Autecology and clonal variability of the marine centric diatom Thalassiosira rotula (Bacillariophyceae) in response to light,temperature and salinity

The influence of 49 combinations of salinity (10–40 S, at 5 S intervals) and temperature (0°–30°C, at 5C° intervals) on the maximum daily division rate (K) and 18 combinations of light intensity (six levels) and temperature (5°, 15°, and 25°C) on photosynthesis, cell division, and chlorophyll a was examined using two clones of Thalassiosira rotula Meunier isolated from the upwelling area of Baja California (clone C8) and from Narragansett Bay, Rhode Islands (clone A8). Physiological differences appear to characterize these to clones with regard to their temperature tolerance (C8 5°–30°C, A8 0°–25°C), maximum growth rate (C8 K=2.9, A8 K=2.4), chlorophyll a content, and in the rates of growth and photosynthesis in response to light intensity and temperature. Optimum salinity for both clones (25–30 S) was generally independent of temperature, while chlorophyll a content decreased with temperature. T. rotula is a cosmopolitan paractic species; experimental studies indicate that it is eurythermal and moderately euryhaline. Comparison of five additional Narragansett Bay isolates of T. rotula reveal minimal spacial or temporal variability in genetically determined physiological characteristics within this local population.     

Saturated uptake kinetics: transient response of the marine diatom Thalassiosira pseudonana to ammonium,nitrate, silicate or phosphate starvation

Changes in the saturated uptake kinetics of the limiting nutrient were followed as Thalassiosira pseudonana (Clone 3 H) batch cultures entered ammonium, nitrate, silicate and phosphate starvation. Cultures starved of ammonium or phosphate developed very high specific uptake capacities over a 24 to 48 h starvation period, due to both decreases in cell quota and increases in uptake rates per cell. In particular, the cell phosphorus quota decreased ca. 8-fold during phosphate starvation and specific uptake rates exceeded 100 d^-1. In contrast, cultures entering nitrate or silicate starvation underwent little or no further cell division, and the uptake capacity declined during starvation. After 24 to 48 h starvation, an induction requirement for uptake of nitrate or silicate was apparent. These responses are consistent with adaptation to the pattern of supply of these nutrients in the field.     

A comparison of Lowry,Bradford and Smith protein assays using different protein standards and protein isolated from the marine diatom Thalassiosira pseudonana

Two spectrophotometric assays for protein commonly used in marine research (Coomassie stain, Bradford; alkaline copper, Lowry) and a more recent assay which has not been applied in this field (bicinchoninic acid, Smith) were compared for homogenates of the marine diatom Thalassiosira pseudonona using bovine serum albumin (BSA) as a standard. When homogenates were prepared by precipitating protein with trichloroacetic acid (TCA) and redissolving in 1 N NaOH, the protein content estimated by the Lowry and Smith assays agreed closely, but was consistently 20% higher than that indicated by the Bradford assay. To determine if this difference was due to the choice of a protein standard, protein from T. pseudonana was purified and compared to BSA, bovine gamma-globulin (BGG), and casein. The reactivity of the purified protein (expressed as the slope of the absorbance vs protein concentration curve) did not differ between cultures grown at high or low irradiance. For the Smith and Bradford assays the reactivity of BSA was not significantly different from algal protein, but for the Lowry assay, algal protein was significantly higher in reactivity than BSA. BGG was not significantly different in reactivity from algal protein for the Lowry and Smith assays, but BGG gave significantly lower absorbances than algal protein in the Bradford assay. These results suggest that BSA is a suitable standard for algal protein in the Bradford assays, while BGG is preferable for the Lowry assay. Either protein standard could be used for the Smith assay. Differences in purified algal protein reactivity compared to BSA could not account for the differences among the assays, nor could interference by chlorophyll a. Precipitating protein with TCA prior to analyses gave lower protein than direct analyses of homogenates for the Lowry and Smith assays, but no differences were found for the Bradford assay. As a result, the Lowry and Smith assays indicated up to 60% greater protein than the Bradford if TCA precipitation was not performed. This may be due to removal of free amino acids and small peptides which are less reactive in the Bradford assay. The 20% higher protein found in the Lowry or Smith vs Bradford assays may be due to different assay sensitivity to small peptides or other compounds which are precipitated along with proteins by TCA. Although the Smith assay is substantially simpler to perform than the Lowry, there appear to be no quantitative differences in the results. It remains unclear which spectrophotometric assay is most accurate, but the Bradford assay is faster and simpler, and is less likely to be affected by non-protein compounds found in marine phytoplankton.     

The diatom Thalassiosira rotula affects reproductive success in the copepod Acartia clausi

Fecundity, egg viability and fecal pellet production are reported for Acartia clausi females collected in the Bay of Naples, Italy, from April to October 1992 and fed either with a diatom (Thalassiosira rotula) or dinoflagellate (Prorocentrum minimum) diet, at food saturated conditions. The diatom diet significantly reduced both egg and fecal pellet production as well as hatching success. Blockage of egg development occurred with both axenic and non-axenic cultures of T. rotula, suggesting that inhibitors were provided by the diatoms and not by the bacteria associated with diatom cultures. Low hatching success was also artificially induced by exposing newly spawned A. clausi eggs to high concentrations of diatom extracts, indicating the presence of deleterious, inhibitory compounds blocking copepod embryogenesis. Fecundity and hatching success diminished significantly with female age. In contrast, female longevity was not significantly modified by food type. The presence of males did not significantly alter fecundity or egg viability. Females continued to produce viable eggs throughout the period of incubation, with and without males, in both food conditions, indicating that remating is infrequent and not necessary to sustain viable egg production in this species. The succession in low and high population densities may therefore be the outcome of variations in survival rates of eggs, rather than reproductive protential perse; such variations may strongly depend on the adult copepod diet.     

Growth and competition of the marine diatoms Phaeodactylum tricornutum and Thalassiosira pseudonana. II. Light limitation

The marine diatoms Phaeodactylum tricornutum (Bohlin) and Thalassiosira pseudonana (Hasle and Heimdal) were grown under both continous illumination and a 14 h light: 10 h dark cycle at light intensities ranging from 1.53×10^-4 to 2.95×10^-1 ly min^-1. Under both photoperiods, T. pseudonana exhibited higher division rates than P. tricornutum at high light intensities, but the reverse was true at all light intensities <3×10^-3 ly min^-1. Comparison of these results with available data on light-limited growth of other planktonic algae suggests that P. tricornutum may be unusually efficient at maintaining its cell division rate at low light intensity. This efficiency may contribute substantially to its success in turbid, nutrient-enriched mass algal culture systems, the only environments in which it is known to attain great numbers.Contribution No. 4086 from the Woods Hole Oceanographic Institution.     

Cellular composition and physiological characteristics of the diatom Thalassiosira weissflogii adapted to cadmium stress

Prolonged exposure of Thalassiosira weissflogii (Grunow) to a sub-lethal concentration of cadmium in continuous culture resulted in the development of cellular characteristics allowing optimal growth in the presence of Cd. Examination of Cd-adapted and unadapted cells was made on steady-state populations growing at the same rate in order to eliminate any effects of differing growth rate on metabolism. Adaptation to Cd stress was manifested as increases in mean cell volume, dry weight, protein: DNA, protein: RNA, protein: carbohydrate, protein nitrogen: total cell nitrogen and carotenoid: chlorophyll a ratios. Subsequent exposure of the cells to Cd over a wide concentration range showed that cellular division rate, carbon photoassimilation and extracellular release of dissolved organic compounds were greatest near the Cd concentration to which the cells had previously been adapted. Enhanced cellular carbon photoassimilation in Cd-adapted cells correlated exactly with increased cellular protein content. The amount of dissolved organic excretion by Cd-adapted cells at the adaptation concentration was the same as that of unadapted cells at the same concentration. Since total carbon photoassimilation was greater in Cd-adapted cells at this concentration, the percentage of carbon excreted was less in these cells.     

Water-soluble extracts of the diatom Thalassiosira rotula induce aberrations in embryonic tubulin organisation of the sea urchin Paracentrotus lividus

Eggs and embryos of the sea urchin Paracentrotus lividus were used as a model to study the effect at the cellular level of potential anti-mitotic compounds extracted from the diatom Thalassiosira rotula. Eggs and embryos incubated in a water-soluble diatom extract, corresponding to 5 × 10⁶ and 10⁷ cells ml⁻¹, were totally blocked (i.e. cell division was blocked) at the one-cell stage. At lower concentrations (2.5 and 1.25 × 10⁶ cells ml⁻¹), the first mitotic division was inhibited in 32 ± 26% and 25 ± 3.5% of the zygotes, respectively, demonstrating the dose-dependent effect of diatom extracts on sea urchin development. Immunofluorescence dyes, specific for DNA and α-tubulin subunits, were used to stain nuclei and microtubules in sea urchin embryos during various phases of development. Images with the confocal laser scanning microscope showed that tubulin was not organised in filaments at the sperm aster and cortex levels, and that the pronuclei were not fused in embryos incubated soon after fertilisation with water-soluble diatom extracts corresponding to 10⁷ cells ml⁻¹. At lower diatom-extract concentrations (4 × 10⁶ cells ml ⁻¹), fusion of the pronuclei occurred but the mitotic spindle was not formed. Microtubules were clearly de-polymerised and the chromatin appeared globular and compacted at the centre of the cell. A similar structure was observed for sea urchin embryos incubated with 0.1 mM colchicine, a potent anti-mitotic compound. When sea urchin embryos were incubated in water-soluble diatom extracts at different times prior to the first mitotic division, microtubules appeared de-polymerised at each step, from pronuclear fusion to telophase, and cell division was blocked. At the histological level, embryos incubated with 4 × 10⁶ cells ml⁻¹ diatom extract showed nuclear fragmentation without cytokinesis. The possible use of sea urchin embryos as a bioassay to test for other unknown compounds with cytotoxic activity in phytoplankton species is discussed. Received: 7 May 1998 / Accepted: 9 December 1998     

Light absorption by a marine diatom: experimental observations and theoretical calculations of the package effect in a small Thalassiosira species

The relationship between in vivo light absorption efficiency of whole cells and in vitro absorption efficiency of algal pigments has been examined experimentally in the marine diatom Thalassiosira sp. In vitro absorption spectra were obtained for cells disrupted by either ultrasonic treatment or high-pressure shearing stress in a low-temperature (-40°C) pressure cell. A dimensionless measure of the magnitude of the package effect (Q _a ^*), calculated from the ratio of whole-cell to disrupted-cell absorption, ranged from about 0.5 at the blue absorption peak of chlorophyll a (λ=435 nm) to 0.7 at the red chlorophyll a peak (λ=670 nm) to 1.0 at the absorption minimum (λ=600 nm). Cell diameter was found to be an inappropriate measure of size for assessing the magnitude of the package effect. Instead, the effective optical diameter for calculation of intracellular self-shading was found to be less than the cell diameter. This observation is consistent with the fact that most algal pigments are contained within chloroplasts, and that chloroplast volume is necessarily smaller than cell volume.     

Silicon and the ecology of marine plankton diatoms. I. Thalassiosira pseudonana (Cyclotella nana) grown in a chemostat with silicate as limiting nutrient

The small marine plankton diatom Thalassiosira pseudonana Hasle and Heimdal (Guillard's clone 3H) was grown in chemostats with silicate as the limiting nutrient. The calculated maximum growth rates were comparable to those previously reported for this species. The silica content of the diatom shells varied with the growth rate. As the growth rate approached zero, there were still measurable quantities of residual reactive silicate in the medium. In one of the two chemostats used, silicate assimilation by the cells was inefficient due to some unknown internal or external factor. In the other chemostat, statistically calculated half-saturation constants of growth were in the range of 0.5 to 0.8 g—at Si/l, depending on which kind of correction was made for residual silicate. Half-saturation constants of steady-state mean silicate uptake per cell and hour, calculated in a similar fashion, were in the range of 1.4 to 2.6 g—at Si/l. These results indicate that the silicate concentrations causing a reduced silicate uptake by this species in nature do not necessarily result in a correspondingly reduced growth rate. Growth in coastal waters is likely to become seriously limited by a shortage of silicate only when most of the silicate originally present has been removed in the course of a diatom bloom.     

Genetic variability in the cosmopolitan deep-water ophiuran Ophiomusium lymani

A population of the cosmopolitan deep-sea ophiuran Ophiomusium lymani was studied by gel electrophoresis, and proved to be highly variable genetically; about 53% of the 15 loci studied are polymorphic, and the average individual is heterozygous at about 17% of the loci. This is approximately the same genetic variability displayed by other species, belonging to other phyla or classes, from the same deep-sea trawl. A similarly high level of genetic variation occurs in deep-sea organisms in general, and in a shallow-water tropical species. Both the deep-sea and the tropics are trophically stable environments. On the other hand, low genetic variabilities have been found in marine species from trophically unstable environments. These data suggest that any phylogenetic effects on genetic variability are secondary, and that the trophic regime may be of major importance in determining genetic strategies of adaptation.This study is no. 5 of a series of reports on the results of Expedition Quagmire.     

Genetic variability and differentiation of sporophytes and gametophytes in populations of Gelidium arbuscula (Gelidiaceae: Rhodophyta) determined by isozyme electrophoresis

Genetic differentiation and genetic variability of sporophytic and gametophytic populations of Gelidium arbuscula (Bory) from three localities sampled in 1989 and 1990 in the Canary Islands (Spain) were examined by isozyme electrophoresis. Twenty-three to 29 putative alleles corresponding to 22 gene loci, were compared. High deviations in Hardy-Weinberg equilibrium, and significant differences between allelic frequencies of sporophytic and gametophytic subpopulations at the same locality were found, suggesting a predominant asexual reproduction of G. arbuscula. The genetic variability (percentage of polymorphic loci, mean number of alleles per locus and average gene diversity) of haploid subpopulations was lower than that of diploid subpopulations at all three localities, being the lowest described for seaweeds. No correlation between genetic and geographical distance was found. The high genetic differentiation coefficient between all subpopulations suggests a very reduced genetic flow between subpopulations of the same and of different localities. These results suggest that the genetic structure of the populations of G. arbuscula from the Canary Islands is due to a founder-effect combined with a predominance of asexual reproduction. This is the first report comparing allelic frequencies between sporophytic and gametophytic subpopulations of seaweeds.     

Genetic differentiation in the brackish-water ostracod Cyprideis torosa

An electrophoretic survey of allozyme variation suggests that two separate populations of Cyprideis torosa (Jones) can be distinguished, one from the coast of England and the Netherlands, the other from the south coasts of the Baltic Sea. Gene flow between these two populations is limited and asymmetrical and seems to be more intense westwards than eastwards. On the other hand, particular local populations within the range of either population, although inhabiting geographically isolated estuaries, lagoons etc., are genetically similar to each other to such a degree that they should be treated as sets of subpopulations rather than as separate units. It is possible that in C. torosa a similar population structure occurs everywhere along the main migration routes of aquatic birds, which have clearly participated in the passive dispersal of this ostracod. Clines are observed in the frequencies of some alleles. Phosphogypsum pollution may be the cause of deviation from Hardy-Weinberg equilibrium in a local population.     

Genetic variation in marine teleosts: High variability in habitat specialists and low variability in habitat generalists

Genetic variation was reviewed in 106 species of marine teleosts. Two heterozygosity estimates were used, one including all protein and enzyme loci and a second excluding the non-enzymatic protein loci. Mean heterozygosities are 0.055±0.036 based on all loci in 106 species and 0.060+0.038 based on enzymatic loci in 89 species. A significant negative correlation was noted between heterozygosity and the proportion of general protein loci included in the estimate. A comparison was made of heterozygosities among taxonomic orders and families, life zones, reproductive mode, geographical range and size. High levels of genetic variation are found in Clupeiformes, Atheriniformes, Pleuronectiformes, temperate pelagic, tropical, intertidal-sublittoral and wide-range species. Low levels of genetic variation are found in Gadiformes, Scorpaeniformes, temperate demersal, polar, and narrowrange species. The most striking differences in heterozygosities are between temperate demersal flatfishes and temperate demersal round fishes. It is suggested that much of the data can be explained by a habitat specialist-generalist model, with high heterozygosities in specialists and low heterozygosities in generalists, but that this is only one of a mosaic of factors which influence genetic variation.     

Genetic experssion of temperature tolerance in the copepod Eurytemora affinis in different salinity and temperature environments

The genetic expression of temperature tolerance in Eurytemora affinis Poppe in different environments was investigated by testing temperature tolerances of broods in pairs of salinity and temperature environments. Three methods were used to identify interaction between genotype and environment: (1) the correlation between mean tolerances of halves of broods grown in the two environments, (2) brood by environment interaction and (3) heritabilities in the two environments. All correlations between means of half broods were positive, only one of six brood by environment interactions was significant and the heritability estimates were not markedly different. Thus, the variability in temperature tolerance in different salinities and in different temperatures seems to be due to the same genes; and selection pressure on temperature tolerance is in the same direction with seasonal changes in temperature and salinity.     

Feeding deterrence properties of apo-fucoxanthinoids from marine diatoms. II. Physiology of production of apo-fucoxanthinoids by the marine diatoms Phaeodactylum tricornutum and Thalassiosira pseudonana,and their feeding deterrent effects on the copepod Tigriopus californicus

The marine diatoms Phaeodactylum tricornutum and Thalassiosira pseudonana have been shown to produce apo-fucoxanthinoid compounds which act as feeding deterrents against the harpacticoid copepod Tigriopus californicus. The amounts and types of apo-fucoxanthinoids produced were species specific. Th. pseudonana produced small quantities of apo-12-fucoxanthinal and apo-13-fucoxanthinone only during senescence, while P. tricornutum produced much greater quantities of these two compounds during both log and senescence phases, in addition to producing a third compound, apo-10-fucoxanthinal, only during senescence. For both species, production of apo-fucoxanthinoids increased as the cells entered senescence phase due to phosphate limitation. The amounts of apo-fucoxanthinoids necessary to reduce feeding in T. californicus by 50% ranged from 2.22 to 20.2 ppm. This range was approximately 1000 times lower than the total apo-fucoxanthinoid concentration in P. tricornutum. The amounts of apo-fucoxanthinoids necessary to cause a 50% mortality in a population of T. californicus ranged from 36.8 to 76.7 ppm. Thus, these compounds are present in concentrations which may have ecological significance in the control of bloom formation and grazing. The production of apo-fucoxanthinoids may be a phenomenon common to many diatoms, particularly as they enter senescence due to nutrient limitation.     

Genetic differentiation ofMytilus edulis in eastern North America

There is significant differentiation at five polymorphic loci ofMytilus edulis among certain geographical areas of the Atlantic coast of North America. Non-metric multidimensional numerical methods distinguished three population groups: (I) populations south of Cape Cod, (II) populations throughout the Gulf of Maine, Gulf of St. Lawrence, areas of both southern and northern Newfoundland, and southern Hudson Bay, and (III) populations in southeastern Nova Scotia, northern Newfoundland and Hudson Strait, Quebec. Each subset consists of populations that exhibit characteristic multilocus, multiple allele genotypes. Populations in Groups II and III are spatially interdigitated among each other. At least one geographical area of mixing between genetically distinct populations occurs in northeastern Newfoundland. There is no evidence for interbreeding among genetically distinct individuals in mixed populations, suggesting the possibility that populations in the Atlantic Canadian Provinces and areas of northern Canada may consist of two distinct species.     

Genetic variability in bloodworm (Glycera dibranchiata) populations in the Gulf of Maine

Genetic structure of eight Northwest Atlantic populations of the marine polychaeteGlycera dibranchiata Ehlers was examined with starch gel electrophoresis. Samples were collected during summer and fall 1981, and seven polymorphic and four monomorphic loci were consistently scored. Average heterozygosity (0.126) and percent polymorphic loci (59.3) were comparable to the averages reported for marine invertebrates. Minimum genetic distances between populations ranged from 0.003 to 0.093, levels typically associated with local populations of the same species in other taxa. Based on these data, inter- and intra-estuarine migration and gene flow appear to be low. Only two populations, separated by 13 km along the same river in New Brunswick, Canada, were not genetically different from each other. These findings may have relevance for management strategies in bloodworms.     

Geographic variation,niche breadth and genetic differentiation at different geographic scales in the mussels Mytilus californianus and M. edulis

Genetic differentiation was investigated in the marine mussels Mytilus californianus Conrad and M. edulis Linn. from the west coast of North America. In allopatry with M. californianus, M. edulis occurs throughout the intertidal zone; however, in microgeographic sympatry its ecological range is restricted to above the M. californianus mussel bed and to patches of substratum opened by natural disturbances within the bed. Over the same geographic scale, the broader-niched M. edulis shows greater among-locality genetic difference and greater levels of polymorphism than M. californianus at two enzyme loci. Genetic differentiation on a geographic basis was investigated in M. californianus at a single rock (on a scale of meters), on an island (on a scale of kilometers), throughout a strait (on a scale of 10² kilometers), and along the west coast of North America (on a scale of 10³ km). Differentiation was minimal over the west coast, and could be explained by microhabitat differences in a local area. The minimal differentiation in west coast mussels relative to strong geographic differentiation of M. edulis on the east coast of North America may be related to the steeper latitudinal thermal gradient on the east coast. Local ecologically related microgeographic variation can result in biased and misleading estimates of genetic heterogeneity but microgeographic variation at enzyme loci may not be due to selection at the loci investigated or may even be due to the breeding structure of the mussel populations, as suggested by Tracey et al. (1975).     

Genetic differentiation independent of intertidal gradients in the pulmonate limpet Siphonaria kurracheensis

The intertidal limpet Siphonaria kurracheensis (Reeve, 1856) has a bimodal vertical distribution of abundance on rocky shores at Rottnest Island, Western Australia. An electrophoretic study of 5 polymorphic enzymes revealed no consistent genetic differences between adults high and low on the shore. Contrasting with this absence of a detectable genetic response to the steep environmental gradients in the intertidal zone, there were genetic differences among low-shore adults from different sites, and between adults and recruits. This genetic differentiation could be due to either localized selection or temporal variation in the genetic makeup of recruits.Communicated by G. F. Humphrey, Sydney