THE GLUCOSE TRANSPORT SYSTEM OF AMPHORA COFFEAEFORMIS (BACILLARIOPHYCEAE)1

Amphora coffeaeformis (Ag.) Kütz. var. perpusilla (Grun.) Cleve took up glucose by an inducible transport system. The system was induced by d -fructose, d -mannose, as well as glucose. Some d -pentoses also induced a glucose uptake system but it may not be the same one as that induced by hexose. d -fructose, d -mannose and 2-deoxy-d -glucose inhibited 2 mM glucose uptake at equimolar concentration, but d -pentoses did not. The uptake system decayed in ca. 5 h in the absence of glucose. The half-saturation constant for uptake, K₈ was ca. 0.1 mM glucose with a maximum uptake rate, V_max= 0.4 nmol/10⁶ cells-min^?1.     

ISOLATION AND PHYSIOLOGICAL STUDIES ON THREE ISOLATES OF AMPHORA (BACILLARIOPHYCEAE)1

Three clones of the diatom Amphora were euryhaline, able to grow autotrophically at 160 lx (0.001 ly/min) and heterotrophically on glucose and fructose. Furthermore 2 clones grew on glutamate and feast extract. Light-limited growth of individual clones was stimulated by glycerol, galactose, lactate, acetate, aspartate and asparagine, although mannose torn inhibitory at low and high light levels. The half-saturation constant for growth of A. coffeaefomis var. perpusilla Grunow (Cleve) with glucose was 25 μM. Heterotrophic growth rate of this organism became saturated with respect to glucose at 0.5 mM.     

PROTEINACEOUS AND IMMUNOCHEMICAL DISTINCTIONS BETWEEN THE OVAL AND FUSIFORM MORPHOTYPES OF PHAEODACTYLUM TRICORNUTUM (BACILLARIOPHYCEAE)1

Two morphotypes, fusiform and oval, were isolated from a single clone of the diatom Phaeodactylum tricornutum Bohlin and maintained as subclones by culturing in liquid and solid substrates, respectively. Salinity of the medium, from brackish to marine, had no effect on expression of the phenotypes. The oval cell is generated endogenously within a “transformed”fusiform cell upon transfer from liquid medium to agar plates. With the light microscope, normal and “transformed”fusiform cells, prior to giving rise to oval cells, can be discriminated by means of their staining response to toluidine blue. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of protein extracts from lysed cells revealed slight differences in polypeptide composition between fusiform and oval types. A phenotype-restoration experiment from oval to fusiform demonstrated that the oval type readily reestablished not only fusiform morphology but also the protein pattern characteristic for the fusiform type. Immunochemical analyses (western blots) using antisera raised against whole and lysed cells of both morphotypes revealed antigenic alterations of the oval morphotype. Several antigenic determinants restricted mainly to the surface of oval cells were detected. Results indicate that environmentally induced phenotypes of Phaeodactylum may be not only the consequence of specific gene expression but also the result of significant, general post-translational modifications.     

COMPARATIVE ULTRASTRUCTURE OF THE VALVES OF SOME CYCLOTELLA SPECIES (BACILLARIOPHYCEAE)1

Cyclotella atomus Hust., C. meneghiniana Kütz., C. comta (Ehr.) Kütz., C. antiqua W. Sm., C. michiganiana Skv., C. ocellata Pant., C. glomerata Bachmann, C. pseudostelligera Hust., and C. stelligera (Cleve et Grun.) V. H., were selected for ultrastructure investigations on the basis of their ready availability. Specimens were examined employing both transmission and scanning electron microscopy. Strutted processes, labiate processes, spines and other special features were surveyed in all species. On the basis of these observations the species of Cyclotella observed seem to be in one of 3 morphological groups, the meneghiniana group, the comta group, and the stelligera group.     

THE ULTRASTRUCTURE AND CYTOCHEMISTRY OF RESTING CELL FORMATION IN AMPHORA COFFAEFORMIS (BACILLARIOPHYCEAE)1

The ultrastructure of logarithmic-growing cells and of resting cells in laboratory cultures of Amphora coffaeformis (Ag.) Kütz. isolated from deep ocean water was examined using electron and light microscopy. The acid Phosphatase activity, chlorophyll a and lipid content were assessed at weekly intervals of resting cell formation during cold-dark treatment, simulating deep ocean water. Approximately 4 wk are required to complete resting cell formation. During the first week, the cytoplasm undergoes extensive transformation and lysosomal activity is observed. Large vacules decrease in size and many small ones develop, the mitochondria become fewer and one or more massive mitochondria appear possibly by fusion of smaller ones; the cytoplasm becomes densely granular. During the second and third week, the cytoplasm continues to contract, lipid bodies begin to develop and the plastid becomes densely stained. At the fourth week, the mature resting cell is formed containing one or more massive mitochondria, a well-formed plastid, and granular cytoplasm containing occasional lipid droplets. There is no change in frustule morphology and the cytoplasm does not produce a protective layer. The variation in chemical constituents correlates with microscopic structure of the cells. The fine structure of cells during growth resumption when exposed to light at 25 c is presented. Previous reports of viable, chlorophyll-containing cells at great depths in the ocean may be explained by the results reported in this paper.     

OBSERVATIONS OF A TUBE-DWELLING DIATOM: NAVICULA HAMULIFERA (BACILLARIOPHYCEAE)1

Colonies of the tube-dwelling diatom Navicula hamulifera Grunow living on mangrove prop roots in Indian River, Florida and at La Parguera, Puerto Rico, were studied using light and electron microscopy. Observations of the tube morphology and cell structure of this diatom from fresh samples and cultures are described, as well as the ultrastructural morphology of its frustule. The formation of tubes by this diatom is reported for the first time. Comparisons are made with the closest species; Navicula delognei V.H. and Navicula pseudoco moides Hendey.     

OBSERVATIONS OF A TUBE-DWELLING DIATOM: NAVICULA HAMULIFERA (BACILLARIOPHYCEAE)1

Colonies of the tube-dwelling diatom Navicula hamulifera Grunow living on mangrove prop roots in Indian River. Florida and at La Parguera, Puerto Rico, were studied using light and electron microscopy. Observations of the tube morphology and cell structure of this diatom from fresh samples and cultures are described, as well as the ultrastructural morphology of its frustule. The formation of tubes by this diatom is reported for the first time. Comparisons are made with the closest species; Navicula delognei V.H. and Navicula pseudocomoides Hendey.     

EFFECT OF SILICATE LIMITATION ON VALVE MORPHOLOGY IN THALASSIOSIRA AND COSCINODISCUS (BACILLARIOPHYCEAE)1

Coscinodiscus radiatus Ehrenb. and Thalassiosira eccentrica (Ehrenb.) Cleve were grown in a silicate-limited chemostat at silicate concentrations below 1 μg-atoms · l^?1. The resulting abnormal valves of C. radiatus lacked a thickened ring around the foramina; their pore membranes were thinner and their loculi shallower than those in normal cells. Abnormal valves of T. eccentrica had a fasciculate areolae pattern; they lacked a silica covering over the foramina and some tangential areolae walls. Neither abnormal valve could be termed a new species.     

PSEUDO-NITZSCHIA AS A GENUS DISTINCT FROM NITZSCHIA (BACILLARIOPHYCEAE)1

Morphologic, diagnostic characters of the subgenus Nitzschia, genus Nitzschia Hassall 1845, and the marine planktonic genus Pseudo-nitzschia H. Peragallo in H. & M. Peragallo 1900 were compared. Colony formation by overlap of cell ends; weakly silicified, shallow, and flattened valves; an extremely eccentric raphe, not elevated above the general level of the valve; lack of con-opea; and striated girdle bands characterize Pseudo-nitzschia as a natural group and a genus separate from Nitzschia.     

THE EFFECTS OF TEMPERATURE ON THE PHOTOSYNTHETIC PARAMETERS AND RECOVERY OF TWO TEMPERATE BENTHIC MICROALGAE,AMPHORA CF. COFFEAEFORMIS AND COCCONEIS CF. SUBLITTORALIS (BACILLARIOPHYCEAE)1

Temperature and irradiance are the most important factors affecting marine benthic microalgal photosynthetic rates in temperate intertidal areas. Two temperate benthic diatoms species, Amphora cf. coffeaeformis (C. Agardh) Kütz. and Cocconeis cf. sublittoralis Hendey, were investigated to determine how their photosynthesis responded to temperatures ranging from 5°C to 50°C after short‐term exposure (1 h) to a range of irradiance levels (0, 500, and 1,100 μmol photons · m^?2 · s^?1). Significant differences were observed between the temperature responses of maximum relative electron transport rate (rETRmax), photoacclimation index (E_k), photosynthetic efficiency (α), and effective quantum yield (ΔF/F_m’) in both species. A. coffeaeformis had a greater tolerance to higher temperatures than C. sublittoralis, with nonphotochemical quenching (NPQ) activated at temperatures of 45°C and 50°C. C. sublittoralis, however, demonstrated a more rapid rate of recovery at ambient temperatures. Temperatures between 10°C and 20°C were determined to be optimal for photosynthesis for both species. High temperatures and irradiances caused a greater decrease in ΔF/F_m’ values. These results suggest that the effects of temperature are species specific and that short‐term exposure to adverse temperature slows the recovery process, which subsequently leads to photoinhibition.     

OBSERVATIONS ON NAVICULA THALLODES (BACILLARIOPHYCEAE), A BLADE-FORMING DIATOM FROM THE BERING SEA1

A thallus-forming diatom, Navicula thallodes Proschkina-Lavrenko, previously known only from the original collection at Bering Island (U.S.S.R.), has been found at Amchitka Island in the Aleutians, Alaska. The most remarkable observation of the present report is that N. thallodes may form blades up to 50 cm long, which to our knowledge is the greatest length reported for a colonial diatom. SEM observations of this diatom are presented for the first time.     

IDENTIFICATION OF CULTURED PSEUDO-NITZSCHIA (BACILLARIOPHYCEAE) USING SPECIES-SPECIFIC LSU rRNA-TARGETED FLUORESCENT PROBES1

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.     

CORRELATED EVOLUTION OF GENOME SIZE AND CELL VOLUME IN DIATOMS (BACILLARIOPHYCEAE)1

A correlation between genome size and cell volume has been observed across diverse assemblages of eukaryotes. We examined this relationship in diatoms (Bacillariophyceae), a phylum in which cell volume is of critical ecological and biogeochemical importance. In addition to testing whether there is a predictive relationship across extant species, we tested whether evolutionary divergences in genome size were correlated with evolutionary divergences in cell size (using independent contrasts). We estimated total DNA content for 16 diatom species using a flow cytometer and estimated cell volumes using critical dimensions with scaling equations. Our independent contrast analyses indicated a significant correlated evolution between genome size and cell volume. We then explored the evolutionary and ecological implications of this evolutionary relationship. Diatom cell volume is an important component of the global carbon cycle; therefore, understanding the mechanisms that drive diatom genome evolution has both evolutionary and ecological importance.     

CARBONIC ANHYDRASE IN THE MARINE DIATOM THALASSIOSIRA WEISSFLOGII (BACILLARIOPHYCEAE)1

The zinc metalloenzyme carbonic anhydrase plays a critical role in inorganic carbon acquisition in marine diatoms, thus conferring on zinc a key role in oceanic carbon cycling. As a first step in determining the location and function of carbonic anhydrase (CA) in Bacillariophyceae, we purified and partially sequenced CA from T. weissflogii (Gru) Fryxell et Hasle (TWCA1) and cloned the corresponding cDNA (twca1). The twca1 sequence is different from other known algal carbonic anhydrase genes, and encodes a protein of roughly 34 kDa. The amino terminal amino acids sequenced from purified TWCA1 are 72 residues downstream of the putative starting methionine predicted by twca1. This difference may be due to the presence of a short-lived signal sequence designed to guide the enzyme to the correct cellular location. The absence of any homology between TWCA1 and previously sequenced CAs from Chlorophyceae may indicate either convergent evolution or that carbon acquisition represents a fundamental physiological difference among algal phyla.     

GROWTH RESPONSES OF THE DIATOM,CYCLOTELLA CRYPTICA (BACILLARIOPHYCEAE), TO GIBBERELLIC ACID1

The plant hormone, gibberellic acid (GA), stimulated growth of a marine diatom, Cyclotella cryptica Reimann, Lewin and Guillard. Four concentrations of GA (5 × 20 × 25 × and 35 × 10^?6 g/mL) were added to axenic cultures of C. cryptica. Changes in cell densities, measured by cell counts and turbidimetric readings, confirmed that GA at 20 × 10^?6 g/mL produced maximum stimulation. There was an increase in the total number of cells produced and a shorter lag phase of growth at this concentration. Coulter counter measurements of cell size, as well as ocular micrometer measurements, indicated there was no significant variation in cell volumes of GA grown cells over that of the controls.     

PHOTOPHYSIOLOGICAL RESPONSES OF FRAGILARIOPSIS CYLINDRUS (BACILLARIOPHYCEAE) TO NITROGEN DEPLETION AT TWO TEMPERATURES1

The photosynthetic efficiency and photoprotective capacity of the sea‐ice diatom, Fragilariopsis cylindrus (Grunow) W. Krieg., grown in a matrix of nitrogen repletion and depletion at two different temperatures (?1°C and +6°C) was investigated. Temperature showed no significant effect on photosynthetic efficiency or photoprotection in F. cylindrus. Cultures under nitrogen depletion showed enhanced photoprotective capacity with an increase in nonphotochemical quenching (NPQ) when compared with nitrogen‐replete cultures. This phenomenon was achieved at no apparent cost to the photosynthetic efficiency of PSII (F_V/F_M). Nitrogen depletion yielded a partially reduced electron transport chain in which maximum fluorescence (F_M) could only be obtained by adding 3‐(3,4‐dichlorophenyl)‐1,1‐dimethylurea (DCMU). reoxidation curves showed the presence of Q_B nonreducing PSII centers under nitrogen depletion. Fast induction curves (FICs) and electron transport rates (ETRs) revealed slowing of the electrons transferred from the primary (Q_A) to the secondary (Q_B) quinone electron acceptors of PSII. The data presented show that nitrogen depletion in F. cylindrus leads to the formation of Q_B nonreducing PSII centers within the photosystem. On a physiological level, the formation of Q_B nonreducing PSII centers in F. cylindrus provides the cell with protection against photoinhibition by facilitating the rapid induction of NPQ. This strategy provides an important ecological advantage, especially during the Antarctic spring, maintaining photosynthetic efficiency under high light and nutrient‐limiting conditions.     

CHRONIC EFFECTS OF ULTRAVIOLET-B RADIATION ON GROWTH AND CELL VOLUME OF PHAEODACTYLUM TRICORNUTUM (BACILLARIOPHYCEAE)1

Cultures of the temperate estuarine diatom, Phaeodactylum tricornutum Bohlin (NEPCC Clone 31), were grown under ambient intensities of ultraviolet-A radiation (UVAR), photosynthetically active radiation (PAR), and various intensities of ultraviolet-B radiation (UVBR; 290–320 nm). Growth rates and cell volumes were monitored for 36 d. UVBR decreased growth rates and increased cell volumes. Sensitivity of growth to UVBR increased with time. Growth rates of P. tricornutum decreased with increasing ratios of UVBR:UVAR + PAR.     

OBSERVATIONS ON CHAETOCEROS BUCEROS (BACILLARIOPHYCEAE), A RARE TROPICAL PLANKTONIC SPECIES COLLECTED FROM THE MEXICAN PACIFIC1

Examination of net plankton samples from the Gulf of Tehuantepec (Mexican Pacific) yielded a rare tropical species of the genus Chaetoceros, C. buceros Karsten. This species is conspicuous because of its relatively large size, type of chain formation, and shape of the terminal setae. Electron microscopic study revealed other interesting characteristics: the intercalary values are more lightly silicified than the terminal valves and show a pattern of costae radiating form the center with various annuli (“central hyaline fields”) in the center of the valve face; both kind of valves are perforated by small poroids. Terminal valves possess several (21–30) slitlike and randomly oriented rimoportulae. Important morphological differences exist between the intercalary and the terminal setae. Morphological characters, comparision with related species, and the systematic position are discussed. A new section of the subgenus Hyalochaete, Conspicua, is proposed to include C. buceros.