Isolation of the major basic nuclear protein and its localization on chromosomes of the dinoflagellate,Oxyrrhis marina

Basic nuclear proteins were extracted from isolated nuclei of Oxyrrhis marina. The HPLC pattern of the extract showed a single major peak, which consisted of a major band with an apparent molecular mass of 23 kDa on an SDS-PAGE gel. We designated this protein as Np23 because of its apparent molecular mass. The amino acid composition of this protein revealed its extremely basic nature with a high lysine content. Polyclonal antibodies were raised against Np23. Immunofluorescence microscopy showed that Np23 was localized within the nucleus of dividing and non-dividing cells as well, and immuno-electron microscopy showed that the protein was localized only on chromosomes. These data established that Np23 is the major basic chromosome protein of Oxyrrhis marina.     

Identification of the nuclear matrix and chromosome scaffold in dinoflagellate Crypthecodinium cohnii~1

Dinoflagellate is one of the primitive eukaryotes,whosenucleus may represent one of the transition stages fromprokaryotic nucleoid to typical eukaryotic nucleus.Usingselective extraction together with embeddment-free sectionand whole mount electron microscopy,a delicate nuclearmatrix filament network was shown,for the first time,indinoflagellate Crypthecodinium cohnii nucleus.Chromosomeresidues are connected with nuclear matrix filaments to forma complete network spreading over the nucleus.Moreover,we demonstrated that the dinoflagellate chromosome retainsa protein scaffold after the depletion of DNA and solubleproteins.This scaffold preserves the characteristic mor-phology of the chromosome.Two dimensional elec-trophoreses indicated that the nuclear matrix and chromo- some scaffold are mainly composed of acidic proteins.Ourresults demonstrated that a framework similar to the nuclearmatrix and chromosome scaffold in mammalian cells appearsin this primitive eukaryote,suggesting that these structuresmay have been originated from the early stages of eukaryoteevolution.     

Dehydrodinosterol,dinosterone and related sterols of a non-photosynthetic dinoflagellate, Crypthecodinium cohnii

The heterotrophic dinoflagellate Crypthecodinium cohnii contained the 4α-methyl sterols, dinosterol, dehydrodinosterol (4α,23,24-trimethylcholesta-5,22-dien-3β-ol) and the tentatively identified 4α,24-dimethyl-cholestan-3β-ol and 4α,24-dimethylcholest-5-en-3β-ol. The major 4-demethyl sterol was cholesta-5,7-dien-3β-ol which was accompanied by a smaller amount of cholesterol and traces of several other C₂₇,C₂₈ and C₂₉ sterols. In addition, a 3-oxo-steroid fraction was isolated and the major component identified as dinosterone (4α,23,24-trimethylcholest-22-en-3-one). The possible biosynthetic relationships of these compounds are discussed.     

An enigmatic GAPDH gene in the symbiotic dinoflagellate genus Symbiodinium and its related species (the order Suessiales): possible lateral gene transfer between two eukaryotic algae, dinoflagellate and euglenophyte

A group of unicellular eukaryotic algae, the dinoflagellates, are known to possess two types of gene for glyceraldehyde-3-phosphate dehydrogenase (GAPDH). An enzyme encoded by one type of gene possibly plays a key role in the glycolytic pathway of the cytosol and the other in the Calvin cycle of plastids. In the present study, an additional type of GAPDH gene (GapC3) was found in the symbiotic dinoflagellates, Symbiodinium spp. and their related species, Gymnodinium simplex and Polarella glacialis, all of which belong to the order Suessiales. Since no intracellular translocation signal is found at both amino- and carboxy-termini of its deduced amino acid sequence, the protein is predicted to function in the cytosol. However, it may not be involved in glycolysis due to the presence of an amino acid signature that allows binding for NADP⁺. It is likely that dinoflagellate species, other than Suessiales investigated in this study, lack this type of GAPDH. Phylogenetic analysis placed GapC3 from the Suessialean species firmly in the clade composed of GAPDH from spirochetes, euglenophytes (cytosolic type) and kinetoplastids (glycosomal type). Specifically, this enigmatic GAPDH gene in dinoflagellates was closely related to its cytosolic counterpart in euglenophytes. It has been previously reported that plastid-targeted (Calvin cycle) GAPDH genes of the dinoflagellates Pyrocystis spp. and that of the euglenophyte Euglena gracilis also seem to share a common ancestor. It appears highly likely that at least two genes (cytosolic and plastid-targeted GAPDH genes) have been laterally transferred between these two eukaryotic algal groups.     

Implications of life-history transitions on the population genetic structure of the toxigenic marine dinoflagellate Alexandrium tamarense

Genotypic or phenotypic markers for characterization of natural populations of marine microalgae have typically addressed questions regarding differentiation among populations, usually with reference to a single or few clonal isolates. Based upon a large number of contemporaneous isolates from the same geographical population of the toxigenic species Alexandrium tamarense from the North Sea, we uncovered significant genetic substructure and low but significant multilocus linkage disequilibrium (LD) within the planktonic population. Between the alternative molecular genotyping approaches, only amplified fragment length polymorphism (AFLP) revealed cryptic genetic population substructure by Bayesian clustering, whereas microsatellite markers failed to yield concordant patterns. Both markers, however, gave evidence for genetic differentiation of population subgroups as defined by AFLP. A considerable portion of multilocus LD could be attributed to population subdivision. The remaining LD within population subgroups is interpreted as an indicator of frequency shifts of clonal lineages during vegetative growth of planktonic populations. Phenotypic characters such as cellular content and composition of neurotoxins associated with paralytic shellfish poisoning (PSP) and allelochemical properties may contribute to intra- or inter-annual differentiation of planktonic populations, if clonal lineages that express these characters are selectively favoured. Nevertheless, significant phenotypic differentiation for these characters among the genetically differentiated subgroups was only detected for PSP toxin content in two of the four population subgroups. By integrating the analysis of phenotypic and genotypic characteristics, we developed a conceptual population genetic model to explain the importance of life-cycle dynamics and transitions in the evolutionary ecology of these dinoflagellates.     

Genome size and determination of DNA content of the X chromosomes, autosomes, and germ line-limited chromosomes of Sciara coprophila

The unique chromosome biology of the fungus fly Sciara coprophila has fascinated investigators for over 80 years. Male meiosis exhibits a monopolar spindle, nonrandom segregation of imprinted chromosomes and nondisjunction of the X chromosome. The unusual mechanism of sex determination requires selective elimination of X chromosomes in embryogenesis. Supernumerary (L) chromosomes are also eliminated from the soma during early cleavage divisions. Distinctive DNA puffs on the larval salivary gland chromosomes are sites of DNA amplification. As a foundation for future genome studies to explore these many unusual phenomena, we have used DNA-Feulgen cytophotometry to determine genome size from hemocyte nuclei of male (X0) and female (XX) larvae and adults. The DNA content of the X chromosome is approximately 0.05 pg DNA and the autosomal complement is approximately 0.45 pg DNA. Measurements of DNA levels for individual sperm from adults showed that the DNA contribution of the germ line-limited (L) chromosomes constitutes as much as 35% of the DNA of the male gamete. A parallel study using Sciara ocellaris, a related species lacking L chromosomes, confirmed the presence of two X chromosomes in the sperm of this species.     

Population genetic structure and connectivity of the harmful dinoflagellate Alexandrium minutum in the Mediterranean Sea

The toxin-producing microbial species Alexandrium minutum has a wide distribution in the Mediterranean Sea and causes high biomass blooms with consequences on the environment, human health and coastal-related economic activities. Comprehension of algal genetic differences and associated connectivity is fundamental to understand the geographical scale of adaptation and dispersal pathways of harmful microalgal species. In the present study, we combine A. minutum population genetic analyses based on microsatellites with indirect connectivity (C(i)) estimations derived from a general circulation model of the Mediterranean sea. Our results show that four major clusters of genetically homogeneous groups can be identified, loosely corresponding to four regional seas: Adriatic, Ionian, Tyrrhenian and Catalan. Each of the four clusters included a small fraction of mixed and allochthonous genotypes from other Mediterranean areas, but the assignment to one of the four clusters was sufficiently robust as proved by the high ancestry coefficient values displayed by most of the individuals (>84%). The population structure of A. minutum on this scale can be explained by microalgal dispersion following the main regional circulation patterns over successive generations. We hypothesize that limited connectivity among the A. minutum populations results in low gene flow but not in the erosion of variability within the population, as indicated by the high gene diversity values. This study represents a first and new integrated approach, combining both genetic and numerical methods, to characterize and interpret the population structure of a toxic microalgal species. This approach of characterizing genetic population structure and connectivity at a regional scale holds promise for the control and management of the harmful algal bloom events in the Mediterranean Sea.     

Protoperidinium tricingulatum sp. nov. (Dinophyceae), a new motile form of a round,brown, and spiny dinoflagellate cyst

A small, broadly ovoidal and heterotrophic dinoflagellate containing round, brownish, and spiny cyst was found in the water column of Huibertsplaat in the Wadden Sea off the coast of the Netherlands. This dinoflagellate had these conspicuous morphological characters: a five‐sided first apical plate (1′), only three cingular plates, and an extremely small first antapical plate. Based on these morphological features, Protoperidinium tricingulatum Kawami, vanWezel, Koeman et Matsuoka is described as a new species. The flagellar pore of P. tricingulatum is covered with a small fin, which rises from the left side of the right sulcal plate to the large V‐shaped posterior sulcal plate. This feature suggests that P. tricingulatum is assigned to the Abé's Monovela Group. The cyst stage of P. tricingulatum was positively linked to the vegetative stage by comparison of the ribosomal 5.8S rDNA, internal transcribed spacers (ITS1 and ITS2). Living cysts of P. tricingulatum are round, brownish, and covered with many slender spines bearing capitate or cauliforate distal ends. The cyst also possesses a theropylic archeopyle formed by a slit corresponding to parasutures between three apical and two apical intercaraly plates. These morphological characters indicate that this species is morphologically related to two dinoflagellate cyst‐genera Islandinium and Echinidinium.     

The effects of continuous illumination on the function and structure of Micrasterias torreyi Bail. (Conjugatophyceae)

Abstract. The division rate of Micrasterias torreyi cells grown under continuous illumination first accelerated but soon slowed down, and the cells lost their ability to divide after about 1 month. During the treatment the cells became pale green, the pyrenoids became fewer in number and defects appeared in the chloroplasts. After 1 month, the cells also soon died, even when subjected to intermittent illumination. The most striking structural alterations were found in the chloroplasts: the starch granules lost their typical structure, the lamellae were damaged and numerous electron dense precipitates appeared in the chloroplasts. The precipitates were similar to those formed in cells treated with supraoptimal external calcium concentrations and X-ray microanalysis showed that the precipitates were rich in calcium in both cases. The results suggest that light controls and activates the Ca²⁺ uptake in the plasma membrane as well as in the chloroplast envelope, that the large sized chloroplasts of Micrasterias are effective in regulation of cytoplasmic Ca²⁺ concentration, and that the injuries caused by continuous illumination may be largely due to the accumulation of Ca²⁺ in the chloroplasts.     

Nutritional enrichment of a microbial community: The effects on activity, elemental composition, community structure and virus production

Abstract Viruses are active members of the microbial community in natural waters but little is known about the factors that regulate their activity and production. In this study we have investigated the effects of increased availability of organic nutrients and inorganic phosphate on activity, elemental composition, community structure and virus production in a natural bacterial community. The fraction of active cells in the community as estimated from microautoradiography of cells assimilating ³H-labeled thymidine ranged from 0–22%, but changes in the elemental composition of the cells indicated that more than 90% of the cells were active. The increase in carbon and energy availability stimulated virus production more than bacterial biomass production, while the increase in phosphate availability stimulated biomass production rather than virus production. A decrease in morphological diversity of the bacterial community was paralleled by a reduction in the virus-to-bacteria ratio (VBR) but the relationship between bacterial diversity and viral activity is uncertain. Our general conclusion is that nutrient availability, in addition to the bacterial activity, also affects the viral activity, and that both of these may affect the structure and diversity of the bacterial community.     

Variation in the distribution of a genome-specific DNA sequence on chromosomes reveals evolutionary relationships in the Triticum and Aegilops complex

 The present study analyzed the distribution pattern of the Ae. speltoides–derived repetitive clone pGc1R-1 in the Triticum/Aegilops complex. Fluorescence in situ hybridization analysis showed that clone pGc1R-1 is a S-genome-specific repetitive sequence that hybridized to the S-genome of three species in the section Sitopsis, Aegilops speltoides (S), Ae. longissima (S^l), and Ae. sharonensis (S^sh), but not to Ae. bicornis (S^b) and Ae. searsii (S^s), nor to any other diploid Aegilops species. This clone also hybridized to the very closely related G-genome of T. timopheevii subsp. armeniacum and T. timopheevii ssp. timopheevii, but not to the B-genome of T. turgidum and T. aestivum. Hybridization also was observed in the polyploid Aegilops species, Ae. kotschyi (U^kS^k), Ae. peregrina (U^pS^p), and Ae. vavilovii (X^vaD^vaS^va). Large inter- and intraspecific variations were observed. Our results confirm that the S genome is related more to the S^l and S^sh genomes than to the S^b and S^s genomes; there is a greater affinity between the G and S genomes than between the B and S genomes. Mechanisms to account for the variation in the FISH pattern with different genomes include sequence amplification and deletion. Variation in the distribution of this genome-specific DNA sequence, pGc1R-1, on chromosomes can be used to reveal evolutionary relationships in the Triticum and Aegilops complex. Received April 10, 2002; accepted July 12, 2002 Published online: November 28, 2002 Address of the authors: Peng Zhang, Bernd Friebe (e-mail: friebe@ksu.edu), Bikram S. Gill, Wheat Genetics Resource Center, Department of Plant Pathology, 4024 Throckmorton, Plant Sciences Center, Kansas State University, Manhattan, KS 66506-5502, USA.     

Fine structure observations on the chromosomes in the spermatids of the Ascidian,Boltenia villosa

Electron micrographs of the relatively young spermatids of the Ascidian, Boltenia villosa, reveal that its chromosomes exist in the form of 120–180 Å fibers which are easily seen to be composed of two subunits of about 50–70 Å each. Fibers of the diameter of such subunits have never been seen as single entities without a mate. Subunits in pairs seen imbedded in areas of fused chromosomes only measure 30–40 Å, probably due to the loss of some protein. It is pointed out that the chromatin goes into the sperm as chromosomes in two-chromatid state, and that each chromatid probably contains only one Watson-Crick double helix.     

Mosaic structure of the DNA molecules of the human chromosomes 21 and 22

We calculated nucleotide distribution curves along the DNA molecules of the human chromosomes 21 and 22, their correlations in more than 10000 equidistant positions, and subjected the correlations to cluster analysis. The cluster analysis demonstrated that both DNA molecules were composed of two types of segments exhibiting qualitatively different correlations. The segments differed most in the correlation of the distribution curves of cytosine and guanine, which was very high in type I segments but weak in type II segments. The type I and II segments also significantly differed in the correlations of the distribution curves of adenine with thymine. In addition, adenine strongly anticorrelated with cytosine but this anticorrelation was uniform along both chromosomes and, therefore, it did not contribute to the distinction of the two types of segments. The segments were up to 100 kbp long but they had nothing in common with isochores. Building blocks of the mosaic structure of the DNA molecules of the human chromosomes 21 and 22 are very similar but different in several interesting aspects from those of E. coli.     

Karyotype structure, supernumerary chromosomes and heterochromatin distribution suggest a pathway of karyotype evolution in Dactylorhiza (Orchidaceae)

The relationships between Dactylorhiza romana and D. saccifera from southern Italy were analysed. These two species, both with 2 n = 2 x = 40 chromosomes and belonging to different sections of the genus, were distinguishable on the basis of karyotype structure and heterochromatin amounts and distribution. Their C-banded karyotypes differed considerably. D. saccifera showed most chromosomes with banded regions in the short arms, whereas in D. romana the bands were located mostly at telomeric regions of longer arms. Several individuals of D. romana had one or two very large heterochromatic supernumerary chromosomes. Based on evidence resulting from karyotype structure and heterochromatin distribution in the two species and on the genetic distances derived from the comparison of ITS sequences, it is suggested that D. romana represents a primitive form with respect to D. saccifera and is a possible intermediate step in the evolution of the genus Dactylorhiza from the 42-chromosome Orchis group. © 2002 The Linnean Society of London, Botanical Journal of the Linnean Society , 138 , 85–91.