ICE‐BINDING PROTEINS FROM SEA ICE DIATOMS (BACILLARIOPHYCEAE)1

Sea ice diatoms thrive under conditions of low temperature and high salinity, and as a result are responsible for a significant fraction of polar photosynthesis. Their success may be owing in part to secretion of macromolecules that have previously been shown to interfere with the growth of ice and to have the ability to act as cryoprotectants. Here we show that one of these molecules, produced by the sea ice diatom Navicula glaciei Vanheurk, is a ～25 kDa ice‐binding protein (IBP). A cDNA obtained from another sea ice diatom, Fragilariopsis cylindrus Grunow, was found to encode a protein that closely matched the partially sequenced N. glaciei IBP, and enabled the amplification and sequencing of an N. glaciei IBP cDNA. Similar proteins are not present in the genome of the mesophilic diatom Thalassiosira pseudonana. Both proteins closely resemble antifreeze proteins from psychrophilic snow molds, and as a group represent a new class of IBPs that is distinct from other IBPs found in fish, insects and plants, and bacteria. The diatom IBPs also have striking similarities to three prokaryotic hypothetical proteins. Relatives of both snow molds and two of the prokaryotes have been found in sea ice, raising the possibility of a fungal or bacterial origin of diatom IBPs.     

PHYSIOLOGY OF SEA ICE DIATOMS. I. RESPONSE OF THREE POLAR DIATOMS TO A SIMULATED SUMMER-WINTER TRANSITION1

Three axenic polar sea ice diatom cultures were subjected to a 30 day simulated summer-winter transition in which light and temperature were decreased and salinity was increased to mimic seasonal changes previously reported for ice-covered polar seas. The diatoms responded to these changes by a reduction in cellular metabolism as indicated by: 1) A decline in growth rate and photosynthetic rate; 2) a decrease in cellular ATP; and 3) the storage and subsequent utilization of endogenous carbon reserves. In addition, heterotrophic potential of the three clones increased by as much as 60-fold. In some cases, the decrease in light intensity characteristic of the onset of polar winter was alone sufficient to trigger these physiological changes.     

A MORPHOLOGICAL AND TAXONOMIC CONSIDERATION OF TABELLARIA (BACILLARIOPHYCEAE) FROM THE NORTHCENTRAL UNITED STATES1,2

The taxonomy, distribution, and ecology of the diatom genus Tabellaria Ehr. from a portion of the northcentral United States were studied from collections made from August 1968 to October 1971. Using the transmission electron microscope with carbon replicas of the frustule surfaces, taxonomically significant characters were identified and related to those visible with the light microscope, four morphologically distinguishable taxa were recognized. The frustular morphology of T. fenestrata (Lyngbye) kütz. em. Knudson var. fenestrata was confirmed by both light and electron microscopy. T. flocculosa (Roth) Kütz. var. flocculosa is composed of 3 morphologically distinguishable strains. One of which is sufficiently different from the nominate variety to be described as T. flocculosa var. linearis var. nov. Also, a planktonic ecotype of one of the strains was identified.     

BRACKISH WATER AND FRESHWATER SPECIES OF THE DIATOM GENUS SKELETONEMA. II. SKELETONEMA POTAMOS COMB. NOV.1

Electron microscope investigations of the siliceous frustule show that the diatom described by Hustedt as Stephanodiscus subsalsus (A. Cleve) Hust. is not Skeletonema subsalsum (A. Cleve) Bethge (Melosira subsalsa A. Cleve) but is Microsiphona potamos Weber. This species is so similar to Skeletonema costatum (Grev.) Cleve and Skeletonema subsalsum that the combination Skeletonema potamos (Weber) Hasle is suggested. Present records classify Skeletonema potamos as a freshwater species of lakes and rivers. In Sandusky Bay, Lake Erie (U.S.A.) and in River Wümme, a tributary of the River Weser (Germany) it grows with Skeletonema subsalsum. In nature, and when grown in cultures at a salinity of 0%, the processes are extremely short; when grown at salinities of 2% or more, the processes are much longer.     

MORPHOLOGY OF THE MARINE DIATOM NITZSCHIA NAVIS-VARINGICA, SP. NOV. (BACILLARIOPHYCEAE), ANOTHER PRODUCER OF THE NEUROTOXIN DOMOIC ACID

A new marine diatom, Nitzschia navis-varingica , sp. nov., isolated from Vietnamese waters, is described by light, transmission, and scanning electron microscopy, including thin sectioning. The new species has been found to produce the neurotoxin domoic acid (DA), better known from several species of Pseudo-nitzschia Peragallo and one species of Amphora Ehrenberg. Production of DA is therefore more widespread among diatoms than previously thought. Taxonomically, the genus Nitzschia Hassall is exceptionally difficult, with about 900 described taxa. Grunow (in Cleve and Grunow 1880 divided the genus into 24 sections, and this system is still used with modifications. Nitzschia navis-varingica , sp. nov. fits best into a group of sections that includes Dubiae, Bilobatae , most of the Lanceolatae , and Lineares , all sensu Grunow, as the cell is slightly indented in the middle in girdle view and has a moderately eccentric raphe and a weak longitudinal fold on the valve. Many species within these sections have features similar to N. navis-varingica , but no species seems to be identical. Because both Pseudo-nitzschia and Nitzschia belong to the family Bacillariaceae, it seems reasonable to look for further producers of DA in this family, including freshwater species, which mainly comprise species within the sections Dubiae, Bilobatae, Lanceolatae , and Lineares.     

LIGHT AND NUTRIENT LIMITATION OF SEA-ICE MICROALGAE (HUDSON BAY,CANADIAN ARCTIC)1

Changes in the biochemical composition of sea-ice microalgae (southeastern Hudson Bay, Canadian Arctic) were used to assess the light and nutrient status of cells growing at the ire-water interface. These changes allowed us to test the hypothesis that ire algae are limited by light at the beginning of their growth season and become periodically limited by nutrients as the season progresses. During the vernal growth season, three patterns of variation in cellular components were found in response to changes in environmental conditions. 1) Chlorophylls a and c, ATP, carbohydrate, and carbon followed the seasonal increase in under-ice irradiance, which was mainly mused by melting of the snow cover. 2) Dissolved and biogenic silicon underwent periodic variations, which were coupled to the fortnightly neap-spring cycle of tidal mixing. 3) Cellular contents of free amino acids, protein, and total nitrogen remained relatively constant during the season. An early decrease in intracellular chlorophylls a and c suggests that ire algae did respond to small changes in solar irradiance by changing the pigment composition of their photosynthetic units. Seasonal increases in ATP, carbohydrate, and total carbon indicate light limitation in April, followed in May by a period of excess irradiance and/or nutrients in short supply. The seasonal increase in ATP and the high values of the ratio free amino acids: protein show that neither phosphorus nor nitrogen limited algal growth at the ire-water interface. In May, higher values of carbohydrate: protein, carbon: nitrogen, carbon: chlorophyll a, and also carbon: silicon and ATP: silicon indicate that the ice algae became silicon-deficient in their natural environment. Following a period of light limitation, at the beginning of the season, ice-algal growth became silicon-limited, when in situ irradiance and the accumulated algal biomass were high and the tidally-driven nutrient supply was not strong enough to satisfy algal nutrient requirements.     

THALASSIOSIRA ANDAMANICA SP. NOV. (BACILLARIOPHYCEAE), A NEW DIATOM FROM THE ANDAMAN SEA (THAILAND)

A new marine diatom, Thalassiosira andamanica, is described from light and electron microscopy. The specimens were collected in the vicinity of Phuket Marine Biological Center, Thailand, and later brought into clonal culture. Thalassiosira andamanica possess a rimoportula with a pronounced outer extension, one marginal ring of fultoportulae, and three rings of fultoportulae on the valve face. Cells are united into colonies by a single thread secreted through a central fultoportula. Marginal fultoportulae extensions are shortest on the inside of the valve. The areolae are arranged in sectors, and the valve margin is ribbed with approximately 38 ribs in 10 μm. The valvocopula and copula have rows of pores, four to six pores in 1 μm. Apparently, the pleurae are hyaline. Experiments with a clonal culture isolated at Phuket, Thailand, showed that growth (cell divisions·24 h⁻¹) was reduced for cultures grown at 14° C compared to those grown at 19°, 24°, or 30° C. The maximum growth rate (2.2 divisions·24 h⁻¹) was at 30° C. Thalassiosira andamanica is compared with morphologically similar taxa. On the basis of morphological features and the response to different temperature regimens, it is concluded that this taxon must be recognized as a new warm-water species. In addition, T. andamanica does not clearly belong to any of the two subgroups of species of Thalassiosira. To accommodate the morphological characteristics of T. andamanica, the establishment of a possible third subgroup is discussed.     

SEA ICE MICROBIAL COMMUNITIES. V. THE VERTICAL ZONATION OF DIATOMS IN AN ANTARCTIC FAST ICE COMMUNITY1

A distinct vertical zonation was observed among diatoms in a bottom congelation ice community at McMurdo Sound, Antarctica during the 1981 spring bloom. The bottom 20 cm of ice collected in December from four stations with variable snow cover was subdivided into 5 cm sections for analysis of algal distribution. Algal abundance was inversely related to the depth of snow cover, and generally decreased with increasing distance above the ice-water interface. Most diatoms, including the dominant species Nitzschia stellata Manguin, Amphiprora kufferathii Manguin and Fragilaria islandica var. adeliae Manguin showed peak abundance in the bottom 10 cm of the ice, where the proportion of living to empty cells was also highest. Two species, however, an Auricula Castracane sp. and Navicula glaciei van Heurck, reached highest concentrations at depths 10–20 cm above the ice-water interface. We considered two factors as contributing to the observed vertical zonation: (1) successive blooms at the ice-water interface become spatially stratified within the ice by further accretion below; (2) a differential growth of species occurs along physicochemical gradients within the ice column. A comparison of early versus late season profiles suggests the latter mechanism may prevail once ice accretion has ceased.     

PRESCOTTIELLA,A NEW GENUS OF ASYMMETRICAL DESMIDS (CHLOROPHYCEAE)1

Bipolar asymmetry has been considered a morphological characteristic sufficient for differentiation of genera among the Desmidiaceae. Therefore, Micrasterias sudanensis Grönbl., Prowse & Scott, the only species of Micrasterias showing such asymmetry, is made the type of a new genus, Prescottiella, gen. nov.     

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.     

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.     

EFFECT OF HYPEROXIA ON THE GROWTH AND PHOTOSYNTHESIS OF POLAR SEA ICE MICROALGAE1

Sea ice algal communities are naturally exposed to very high concentrations of dissolved oxygen, which are likely to lead to increasing stress levels and declines in productivity. To test this hypothesis, cultures of Fragilariopsis cylindrus (Grun?) Hasle, Pseudo‐nitzschia sp., Fragilariopsis curta (Van Heurch), Porosira glacialis (Grunow), and Entomoneis kjellmannii (Cleve) from Antarctic sea ice and Nitzschia frigida from Arctic sea ice were exposed to elevated dissolved oxygen levels, and their growth, maximum quantum yield, relative maximum electron transport rate, and photosynthetic efficiency were measured. At oxygen concentrations equivalent to approximately four times air saturation (89% oxygen), the growth rate and maximum quantum yield were significantly reduced in all taxa. When the oxygen concentration was regularly allowed to drop, the effect on growth and quantum yield was reduced. At lower dissolved oxygen concentrations (52%), the declines in growth and quantum yield were reduced but were still mostly significantly different from the controls (21% oxygen). It is likely that the generation of excess active oxygen radicals in the presence of free oxygen is responsible for most of the decline in growth, maximum quantum yield, relative maximum electron transport rate, and photosynthetic efficiency in all species.     

ASTEROMENIA (RHODYMENIACEAE,RHODYMENIALES), A NEW RED ALGAL GENUS BASED ON FAUCHEA PELTATA1

Asteromenia gen. nov. (Rhodymeniales, Rhodophyta) is proposed with a single species, Asteromenia peltata (W. R. Taylor) comb. nov. (basionym: Fauchea peltata W. R. Taylor). Thalli of the proposed new genus are stipitate with dorsiventral, peltate blades that are initially circular in shape but with age become stellate with ligulate arms. Internally, the blades have a polystromatic medulla of large, hyaline cells, grading into a cortex of smaller, pigmented cells. Clusters of translucent cells occur on the dorsal surface of the blade. Tetrasporangia are formed by transformations of intercalary midcortical cells. Mature tetrasporangia have cruciately arranged spores and are densely aggregated in the cortex, mostly on the ventral surface, but occasional tetrasporangia also arise on the dorsal surface. Carpogonial branches are four-celled and arise on inner cortical cells. Auxiliary cells are borne on auxiliary mother cells attached to supporting cells of the carpogonial branches. Cystocarps are protuberant, with well-developed, ostiolate pericarps that often have extended, proboscis-like necks. The new genus differs from the previously described peltate or dorsiventral taxa in the Rhodymeniaceae by its polystromatic medulla (Maripelta and Sciadophycus have a monostromatic medulla), intercalary tetrasporangia formed in an unmodified cortex, and four-celled carpogonial branches (Halichrysis, as typified by H. depressa (J. Agardh) F. Schmitz, has terminal tetrasporangia in nemathecia and three-celled carpogonial branches).     

SEXUAL REPRODUCTION,MATING SYSTEM,AND PROTOPLAST DYNAMICS OF SEMINAVIS (BACILLARIOPHYCEAE)1

Cell division, the mating system, and auxosporulation were studied in the marine epipelic diatom Seminavis cf. robusta Danielidis & D. G. Mann. The interphase protoplast contains two girdle‐appressed chloroplasts, each with an elongate bar‐like pyrenoid, and also a central nucleus, located in a bridge between two vacuoles. Before cell division, the chloroplasts divide transversely and translocate onto the valves. The nucleus relocates to the ventral side for mitosis. After cytokinesis and valve formation, the chloroplasts move back to the girdle, showing a constant clockwise movement relative to the epitheca of the daughter cell. Seminavis cf. robusta is dioecious, and sexual reproduction is possible once cells are less than 50 μm. In crosses of compatible clones, gametangia pair laterally, without the formation of a copulation envelope, and produce two gametes apiece. The intensity of sexualization increases as cells reduce further in size below the 50‐μm threshold. At plasmogamy, the gametangia dehisce fully and the gametes, which were morphologically and behaviorally isogamous, fuse in the space between the gametangial thecae. The auxospore forms a transverse and longitudinal perizonium. After expansion is complete, there is an unequal contraction of the protoplast within the perizonium, creating the asymmetrical shape of the vegetative cell. Apart from this last feature, almost all characteristics exhibited by the live cell and auxospores of Seminavis agree with what is found in Navicula sensu stricto, supporting the classification of both in the Naviculaceae. Haploid parthenogenesis and polyploid auxospores were found, lending support to the view that change in ploidy may be a significant mechanism in diatom evolution.     

SYNARTHROPHYTON,A NEW GENUS OF CORALLINACEAE (CRYPTONEMIALES,RHODOPHYTA) FROM THE SOUTHERN HEMISPHERE1

Synarthrophyton gen. nov. is described based on southern Australian material of Melobesia patena Hooker fils & Harvey, which shows features intermediate between Mesophyllum Lemoine and Lithothamnium Philippi. It has the thin-walled rounded epithallial cells and coaxial hypothallium characteristic of Mesophyllum and the type of procarp and male structures common to Lithothamnium. Synarthrophyton is unique within the subfamily Melobesioideae (J. E. Areschoug) Yendo because it displays secondary pit connections. Details of the vegetative and reproductive morphology of S. patena (Hooker fils. & Harvey) comb. nov. are given and its taxonomic position in the Corallinaceae Lamouroux discussed.     

A PRELIMINARY INVESTIGATION OF THE PHYLOGENETIC RELATIONSHIPS AMONG THE FRESHWATER,APICAL PORE FIELD-BEARING CYMBELLOID AND GOMPHONEMOID DIATOMS (BACILLARIOPHYCEAE)1

Relationships among the apical pore field-bearing diatom genera Brebissonia, Cymbella, Gomphonema, Gomphoneis, Didymosphenia, Gomphocymbella, and Reimeria and their close ally Encyonema were evaluated using cladistics. These biraphid genera are a polythetic group defined by chloroplast number and type, mode of sexual reproduction, valve orientation and presence of apical pore fields. Character states of valve as well as non-siliceous features were described and then polarized using the outgroup method, with the naviculoid genera Anomoeoneis and Placoneis serving as outgroups. The cladistic analysis suggests two groups of terminal taxa, corresponding to to a predominantly cymbelloid lineage and a gomphonemoid lineage. Didymosphenia, previously thought to be closely allied to gomphonemoid diatoms, is shown to be more closely related to Cymbella. Gomphonema appears to be without distinguishing characteristics. Relationships of the other genera are described. The systematic position of small, doubly-punctate Gomphonema species is discussed.     

LOCATION AND EXPRESSION OF FRUSTULINS IN THE PENNATE DIATOMS CYLINDROTHECA FUSIFORMIS, NAVICULA PELLICULOSA, AND NAVICULA SALINARUM (BACILLARIOPHYCEAE)

A detailed immunocytochemical and biochemical study of the location and expression of frustulins, a family of proteins associated with the frustules of diatoms, has been performed for Cylindrotheca fusiformis Reimann et Lewin, Navicula pelliculosa (Brébisson et Kützing) Hilse, and Navicula salinarum (Grunow) Husted. Immunocytochemistry revealed that frustulins, which share homologous epitopes but are different in size, were predominantly located in the organic casing. Based on timed immunolocalization experiments and Western blotting analysis of cell extracts obtained sequentially after repleting silicate to Si-synchronized cells, the continuous presence of the frustulins in the mature and parental organic casing of the examined species was observed. The frustulins of N. pelliculosa appeared as proteins similar to those of C. fusiformis, sharing identical epitopes. The extractions, however, yielded a markedly lower abundance of frustulins in N. pelliculosa. Peak concentrations of extracted frustulins appeared to be expressed just ahead of the silicification process in C. fusiformis, whereas the level of expression in N. pelliculosa increased along with maturation of the new valves. For N. salinarum, the presence of the frustulins could not be confirmed properly by Western blotting, most probably because of the small sample volumes, inefficient extraction, and a lower amount of homologous frustulins in the casing of this species. It is concluded that the frustulins of these species are not associated with the silicalemma of the newly formed silica deposition vesicles and therefore do not seem to be involved in the silicification process itself. Overall, the results imply a structural role of the frustulins in the casing of diatoms rather than a regulation of the silicification process.     

ULTRASTRUCTURE OF VEGETATIVE ORGANIZATION AND CELL DIVISION IN THE UNICELLULAR RED ALGA DIXONIELLA GRISEA GEN. NOV. (RHODOPHYTA) AND A CONSIDERATION OF THE GENUS RHODELLA1

This study suggests that the genus Rhodella be restricted to that set of features currently observed only in Rhodella maculata Evans and Rhodella violacea (Korn-mann) Wehrmeyer, that a new genus Dixoniella be established to accommodate the unicellular red alga, Rhodella grisea (Geitler) Fresnel, Billard, Hindák et Pekár-ková, and that Rhodella cyanea Billard et Fresnel be further studied for probable reclassification. These conclusions are based on ultrastructural comparisons of Dixoniella grisea with published information on the genus Rhodella. The presence of thylakoids in the pyrenoid, a peripheral encircling thylakoid in the chloroplast, a dictyosome/nuclear envelope association, and the lack of a specialized pyrenoid/nucleus association in D. grisea separate this alga from the genus Rhodella. Cell division in D. grisea is not demonstrably different from that in Rhodella, although the unusually well-defined material of the presumptive microtubule organizing center (MTOC) made it possible to follow the development and behavior of the MTOC to a greater degree than in previously studied red algal cells. The surprising amount of conformity in cell division characters between D. grisea and the genus Rhodella prompted a comparison of cell division characteristics in all red algal unicells studied to date. All unicells show a remarkable degree of similarity except for differences in interzonal spindle length, dissimilarities in size of the nucleus-associated organelle (NAO), and the unusual NAO of Porphyridium purpureum (Bory) Drew et Ross.