Immunological characterization of nitrogenase in the filamentous non-heterocystous cyanobacterium Oscillatoria limosa

The filamentous non-heterocystous cyanobacterium Oscillatoria limosa was subjected to Western blot analyses using two antisera raised against the small subunit (Fe-protein) of the nitrogenase complex. Two polypeptides were recognized in nitrogen-fixing cultures irrespective of the antiserum used while no bands were detectable in nitrate-grown cultures. The apparent molecular weights of the two polypeptides were approximately 40.5 and 39.5 kDa respectively, with the former, probably an inactive form, dominating. In situ immunogold electron microscopy was used to reveal the cellular and subcellular localization on the Fe-protein. All cells of the trichomes of nitrogen-fixing O. limosa showed a dense label. The label was homogeneously distributed throughout the cytoplasm including the thylakoid area. Nitrate-grown cultures contained a very low label.Abbreviations SDS sodium dodecyl sulfate - PAGE polyacrylamide gel electrophoresis This study was supported by the Swedish Natural Science Research Counsil and the M. and M. Wallenberg Fund (to B. Bergman). We are grateful to Dr. S. Nordlund (University of Stockholm, Sweden) for providing us with the antiserum of Rhodospirillum rubrum nitrogenase and to Drs. S. Reich and P. Böger (University of Konstanz, FRG) for the antiserum of Anabaena variabilis. Skilful technical assistence by K. Östlund and E. Danielsson is gratefully acknowledged. We would also like to thank M. Villbrandt (University of Oldenburg, FRG) for providing cultures of Oscillatoria limosa and Dr. P. Lindblad for valuable discussions and suggestions.To whom correspondence should be addressed.     

Lokalisierung von rRNA-Genorten und Sekundäreinschnürungen beiVicia narbonensis undVicia sativa

Sites of 18/25S RNA genes and those of secondary constrictions have been located in metaphase chromosomes ofV. narbonensis andV. sativa by RNA/DNA in situ hybridization and Feulgen staining. InV. narbonensis the rRNA genes are located in median position on one pair of chromosomes, which is the shortest of all in the genome. InV. sativa rRNA genes are located in two pairs of chromosomes. The two heterologous sites differ markedly in the level of labeling. Strong labeling is found in a submedian position of a short pair of chromosomes. Weaker labeling is found in a median position on the longest pair of chromosomes. InV. narbonensis andV. sativa the position of the grain clusters correlate with the position of the secondary constrictions in chromosomes stained by Feulgen. The implications with respect to karyograms ofV. narbonensis andV. sativa known from the literature are discussed.

     

Zwergstrauchige Arten der FlechtengattungCaloplaca

The frutescent species of the lichen genusCaloplaca are usually united in sect.Thamnoma, but they do not form a natural group. They are derived from different species groups within sect.Gasparrinia from different parts of the world, presumable from species having scleroplectenchymes in cortex and medulla. The algal cells are concentrated between the scleroplectenchymatic strands in large and dense groups, from where medullary plectenchyme extends to the cortex and forms characteristic pseudocyphellae there.—Most of the species seem to be ornithocoprophilous; they grow on rocks along marine coasts where much fog is induced by cold currents.—Caloplaca cribrosa is endemic in Tasmania and New Zealand,C. regalis and the doubtfulC. ambitiosa belong to the antarctic element.C. fragillima from central Chile seems to be propagated by thallus fragments.C. coralloides andC. thamnodes are endemic to California and Baja California respectively.C. cladodes from the Rocky Mountains deviates in many characteristics from the other species i.a. by it different ontogenetic development, reduced spore septum, and cementing amyloid polysaccharides within the scleroplectenchymatic strands. The African species are characterized by their distinctly dorsiventral lobes and usually possess oil cells in some of the paraphyses.Caloplaca bonae-spei, C. fragillima andC. thamnodes are new to science.

     

Haustorienbefall und inäquale Teilungen desNostoc-Phycobionten vonLempholemma botryosum (Lichinaceae)

In fully developed parts of thalli ofLempholemma botryosum a relatively high number of algal cells is attacked by haustoria. They differ specifically in form and effect from haustoria of otherLempholemma species. Cells of theNostoc phycobiont attacked by hyphae developing into haustoria may divide to give inequal besides equal daughter cells.

     

Simultaneous heterolactic and acetate fermentation in the marine cyanobacterium Oscillatoria limosa incubated anaerobically in the dark

The marine nitrogen-fixing cyanobacterium Oscillatoria limosa, strain 23 (Oldenburg) was investigated with respect to its dark anaerobic metabolism. As soon as the cells were incubated anaerobically in the dark, they started to ferment. Glycogen was presumably degraded via the heterolactic fermentative pathway. Glycogen-glucose was degraded to equimolar amounts of lactate, ethanol and carbon dioxide. The disaccharide trehalose, which serves as an osmoprotectant in O. limosa, was also catabolized. Most probably, this compound was fermented almost exclusively to acetate. Some hydrogen was produced as well. In the presence of elemental sulfur, fermentative hydrogen production ceased and sulfide was produced instead. The presence of elemental sulfur had no effect on the amounts and ratios of the fermentation products produced.     

Diurnal and seasonal variations of nitrogen fixation and photosynthesis in cyanobacterial mats

In situ measurements of nitrogenase activity and photosynthesis were performed simultaneously in cyanobacterial mats of intertidal sand flats of the Southern North Sea. Two types of cyanobacterial mats, which differed in species composition and biomass content, were investigated. The measurements were done monthly during 3 years to detect seasonal variations of nitrogen fixation and photosynthesis. Diurnal variations were investigated as well. The results showed that (i) freshly colonized sediment with the cyanobacteriumOscillatoria limosa as the dominant organism revealed the highest specific nitrogenase activities (ii) nitrogenase activities were highest in spring and summer, when mat development was initiated and (iii) diurnal fluctuations of nitrogenase activity indicated that it occurred temporally separated from oxygenic photosynthesis.     

Über den Verzweigungsvorgang beiPsilotum undSelaginella mit Anmerkungen zum Begriff der Dichotomie

After a critical evaluation of the concept of dichotomous branching in Cormophytes the shoot apical meristems ofPsilotum triquetrum andSelaginella speciosa are described. InPsilotum only the terminal meristems of the cryptophilic shoots have a three sided apical cell. Those of the photophilic shoots lack a typical apical cell.Selaginella has a two sided apical cell. The process of branching is independent from apical cells. It is due to an equal or unequal fractionation of the initial zone of the shoot apex which embraces all tissues above the leaf producing zone of the apical meristem.

Herrn Univ.-Prof. Dr.Walter Leinfellner zum 70. Geburtstag gewidmet.     

An ultrastructural comparative study of the sperm of<Emphasis Type="Italic"> Hyla pseudopseudis</Emphasis>,<Emphasis Type="Italic"> Scinax rostratus</Emphasis>, and<Emphasis Type="Italic"> S. squalirostris</Emphasis> (Amphibia: Anura: Hylidae)

We make detailed comparisons of the ultrastructure of the spermatozoon among three species of the family Hylidae, Hyla pseudopseudis, Scinax rostratus, and S. squalirostris. The acrosome complex consists of two conical structures covering the nuclear rostrum, the acrosome vesicle, and the subacrosomal cone. The nucleus has a moderately condensed chromatin with a conical shape in longitudinal sections and a circular shape in cross-sections. In H. pseudopseudis, mitochondria are numerous and circular, and in S. rostratus and S. squalirostris there are fewer mitochondria that are more elongate in longitudinal and transverse sections. In H. pseudopseudis, the mitochondrial collar starts adjacent to the distal centriole, occupying the whole midpiece, whereas in both Scinax species the mitochondrial collar starts only at the posterior one-third of the midpiece. In both Scinax species, the presence of juxta-axonemal fiber, axial sheath, and axial fiber in the tail are seemingly plesiomorphic characters, widespread among bufonoid frogs. In H. pseudopseudis, however, the absence of axial fiber and axial sheath seems to be derived from the typical bufonoid condition. The differences between Hyla and Scinax sperm endorse the separation of the two genera and suggest that sperm ultrastructure can be a useful tool to investigate relationships at the intrafamily level.     

Constant and variable parts of the 155-bp centromeric repeat in Camptochironomus

Centromeres in dipteran insects belonging to the subgenus Camptochironomus (C. tentans and C. pallidivittatus) contain tandem repeats of a 155-bp repeat. A special structural feature of the 155-bp unit is a region with two palindromes connected by a short piece of DNA with only AT base pairs, which has at least a superficial similarity to a functionally important part of the Saccharomyces cerevisiae centromere. As a parameter for functional importance we have measured frequencies of mutations along the unit in samples of repeats from the two species. We find that it consists of about equal parts of highly conserved and considerably less-conserved DNA. The palindromes are localized in the conserved part of the repeat. Correspondence to: J.-E. Edström     

Vergleichende Karyologie der Gräser-SubtribenAristaveninae undAirinae (Poaceae — Aveneae)

The chromosome numbers of nearly all species of the grass subtribesAristaveninae andAirinae from Europe and northern Africa are presented. Among theAristaveninae the genusAristavena has 2n = 14 chromosomes, whereasDeschampsia forms a polyploid series with the basic number x = 13. In the subtribeAirinae the basic number x = 7 predominates.Avenella includes a polyploid series up to dekaploidy, whilst the lowest diploid value so far known in grasses — caused by descending dysploidy — exists in the annual generaAiropsis andPeriballia with 2n = 8.From both subtribes 12 different karyotypes are described and depicted as idiograms. The basic karyotypes ofCorynephorus, Periballia andVahlodea differ from each other by different chromosome length. SAT-chromosomes in theAirinae vary somewhat. Some marker chromosomes eludicate phylogenetic relationships. Amphiplasty appears in various genera and was studied particularly in the amphidiploidAira caryophyllea. Karyological and genomatic trends are considered in relation to evolutionary strategies of annuals and perennials.The nuclear DNA content of some species has been determined cytophotometrically. In subtribeAirinae a positive correlation exists between chromosome volume, pollen diameter, and DNA content. A comparison of the duration of microsporogenesis and microgametogenesis in annual and perennial species with their nuclear DNA content has shown that a primary nucleotypic influence is not recognizable.

     

Über denChlorella-Phycobionten vonTrapelia coarctata

Trapelia coarctata is lichenized withChlorella saccharophila var.ellipsoidea; this is in accordance with one of two former statements. The cells of the isolated alga may be covered individually by a gelatinous envelope; they also can be embedded in confluent mucilage. The course of succedanous divisions leading to the formation of autospores starts with the appearance of a second, new pyrenoid and goes on with bipartioning of the chloroplast, nuclear division and cytokinesis. Starch grains identical to those in the stroma surround the pyrenoid more or less loosely and not in the form of saucer-shaped parts constituting a coherent shell.

     

Die Lage des Chromatophors in Beziehung zur Systematik vonCymbella-Arten (Bacillariophyceae)

InCymbella the position of the chromatophore corresponds to the taxonomic differentiation into sections. The pyrenoid always shows the same right hand — left hand inclination.

     

Entwicklungsgeschichtliche Eigentümlichkeiten einigerAchnanthes-Arten (Diatomeae)

InAchnanthes lanceolata pairing occurs by chance in regard to the arrangement of the mother cells, inA. minutissima it is accomplished selectively: contact at the sides occupied by the nucleus is favoured very strongly. In contrast toA. exilis, pairing inA. minutissima occurs by detachment of both partner cells from their stalks, whileA. lanceolata has no stalks at all. The isogamous gamete fusion inA. lanceolata should be understood as a modification of the anisogamous behavior inA. minutissima andA. exilis. For the three species the same constant orientation of the primary cell in relation to the substratum and to the mother thecae is characteristic. The first epivalva always is the raphe valve. InA. lanceolata in connection with the property of the primary cell possessing two chromatophores resp. pyrenoids instead of one, the transapical heteropolarity of the rapheless valve, a specific character, changes to isopolarity. It seems that a sort of induction from the pyrenoids towards the formation of the rapheless valve is at work. InA. lanceolata the pseudoraphe shows a structure hitherto unknown.A. lanceolata fo.ventricosa Hust. is to be eliminated. Var.rostrata (Östrup)Hust. is a good taxon; two new differential characters are found.

     

Floral development and systematics ofCistaceae

The floral development of representatives of six genera ofCistaceae has been studied. Calyx development involves the formation of a ring primordium in several taxa. Androecium development in species with intermediate or higher stamen numbers starts with the formation of a ring meristem on which the stamens are initiated in a centrifugal direction. In many taxa five alternipetalous leading stamen primordia can be observed. In the apetalous (cleistogamous) flowers ofTuberaria inconspicua androecium development appears to be unordered; this is probably due to the lack of petals. InLechea intermedia (also cleistogamous) the corolla is trimerous and three complex stamen primordia are produced, which give rise either to one or three stamens. Relationships withinCistaceae are discussed. Floral development inCistaceae is compared with that in otherMalvanae. Among the eight families ofMalvanae from which information on floral development is availableCochlospermaceae andBixaceae exhibit the greatest similarities toCistaceae. InCistaceae the leading stamen primordia are alternipetalous. InBixa the same condition seems to be present. InMalvales s. str. mostTiliaceae also show earliest stamen initiation in alternipetalous sectors, whereas the stamens of the innermost alternipetalous position are retarded early or even suppressed inSterculiaceae, Bombacaceae, andMalvaceae. WithinMalvales s. str. the diversity of androecial developmental patterns seems to decrease inBombacaceae andMalvaceae due to increasing synorganization in the mature androecium. The derivation of polyandry inMalvanae from diplo- or obdiplostemony is discussed by comparison with the sister clades ofMalvanae as shown in recentrbcL studies (i.e.Sapindales, Rutales, the glucosinolate producing clade, andMyrtales).     

Larval mortality of pine sawflies [Hymenoptera: Diprionidae] in relation to pollution level: A field experiment

Larval mortality ofNeodiprion sertifer (Geoffroy),Diprion pini (L.) andGilpinia pallida (Klug) were studied in field experiments around a factory complex in southwestern Finland. Larval colonies were transferred on the shoots of Scots pines growing at different distances from the emission source. Larval mortality was highest near the factories. InN. sertifer, larval mortality caused by the nuclear polyhedrosis virus was higher and cocoon mortality caused by parasitoids was lower near the pollutant source. The most abundant parasitoid species wereSynomelix scutulata (Hartig) andLamachus eques (Hartig). 16–67% of theN. sertifer, 0–5% of theD. pini and the 73–100% ofG. pallida cocoons contained parasitoids oviposited during the larval period of the sawflies.      

Comparison of the ribosomal RNA genes in four closely relatedCucurbitaceae

Cucurbitaceae are characterized by a high copy number for nuclear ribosomal RNA genes. We have investigated the genomic ribosomal DNA (rDNA) of four closely related species of this family with respect to structure, length heterogeneity, and evolution. InCucumis melo (melon) there are two main length variants of rDNA repeats with 10.7 and 10.55kb.Cucumis sativus (cucumber) shows at least three repeat types with 11.5, 10.5, and 10.2kb.Cucurbita pepo (zucchini) has two different repeat types with 10.0 and 9.3kb. There are also two different repeat types inCucurbita maxima (pumpkin) of about 11.2 and 10.5kb. Restriction enzyme mapping of the genomic rDNA of these four plants and of cloned repeats ofC. sativus shows further heterogeneities which are due to methylation or point mutations. By comparison of the restriction enzyme maps it was possible to trace some evolutionary events in the family ofCucurbitaceae. Some aspects of regulation and function of the middle repetitive rRNA genes (here between 2000 and 10000 copies) are discussed.     

Gloeomonas oderChlamydomonas? Elektronenmikroskopische Untersuchungen anGloeomonas simulans

The fine structure ofGloeomonas simulans Fott (1957) was studied electron microscopically to ascertain whether it belongs to the genusChlamydomonas rather than toGloeomonas. Most cytoplasmic elements and the cell wall do not differ from otherChlamydomonadaceae but its flagellar rootlet system is unique: Each of the two flagella has an accessory basal body; its basis is accompanied by two inner and two outer bands which are connected distally (one inner and one outer band on each side) resp. proximally (the two outer bands); the latter form a long (up to 3–5 µm) connecting band between the two flagellar bases. The nucleus contains fibrillar bundels.—The unique flagellar rootlet system seems to provide a better basis for the generic classification ofGloeomonas than the position of the contractile vacuoles or the size of the apical papilla, and strongly suggests the exclusion ofG. simulans fromChlamydomonas.

Zweiter Beitrag einer vonEttl (1965a) begonnenen Publikationsreihe.     

Stomata and plasmodesmata

Summary In developing epidermal tissue ofPhaseolus vulgare L. complete plasmodesmatal connections occurred between guard cells and epidermal cells and between sister guard cells of a stoma but they were not seen in fully differentiated tissue. However, incomplete, aborted plasmodesmata were occasionally seen in the common guard/epidermal cell wall, usually connected to the epidermal cell protoplast, in mature tissue. Plasmodesmatal connections between neighbouring epidermal cells were commonly observed in tissue at all stages of development. In all locations, the plasmodesmata were usually unbranched occurring singly or in small pit fields; very rarely branched, incomplete plasmodesmata were also seen in the wall between mature guard and epidermal cells. The significance of these findings were related to stomatal functioning and to the development of plasmodesmata in general.     

Zellteilung und Bildung von Innenschalen beiHantzschia amphioxys undAchnanthes coarctata

Before cytokinesis, the identically constructed chromatophores ofHantzschia amphioxys andAchnanthes coarctata are transformed into less effigurated bodies. In normal cytokinesis, the course of mitosis, chromatophore division, and cleavage furrowing are exactly synchronized. The division of the chromatophore appears as a passive process, i.e. intersection by the cleavage furrow. In inequal cell divisions before the formation of inner valves cytokinesis can take place without chromatophore division. Once chromatophore division without mitosis and cytokinesis was observed. InHantzschia there are three types of inner valve formation, inAchnanthes coarctata only two. The inner valves develop under unfavorable growth conditions, the cells possessing them, however, are not resting spores as in some other diatoms. InHantzschia, auxospore formation is suppressed under the cultural conditions used, the cells multiply intensely without diminution.

     

The behaviour of kinetochore microtubules during meiosis in the fungusSaprolegnia

Summary InSaprolegnia, kinetochore microtubules persist throughout the mitotic nuclear cycle but, whilst present at leptotene, they disappear coincidently with the formation of synaptonemal complexes at pachytene and reform at metaphase I. In some other fungi chromosomal segregation is random in meiosis and non-random in mitosis. The attachment of chromosomes to persistent kinetochore microtubules in mitosis, but not meiosis, inSaprolegnia provides a plausible explanation for such behaviour. At metaphase I each bivalent is connected to the spindle by 2 laterally paired kinetochore microtubules whereas at metaphase II (as in mitosis) each univalent bears only one kinetochore microtubule, thus showing that all kinetochores are fully active at all stages of meiosis.