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.     

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.     

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.     

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.     

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.     

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.     

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.     

PHYLOGENETIC AFFINITIES OF CRYSONEPHELE PALUSTRIS (CHRYSOPHYCEAE) BASED ON INFERRED NUCLEAR SMALL-SUBUNIT RIBOSOMAL RNA SEQUENCE

The nuclear small-subunit ribosomal DNA sequence was determined from the colonial chromophytic alga Chrysonephele palustris Pipes, Tyler, et Leedale using polymerase chain reaction methods. The inferred ribosomal RNA sequence was included in a multiple alignment, containing heterokont chromophytes, and subjected to molecular systematic analyses in order to determine the phylogenetic relationships of this alga . Chrysonephele palustris grouped within the Chrysophyceae and not in an intermediate position between the Chrysophyceae and Eustigmatophyceae, suggesting that Chrysonephele is not a phylogenetic link between these algal classes .     

MERCURY AND TEMPERATURE INTERACTIONS ON THE GROWTH RATES OF THREE SPECIES OF FRESHWATER PHYTOPLANKTON1,2

Cultures of freshwater algae, representing three algal divisions, Synura petersenii Korshikov; Chlamydomonas sp.: and Nitzschia sp., were subjected to four different mercury-temperature shock interactions to demonstrate synergistic effects between mercury and temperature. Algal growth, measured by temporal changes in vivo fluorescence of chlorophyll was used to ascertain the effects. Mercury addition and temperature shock had various inhibitory effects on algal growth. Prior environmental conditions influence the effect of subsequent treatments. Growth of S. petersenii was severely inhibited by mercury in all experiments. Control cultures would not grow at 30 C and died when shacked at this temperature. Chlamydomonas sp. cultures initially inhibited by mercury were able to recover under most conditions after a period of reduced growth. Nitzschia sp. was resistant to mercury except when simultaneously shocked with temperature. Mercury analyses showed that Nitzschia cells at 25 C and 30 C contained a high percentage of the mercury initially added to the cultures. There was a significant loss of mercury at the end of each experiment from all cultures, probably due to volatilization.     

PHOSPHATE AND SILICATE GROWTH AND UPTAKE KINETICS OF THE DIATOMS ASTERIONELLA FORMOSA AND CYCLOTELLA MENEGHINIANA IN BATCH AND SEMICONTINUOUS CULTURE1

Information on the nutrient kinetics of Asterionella formosa Hass. and Cyclotella meneghiniana Kutz. under either phosphate or silicate limitation was obtained for use in a Monod model and in a variable internal stores model of growth. Short-term batch culture growth experiments were fit to the Monod model and long-term semicontinuous culture experiments and short-term uptake experiments were fit to the variable internal stores model. Mathematical analysis indicates that the parameters of the 2 models may be expressed in terms of each other at steady state. The qualitative results of both batch and steady state culture methods agree. For limiting phosphate experiments. A. formosa is better able to grow at low PO₄-P concentrations than C. meneghiniana, as shown by its lower K for PO₄-P limited growth. The k_Q of A. formosa compared to C. meneghiniana found in long-term semicontinuous culture indicates that A. formosa is almost an order of magnitude more efficient at using internal phosphate for growth. The qualitative results under silicate-limited growth of C. meneghiniana is less than that of A. formosa. The k_Q from semicontinuous culture experiments indicates that C. meneghiniana is the more efficient at using internal silicate for growth. Nutrient uptake experiments showed more variability from a Michaelis-Menten relationship than short-term growth experiments. There were no significant differences between the 2 species in half saturation constants for either phosphate or silicate uptake. We observed a marked dependence of the coefficient of luxury consumption (R) of phosphate on the steady state growth rate. A. formosa has a higher R than C. meneghiniana.     

OLSTHODISCUS LUTEUS (CHRYSOPHYCEAE) II. FORMATION AND SURVIVAL OF A BENTHIC STAGE1

Motile unicells of Olisthodiscus lutheus Carter aggregated to form encapsulated masses of nonmotile cells in a benthic stage throughout a temerature range of 15–30°C at salinities o f 10–50%. Motile cells were released from beneathic masses at 10–30°C but at 5°C, cells were not motile and at 0°C cells lysed. Exposure of benthic masses of I day to 8 wk to temperatures of 0–30C in lighted growth chambers resulted in mortality to cells kept below 10°C and normal growth at higher temperatures. Benthic stage cells kept tn darkness at the same temperatures exhibited mortality in all but those at 5 and 10°C. Cells at these thmoeratures remained viable 15 wk in continual darkness. Comparison of cell morphology of matile and benthic stage O. luteus to other Olisthodiscus species and the ecological implications o fthe benthic stage are discussed.