterial strains is so far the only way to induce the MG effect, which suggests that for MG direct contact between Ulva and the bacterial strain is necessary.

Effects of cadmium on growth and superoxide dismutase activity of the marine microalga tetraselmis gracilis (prasinophyceae)1

Abstract: Marine planktonic algae are frequently exposed to metallic contaminants. Because heavy metals can be assimilated and accumulated by algal cells, they can then be transferred to higher trophic levels of food chains. We studied the effects of cadmium on protein production and the growth of the marine prasinophyte Tetraselmis gracilis (Kylin) Butcher. By means of toxicological assays, we estimated the LC{sub 50} of cadmium as 3.2 ppm and 1.8 ppm after 48 h and 96 h of exposure to this heavy metal, respectively. The growth of curves and survival percentages of cell cultures in the presence of cadmium were determined, and a proportional reduction of both parameters with increasing metal concentrations of cadmium, T. gracilis contained high levels of superoxide dismutase (SOD) activity, one of the main enzymes of the cell`s antioxidant defense mechanism. Under these growth conditions, total SOD activity in crude extracts was increased by 41% (at 1.5 ppm) and 107% (at 3.0 ppm). Assays of SOD activity in nondenaturing polyacrylamide gels also showed a similar induction by cadmium. These results show that cadmium has potentially toxic properties since it significantly inhibited the growth of T. gracilis at low concentrations and promoted by induction of SOD activity, suggestivemore » of an oxidative stress state. Besides being the first report of SOD in T. gracilis, this work describes experimental evidence of SOD induction by cadmium in this species. 56 refs., 4 figs., 1 tab.« less

Differentiation of ulva mutabilis (chlorophyta) gametangia and gamete release are controlled by extracellular inhibitors1

Abstract: Blade cells of Ulva mutabilis Foyn (Chlorophyta) excrete regulatory factors into their cell walls and into the environment. These factors are essential for the maintenance of the vegetative state. “Sporulation inhibitor-1a” (SI-1a) is a glycoprotein that was isolated from the culture medium of axenic Ulva growing as an undifferentiated callus. This protein was unusually stable to denaturing treatments and showed an extremely high apparent molecular mass (Mr) of 1–4 × 107 daltons estimated by size exclusion chromatography. The glycosylation was not essential for activity. SI-1a suppressed gametogenesis completely at concentrations lower than 10−14 M. When Ulva developed normally in the presence of their symbiotic bacteria, smaller forms of SI-1 accumulated in the medium (104–106 daltons). Sporulation inhibitors of the same size spectrum and with similar properties were also extracted from crude cell walls of nonaxenic Ulva. A class of different nonprotein sporulation inhibitors (SI-2) of very low Mr and yet unknown structure was isolated from the inner space between the two blade cell layers. Excretion of all SI-1 forms decreased with maturation of the thallus, whereas the overall concentration of SI-2 in the thallus stayed constant throughout the life cycle. The SI-2 affected different Ulva species whereas SI-1 was species-specific. Gametogenesis was induced upon removal of both Sporulation inhibitors from small single-layered fragments of mature blades. After a “determination phase” of 23–46 h, dependent on the time of induction within the cell cycle, the cells became irreversibly committed to differentiation and were no longer susceptible to SI-1 or SI-2. Subsequently, during a 28-h “differentiation phase,” 16 progametes were formed by synchronous genome doublings and cell divisions and differentiated into mature gametes. These became motile and were released from the gametangia when the concentration of a “swarming inhibitor” of low Mr, excreted mainly during the “determination phase,” declined below a threshold concentration. The biochemical properties of these regulatory factors and their effects on gametogenesis and gamete release are described.

Why do dinoflagellates have lower growth rates?1

Abstract: Dinoflagellates have substantially lower growth rates than other taxa of similar size. These low growth rates have been suggested to reflect the lower chlorophyll a to carbon ratio (Chl a:C) in dinoflagellates, but that speculation has never been widely tested. This study tests if the variations in growth rates among taxa are related to differences in Chl a:C using published data. I collected 92 data entries from the literature representing 31 species, mostly from two divisions (Chrysophyta and Pyrrophyta), and found a significant relation (r2= 0.39) between growth and Chl a:C. Since Chl a:C is almost independent of C content, I also developed a growth model using both C and Chl a:C. Together, the two variables explain 68% of variation in algal growth. However, a further 6.4% of the variance in growth can still be attributed to phyletic differences. Low Chl a:C is only a partial explanation for the low growth rates of the dinoflagellates.

Physiological characteristics of spirulina platensis (cyanobacteria) cultured at ultrahigh cell densities

Abstract: Photosynthetic activity and growth physiology of Spirulina platensis (Nordstedt) Geitler cultures maintained at ultrahigh cell densities (ie above 100 mg chlorophyll-L−1) in a newly designed photobioreactor were investigated Nitrogen (NaNO3) in standard Zarouk medium was characterized as a major nutrient-limiting factor in such cultures The effect of ultrahigh cell density on photoinhibition of photosynthesis, as reflected by chlorophyll fluorescence and photosynthetic oxygen evolution, was studied: elevating the population density may arrest photoinhibition induced by high photon flux density, as well as low temperature The relationship between incident irradiance and oxygen production rate was linear in situ for cultures at the optimal cell density, indicating that light limitation rather than light saturation or photoinhibition is the dominant condition outdoors in cultures of ultrahigh cell densities In contrast with other reports, the extent of biomass loss at night due mainly to dark respiration was found to be relatively small when cell density was optimal, exerting only a minor effect on overall net productivity Measurements of oxygen consumption at night revealed low rates of respiration, which may be explained by the low value of the volumetric mass transfer coefficient (KLa) of oxygen Hence, reduced oxygen tension may play a role in preventing full expression of the respiratory potential in ultrahigh cell density cultures in which photoadaptive strategy may explain cell composition Ultrahigh cell densities optimized with respect to the intensity of the light source, the length of the light path, and the extent of stirring represent the key for obtaining high output rates of cell mass and some natural products

IDENTITY OF THE ENDOSYMBIONT OF PERIDINIUM FOLIACEUM (PYRROPHYTA): ANALYSIS OF THE rbcLS OPERON1

Abstract: Extant chromophytic algae have been suggested to have originated via the engulfment of a photo synthetic alga by a colorless protist. The dinoflagellate Peridinium foliaceum (Stein) Biecheler contains a reduced chlorophyll c–containing endosymbiont and, thus, represents an evolutionary intermediate stage in the establishment of chloroplasts. Although the exact phylogenetic relationship of the symbiont to extant algal species is unknown, it had been suggested that the P. foliaceum symbiont was either a diatom or a chrysophyte. Identification of the closest living relative of the P. foliaceum symbiont would provide a free-living model system with which the photosynthetic symbiont could be compared. Nucleotide sequence analysis of rbcL and rbcS (encoding the large and small subunits ofribulose-1,5-bisphosphate carboxylase/oxygenase) by the P. foliaceum symbiont was performed to provide insights into its identity. Cloned restriction fragments from a chloroplast DNA library were screened, and clones encoding the rbcLS operon were sequenced. Parsimony phylogenetic analysis was performed for each gene. Our data strongly suggest that the symbiont originated from a photosynthetic diatom.

GENETIC CHARACTERIZATION OF STRAINS OF CYANOBACTERIA USING PCR‐RFLP OF THE cpcBA INTERGENIC SPACER AND FLANKING REGIONS1

Abstract: 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 Kutzing, 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.

Characterization of nitrogen uptake by natural populations of Aureococcus anophagefferens (Chrysophyceae) as a function of incubation duration, substrate concentration, light, and temperature

Abstract: Nitrogen uptake studies were conducted during an aestival “brown tide” bloom in Shinnecock Bay, Long Island, New York. The same station was sampled in late July and mid-August 1995 when Aureococcus anophagefferens composed >90% and 30–40% of the total cell density, respectively. Experiments were designed to examine the effect of incubation duration on the uptake kinetics, and the effect of light and temperature dependencies of NH4+, urea, and NO3− uptake. Maximum specific uptake rates (V'max) decreased in the order NH4+, urea, NO3− and were nonlinear with time for NH4+ and urea, both of which exhibited an exponential decline between 1 and 10 min and then did nut significantly change for 60 min. Nitrogen uptake kinetic experiments exhibited a typical hyperbolic response for urea and NO3−. Half-saturation constants. (Ks) were calculated to he 0.03 and 0.12 μmol · L−1 for urea and NO3−; respectively, but could not be calculated for NH4+ under these experimental conditions. Nutrient uptake rate versus, irradiance (NI) experiments showed that maximum uptake rates occurred at ≤% of incident irradiance on both sampling dates and that values of V′max-cell (NH4+) were on average 30% greater than V′max-cell (urea). A7°–9°C temperature decrease in incubation temperature between the two NI experiments in August resulted in a 30% decrease in V′max-cell(NH4+), no change in V′max-cell(urea), and a 3–4-fold decrease in calculated Klt values for both NH4+ and urea. The results from these experiments demonstrate that A. anophagefferens has a higher affinity for NH4+ and urea than for NO3− and that this particular species is adapted to use these substrates at low irradiances and concentrations. The data presented in this study are also consistent with the hypothesis that A. anophagefferens may be an oceanic clone that was displaced by an anomalous oceanographic event.

Identification of the toxic dinoflagellates alexandrium catenella and a. tamarense (dinophyceae) using dna probes and whole‐cell hybridization1

Abstract: Fluorescent DNA probes (cCAT-F1 and cTAM-Fl) complementary to the 3′ end of ribosomal RNA (rRNA) internal transcribed spacer 1 sequences (ITS 1: positions 154–176) of toxic species of Alexandrium catenella (Whedon and Kofoid) Taylor and A. tamarense (Lebour) Taylor were applied to various cultures of the genus Alexandrium and several other phytoplankters using whole-cell fluorescent in situ hybridization. cCAT-F1 and cTAM-F1 reacted with targeted strains of A. catenella (catenella type) and A. tamarense (tamarense type), respectively, and did not react with isolates of A. affine (Inoue et Fukuyo) Balech, A. fraterculus (Balech) Balech, A. insuetum Balech, A. lusitanicum Balech, A. pseudogonyaulux (Biecheler)Horiguchi ex Yuki et Fukuyo comb. nov., nor isolates of Prorocentrum micans Ehrenberg, Amphidinium carterae Hulburt, Heterocapsa triquetra (Ehrenberg) Stein, Gymnodinium mikimotoi Miyake et Kominami ex Oda, Skeletonema costatum (Greville) Cleve, Heterosigma akashiwo (Hada) Hada, and Chattonella antiqua (Hada) Ono. DNase I and RNase A treatment showed that probes hybridized to ribosomal DNA, not rRNA. Probes were localized at the bottom of the U-shaped nucleus, a region that corresponds to the nucleolus. The probes are highly specific for particular strains of A. catenella and A. tamarense and are applicable for identifying these species collected from cultured and possibly natural populations.

Identification of the class prymnesiophyceae and the genus phaeocystis with ribosomal rna–targeted nucleic acid probes detected by flow cytometry1

Abstract: Target regions specific for the class Prymnesiophyceae and the genus Phaeocystis (Har.) Lag. were identified from 18S ribosomal RNA coding regions, and two complementary probes were designed (PRYMN01 and PHAEO01). Detection of whole cells hybridized with these probes labeled with fluorescein isothiocyanate was difficult using epifluorescence microscopy because autofluorescence of the chlorophylls seriously interfered with the fluorescence of the probes. In contrast, flow cytometry proved very useful to detect and quantify the fluorescence of the hybridized cells. Hybridization conditions were optimized, especially with respect to formamide concentration. Both probes were tested on a large array of both target and nontarget strains. Positive and negative controls were also analyzed. Specificity was tested by adding a competing nonlabeled probe. Whereas probe PHAEO01 seems to have good specificity, probe PRYMN01 appeared less specific and must be used with stringent positive and negative controls.

Fatty acid and lipid composition of the acidophilic green alga chlamydomonas sp.

Abstract: The lipid and fatty acid compositions of Chlamydomonas sp. isolated from a volcanic acidic lake and C. reinhardtii were compared, and the effects of pH of the medium on lipid and fatty acid components of Chlamydomonas sp. were studied. The fatty acids in polar lipids from Chlamydomonas sp. were more saturated than those of C. reinhardtii. The relative percentage of triacylglycerol to the total lipid content in Chlamydomonas sp. grown in medium at pH 1 was higher than that in other cells grown at higher pH. A probable explanation might be that Chlamydomonas sp. has two low pH adaptation mechanisms. One mechanism is the saturation of fatty acids in membrane lipids to decrease membrane lipid fluidity, and the other is the accumulation of triacylglycerol, as a storage lipid, to prevent the osmotic imbalance caused by high concentrations of H2SO4.

Sporeling coalescence and intraclonal variation in gracilaria chilensis (gracilariales, rhodophyta)

Abstract: This study evaluates the hypothesis that spore coalescence may cause intraclonal variation. Spore coalescence might allow the occurrence of unitary thalli that in fact correspond to genetically different, coalesced individuals. Plant portions simultaneously derived from these chimeric individuals may exhibit dissimilar growth responses even when incubated under similar abiotic conditions. Testing of the hypothesis included various approaches. Transmission electron microscopy observations of early stages of sporeling coalescence indicated that polysporic plantlets were formed by groups of spores and their derivatives. Even though adjacent cells in two different groups may fuse, these groups maintained an independent capacity to grow and form uprights. Laboratory-grown plantlets showed a significant correlation between the initial number of spores and the total number of erect axes differentiated from the sporeling. Construction and growth of bicolor individuals indicated the chimeric nature of the coalesced individuals. Coalesced, bicolor holdfasts had green and red cells, which subsequently produced green and red uprights, respectively. Individuals fronds were also chimeric, as indicated by the production of green and red branchlets from single, red uprights. The existence of mixed tissues was further substantiated by random amplified polymorphic DNA analysis. The banding pattern produced by branchlets of a unisporic thallus was consistently monomorphic, whereas the patterns produced by the polysporic thallus were polymorphic. Growth rates of polysporic thalli had larger data dispersal and variation coefficients than oligosporic or monosporic thalli. Therefore, all results support the original hypothesis and suggest that coalescence might be ecologically more important than previously thought.

Immunofluorescence in phytoplankton research: applications and potential

Abstract: During the last decade, immunofluorescence has become an important tool in studies of the systematics, biogeography, physiology, and ecology of freshwater and marine phytoplankton. The technology originated in the medical sciences (e.g. Coons et al. 1941, 1955) and was gradually applied to soil bacteria (Fliermans et al. 1974, Josserand and Cleyet-Mare1 1979) and bacteria from aquatic environments (Fliermans and Schmidt 1977, Gates and Pham 1979, Ward and Perry 1980, Baker and Mills 1982, Dahl and Laake 1982, Yoshioka et al. 1982, Ward and Carlucci 1985, Currin et al. 1990). Applications of immunology to phytoplankton originated over two decades ago with the innovative studies of Bernhard et al. (1969), who attempted to differentiate phytoplankton species using antibodies. Modern immunological studies of phytoplankton are more diverse in their applications, but the same principles of using antibodies to label and visualize target cells or cellular constituents still apply. Coupled with the power and sensitivity of fluorescence, immunological techniques open the door to physiological and ecological investigations that formerly were not possible. This is especially true in studies that focus on a single species within a complex field assemblage of organisms and detritus. Applications of immunofluorescence include identification and enumeration of phytoplankton species, localization and quantification of cell constituents (enzymes, toxins, structural proteins, and polysaccharides), and labeling of cells for better quantification of grazing rates and mixotrophic or heterotrophic potential. Given the rapid pace of development in this field, it seems appropriate at this time to review the methods, applications, and potential of immunofluorescence as a tool in phytoplankton research, in hopes that such an overview will help to sustain and focus future investigations.

Gradient analysis of diatom assemblages in western kentucky wetlands

Abstract: 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.