Ultrastructure of the flagellar apparatus in the green flagellateSpermatozopsis similis

The ultrastructure of the flagellar apparatus of the naked, biflagellate green algaSpermatozopsis similis Preisig & Melkonian has been studied in detail using an absolute configuration analysis. The two basal bodies are displaced by 350 nm in the 1/7 o'clock direction and do not overlap proximally. They are interconnected by a principal distal connecting fibre consisting of a bundle of 5–8 nm filaments and possibly two proximal striated connecting fibres. The flagellar root system is cruciate (5-2-5-2 or 4-2-4-2 system) and contains a prominent continuous system I fibre overlying the two opposite two-stranded roots. A system II fibre is absent. Pronounced structural differences have been observed in the flagellar apparatus ultrastructure at two types of flagella orientation: During backward swimming basal bodies are parallel, the distal connecting fibre is extremely contracted; during forward swimming basal bodies assume various angles (from 20° to 180°) and the connecting fibre is about five times longer compared to the contracted state. The function of the connecting fibre as a contractile organelle and the mechanism of its contraction are discussed. On the basis of the flagellar apparatus ultrastructure,Spermatozopsis similis is related toChlamydomonas-type green algae.     

Flagellar apparatus ultrastructure inMesostigma viride (Prasinophyceae)

The ultrastructure of the flagellar apparatus ofMesostigma viride Lauterborn (Prasinophyceae) has been studied in detail with particular reference to absolute configurations, numbering of basal bodies, basal body triplets and flagellar roots. The two basal bodies are interconnected by three connecting fibers (one distal fiber = synistosome, and two proximal fibers). The flagellar apparatus shows 180° rotational symmetry; four microtubular flagellar roots and two system II fibers are present. The microtubular roots represent a 4-6-4-6-system. The left roots (1s, 2s) consist of 4 microtubules, each with the usual 3 over 1 root tubule pattern. Each right root (1d, 2d) is proximally associated with a small, but typical multi-layered structure (MLS). The latter displays several layers corresponding to the S₁ (the spline microtubules: 5–7), and presumably the S₂—S₄ (the lamellate layers) of the MLS of theCharophyceae. At its proximal origin (near the basal bodies) each right root originates with only two microtubules, the other spline microtubules being added more distally. The structural and positional information obtained in this study strongly suggest that one of the right roots (1d) ofMesostigma is homologous to the MLS-root of theCharophyceae and sperm cells of archegoniate land plants. Thus the typical cruciate flagellar root system of the green algae and the unilateral flagellar root system of theCharophyceae and archegoniates share a common ancestry. Some functional and phylogenetic aspects of MLS-roots are discussed.Dedicated to Prof. DrLothar Geitler on the occasion of his 90th birthday.     

Fine structure of the flagellar apparatus of gametesin situ and motile zygotes of the green algaUlothrix flacca var.Roscoffensis (Ulotrichales) (Chlorophyta)

Summary This fine structural study ofUlothrix flacca (Dillw.) ThuretRoscoffensis variety (Berger-Perrot), a marineUlothrix, describes in detail the flagellar apparatus configuration of gametesin situ in the gametangia and in motile zygotes. The gametes's flagellar apparatus shows two basal bodies overlapping at their proximal end at a 30° angle, in an 11/5 o'clock configuration or with a counterclockwise absolute orientation. The basal bodies are interconnected by a non-striated band or capping plate. They are wrapped in their proximal part by an electron-dense sheath and obtured by a bilobed terminal cap. A cruciate microtubular root system having a 4-2-4-2 alternation pattern is present. A striated microtubule associated component (S.M.A.C.) or system I fibres accompany the two membered root R₂. The system II fibres or rhizoplasts along with striated bands associated to the microtubular roots, were not observed and are presumed to be absent.In the motile zygotes, the basal bodies are paired in a cruciate pattern. During the fusion process, two basal bodies, one of each pair, slide in a face to face position with a slight displacement into the 11/5 o'clock direction; the other two make a 30° counterclockwise rotation, thus making a 60° angle between the two basal bodies of each pair instead of 30° in the gamete.After comparison with the flagellar apparatus of other green alga gametes, it is concluded that the taxonomic affinities ofUlothrix flacca var.Roscoffensis, lie with theUlvophyceae sensuStewart andMattox 1978.Abbreviations CP capping plate - ER endoplasmic reticulum - G Golgi body - LG lipid globule - M mitochondria - MS presumed mating structure - N nucleus - R ₂,R ₄ microtubular roots - SH sheath - SMAC striated microtubule associated component - TC terminal cap - V vacuole - Ve vesicles in the anterior papilla - 1, 2, 1, 2 basal bodies numerotation     

Comparative cytology and taxonomy of theUlvaphyceae II. Ulvalean characteristics of the stephanokont flagellar apparatus ofDerbesia tenuissima

Summary The stephanokont flagellar apparatus of the zoospores ofDerbesia tenuissima (De Not.) Crouan is examined and compared to the flagellar apparatuses of other green algae. The flagella ofDerbesia are attached to two of three bands which lie at the junction of the body and papilla. Serial longitudinal and cross sections reveal that the basal bodies are attached to the bands along their sides and at their proximal ends. The bands are not striated in any plane. The lack of striation in the bands and the partial covering of the proximal end of the basal bodies by one of the bands closely resemble the type of flagellar connection system described as the Bryopsis-type byMelkonian (1980). Zoospores of ulvalean green algae also possess these features, suggesting that green siphons are phylogenetically related to theUlvales. It is proposed that green siphons be tentatively classified in theUlvaphyceae rather than in theChlorophyceae orCharophyceae.This work supported by NSF Grant DEB 78-03554.     

Stable expression of hepatitis B surface antigen gene in Dunaliella salina (Chlorophyta)

A stable transformation system for theexpression of foreign genes in theunicellular marine green alga Dunaliella salina was established. Amongfive antibiotics, 60 g mL^-1chloramphenicol completely inhibitedgrowth. Of five promoters tested, theubiquitin-$Ohgr; promoter yielded thehighest -glucuronidase (GUS) activity.The hepatitis B surface antigen (HBsAg)gene was introduced into the cells by usingelectroporation. PCR and Southern blotanalysis amd it was shown that the gene wasintegrated into the genome. The stableexpression of HBsAg protein was confirmedby enzyme-linked immunosorbent assay(ELISA) and Western blot analysis. Theintroduced DNA and HBsAg expression weremaintained stable for at least 60generations in medium devoid ofchloramphenicol. This is an important steptoward the production of useful foreignproteins in the alga.     

Synthesis of an H type 2 and a Y (Ley) glycoside from thioglycoside intermediates

The trisaccharide 2-(p-trifluoroacetamidophenyl)ethyl 2-acetamido-2-deoxy-4-O-[2-O-(-l-fucopyranosyl)--d-galactopyranosyl]--d-glucopyranoside 1 and the tetrasaccharide 2-(p-trifluoroacetamidophenyl)ethyl 2-acetamido-2-deoxy-3-O-(-l-fucopyranosyl)-4-O-[2-O-(-l-fucopyranosyl)--d-galactopyranosyl]--d-glucopyranoside 2 were synthesized. Thioglycosides, suitably protected, activated directly with methyl trifluoromethanesulfonate or dimethyl(methylthio)sulfonium tetrafluoroborate or activated after bromine treatment with halophilic reagents, were used as glycosyl donors in the construction of the glycosidic linkages.Abbreviations DMTSB dimethyl(methylthio)sulfonium tetrafluoroborate - Phth phthaloyl - MBn p-methoxybenzyl - ClBn p-chlorobenzyl     

Glycosidations with thioglycosides activated by sulfuryl chloride/trifluoromethanesulfonic acid: synthesis of a human blood group B trisaccharide glycoside

The trisaccharide 2-(p-trifluoroacetamidophenyl)ethyl 2-O-(-l-fucopyranosyl)-3-O-(-d-galactopyranosyl)--d-galactopyranoside, corresponding to the human blood group B determinant, was synthesized. Thioglycosides activated by sulfuryl chloride/trifluoromethanesulfonic acid were used as glycosyl donors in the construction of the three glycosidic linkages.     

Growth and ultrastructure of the marine unicellular alga <Emphasis Type="Italic">Dunaliella salina</Emphasis> (Chlorophyta) after chronic selenium intoxication

The effects of selenium (0.01, 0.5, 1, 5 and 10 mg/liter) on the growth and ultrastructure of the microalga Dunaliella salina were investigated following its transfer into clean water. Selenium concentrations of 5 and 10 mg/liter were toxic to D. salina, and reinoculation of microalga into clean water did not prevent it from total mortality. When reinoculated from medium with 0.01 mg Se/liter, the cell population density of D. salina was restored in 14 days. The number of ultrastructural alterations in cells was the same as in the control, while the excretory activity of microalga between days 4 and 10 of this experiment was higher. Cell population growth of D. salina transferred from 0.5 and 1 mg Se/liter was lower than in the control. No ultrastructural defects were observed in microalga reinoculated from medium with a selenium concentration of 0.5 mg/liter and the excretion level corresponded to that at 0.01 mg/liter. Various types of ultrastructural damage were found in microalga from medium with 1 mg Se/liter, which was previously reported to be threshold for D. salina; however, the number of cell injuries decreased with increasing time in clean medium. Excretory activity was decreased at the beginning of experiment; but after 7 days, it was restored to the control level. Though there were no ultrastructural alterations in microalgal cells from medium with 0.5 mg Se/liter, we assume that they had molecular defects that could inhibit the cell population growth. The study of microalgae following their reinoculation from medium containing toxicants into clean medium can be a useful method for evaluating algal survival after toxic exposure.     

Regulation of biosynthesis of aspartate family amino acids in the photosynthetic bacterium Rhodopseudomonas palustris

The four amino acids of the aspartate family (l-lysine, l-methionine, l-threonine, and l-isoleucine) are produced in bacteria by a branched biosynthetic pathway. Regulation of synthesis of early common intermediates and of carbon flow through distal branches of the pathway requires operation of a number of subtle feedback controls, which are integrated so as to ensure balanced synthesis of the several end products. Earlier studies with nonsulfur purple photosynthetic bacteria were instrumental in revealing the existence of alternative regulatory schemes, and in this communication we report on the control pattern of a representative of this physiological group not previously investigated, Rhodopseudomonas palustris. The results obtained from study of the properties of four key regulatory enzymes of the aspartate family pathway (-aspartokinase, homoserine dehydrogenase, homoserine kinase, and threonine deaminase) and of the effects of exogenous amino acids (i. e., the end products) on growth of the bacterium indicate that the control schema in Rps. palustris differs substantially from the schemes described for other Rhodopseudomonas species, but resembles the regulatory pattern observed in Rhodospirillum rubrum.Abbreviations A absorbancy - AK -aspartokinase - ASA aspartate -semialdehyde - DTT dithiothreitol - HS l-homoserine - HSDH homoserine dehydrogenase - HSK homoserine kinase - I l-isoleucine - KU Klett-Summerson photometer units - L l-lysine - M l-isoleucine - KU Klett-Summerson photometer units - L l-lysine - M l-methionine - ME -mercaptoethanol - PABA p-aminobenzoic acid - T l-threonine - TD threonine deaminase - RCV synthetic growth medium (see text) - YP agar medium containing 0.3% yeast extract, 0.3% peptone, and 1.5% agar - Y2T synthetic growth medium (see text)     

Salmonella typhimurium LT2 metF operator mutations

Summary Using an Escherichia coli lac deletion strain lysogenized with lambda phage carrying a metF-lacZ gene fusion (Flac), in which -galactosidase levels are dependent on metF gene expression, cis-acting mutations were isolated that affect regulation of the Salmonella typhimurium metF gene. The mutations were located in a region previously defined as the metF operator by its similarity to the E. coli metF operator sequence. Regulation of the metF gene was examined by measuring -galactosidase levels in E. coli strains lysogenized with the wild-type Flac phage and mutant Flac phage. The results suggest that the mutations disrupt the methionine control system mediated by the metJ gene product, but not the vitamin B₁₂ control system mediated by the metH gene product. The results also demonstrate that negative control of the metF gene by the metH gene product and vitamin B₁₂ is dependent on a functional metJ gene product.Abbreviations Ap ampicillin - dNTP deoxyribonucleoside triphosphates - GM glucose minimal - Km kanamycin - L-agar Luria agar - LM lactose minimal - SAM s-adenosyl-L-methionine - TPEG phenylethyl -D-thiogalactoside - X-gal 5-bromo-4-chloro-3-indolyl -D-galactopyranoside - [] designates plasmid-carrier state - :: novel joint     

Subcellular distribution of terminal α-D- and β-D-galactosyl residues in Ehrlich tumour cells studied by lectin-gold techniques

We have studied by high resolutionin situ light and electron microscopic lectin-gold techniques the subcellular distribution of -d-Gal residues using theGriffonia simplicifolia I-B₄ isolectin and compared it with that of -d-Gal residues as detected with theDatura stramonium lectin in Ehrlich tumour cells grown as ascites or monolayer. The microvillar but not the smooth plasma membrane regions were labelled with theGriffonia simplicifolia I-B₄ isolectin whereas both plasma membrane regions were equally well labelled with theDatura stramonium lectin. Elements of the endocytotic/lysosomal system such as coated membrane invaginations and vesicles, early and late endosomes and secondary lysosomes were positive for both -d-Gal and -d-Gal residues. A particular feature of Ehrlich tumour cells is an elaborate tubular membrane system located in the pericentriolar region which is labelled throughout by both lectins and represents part of the endosomal system. In the Golgi apparatus labelling with both lectins was observed to commence in trans cisternae which is indirect evidence for a joint distribution of the sequentially acting 1,4 and 1,3-galactosyl-transferases.     

Revision der GattungLoxonia (Gesneriaceae)

Within the genusLoxonia Jack, currently regarded as monotypic, three species are recognized:L. hirsuta Jack (Sumatra, Mentawai-Islands, Java, Borneo, Anambas-Islands, Malay Peninsula),L. discolor Jack (Sumatra) andL. burttiana A. Weber, spec. nova (Borneo). [Key with English translation p. 203.] There is evidence thatL. discolor is the most primitive species within the genus, the two others being derived from it.

Teil V der Beiträge zur Morphologie und Systematik derKlugieae undLoxonieae (Gesneriaceae).     

Experimentelle und biometrische Untersuchungen über die systematische Relevanz von Samen- und Fruchtmerkmalen anArabis glabra var.glabra und var.pseudoturritis (Brassicaceae)

Arabis glabra (L.)Bernh. var.pseudoturritis (Boiss. & Heldr.)Fiori (till now treated as a separate Mediterranean species:Arabis pseudoturritis Boiss. & Heldr.) differs fromA. glabra var.glabra only in the larger ± pleurorhizal seeds with a ± conspicuous wing that are arranged in one row in each loculus of the (sometimes longer) pods, the pedicels of the siliquae being shorter than in var.glabra. These closely correlated characters are of little systematic relevance according to biometrical and statistical analyses of both taxa and their experimental hybrids. Furthermore, there is correspondence in pod septum anatomy and chromosome number (2n = 12). This, the lack of intrinsic crossing barriers and sympatric occurence (outside the Mediterranean area!) in North America confirm the classification of these taxa as varieties withinA. glabra. Obviously, the seed and fruit characters discussed, are of even less value as generic characters for a separate genus Turritis (comprising Turritisglabra L.).

Herrn Univ.-Prof. Dr.Hermann Merxmüller anläßlich der Vollendung seines 60. Lebensjahres gewidmet.     

Natural vitamin E enrichment of Artemia salina fed freshwater and marine microalgae

Three species of microalga, the freshwater Euglena gracilis and the marine Dunaliella salina and Tetraselmis suecica, were compared in terms of vitamin E enrichment and survival and growth of the brine shrimp Artemia salina. The tocopherol content was investigated using HPLC for the post-larval and pre-adult stages of Artemia fed the microalgae and the results compared to the initial content of unfed newly hatched nauplii. There was a markedly higher content of tocopherols (about two-fold) in Artemia fed Euglena. Since this microalga contains other antioxidants such as -carotene, vitamin C and glutathione, bioactive molecules such as PUFA, and the immunostimulant polysaccharide -glucan, it represents a valuable alternative for enriching the diets of Artemia and increase its nutritional value as a food item.     

Zur Ultrastruktur der Blutzellen wirbelloser Tiere

Zusammenfassung Die Hämolymphe vonPsammechinus miliaris (Echinoidea) enthält zwei Hämocytenarten: Die Phagocyten und die Eleocyten. Alle Stadien der aus einer geißeltragenden Jugendform hervorgehenden Phagocyten besitzen gleichartige Feinstrukturen. Diese Zellen können Plasmodien mit breiten, schwingenden Pseudopodienschleiern bilden, mit deren Hilfe Fremdmaterial, aber auch intakte Eleocyten eingefangen und phagocytiert werden. Die Eleocyten treten in zwei Varianten auf: Die roten Eleocyten transportieren in membranbegrenzten, vom Golgi-Apparat stammenden Granula das rote Pigment Echinochrom. Die weißen Eleocyten sind als Melanocyten aufzufassen, deren Melanisierungsprozeß gehemmt ist, solange sie sich frei in der Hämolymphe bewegen. Sie enthalten außer einem pyknotischen Kern nur membranbegrenzte Inklusionen, die eine charakteristische Binnenstruktur aufweisen und vom Golgi-Apparat produziert werden.

---

On the ultrastructure of invertebrate hemocytesVI. On the hemocytes ofPsammechinus miliaris (Echinoidea)

Summary The hemolymph ofPsammechinus miliaris (Echinoidea) contains two cell types: The phagocytes and the eleocytes. The phagocytes originate from flagellated stem cells and exhibit in all stages of their development similar ultrastructural features. They form plasmodial aggregations and possess undulating veil-like pseudopodia, enabling the cells to catch and to ingest not only foreign material, but also intact eleocytes. The eleocytes appear in two variants: In the red eleocytes a red pigment, echinochrome, is localized in membrane-bounded granules, which originate from the Golgi apparatus. The white eleocytes are identical with melanocytes the melanization of which is inhibited while they are free floating in the hemolymph. They contain a pycnotic nucleus and characteristic inclusions which are produced in the Golgi apparatus.

     

Effects of selenium on growth and ultrastructure of the marine unicellular alga <Emphasis Type="Italic">Dunaliella salina</Emphasis> (Chlorophyta)

This study examines the effects of selenium in concentrations of 0.01, 0.5, 1, 5, and 10 mg/liter on the growth and ultrastructure of the microalga Dunaliella salina. Selenium in concentrations of 0.01 and 0.5 mg/liter stimulated cell population growth, while the number of ultrastructural alterations was the same as in the control cells. At a selenium concentrations of 1 mg/liter, cell population growth slightly decreased by the end of the experiment, and there was some increase in the number of cells with damaged organoids and in the number of completely destroyed cells. As well, the excretory function of cell vacuoles was suppressed, and the autophagic activity of these vacuoles was activated to destroy the cytoplasm and nucleus. Concentrations of 5 and 10 mg/liter were toxic to D. salina, suppressing cell population growth and promoting extensive destructive changes. The threshold concentration of selenium for D. salina was 1 mg/liter, which is 1000 times greater than the maximum permissible concentration (MPC). The fact that the microalga was able to survive for several days in this concentration is indicative of its high resistance to selenium.     

The mass culture of Dunaliella viridis (Volvocales,Chlorophyta) for oxygenated carotenoids : laboratory and pilot plant studies

The biology, and hence the mass culture, of Dunaliella viridis closely follows that of Dunaliella salina, which is successfully mass cultured for the production of -carotene. Both algae can grow at extremely high salinities and light intensities. They co-exist in the coastal salt lake, Hutt Lagoon, Western Australia. In contrast to D. salina, D. viridis does not accumulate large amounts of -carotene, producing only up to 0.7% of mixed carotenoids (lutein, zeaxathin, other oxygenated carotenoids and -carotene), compared to D. salina's ca 10% dry wt of mainly -carotene. However, in laboratory experiments, D. viridisgrew much faster and to much higher cell densities than D. salina, and attained levels of mixed carotenoids similar to those of D. salina (ca 13 mg L^–1 carotenoid). Preliminary experiments in outdoor ponds were much less promising. Harvesting by chemical flocculation was as effective as with D. salina, but extraction of carotenoids directly into vegetable oil proved inefficient. When incorporated into feed, caretonoids derived from D. viridis pigmented hen eggs acceptably. Extrapolating from laboratory results, and using costing derived from D. salina technology, the cost of production of mixed oxygenated carotenoids from D. viridis was similar to that for the production of -carotene from D. salina, at ca $A500 kg^–1.     

Ultrastructure des cellules germinales au cours du développement embryonnaire du Lézard vivipare (Lacerta vivipara Jacquin)

Résumé Les gonocytes primaires sont relativement pauvres en polysomes et l'ergastoplasme granulaire est très réduit. Le reticulum endoplasmique de type lisse se développe au cours du développement embryonnaire. L'appareil de Golgi est bien représenté et localisé au niveau de la calotte juxtanucléaire mitochondriale. Les liposomes cytoplasmiques sont très nombreux.Cette étude précise la structure du nucléole «annulaire» et de la «masse paranucléolaire» observés en microscopic photonique. Des modifications nucléolaires sont constatées au cours du développement embryonnaire. Certains aspects ultrastructuraux sont vraisemblablement en rapport avec le déplacement autonome des gonocytes. La signification physiologique du nucléole «annulaire» et de la «masse paranucléolaire» est envisagée.

---

Ultrastructural study on the primordial germ cells during embryonic development of Lacerta vivipara Jacquin

Summary The primordial germ cells of Lacerta vivipara have relatively few free polysomes and little ergastoplasm. Smooth endoplasmic reticulum increases during embryonic development. The Golgi apparatus is well developed and lies close to the mitochondrial juxtanuclear cap. This study shows the ultrastructure of the ring-shaped nucleoli and the masse paranucléolaire. Modifications in nucleolar structure are observed during embryonic development. Some ultrastructural features are probably related to the ameboid movement of the primordial germ cells. The physiological meaning of the ring-shaped nucleoli and the masse paranucléolaire is considered.

Avec la collaboration technique de Mme. M. Hubert.     

Callus proliferation from the protoplasts of embryogenic cells of Quercus serrata

Embryogenic culture was induced from the immature embryos of Quercus serrata using Marashige and Skoog's medium (MS) containing 0.1 M each of 2,4-d and BAP, and subcultured for seven months before isolation of protoplasts by using 1% Cellulase RS in 0.6 M mannitol solution. Efficient colony formation was obtained when protoplasts were cultured in a liquid MS medium containing 0.6 M mannitol, 3% sucrose and combination of 0.1 M or 1 M each of 2,4-d and BAP. Excluding ammonium nitrate from the MS medium resulted in the decrease of the percentage of colony formation. From colonies, both agar culture and liquid culture were sustained in the MS media without mannitol containing no plant growth regulator, or containing 0.1 M of BAP in combination with 0.1 M or 1 M of 2,4-d.Abbreviations BAP 6-benzylaminopurine - 2,4-d 2,4-dichlorophenoxyacetic acid - MS medium after Murashige & Skoog (1962).     

Haustoria inUrocystis (Tilletiales)

Haustoria of severalUrocystis spp. have been investigated by transmission electron microscopy. The haustoria are botryose and have an extrahaustorial matrix with vesiclelike bodies. The extrahaustorial membrane shows high ATPase activity in contrast to the haustorial plasmalemma. In walled off haustoria the haustorial plasmalemma stains more intensely than the extrahaustorial membrane. The vesicle-like bodies are ATPase negative. The role of the vesicle-like bodies is discussed.Dedicated to Prof. DrLothar Geitler on the occasion of the 90th anniversary of his birthday. Part 55 of a series Studies inHeterobasidiomycetes.