HETEROTHALLIC SEXUALITY AND DENSITY DEPENDENT ENCYSTMENT IN THE CHRYSOPHYCEAN ALGA SYNURA PETERSENII KORSH.1

Sexual reproduction of the common planktonic chrysophyte Synura petersenii is described from observations made on clonal isolates grown in defined culture. Sexual fusion was isogamous and heterothallic, with cells of normal appearance from compatible clones serving as hologametes. No special culture conditions were required to induce sexual behavior; actively growing cell populations appeared to be continually receptive to mating when mixed with a sufficient number of cells from a compatible clone. A single, bipolar mating group was documented containing five of the seven clones tested. Zygotic statospores were found to be binucleate and to contain 4 chloroplasts at maturity. Production rates of zygospores were low for even the most highly compatible clones, with batch culture yields ranging from 1-20% of final cell density under the culture conditions utilized. Six of the clones tested were also capable of very low frequency (0.001-0.01%) homothallic statospore production but the reproductive significance of these cysts remains enigmatic. The dynamics of sexual encystment suggest that the process proceeds during periods of active population growth and is density dependent. Based on the characteristics of cyst induction and encystment dynamics, it is concluded that chrysophycean flagellates may have a perennation strategy quite different from that of the majority of planktonic diatoms, dinoflagellates, and green algae for which resting cyst production requires an exogenous trigger usually associated with physiological stress and periods of negative growth.     

VIRUS-LIKE PARTICLES IN HYDRURUS (CHRYSOPHYCEAE)1,2

Virus-like particles (VLPs) have been found in cells of the colonial, chrysophycean alga Hydrurus foetidus (Vill.) Trev. The infected cells, probably zoospores, were discovered in material fixed directly in the field. The VLPs appear either hexagonal or pentagonal when sectioned and are therefore presumed to be icosahedra, They are 50–60 nm diam. and demonstrate distinct trilaminar coals. The VLPs were observed in the cytoplasm, but not in the chloroplast or nucleus.     

A STATISTICAL CHARACTERIZATION OF GROWTH AMONG CLONES OF SYNURA PETERSENII (SYNUROPHYCEAE)1

Each of four clones from the Synura petersenii complex was grown at different pHs (5.5, 6.5, 7.5, 8.5) in batch culture experiments. Growth response curves and exponential growth rates were compared among clones and pH treatments in order to examine growth trend variation among the clonal groups. The clones were isolated from geographically distant North American localities. The clonal groups represented distinct mating types, an isolate and its subisolate, and S. petersenii- and S. glabra-like scale morphologies. No consistent relationship existed between growth response curve, and culture medium pH. Additionally, the trends across time differed according to clone and pH combination. Pairwise comparisons of linear trends from transformed growth response curves indicated two distinct clonal associations. Although the clonal associations corresponded with the final cell density of the cultures, growth response curves did not correspond with mating type, the parent-isolate and subisolate, or scale morphology. Clones with glabra-like scales had greater growth rates than the clone with petersenii-like scales. The conflicting results generated from growth response curve and growth rate analyses support the concept that S. petersenii and S. glabra form a highly variable, homogeneous grouping.     

PARTIAL CHARACTERIZATION OF THE CHLOROPLAST GENOME FROM THE CHROMOPHYTIC ALGA SYNURA PETERSENII (SYNUROPHYCEAE)1

Hoechst dye 33258-CsCl density gradients were used to isolate two satellite DNA species from Synura petersenii Korsh. sensu lato, a member of the Synurophyceae. One satellite DNA was identified as the chloroplast genome. The chloroplast genome is the smallest (91.5 kb) published for any chromophyte and approximates the size of the smallest functional chlorophyte chloroplast genome (Codium fragile, 89 kb). The second satellite DNA was small (34.5 kb), and its origin is undetermined. The potential of using the S. petersenii chloroplast genome in comparative studies for evaluating organellar evolution and algal systematics is discussed.     

SILICEOUS SCALE PRODUCTION IN CHRYSOPHYTE AND SYNUROPHYTE ALGAE. I. EFFECTS OF SILICA-LIMITED GROWTH ON CELL SILICA CONTENT,SCALE MORPHOLOGY,AND THE CONSTRUCTION OF THE SCALE LAYER OF SYNURA PETERSENII1

The cells of synurophyte flagellates (algal class Synurophyceae, formerly included in the Chrysophyceae) are enclosed within a regularly imbricate layer of ornamented siliceous scales. Scale morphology is of critical taxonomic importance within this group of algae, and the scales are valuable indicator microfossils in paleolimnological studies. The data presented here demonstrate that scale morphology and the integrity of the scale layer can exhibit extreme variability in culture as a function of the cellular quota of silica under silica-limited growth. Silica-limited, steady-state populations of the colonial flagellate Synura petersenii Korsh. were maintained over a range of specific growth rates (μ= 0.11–0.69 days^?1) and silica cell quotas (Q_si= 0.13–2.40 pmoles Si · cell¹). Scale morphology and the organization of the scale layer became increasingly aberrant as silica stress increased. Under severe stress, scale deposition was completely suppressed so that cells appeared scale-free. This depression of scale deposition was reversible; populations of silica-starved, scale-free cells rapidly regenerated new scale layers when placed in batch culture and spiked with dissolved silica. During recovery from silica stress, cell division was repressed for 24 h while mean cell silica quota increased 25-fold. The first new scales appeared within 2 h after the silica addition, and development of the new scale layer proceeded in an approximately synchronous manner, residting in normal scale layers on virtually all cells after 48 h of recovery in Sirich medium. Silica content of silica-replete Synura cells is comparable to freshwater diatoms of siynilar size, but Synura has much greater potential quota variability than diatoms and no apparent threshold silica requirement. Silica-limited growth kinetics and competition between diatoms and Synura for silica are discussed. The results suggest that morphological variability of siliceous scales in natural populations of synurophyte flagellates may result from silica stress and that the experimental approach developed here has great potential value as a means for circumscribing ecotypic variation in scale morphology. Results also demonstrate that scale production can be uncoupled from cell division, suggesting that cell cycle regulation of silica biomineralization in the Synurophyceae may be fundamentally different from that of diatoms (algal class Bacillariophyceae). This experimental system has application in the future study of the intracellular membrane systems and the regulatory processes involved in silica biomineralization.     

TWO NEW SPECIES OF PSEUDOKEPHYRION (CHRYSOPHYCEAE) IN THE PLANKTON OF A LAKE1

Two new species of Pseudokephyrion are described from Little Aurora Lake, Ontario, Canada. The distribution of both P. taeniatum sp. nov. and P. auroreum sp. nov. is restricted, for reasons which are not clear; neither species was found in samples from lakes adjacent to Little Aurora Lake. All lakes in the vicinity are very dilute, oligotrophic and acidic.     

CHRYSOAMPHIPYXIS GEN. NOVA A NEW GENUS IN THE STYLOCOCCACEAE (CHRYSOPHYCEAE) 1

Chrysoamphipyxis Canadensis gen. Et sp. nov. a new loricate, shizopodial chrysophyte was discovered growing as an epiphyte on filamentous algae in an Ontario pond, Lorica structure includes some features of both Chrysoamphitrema and Chrysopyxis and it can be inferred that Chrysoamphipyxis occupics the same phylogenetic position in the stylococcaceae relative to Chrysoamphitrema and Porostylon as Chrysopyxis does to Lagynion and Stylococcus.     

NOVEL PROTEINS COMPRISING THE STRAMENOPILE TRIPARTITE MASTIGONEME IN OCHROMONAS DANICA (CHRYSOPHYCEAE)1

Two‐dimensional (2‐D) protein analysis of the mastigoneme fraction of the chromophyte alga Ochromonas danica E. G. Pringsh. showed the presence of several component proteins of the tubular mastigoneme. Adding to the reported gene Ocm1, three new genes (Ocm2, Ocm3, and Ocm4) belonging to the Ocm gene family were isolated using degenerate primers designed from predicted Ocm1 amino acid sequences. The predicted polypeptides encoded by Ocm2, Ocm3, and Ocm4 were smaller in size than Ocm1. However, they shared four highly conserved, cysteine‐rich, epithelial growth factor (EGF)‐like motifs, potentially involved in protein–protein interaction. In addition, Ocm2, Ocm3, and Ocm4 showed homology to the SIG protein family in the centric diatom Thalassiosira weissflogii (Grunow) Fryxell et Hasle, which is up‐regulated during early stages of sexual reproduction. Immunofluorescence analysis with a polyclonal antibody against the partial amino acid sequences of Ocm2, Ocm3, and Ocm4 showed that Ocm2 and Ocm3 were located in the basal segment region of mastigonemes attached on the surface of the anterior flagellum, and that Ocm4 was located within the tubular shaft portion similar to Ocm1.     

BIOMINERALIZATION OF THE STALKS OF ANTHOPHYSA VEGETANS (CHRYSOPHYCEAE)1

Mineralized stalks of the chryophyte Anthophysa ve-getans (O. F. Müller) Stein were examined by electron microscopy, and the composition of the stalks was determined by energy dispersive x-ray microanalysis. Colonies grown in water from a local lake (Dowdy Lake) had mineralized stalks that were colorless and composed mainly of spherules of calcium phosphate up to 0.7 μm in diameter. The addition of 0.1 mM MnCl₂ to the culture medium resulted in deeply orange-brown stalks mineralized primarily with minute granules of a manganese compound. The addition of 0.1 mM FeCl₃ to the culture media resulted in light yellow stalks mineralized mainly with small granules of an iron compound. The addition of 0.1 mM MgCl₂ or KCl resulted in stalks that were similar in appearance and composition to the colonies grown in culture medium alone. These results show that the composition of the mineralized stalks of A. vegetans is dependent on the elements present in the culture medium. This is also the first time that calcium phosphate has been reported as a mineralization product in the chrysophytes.     

MITOSIS AND CELL DIVISION IN HYDRURUS FOETIDUS (CHRYSOPHYCEAE)1

Mitosis and cell division have been examined ultrastructurally in the vegetative cells of Hydrurus foetidus (Vill) Trev. and found to resemble that of Ochromonas in two important aspects. First, the rhizoplast acts as the spindle organizing body and second, the spindle elongates considerably during anaphase. It differs from Ochromonas in that there is no movement of the basal bodies and flagella towards the poles. Moreover, the nuclear envelope remains relatively intact throughout early stages of mitosis, with gaps developing at the poles during prophase to permit entry of spindle microtubules. Disruption of the nuclear envelope does not occur in the equatorial plane until late anaphase. The spindle persists into telophase and is bent towards the posterior of the cell by the ingrowing edge of the cleavage furrow. Persistence of the spindle and lack of Ochromoms-type cell elongation may be related to the constricting presence of the sheath during cell division—a completely different strategy to that adopted by the green algae under conditions of similar constraint.     

GROWTH RATES OF PSEUDOPEDINELLA PYRIFORME (CHRYSOPHYCEAE) IN RESPONSE TO 75 COMBINATIONS OF LIGHT,TEMPERATURE AND SALINITY1

Clonal cultures were established of single cells of Pseudopedinella pyriforme Carter isolated from the Chesapeake Bay. Axenic cultures were incubated at 5, 10 and 15 C; in 2 1/2, 5, 7 1/2, 10 and 15‰; and at 143, 285, 428, 571 and 714 μW/cm² white light. Maintainable growth rates were calculated from absorbance readings taken for 3 to 5 days from acclimated cultures. The maximum number of doublings per day was 0.9 at 2 1/2‰ salinity, 571 μW/ cm² and 15 C. A salinity of 5 ‰ produced the fastest growth at most light intensities up to 428μW/cm² and temperatures of 5, 10 and 15 C.     

ULTRASTRUCTURE OF CYST FORMATION IN OCHROMONAS TUBERCULATA (CHRYSOPHYCEAE)1

The cysts (statospores) of Ochromonas tuberculata Hibberd are produced within a cytoplasmic silica deposition vesicle (SDV) whose membrane (silicalemma) appears to be formed by the coalescence of golgi vesicles. Silica is first deposited as small nodules and the collar and spines develop by centrifugal growth only after a complete but still thin wall has been laid down. Small vesicles appear to be attached to the SDV only in the region overlying the developing collar; a cap of radially arranged, moderately electron-dense material occurs at the tip of the growing spines. The cyst pore is formed at the anterior end of the flagellate cell, by lack of silica deposition over a small region of the SDV and rupture of the SDV and other membranes crossing this region. When the cyst wall is complete, an extracystic plug is formed in the pore, resulting in the loss of some extracystic cytoplasm and the plasmalemma, and the inner SDV membrane becomes the functional plasmalemma. The plug develops first by coalescence with the cell membrane of golgi-derived vesicles containing dense but apparently nonsiliceous spicules surrounded by amorphous material. During later stages of plug formation only fibrous material is deposited, some of which may be extruded through the pore forcing out some of the spiculate component. Scanning electron micrographs of the mature wall show it is smooth except for the concentrically wrinkled inner face of the flared collar and that the real pore diameter is only ca. half that of the collar. At germination the plug completely disappears in an unknown way and a single cell, similar to a normal vegetative cell emerges through the pore. Chrysophycean cyst formation generally resembles cell wall formation in diatoms, but differs in some details.     

THE OCCURRENCE OF CHLOROPHYLLS C1 AND C2 IN THE CHRYSOPHYCEAE1

We analyzed 34 strains representing 25 species of Chrysophyceae for chlorophylls c₁ and c₂ using thin-layer chromatography. Most organisms had both chlorophylls c₁ and c₂ in addition to chlorophyll a but 17 strains of 9 species of Synura and Mallomonas possessed only chlorophylls a and c₁. These are the first chlorophyll c-bearing algae which lack chlorophyll c₂. We postulate that at least some of the silica-scaled algae including Mallomonas and Synura may be distinct from other Chrysophyceae based upon pigmentation and other characters described in the literature.     

SEASONAL PERIODICITY OF CHRYSOPHYCEAE AND SYNUROPHYCEAE IN A SMALL NEW ENGLAND LAKE: IMPLICATIONS FOR PALEOLIMNOLOGICAL RESEARCH1

The seasonal periodicity of taxa of Chrysophyceae and Synurophyceae from a small New England lake is described for the period September 1983 through June 1988. We found 51 taxa, including 29 that accounted for over 10% of the total in at least one collection. The taxa were fitted into one of five seasonal patterns. Patterns I and II represented taxa restricted to warm (pattern I) or cold (pattern II) months, respectively. Pattern HI represented organisms that began growth in the summer, persisted through autumn and disappeared with the onset of an ice cover. Pattern IV was an extension of pattern III, in which the taxon remained in the plankton throughout the winter and disappeared soon after ice out. Species without a clear seasonal pattern were grouped as pattern V. The seasonal periodicity of the flora, as examined with ordination analyses, was found to remain remarkedly similar during the 58–month study. Except for episodes of low pH during spring snow melt and unseasonally warm or cold weather, sample scores followed a fairly consistent pattern along the first and second primary axes. Water temperature, specific conductance, and pH were important variables that controlled changes in the species composition during the course of a given year. The flora was used to develop an inference model for water temperature. According to the analyses, the remains of a surface sediment sample represented a flora that grew primarily during the late autumn period at 7.6 ° C. Ways in which seasonal data could be utilized to improve paleolimnological inference work are discussed.     

OLISTHODISCUS LUTEUS (CHRYSOPHYCEAE) I. EFFECTS OF SALINITY AND TEMPERATURE ON GROWTH. MOTILITY AND SURVIVALS1, 2

The effect of salinity and temperature on Olisthodiscus luteus Carter has been examined to across the relative importance of these factory on dynamics of natural population. A salinity range 2–50% was observed with increased tolerance to low salinity (<5%.) at higher temperature (20–30°C). Slinities at 4–5%. Had densities of 10³ cells/ml^?1, and growth >0.5 division day^?1 at temperature of 15–30°C higher salinities (5–50%.) variable but distinct optima for density, growth and motility were observed 5, 10 and 30°C. Density and motility showed no clear optima from 10–10%.15–25°C where maximum growth rates >1.0 division/day^?1 were common. Temperature increased from (0.5–1.9 division. Day^?1) and increases of three orders of magnitude (10^2?10³) for maximum densities. Temperature optima 20°C for growth 5–35%. And 25°C for >40%. were observed. The implications of these findings to natural populations of O. luleus are discussed.     

CHARACTERIZATION OF THE LIGHT-HARVESTING COMPLEX OF GIRAUDYOPSIS STELLIFER (CHRYSOPHYCEAE) AND EFFECTS OF LIGHT STRESS1

The light-harvesting complex (LHC) of Giraudyopsis stellifer Dangeard was isolated on a sucrose density gradient after digitonin treatment, and the pigment composition was analyzed by reverse-phase high-pressure liquid chromatography. The LHC is a chlorophyll (Chl) a/c/ fucoxanthin/violaxanthin complex, depleted of β-carotene, comparable to the LHC of Fucophyceae. The excitation transfer from Chl c and fucoxanthin to Chl a is efficient in whole cells. Immunological reactions indicate a close relationship between Chrysophyceae and Fucophyceae. The immunocytochemical labeling confirms the lack of segregation of the LHC in the appressed membranes of the three associated thylakoids and its localization in the intrapyrenoid thylakoid. The violaxanthin-antheraxanthin-zeaxanthin cycle is operative in the cells and efficiently protects photosystem II reaction centers against photoinhibition.     

KINETICS OF SILICON-LIMITED GROWTH IN THE FRESHWATER DIATOM ASTERIONELLA FORMOSA1,2

Growth rates of two clones of the freshwater planktonic diatom Asterionella formosa Hass. were measured under conditions in which external silicon concentrations controlled growth. Clone AfOH2 from Lake Ohrid, Yugoslavia, had a higher maximum growth rate (μ_max= 1.11 doublings/day) and apparent half-saturation constant (K_si] + Si_o= 1.93 μM Si) than clone L262 from Lake Windermere, England. (μ_max= 0.61 doublings/day; K_si+ Si_o= 1.09 μM Si). K_lim, the silicon concentration at μ= 0.9 μ_max, is 13.8 μM Si for clone AfOH2 and 6.5 μM Si for clone L262. These values agree well with published field observations showing A. formosa populations decreasing below 0.5 mg/l SiO₂ (= 8.4 μM Si). Calculations of yield gave a range of 0.5–1.5 μM Si/10⁶ cells for clone AfOH2 and 0.6–1.9 μM Si/10⁶ cells for clone L262.     

CHARACTERIZATION OF NITROGEN UPTAKE BY NATURAL POPULATIONS OF AUREOCOCCUS ANOPHAGEFFERENS (CHRYSOPHYCEAE) AS A FUNCTION OF INCUBATION DURATION,SUBSTRATE CONCENTRATION,LIGHT, AND TEMPERATURE1

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. (K_s) 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 K_lt 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.