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.     

VALVE MORPHOGENESIS IN THE PENNATE DIATOM NAVICULA CUSPIDATA1

The deposition of siliceous valves during asexual reproduction of the pennate diatom, Navicula cuspidata Kütz., is described with emphasis on the cytoplasmic components involved. The events accompanying valve secretion are similar to those already known from other pennate species. After mitosis, the microtubule centre (MC) moves to the center of the cleavage furrow where silica deposition is initiated inside a tubular silicalemma, and it remains associated with the prospective central nodule during valve growth. Microtubules (MTs), emanating from the MC, run parallel to the prospective raphe and together with the raphe fibres, appear to be involved with raphe development. Multiple raphe fibres occupy the maturing raphe fissure, in contrast to the single fibre of Pinnularia viridis, P. maior and Hantzschia amphioxys. The fibers exhibit a periodic substructure and are often opposed to the silicalemma where they may inhibit silica deposition and control the shaping of the raphe fissure. In contrast with the above species, in N. cuspidata MTs are clustered strictly opposite the raphe and lose their association with the MC which degenerates before the valves are mature. The primary role of MTs may be the stabilization of the cytoplasmic region where initial silicification occurs. Mitochondria and endoplasmic reticulum are not involved in molding valve growth in this species. Evidence for vesicle involvement in silica transport and deposition was limited. The possible contributions provided by comparative studies on the ultrastructure of valve morphogenesis towards elucidating the control of valve formation and the taxonomy of diatoms are discussed briefly.     

VALVE AND BAND MORPHOLOGY OF SOME FRESHWATER DIATOMS. II. INTEGRATION OF VALVES AND BANDS IN NAVICULA CONFERVACEA VAR. CONFERVACEA12

Chemically cleaned and critical-point dried cells of a clonal culture were examined with scanning electron microscopy. Cells form filaments by valve-to-valve connections maintained by organic material which adheres to the central area of the valve face. Bending of filaments is probably restricted to some extent by the articulation of overlapping spatulate marginal spines with an adjacent underlapping set of much shorter spines (ridges), and with the mantle edge itself. Cell division results in three possible spine patterns for each cell: a set of overlapping and a set of underlapping spines; no overlapping sets of spines (two underlapping); or two sets of overlapping spines (no underlapping). Each filament inherits cells with spine set patterns in the ratio of 2 (with 1 set overlapping): 1 (with no sets overlapping): 1 (with 2 sets overlapping). Valvocopulae are shaped similarly to pleurae except that the partes exteriores of the valvocopulae are wider. The pars interior of both is delimited by an advalvar row of pores continuous around the cell apex. The pars exterior also has a row of pores, but it is median in the valvocopula and first pleura and does not continue around the cell apex. The valvocopulae always underlap the mantle and the pleurae always underlap their preceding band. The ends of both appeared attached, but may become free in acid-cleaned preparations. Bands alternate with each other so that the ends of the valvocopula attach to the first continuous apical portion of the first pleura; the ends of the first pleura attach in that same fashion to the second pleura but at the opposite apex; and all subsequent pleurae alternate in the same fashion with up to at least 13 pleurae/epicingulum. The continuous apical portion of each band is elevated so that a functional (but not structural) ligula is formed, with the continuous apical portion of alternate bands becoming adjacent and underlapping each other only in this region. The valvocopulae in a single cell, or of adjacent cells, may have their continuous apical ends on the same or on opposite apices. It is recommended that N. confervacea var. peregrina (W. Sm.) Grun. be merged with the nominate variety.     

GROWTH OF A MARINE DIATOM AND A HAPTOPHYCEAN ALGA ON PHENYLALANINE OR TYROSINE SERVING AS SOLE NITROGEN SOURCE1

The marine diatom Navicula incerta Hustedt and the haptophyte Isochrysis galbana Parke, both accustomed to grow on nitrate, were tested for their adaptive capacity for growth in axenic culture on 0.025–2.0 mM l -, d -, and dl -isomers of phenylalanine and tyrosine as sole nitrogen sources. The diatom showed immediate and excellent growth (superior to nitrate) on l -phenylalanine and l -tyrosine but required prolonged periods of adaptation before growing on the d -isomers, which growth was considerably slower. Isochrysis showed good growth on l -phenylalanine after short adaptation, fair growth on low but poor growth on high concentrations of l -tyrosine and no growth on the d -isomers of both amino acids. The racemate (dl -mixture) of each amino acid produced growth responses, from both algae, inferior to those of the corresponding l -isomer, indicating competitive inhibition from the d -isomer in the presence of equal amounts of its counterpart. Increasing tyrosine concentration was beneficial to growth of Navicula but markedly inhibitory or toxic to that of Isochrysis. Phenylalanine concentration appeared to benefit growth of both algae. Ecological considerations suggest that N. incerta may be habituated to utilize both aromatic amino acids in its native milieu.     

OBSERVATIONS ON PHOTOHETEROTROPHY IN A MARINE DIATOM1

An unidentified species of the diatom genus Cocconeis has been isolated from the sediments of Biscayne Bay, Florida. The organism is capable of utilizing a range of organic substrates, including lactate, in the light but not in the dark, as shown by growth studies. Information is included on changes in cell carbon, nitrogen, and chlorophyll a during growth in batch culture. Data obtained on the kinetics of uptake of lactate and glucose raise questions on the possible ecological significance of photoheterotrophy among marine microalgae, particularly those in estuaries.     

DIATOM MINERALIZATION OF SILICIC ACID IV. KINETICS OF SOLUBLE Si POOL FORMATION IN EXPONENTIALLY GROWING AND SYNCHRONIZED NAVICULA PELLICULOSA1

Silicic acid taken up from the growth medium by Navicula pelliculosa (Bréb.) Hilse was shown to enter at least two compartments: i) soluble pools; ii) insoluble fraction comprised predominantly of the silica frustule. Soluble Si pools were extracted by a variety of agents from cells uniformly labeled for ten generations in medium containing ⁶⁸Ge-Si(OH)₄. 100 C water soluble and 0 C perchloric acid (PCA) soluble Si pools of 680 mM Si·l^?1 and 490 mM Si·l^?1 cell water represented 13 and 9%, respectively, of total, cell Si in exponential growth phase cells. Uniformly labeled cells synchronized by the combined synchronization technique accumulate at the cell cycle stage where silica frustule development is initiated. These cells contain water and PCA soluble pools of 10 nmol Si·10⁶ cells^?1 and-8.8 nmol Si·10⁶ cells^?1, respectively. On addition of Si(OH)₄, a rapid uptake ensues allowing the Si pool to expand 2.5-fold, apparently to provide precursors of the silica frustule.     

LAUDERIA ANNULATA—A MARINE CENTRIC DIATOM WITH AN ELONGATE BILOBED NUCLEUS1

Lauderia annulata Cleve is probably unique among marine centric diatoms in possessing an elongate dumbbell-shaped nucleus. A lobe at, each end of the nucleus lies adjacent to each valve during interphase and each lobe resembles a typical eucaryotic nucleus. The central portion of the thin strand, which passes through the vacuole connecting the two lobes is Feulgen positive and is nuclear membrane bounded. A group of micro tubules occurs in this strand clustered excentrically between, the nuclear membrane and the tonoplast. Evidence for the coordinated functioning of both nuclear lobes is suggested, by the aggregation of chloroplasts around, both lobes when shade-adapted cells are exposed, to intense white or blue (400–500 nm) light.     

TAXONOMY AND FRUSTULAR STRUCTURE OF THE MARINE CENTRIC DIATOM PARALIA SULCATA1

Light and electron microscopy were used to investigate the complex structure of the frustule of Paralia sulcata (Ehrenb.) Cleve. Rimoportulae are reported for the first time in this diatom and two types of linking processes are described. The ease with which the cingulum is lost is explained with regard to its attachment to the valve. Two kinds of heterovalvy were observed and the taxonomic significance of one of these is discussed. The validity of Heiberg's genus Paralia is confirmed and a type slide of the species is designated.     

DIATOM MINERALIZATION OF SILICIC ACID. II. REGULATION OF Si (OH)4 TRANSPORT RATES DURING THE CELL CYCLE OF NAVICULA PELLICULOSA1

Synchronized populations of Navicula pelliculosa (Bréb.) Hilse show a 10-fold increase in Si(OH)₄ transport rate during traverse through the cell division cycle. The transport activity pattern is similar to a “peak enzyme.” Kinetic analysis showed there was a significant change in K_s values, indicating increased “affinity” for Si(OH)₄ as cells neared maximal uptake rates. However, the dramatic changes in transport rate at various cell cycle stages were also reflected by alterations in the V_max, values of the transport process, suggesting a change in the number of functional transport “sites” in the plasma membrane. Cells in the wall forming stage, arrested from further development by Si(OH)₄ deprivation, maintained high transport rates for as long as 7 h. The rates decreased rapidly if protein synthesis were blocked or if Si(OH)₄ was added, the latter allowing the cells to traverse the rest of the cycle. The half-life of the transport activity ranged from 1.0 to 2.2 h when protein synthesis was inhibited at various cell cycle stages and during the natural decline of activity late in the cycle. The transport system appears to be metabolically unstable as is typical for a “peak protein.” The rise in transport rate through the cell cycle did not depend on the presence of Si(OH)₄ in the medium; therefore, the transport system does not appear to be induced by its substrate. The rise in transport is also observed in L:D synchronized cells developing in the presence of Si(OH)₄; neither does the transport system appear to be derepressed. The transport rate was strongly cell cycle-stage dependent; the data appeared to fit the “dependent pathway” model proposed by Hart-well to explain oscillations in enzyme synthesis during the cell cycle.     

FRUSTULAR MORPHOLOGY AND TAXONOMIC AFFINITIES OF NAVICULA COMPLANATOIDES (BACILLARIOPHYCEAE)1

Live and prepared cells of the marine pennate diatom Navicula complanatoides Hust. were examined with light and electron microscopy. It has narrowly lanceolate valves (26–55 μm long, 4–5 μm wide) and girdles 10–24 μm in depth. Striae are parallel at the center of the valve (24–28 in 10 μm), becoming slightly convergent toward the apices. Electron microscopy revealed that the external valve surface presents a longitudinally ribbed appearance (20–28 parallel ribs at its maximum width), whereas internally, rectangular areolae are occluded by ricae. The raphe slit lies in a narrow axial area, and one side of the raphe sternum is deeper and folds over the other, obscuring the internal opening. Internally, the central virga on one side of the raphe and two virgae on the other are somewhat broader. A conspicuous pore (stigma) is present between the two broadened virgae. The girdle consists of valvocopulae, copulae, and pleurae. There are 16–20 bands per cingulum. The valvocopulae and copulae are hollow tube-like structures, with inner and outer portions contrsting in morphology. They decrease in diameter in an abvalvar direction. There are four pleurae. These are flat bands which facilitate overlap of the epicingulum and hypocingulum. Fundamental features of the valve and girdle reveal the distinctness of this species within Navicula. The areolae, external longitudinal ribs, and raphe structure suggest affinities with Pleurosigma, Gyrosigma, and Haslea. It is hypothesized that they share a derived state which indicates a recent common ancestor for these taxa. N. complanatoides and related species of the Naviculae microstigmatacae are distinctive enough to merit their own genus within the Naviculaceae.     

DIATOM MINERALIZATION OK SILICIC ACID. I Si(OH)4 TRANSPORT CHARACTERISTICS IN NAVICULA PELLICULOSA

Silicic acid transport was studied in the photosynthetic diatom Navicula pelliculosa (Bréb.) Hilse using [⁶⁸Ge] germanic acid (⁶⁸Ge(OH)₄) as a tracer of silicic acid (Si(OH)₄). The initial uptake rate of Si(OH)₄ was dependent on cell number, pH, temperature, light and was promoted by certain monovalent cations in the medium. Na⁺ was more effective than K⁺, whereas Li⁺ and NH⁺₄ were ineffective at promoting uptake. Uncouplers and inhibitors of oxidative phosphorylation and of photophosphorylation reduced uptake by 40–99% of control values. Uptake was also especially sensitive to the sulfhydryl blocking agents at 10^?5 M and to the ionophorous compound valinomycin (10^?7 M) which inhibited uptake by 82%. The Si(OH)₄ transport system displayed Michaelis-Menten-type saturation kinetics with kinetic parameters of K_S= 4.4 p. mol Si(OH)₄· 1^?1, V_max= 334 pmol Si(OH)₄· 10⁶ cells^?1· min^?1. Calculations of the acid soluble silicic acid pool size based on 60 s uptake at 20 μM Si(OH)₄ suggested that intracellular levels of Si could reach 20 mM and as much as 5 mM could exist as free silicic acid, representing maintenance of a 250-fold concentration gradient compared with the medium. Efflux from preloaded cells was dependent on temperature and the Si(OH)₄ concentration of the external medium. In the presence of 100 μMM “cold” Si(OH)₄, approximately 30% of the Si(OH)₄ in preloaded cells was exchanged in 20 min. The initial uptake rate of Si(OH)₄ in logarithmic phase cells was constant, but the uptake rate increased in a linear fashion for 6 h in stationary phase cells. These results suggest that the first step in silica mineralization by diatoms is the active transmembrane transport of Si(OH)₄ by an energy dependent, saturable, membrane-carrier mechanism which requires the monovalent cations Na⁺ and K⁺ and is sensitive to sulfhydryl blocking agents. Silicic acid transport activity also appears to be regulated during different growth stages of the diatom.     

BENTHIC MARINE DIATOM ASSOCIATIONS: UPPER FLORIDA BAY (FLORIDA) AND ASSOCIATED SOUNDS1

Models of the diatom associations found in upper Florida Bay and adjoining sounds have been constructed utilizing Q modal factor-vector analysis and ecologic diversity indices (Shannon index, number of species, evenness). Four distinct associations were defined using Q-mode factor-vector analysis. Two associations were epiphytic, occurring on Thalassia testudinum König—Association I was characterized by Cocconeis placentula Ehr., Association III by Cylindrotheca closterium (Ehr.) Reim. & Lewin and Cocconeis placentula Ehr. The other two associations were epipelic, occurring on the carbonate mud substratum—Association II was characterized by Cyclotella striata (Kütz.) Grun., Rhopalodia gibberula (Ehr.) O. Müller and Surirella fastuosa (Ehr.) Kütz., Association IV by Fragilaria crotonensis Kitton and Cyclotella striata (Kütz.) Grun. The majority of the 161 species identified were present in both the epiphytic and epipelic assemblages. Only 33 species were restricted to the epiphyton, and 18 species restricted to the epipelon. The epipelic assemblage was significantly more diverse than was the epiphytic assemblage. A general trend of increased diversity away from terrestrial environs toward more open areas of water in both the epipelon and epiphyton was also found.     

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.     

OSMOREGULATION IN THE MARINE DIATOM CYLINDROTHECA FUSIFORMIS1

A mechanism for the regulation of internal osmolarity in the diatoms (Bacillariophyta) was investigated using the intertidal lestuarian pennate diatom, Cylindrotheca fu-siformis Reimann & Lewin. The intracellular pools of the free sugar, mannose, and its respective polymer, poly-mannose, were found to be in a dynamic equilibrium which rapidly responds to changes in external osmotic pressure. Increasing the salinity shifts the steady state in the direction of free mannose, while decreasing the salinity stimulates polymerization of the free mannose to its polysaccharide. The mechanism requires a response of certain key enzymes to osmotic pressure rather than to levels of any particular ion since increasing the external osmolarity with an organic solute (sorbitol) was as effective in eliciting a response as using inorganic ions.     

DIEL OSCILLATIONS IN THE PHOTOSYNTHESIS-IRRADIANCE RELATIONSHIP OF A PLANKTONIC MARINE DIATOM1

The amplitude of diel oscillations in photosynthesis as a function of irradiance varied with the growth phase in a marine phytoplankton species. The common centric diatom (Bacillariophyta), Ditylum brightwellii (West) Grun., showed strong periodicity in the photosynthesis-irradiance (P-I) relationship, which damped progressively from early to late exponential and stationary phase. These findings suggest that short-term temporal characteristics of phytoplankton production depend on factors which affect growth, and that the amplitude is most enhanced at maximal growth rates likely to be encountered in the natural environment.     

STUDIES ON MARINE PLANKTON DIATOMS. II. RESTING SPORE MORPHOLOGY1

A morphological study of resting spores in five marine planktonic diatom species using electron microscopy indicates that Bacteriastrum delicatulum Cleve and Leptocylindrus danicus Cleve spores bear little resemblance, to their vegetative cells. Detonula confervacea (Cleve) Gran and Stephanopyxis turris (Grev. & Arn.) Ralfs spores have several features in common with their vegetative cells, and Rhizosolenia setigera Brightwell lies between the two extremes. The function of resting spores in relation to diatom life cycles is briefly discussed. Spore formation may be a primitive characteristic in the life cycle and may no longer have significant survival value for the species.     

EFFECT OF BLUE-GREEN LIGHT ON PHOTOSYNTHETIC PIGMENTS AND CHLOROPLAST STRUCTURE IN THE MARINE DIATOM STEPHANOPYXIS TURRIS1

Blue-green light increased the chlorophyll concentration and chloroplast number of cells of Stephanopyxis turris (Grev.) Ralfs, compared to white light controls. Light fields for growth were 400 μW·cm^?2 (12:12 LD cycles). Chlorophyll increased up to 100%/cell, but no change in the ratio of chlorophylls to major carotenoids occurred. The effect was, therefore, not that of complementary chromatic adaptation. At the same time, blue-green light enhanced the photosynthetic fixation of CO₂. At the ultrastructure level, an increase in, and rearrangement of, the thylakoid system occurred.     

VALVE AND BAND MORPHOLOGY OF SOME FRESHWATER DIATOMS. IV. OUTER SURFACE MUCILAGE OF NAVICULA CONFERVACEA VAR. CONFERVACEA1

The development of the mucilage on the outer surface of Navicula confervacea (Kütz.) Grun., a raphed, filamentous diatom, was studied with scanning electron microscopy. This nonstructural cell wall material, present on the surface after critical-point drying and absent after acid cleaning, was of two types: strands and papillae. Strands were associated with the raphe system, areolae, elongated pores of the mantle, and all girdle sutures. Organic papillae were a common feature of valves, valvo-copulae and pleurae, but their origin and distribution could not be explained since they often occurred between the obvious openings in the frustule. Strands from the raphe and areolae may function in attaching terminal cells to a substrate and adjacent cells to each other. Other strands of the girdle arise from sutures during cell enlargement and continue to lengthen and intertwine until the individual frustules within a filament are obscured. Strands from sutures might originate from the advalvar row of pores of the girdle bands since these pores lie along the suture, but direct observation of this was not made. Secretion between, the bands also cannot be ruled out. Although mucilaginous papillae may sometimes occur at random on the entire surface of frustules, there is also a distinct, narrow multiseriate row of them around the edge of valves without marginal spines.     

A NEW SPECIES OF THE DIATOM GENUS EUNOTIA1

A new species of the diatom genus Eunotia is described. The species, E. catillifera Morrow, is distinguished by the presence of fin-like plates, or transapical ridges, which alternate with the striae on the valve faces. This alga appears to be intermediate between the more typical Eunotia species and Semiorbis Patr. This observation suggests that Semiorbis hemicyclus (Ehr.) Patr. in Patrick and Reimer be returned to Eunotia hemicyclus Ralfs in Pritchard and that the use of the generic name Semiorbis be discontinued.