Nuclease Digestion of Chromatin from the Eukaryotic Algae Olisthodiscus luteus,Peridinium balticum,and Crypthecodinium cohnii 1, 2

Chromatin from a uninucleate dinoflagellate, Crypthecodinium cohnii, a binucleate dinoflagellate, Peridinium balticum, and a chromophyte, Olisthodiscus luteus, was examined by nuclease digestion and the results were compared to those from vertebrates. Gel analysis of the products of staphylococcal (micrococcal) nuclease digestion revealed a DNA repeat unit of 220(±5) base pairs for O. luteus and 215(±5) for P. balticum. Limit digestion gave a core particle of 140 base pairs, revealing that these longer repeat sizes are due to longer linker regions. No repeating subunit structure was found upon electrophoresis of digests of C. cohnii nuclei. Examination of the DNA fragments produced by DNAse I digestion of nuclei isolated from P. balticum and O. luteus showed the same ladder of ten base multiples as seen in chromatin from other eukaryotes. Examination of the kinetics of digestion by DNAse II of Peridinium chromatin revealed less susceptibility when compared to DNAse I digestions while 70% of Olisthodiscus chromatin and 35% of C. cohnii chromatin was sensitive to DNAse II. These data, taken together with previous results from Euglena, indicate that while algal chromatin is similar to that of higher eukaryotes in regard to DNAse I and II action, it differs in that the linker DNA is longer. In addition, the Hl-like histone from O. luteus and P. balticum is located in the linker DNA as in higher eukaryotes.     

Chromatin structure in the unicellular algae Olisthodiscus luteus,Crypthecodinium cohnii and Peridinium balticum

Isolated nuclei of the unicellular alga Olisthodiscus luteus, the uninucleate dinoflagellate Crypthecodinium cohnii and the binucleate dinoflagellate Peridinium balticum were lysed and deposited on grids by the microcentrifugation technique. The ultrastructure of the released chromatin fibers was compared to that of mouse liver nuclei. Chromatin from nuclei of Olisthodiscus luteus and the eukaryotic¹ nuclei of Peridinium balticum, appeared as linear arrays of regularly repeating subunits which were identical in size and morphology to mouse nucleosomes. In contrast, the chromatin fibers from Crypthecodinium cohnii nuclei appeared as smoothe threads with a diameter of about 6.5 nm. Nuclear preparations containing mixtures of dinokaryotic and eukaryotic nuclei of Peridinium balticum also contained smooth fibers which most likely originated from the dinokaryotic nuclei. These and other results demonstrating the presence of nucleosomes in lower eukaryotes suggest that the subunit structure of chromatin arose very early in the evolution of the eukaryotic cell.     

RNA Synthesis in Isolated Nuclei of the Dinoflagellate Crypthecodinium cohnii

Atypical eukaryotic RNA polymerase activity was demonstrated in nuclei of Crypthecodinium cohnii, a eukaryote devoid of histones. Nuclei were isolated from growing cultures of this dinoflagellate and assayed for endogenous RNA polymerase (EC 2.7.7.6) activity. There was a biphasic response to Mg²⁺ with optima at ? 0.01 and 0.02 M MgCl₂, but in contrast to other eukaryotic RNA polymerases, this enzyme activity was inhibited by low MnCl₂ concentrations. In the presence of 0.01 M MgCl₂ the optimum (NH₄)₂SO₄ concentration was 0.025 M, a concentration at which the nuclei were lysed. Incorporation of [₃H]UMP into RNA was inhibited by actinomycin D and dependent on the presence of undegraded DNA, and the reaction product was sensitive to ribonuclease and KOH digestion. Omission of one or more ribonucleoside triphosphates greatly reduced the incorporation. Only a slight enhancement of RNA polymerase activity resulted from the addition of various amounts of native and denatured calf thymus DNA. Spermine caused a marked inhibition while spermidine had little effect on RNA synthesis in the nuclei. Under the optimum conditions described in the present paper the nuclei incorporated ? 3 pmoles of [₃H]UMP/muml; DNA at 25 C for 15 min, and ? 80% of this activity was inhibited by the eukaryotic RNA polymerase II inhibitor, α-amanitin (20 m?/ml). A unique situation therefore exists in C. cohnii nuclei, in which absence of histones (a prokaryotic trait) is combined with α-amanitin-sensitive RNA polymerase activity (a eukaryotic trait).     

Nutrition of the American strain of Gyrodinium cohnii

Summary Only the Woods Hole strain of Gyrodinium cohnii has grown to high densities (4 million cells/ml) in 7 days in a chemically defined medium. The other strains were slow-growing, and preferred seawater media enriched with organics.The Woods Hole strain, at the onset, required histidine, but was trained to grow on ammonia (no histidine strain). The no-histidine strain of G. cohnii utilized NH₄, several amino acids and amines, did not utilize nitrates as N-sources. Though NH₄ supported continued growth, addition of histidine and betaine resulted in higher densities.Glucose, glycerol, and acetate were the best carbon sources; the combination of 2 or 3 C-sources yielded better growth than any singly. G. cohnii is euryhaline and withstood high concentrations of Tris buffer, but was sensitive to temperatures below 20° C.Biotin was needed. Thiamine is probably synthesized but at a low rate: absence of thiamine resulted in indefinite continued slow growth; the addition of thiamine gave dense, rapidly dividing cultures. G. cohnii may perhaps serve for assay of thiamine and biotin in seawater.Supported in part by contract NR 104-202 with the office of Naval Research and by grand G-1198 of the National Science Foundation.     

Effects of salinity on cell growth and docosahexaenoic acid content of the heterotrophic marine microalga Crypthecodinium cohnii

The effects of salinity on cell growth and docosahexaenoic acid (DHA) content of three marine microalgal strains, Crythecodinium cohnii ATCC 30556, C. cohnii ATCC 50051 and C. cohnii RJH were investigated. The lag phases of the three strains increased with increasing salinity in Porphyridium medium. The specific growth rate of C. cohnii ATCC 30556 was the highest at 9 g L⁻¹ NaCl while the other two strains had their highest specific growth rates at 5 g L⁻¹ NaCl. The highest cell dry weight concentrations of 2.51 g L⁻¹ and 1.56 g L⁻¹ were achieved at 9 g L⁻¹ NaCl for C. cohnii ATCC 30556 and ATCC 50051, respectively, while the highest dry weight concentration of 2.49 g L⁻¹ was achieved at 5 g L⁻¹ NaCl for C. cohnii RJH. The highest cell growth yield coefficient on glucose was 0.5 g g⁻¹ for both C. cohnii ATCC 30556 and C. cohnii RJH and 0.45 g g⁻¹ for C. cohnii ATCC 50051. All three strains responded to the change of salinity by modifying their cellular fatty acid compositions. At 9 g L⁻¹ NaCl, C. cohnii ATCC 30556 had the highest total fatty acid content and DHA (C22:6) proportion. In contrast, C. cohnii ATCC 50051 and C. cohnii RJH had the highest DHA content at 5 g L⁻¹ NaCl. C. cohnii ATCC 30556 and ATCC 50051 had the highest DHA yield (131.55 and 68.24 mg L⁻¹ respectively) at 9 g L⁻¹ NaCl while C. cohnii RJH had the highest DHA yield (128.83 mg L⁻¹) at 5 g L⁻¹ NaCl. Received 27 May 1999/ Accepted in revised form 27 August 1999     

Quantitation of Absolute Pneumocystis carinii Nuclear DNA Content. Trophic and Cystic Forms Isolated from Infected Rat Lungs are Haploid Organisms

The Pneumocystis carinii carinii DNA content in nuclei of trophic forms and cysts (spore cases) containing 2, 4, or 8 intracystic bodies, were compared using quantitative fluorescence image analysis. The nuclear DNA content was found to be lower than the theoretical limits of Feulgen cytophotometry. Several fluorescent DNA dyes provide brighter staining, but these techniques suffer from nonspecific binding to other cellular components, such as RNA. It was demonstrated that the thick glycocalyx surfaces of trophic forms and the cyst walls of P. carinii organisms, as well as the cell wall of S. cerevisiae, bound all fluorescent dyes tested to varying degrees. Hence in this study, measurements were performed on cells in which the outer surfaces of organisms were first removed with lyticase. Two stains that appeared most specific for DNA, DB181 and 4′,6-diamidino-2-phenylindole (DAPI), were used for quantitations; lower deviations of fluorescence intensities were observed with DB181. Haploid wild type Saccharomyces cerevisiae and cdc-28 temperature-sensitive mutant cells, accumulated at the restrictive temperature (37° C), were used as quantitative internal standards for estimating the absolute nuclear DNA content of P. carinii. Haploid wild type and mutant nuclei stained with DAPI had the same relative fluorescence intensities. The P. carinii nuclear DNA content of trophic forms and individual intracystic bodies (spores), regardless of life cycle stage, were not different. The mean values obtained were 6.9 and 6.7 fg DNA/nucleus with DB181 and DAPI, respectively (approximately 9.26 and 8.99 Mbp nucleotides, respectively). Since these would include 2C (G-2 phase) and S-phase nuclei, a 1C population of nuclei was selected by histogram distributions of DB181-stained nuclei. Almost all nuclei analyzed in all life cycle stages fell within this population. The 1C mean of 6.55 fg DNA/nucleus (median, 6.62 fg DNA/nucleus) was estimated as representing 8.79 Mbp nucleotides, assuming only A-T binding of the dye and taking into account the G+C content of S. cerevisiae and P. carinii. A 4C (G-2-phase diploid nuclei) population was not detected in histograms of DB181- or DAPI-stained nuclei. The P. carinii nuclear DNA content values obtained in this study were similar to those independently obtained by calculating the total DNA in the organism's chromosomes resolved by electrophoretic techniques. Together, the data on total chromosome numbers and the estimated DNA content of those chromosomes, with our quantitation of nuclear DNA content of different life-cycle stages demonstrate that P. carinii carinii isolated from infected rat lungs are haploid organisms.     

Genome analysis of the moss Physcomitrella patens (Hedw.) B.S.G.

A wild-type (WT) strain of the moss Physcomitrella patens (Hedw.) B.S.G., two mutants derived from it (PC22 and P24), and a somatic hybrid, PC22(+)P24, were analysed. Staining of metaphases revealed 54±2 chromosomes in the somatic hybrid and 27 chromosomes in the wild type and the two mutants. Using flow cytometry (FCM), DNA contents were calculated to be 0.6 pg (WT, PC22), 1.2 pg (P24), and 1.6 pg (PC22(+)P24) per nucleus, respectively. Southern hybridization provided evidence for at least one family of highly repetitive DNA and, furthermore, revealed different amounts of repetitive DNA in the four genotypes. However, these sequences cannot account for the 100% increase in the nuclear DNA amount in mutant P24, relative to wild type. In FCM analyses every moss geno-type generated just one single peak of fluorescence, indicating an arrest in the cell cycle during the daytime. Thermal denaturation of wild-type DNA revealed a G+C content of 34.6% for total DNA and 38.6% for plastid DNA. A cDNA library of 1.2 × 10⁶ independent clones was established, from which sequences homologous to cab and rbcS, respectively, were isolated. These genes show significant homologies to those of higher plants, and, likewise, comprise multigene families. No restriction fragment length polymorphisms could be detected between the four moss genotypes using these cDNA probes.This article is based in part on doctoral studies of M.F. and MW at the University of Hamburg, Faculty of Biology     

NUCLEAR BASIC PROTEINS FROM THE BINUCLEATE DINOFLAGELLATE PERIDINIUM FOLIACEUM (PYRROPHYTA)1

Histories of the endosymbiont nucleus of the binucleate dinoflagellate Peridinium foliaceum Stein were prepared from isolated nuclei and analyzed by peptide mapping, ammo acid composition, and two-dimensional gel electrophoresis. Using these criteria, we identified the presence of two HI-like histories and the core histones H3, H2A, H2B, and H4. These histones are similar but not identical to those of the endosymbiont nucleus of the bi-nucleate dinoflagellate Peridinium balticum Levander.     

A SURVEY OF AUXOTROPHY IN FIVE FRESHWATER DINOFLAGELLATES (PYRRHOPHYTA)1

Peridiniopsis polonicum (Wolosz.) Bourrelly requires vitamin B₁₂, and Peridinium limbatum (Stokes) Lemm. requires thiamin for growth. Unlike marine Peridinium species, Peridinium willei Huit.-Kaas, P. volzii Lemm., and P. inconspicuum Lemm. do not display auxotrophy. Peridinium volzii is strongly inhibited by concentrations of biotin above 1 μg L^?1.     

Deoxyribonucleotide sequence divergence betweenStaphylococcus cohnii subspecies populations living on primate skin

Staphylococcus cohnii strains isolated from various primates could be separated into three distinct groups or subspecies on the basis of phenotypic characterization and DNA-DNA hybridization techniques. These included a human-specificS. cohnii subspecies (denoted here as subsp. 1), a widely distributed primateS. cohnii subspecies (subsp. 2), and a Ceboidea (New World monkey)-specificS. cohnii subspecies (subsp. 3). Divergence of the latter subspecies from the other two is great enough to place it in a near (separate)-species status.S. cohnii represents the third example of aStaphylococcus species where DNA divergence has been demonstrated between human and nonhuman primate-adapted populations. The data presented in this report continue to support the hypothesis that at least certain staphylococci have evolved together with their hosts by conjugate evolution.Paper No. 8555 of the Journal Series of the North Carolina Agricultural Research Service, Raleigh, N.C. 27650     

Evaluation of selected features of Staphylococcus cohnii enabling colonization of humans

Based on iron utilization, sensitivity to skin fatty acids, lipolytic and proteolytic activity the potential abilities ofStaphylococcus cohnii strains to colonize humans were evaluated. The investigation included 60 strains that belong to both subspecies,viz. S. cohnii ssp.cohnii andS. cohnii ssp.urealyticus. Strains were isolated from different sources of theIntensive Care Unit and from non-hospital environment. Most of the strains were multiple antibiotic-resistant. Strains of both subspecies revealed a relatively low iron requirement. These strains were capable of utilizing iron bound in oxo acids and from host iron-binding proteins.S. cohnii ssp.urealyticus were more effective in iron uptake thanS. cohnii ssp.cohnii. All investigated strains revealed sensitivity to skin fatty acids, butS. cohnii ssp.urealyticus strains were more resistant. Special features of strains of this subspecies promote colonization of humans.     

<Emphasis Type="Italic">Staphylococcus cohnii</Emphasis> hemolysins — Isolation,purification and properties

A total 355 of Staphylococcus cohnii isolates from hospital environment, patients (newborns), medical staff and from non-hospital environment were tested for hemolytic activity. Ninety-one % of S. cohnii ssp. cohnii and 74.5 % S. cohnii ssp. urealyticus strains exhibited hemolysis synergistic to S. aureus ATCC 25923 strain. Crude preparations of hemolysins of both bacterial subspecies presented δ-hemolysin, but not α- and β-toxin activity. Highly pure hemolysins were obtained by semipreparative SDS-PAGE or by organic solvent extraction from the freeze-dried crude preparations. Native-PAGE and 2D-PAGE showed their high heterogeneity. Molar masses of single hemolysin units estimated by the Tris-Tricine-SDS-PAGE were calculated as 3.47 kDa for S. cohnii ssp. cohnii and 3.53 kDa for S. cohnii ssp. urealyticus.     

A PCR-ELISA Method for Direct Detection of the Oyster Pathogen Haplosporidium nelsoni

A rapid method, utilizing both polymerase chain reaction (PCR) and enzyme-linked immunosorbent assay (ELISA), was developed for detection of oyster MSX disease. The technique included using Haplosporidium nelsoni pathogen-specific PCR primers (based on ribosomal RNA genes), a Chelex resin (for rapid DNA extraction from oyster mantle tissues), and cloned H. nelsoni rRNA plasmid DNA (for use as a capture probe). Digoxigenin was incorporated into the pathogen-specific PCR products, which were captured by the coated probe in a fast hybridization reaction and then detected by ELISA. The sensitivity of PCR amplification on cloned plasmid DNA was 10 fg for detection by stained agarose gel, and increased to 0.01 fg for ELISA. Positive signals were observed in infected oysters using the PCR-ELISA technique. This method may be applicable to early detection of infection. Received April 14, 1998; accepted September 30, 1998.     

Nuclear and plastid DNAs from the binucleate dinoflagellates Glenodinium (Peridinium) foliaceum and Peridinium balticum

The binucleate dinoflagellates Glenodinium (Peridinium) foliaceum Stein and Peridinium balticum (Levander) Lemmermann were found to contain two major buoyant density classes of DNA. The heavier peak (1.730 g/cm3) was derived from the "dinokaryotic" nucleus and the lighter peak (1.706 g/cm3) from the "endosymbiont" nucleus and this allowed for the fractionation of G. foliaceum DNA in CsCl/EtBr density gradients. An initial CsCl/Hoechst Dye gradient removed a minor A-T rich satellite species which was identified as plastid DNA with a size of about 100-106 kb. Analysis of the nuclear DNA by agarose gel electrophoresis and renaturation studies showed that the endosymbiont nucleus lacked amplified gene-sized DNA molecules, however, this nucleus did have a comparatively high level of DNA. The total amount of DNA per cell and the relative contributions of the two nuclei appeared to vary between two strains of G. foliaceum (75 pg/cell in CCAP strain and 58 pg in UTEX strain). The only strain of P. balticum examined contained 73 pg cell. These results are discussed in relation to the status and possible functioning of the endosymbiont nucleus and the idea that these dinoflagellates provide model systems with which to study the evolution of plastids.     

The effects of oils and oil components on algae: A review

Plate structure analysis of the bloom-forming Peridinium of Lake Kinneret (previously identified as P. cinctum fa. westii) established its identity as P. gatunense. Peridinium cinctum was not observed. Material from different years and periods of the bloom, as well as specimens from cultures and P. gatunense from Lago Cristalino (Brazil), were studied using light and scanning electron microscopy. Peridinium gatunense from Lake Kinneret showed slight differences of the plate pattern as compared with specimens from other localities.     

ISOLATION OF ALGAL GENES BY FUNCTIONAL COMPLEMENTATION OF YEAST1

Algal cDNAs were isolated and characterized by functional complementation of yeast auxotrophs. Two cDNA libraries, one derived from the diatom Phaeodactylum tricornutum Bohlin and the other from the dinoflagellate Crypthecodinium cohnii Biecheler, were constructed using the Saccharomyces cerevisiae expression vector pFL‐61. These libraries were used for functional complementation of auxotrophic markers in two yeast strains. Yeast tryptophan auxotrophs, complemented by the P. tricornutum library, contained a plasmid that encoded a two‐domain protein associated with tryptophan synthesis, indole‐3‐glycerol phosphate synthase‐N‐(5′‐phosphoribosyl) anthranilate isomerase. Another cDNA originating from the C. cohnii library rescued S. cervisiae from a defect in adenine biosynthesis. This cDNA encoded a fusion of phosphoribosylamidoimidazole‐succinocarboxamide synthetase and phosphoribosylaminoimidazole carboxylase, which correspond to the yeast ade1 and ade2 genes, respectively. These results demonstrate that heterologous functional complementation can be used to identify algal genes and may provide advantages over other gene discovery methods.     

Replication of Nuclei from Cycling and Quiescent Mammalian Cells in 6-DMAP-TreatedXenopusEgg Extract

Nuclear membrane permeabilization is required for replication of quiescent (G0) cell nuclei inXenopusegg extract. We now demonstrate that establishment of replication competence in G0 nuclei is dependent upon a positive activity present in the soluble egg extract. Our hypothesis is that G0 nuclei lose the license to replicate following growth arrest and that this positive activity is required for relicensing DNA for replication. To determine if G0 nuclei contain licensed DNA, we used the protein kinase inhibitor, 6-dimethylaminopurine (6-DMAP), to prepare egg extracts that are devoid of licensing activity. Intact nuclei, isolated from mammalian cells synchronized in G1-phase (licensed), G2-phase (unlicensed), and G0 were permeabilized and assayed for replication in 6-DMAP-treated and untreated extracts supplemented with [α-³²P]dATP or biotinylated-dUTP. Very little radioactivity was incorporated into nascent DNA in each nuclear population; however, nearly all nuclei in each population incorporated biotin in 6-DMAP extract. The pattern of biotin incorporation within these nuclei was strikingly similar to the punctate pattern observed within nuclei incubated in aphidicolin-treated extract, suggesting that initiation events occur within most replication factories in 6-DMAP extract. However, density substitution and alkaline gel analyses indicate that the incorporated biotin within these nuclei arises from a small number of active origins which escape 6-DMAP inhibition. We conclude that 6-DMAP-treated egg extract cannot differentiate licensed from unlicensed mammalian somatic cell nuclei and, therefore, cannot be used to determine the “licensed state” of G0 nuclei using the assays described here.     

The mesokaryote Gyrodinium cohnii lacks nucleosomes

The dinoflagellate $\text{Gyrodinium}\text{cohnii}$ has a distinct nuclear membrane but apparently lacks histones associated with its chromatin. Approximately 13% of the nuclear DNA is rapidly digested by micrococcal nuclease to acid soluble fragments and not to nucleosomal sized fragments as in the typical eukaryote. Moreover in the electron microscope the chromatin of $\text{G.}\text{cohnii}$ appears as a thin filament of 40–60 Å in width without regularly spaced nucleosomes. These observations support the view that the dinoflagellates exhibit characteristics of both prokaryotes and eukaryotes.     

Über den Einfluß von Rotschlamm auf die Kultur einiger mariner Planktonalgen

Zusammenfassung 1. Rotschlamm ist ein bei der Aluminiumgewinnung aus Bauxit anfallendes Abfallprodukt. Der Antrag eines Industrieunternehmens auf Verklappung von ca. 800 000 t Rotschlamm (Naßgewicht) pro Jahr in der Nordsee gab den unmittelbaren Anlaß zur Aufnahme von Laborversuchen über die Wirkung auf marine Planktonalgen. Als Testformen dienten die DinoflagellatenPeridinium trochoideum, Gymnodinium splendens undProrocentrum micans sowie die DiatomeenCoscinodiscus granii undChaetoceros socialis. Die Algen wurden sowohl im Batch-Verfahren als auch im kontinuierlichen Verfahren nach dem Turbidostatprinzip gezüchtet.2. Rotschlamm wurde den Kulturen im Batch-Verfahren einmalig bei Versuchsbeginn fein suspendiert zugesetzt und je nach Versuchsanordnung entweder der Sedimentation überlassen oder durch regelmäßiges Aufschütteln bzw. vermittels eines Rührwerkes soweit wie möglich in Turbulenz gehalten. Im kontinuierlichen Verfahren wurde den Kulturen während der gesamten Versuchsdauer täglich frischer Rotschlamm zudosiert und der Sedimentation überlassen. Die Menge des zugegebenen Rotschlammes variierte je nach Versuchsanordnung zwischen 0,001 und 50 g(Naßgewicht)/l Seewasser.3. Als Kriterien einer Schädigung dienten die Beeinflussung der Vermehrungsrate der Algen und die maximal erreichbare Zelldichte der Kulturen. Zugleich wurden die letalen Grenzkonzentrationen bestimmt.4. Die Testversuche zeigten, daß die Empfindlichkeit gegenüber Rotschlamm wesentlich von der verwendeten Algenart und von der jeweiligen Versuchsanordnung abhängt. Reversible Anfangsschädigungen wurden im Batch-Verfahren bei 0,005 g/l(Coscinodiscus granii), 0,01 g/l(Gymnodinium splendens), 0,05 g/l(Peridinium trochoideum undProrocentrum micans) und 0,5 g/l(Chaetoceros socialis) nachgewiesen. Irreversible Schädigung führte im kontinuierlichen Verfahren bei täglicher Zudosierung des Rotschlammes in Mengen von 0,01 g/l(Peridinium trochoideum), 0,05 g/l(Coscinodiscus granii) und 0,5 g/l(Prorocentrum micans) zum Absterben der Kulturen.5. Die Anfangsschädigung ist in erster Linie auf mechanische Wirkung unmittelbar nach Zugabe des frischen Rotschlammes zum Meerwasser zurückzuführen. Die Partikel sedimentieren unter lockerer Koagulation und reißen die Algen mit sich. Gealterte Rotschlammpartikel erweisen sich als weniger schädlich. In höheren Konzentrationen kann eine toxische Wirkung hinzukommen (0,5 g/l Rotschlamm beiGymnodinium splendens), die sich jedoch erst nach längerer Versuchsdauer bemerkbar macht.6. Da die verwendeten Testalgen den relativ robusten Planktonformen zugehören, ist bei einer Übertragung der Ergebnisse auf die in See eintretende Gesamtsituation damit zu rechnen, daß viele andere Planktonalgen gegenüber einer Rotschlammverklappung noch empfindlicher reagieren.7. In einer Literaturübersicht werden die von anderen Autoren an repräsentativen Gliedern der Nahrungskette (Algen, Fischnährtiere und Fische) ermittelten wichtigsten Ergebnisse von Rotschlammversuchen kurz referiert und den vorliegenden Ergebnissen gegenübergestellt. In einer gemeinsamen Stellungnahme mehrerer bundesdeutscher Institute wird aus den angeführten Untersuchungen die Schlußfolgerung gezogen, daß einer Einbringung von Rotschlamm in die Nordsee nicht zugestimmt werden kann.

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On the effect of red mud on the culture of some marine plankton algae

Red mud originates from bauxite processing, the first step of aluminium production. A newly constructed plant intends to release about 800 000 tons of this waste product into the southern North Sea. The effect of red mud on the culture of some marine algae was investigated. The dinoflagellatesPeridinium trochoideum, Prorocentrum micans andGymnodinium splendens, and the diatomsCoscinodiscus granii andChaetoceros socialis served as test organisms. They were examined in batch cultures and in continuous cultures (turbidostats) to which 0.001 to 50 g of red mud per 1 sea water was added. Multiplication rates of the algae were chosen as criterion for assessing the influence of red mud. Maximum cell densities of the batch cultures were also determined. When red mud was added to the batch cultures once, at the beginning of the experiments, the test algae exhibited initial reductions in population growth at 0.005 up to 0.5 g of red mud/l sea water, depending on the species. In most experiments they recovered from initial growth-rate reduction and grew to nearly the same cell densities as did the controls. In the continuous cultures, suspended red mud was added daily. After 9 days, this caused irreversible break-down ofPeridinium trochoideum populations at 0.01 g red mud/l/day.Coscinodiscus granii populations were irreversibly damaged at 0.05 g/l/day;Prorocentrum micans, at 0.5 g/l/day. All test algae represent euryplastic forms. Other, more stenoplastic planktonic algae species are likely to be less tolerant to red mud exposure.

     

Karyology and cytophotometric estimation of nuclear DNA content variation in Gracilaria,Gracilariopsis and Hydropuntia (Gracilariales,Rhodophyta)

Chromosome numbers of 1 N=24 were determined for three species of Gracilaria (G. flabelliforme P. Crouan et H. Grouan ex Schramm et Maze, G. mammillaris Montagne and G. tikvahiae McLachlan) and 1 N=32 for two species of Gracilariopsis (G. lemaneiformis (Bory) Dawson, Acleto et Folvik and G. tenuifrons (Bird et Oliveira) Fredericq et Hommersand). Karyotypes for these species exhibit a characteristic size difference between largest and smallest chromosomes. Polyvalents were a common feature of meiotic nuclei. Microspectrophotometry with the DNA-localising fluorochrome DAPI was used to quantify nuclear genome sizes. A 2 C genome size of 0·37–0·40 pg was determined for five species of Gracilaria (G. chilensis Bird, McLachlan et Oliveira, G. flabelliforme, G. mammillaris, G. pacifica Abbott, G. tikvahiae) and 0·33 pg for an isolate of G. verrucosa (Hudson) Papenfuss from Pas de Calais, France. Species of Hydropuntia (H. cornea (J. Agardh) Wynne and H. dentata (J. Agardh) Wynne) and Gracilariopsis (G. lemaneiformis and G. tenuifrons) were found to have slightly larger 2 C genome contents of 0·42–0·47 pg. No intraspecific variation in 2 C genome sizes was found in regional populations of Gracilaria tikvahiae and Gracilariopsis tenuifrons.