Stratification within centrifuged amoeba nuclei

Summary Two species of large, fresh water amoebae were ultracentrifuged and studied with the electron microscope. Emphasis was placed on the stratification of the nucleoplasm, including nucleoli, within the confines of the nuclear envelope during interphase. Three major strata were found in the nuclei of both amoeba species, namely the centripetal nucleoplasm, the middle chromatin stratum, and the centrifugal nucleolar mass. In the highly radioresistant A. proteus, the nucleolar mass separated into a centripetal electron-opaque layer and a centrifugal electron-lucent layer. The latter layer appears to be missing from the radiosensitive P. illinoisensis. The nature of these nucleolar layers and their possible relationship to differences in radiosensitivity between the two species of amoebae is discussed. The contents of the heavier of the two nucleolar layers in A. proteus might be resistant to radiation damage and may possess radiorestorative capacity.Work supported by U.S. Atomic Energy Commission. A part of the work was reported at the 17th Meeting of the Society of Protozoologists, Boulder, Colorado in 1964.     

Experimental studies on the function of the cortical cytoplasmic zone of the preprophase microtubule band

Summary Centrifugation of young seedlings ofTriticum durum andTriticum aestivum for 8–10 hours at 1,500–2,000 x g causes a serious disorder of the spatial organelle relationships in the interphase as well as the preprophase and mitotic subsidiary cell mother cells (SMCs). The nucleus, most organelles and cytoplasm are displaced to the centrifugal end of the cell, while the vacuoles lie at the other end. However, after centrifugation, the preprophase microtubule bands (PMBs) are nucleated and remain at the expected position close to the guard cell mother cells (GMCs). In some elongated SMCs the PMBs become completely separated from the nucleus. The mitotic spindle exhibits variable orientation and is usually formed at some distance from the PMB cortical zone.Cytokinesis in SMCs is spatially highly disturbed and the cell plate shows a variety of unpredictable dispositions, which seem to be determined by: 1. the position of the preprophase-prophase nucleus and the orientation of the mitotic spindle as well as their spatial relationships to the PMB cortical zone, and 2. the space available for cell plate growth. Many of the daughter cells exhibit a highly variable shape and size in different planes. Usually one edge of the cell plate partly or totally joins the anticlinal parent wall adjacent to the PMB cortical zone.In some SMCs ofZea mays andTriticum aestivum, the junction regions of the periclinal walls with the anticlinal ones, lined by the PMB cortical zone in normal SMCs, are detectably thickened after the arrest of mitosis and the prevention of interphase microtubule formation by a prolonged colchicine treatment. In a small number of protodermal cells of the same plants, participating in the development of stomatal complexes, irregular wall bodies or incomplete wall sheets were formed at wall regions lined by the PMB cortical zone.The presented observations are in line with the following hypotheses: 1. the PMB cortical zone interacts with the growing edges of the cell plate attracting it to fuse with the underlying parent wall when the latter approaches the former at a critical distance, and 2. in SMCs particular regions of the PMB cortical zone and/or the adjacent plasmalemma promote the local wall deposition in the absence of microtubules.     

Variations in environmental factors and their effects on biological characteristics of meromictic Lake Abashiri

 To clarify the variations in environmental factors and biological characteristics of eutrophic, brackish, meromictic Lake Abashiri, Japan, we studied long-term variations in concentrations of chloride and nutrients in lake water and characteristics of their supply. This lake has an anoxic layer containing much chloride, nutrients, and sulfide. Although a dense Anabaena bloom appeared in the lake every summer before 1986, the bloom did not appear after upwelling from the bottom layer (a blue tide) occurred in spring 1987. A dense layer of photosynthetic bacteria (Chlorobium sp.) formed at the top of the anoxic layer from 1989 to May 1992. The concentrations of nutrients and chloride in the oxygenic layer were determined by the balance between nutrients from influent rivers, including dissolved inorganic nitrogen, and matter supplied from the anoxic layer, including chloride and dissolved inorganic phosphorus. These influences controlled the dominant phytoplankton species and their biomass. The appearance of an Anabaena bloom in Lake Abashiri is promoted by nitrogen limitation caused by decreased loading from influent rivers and/or by a stable supply of phosphorus from the anoxic layer (no formation of photosynthetic bacterial layer and small upwelling), but the appearance is inhibited by a high chloride ion concentration with large upwelling. The main reason for the formation of a photosynthetic bacterial layer from 1989 to May 1992 was that sunlight reached the top of the anoxic layer because of the increase in transparency due to inhibition of the Anabaena bloom with upwelling. The dense layer of photosynthetic bacteria blocked nutrient diffusion from the anoxic layer to the oxygenic layer by uptake, and decreased the biomass of phytoplankton in the oxygenic layer. Received: January 11, 2002 / Accepted: April 9, 2002     

Diversity and characteristics of terrestrial cyanobacteria near gimhae city,Korea

Cyanobacteria samples were collected from smears and mats growing on the surfaces of buildings, roadside walls, rocks, and monuments near Gimhae City, Korea. These populations were not as thick and dense, nor as diverse, as those found in temperate and tropical regions. Crusts at Gimhae City were dominated by colonial forms of species fromChroococcus, Aphanothece,Aphanocapsa, Gloeofthece,Gloeocapsa, andChroococidiopsis, as well as by filamentous forms from species ofScytonema andTolypothrix. The colonial forms appeared primarily as smears, especially for species ofGloeocapsa andChroococcidiopsis, whereas filamentous forms were the major constituents of mat populations. Invariably, the cells and trichomes of these colonial and filamentous forms were enclosed with a thick, firm, multi-colored laminar sheath, as seen by transmission electron microscopy. Such an association between cell walls and trichomes was stronger in the colonial forms, particularly forGloeocapsa, compared with the filamentous forms, in which the sheath did not detach from the wall even under arid conditions. The organization of cytoplasmic structures, particularly the thylakoid membranes, was least affected by acute dryness.     

APPLICATION OF GAS-LIQUID CHROMATOGRAPHY TO STUDY OF THE ENVELOPE LIPIDS OF HETEROCYSTS12

Gas-liquid chromatography of algal extracts provides a sensitive assay for their content of 1-(O-α-d -glycopyranosyl)-3, 25-hexacosanediol, the principal glycolipid from the envelope of heterocysts of Anabaena cylindrica. Different patterns of lipids were found in the heterocysts of 5 species of Anabaena and Nostoc. Lipids with comparable chromatographic properties were detected in Gloeocapsa.     

Some fine structural observations on the rhizome ofDendroligotrichum (Bryophyta)

Summary The underground portion (rhizome) of the gametophytic axis ofD. dendroides bears anatomical resemblances to a triarch dicotyledonous root. The similarities include: 1. an epidermis producing epidermal appendages; 2. a cortex with endodermoid layer; and 3. a tri-radiate arrangement of the food and water-conducting tissue. Histochemical observations reveal that the entire radial and transverse walls of the endodermoid cells are encrusted with amorphous deposits, probably of polyphenolic nature. Casparian bands are not present as reported by earlier workers. The radial walls exhibit a fine structure of alternating electron-dense and electron-opaque lamellae. In plasmolyzed cells the plasmalemma does not adhere to the radial wall. Plasmodesmatal connections were observed in the radial and outer tangential walls of the endodermoid cells, but not in the inner tangential walls. These features of the endodermoid layer ofD. dendroides are discussed in relationship to the structure and function of the endodermis of vascular plants.     

The Absorption and Fluorescence Spectra of the Cyanobacterial Phycobionts of Cryptoendolithic Lichens in the High-Polar Regions of Antarctica

The algologically pure cultures of the green–brown cyanobacterium Chroococcidiopsissp. and three cyanobacteria of the genus Gloeocapsa, the blue–green Gloeocapsa sp.₁, the brown Gloeocapsa sp.₂, and the red–orange Gloeocapsa sp.₃, were isolated from sandstones and rock fissures in the high-polar regions of Antarctica. These cyanobacteria are the most widespread phycobionts of cryptoendolithic lichens in these regions. The comparative analysis of the absorption and the second-derivative absorption spectra of the cyanobacteria revealed considerable differences in the content of chlorophyll a and in the content and composition of carotenoids and phycobiliproteins. In addition to phycocyanin, allophycocyanin, and allophycocyanin B, which were present in all of the cyanobacteria studied, Gloeocapsa sp.₂ also contained phycoerythrocyanin and Gloeocapsa sp.₃ phycoerythrocyanin and C-phycoerythrin (the latter pigment is typical of nitrogen-fixing cyanobacteria). The fluorescence spectra of Gloeocapsa sp.₂ and Gloeocapsa sp.₃ considerably differed from the fluorescence spectra of the other cyanobacteria as well. The data obtained suggest that various zones of the lichens may be dominated either by photoheterotrophic or photoautotrophic cyanobacterial phycobionts, which differ in the content and composition of photosynthetic pigments.     

The fine structure of a tri-lamellar body in various species ofAnabaena

Summary A tri-lamellar body has been observed either near or adjacent to the crosswalls in 16 out of 20 different isolates ofAnabaena examined in thin sections. These bodies appear to consist of three discoid lamellae approximately 0.3 m in diameter. The outer lamella (closest to the plasma membrane) is separated from the middle lamella by a 12 nm space and is about 8 nm in thickness. The middle and inner lamellae, spaced about 8 nm apart, are approximately 8 nm in thickness. Electron dense granules, interpreted to be -granules, are associated with the inner lamella. In different species, osmiophilic lines 3 nm wide were observed. The osmiophilic lines run at right angles to the lamellae, either between the outer and middle lamellae, between the middle and inner lamellae or between all three lamellae. In some species, osmiophilic lines are absent. Up to six tri-lamellar bodies have been observed in median longitudinal sections. Pores 20 nm in diameter and 60 nm apart were observed in layer 2 of the cell wall of all the species ofAnabaena examined. All species which had tri-lamellar bodies also had wall pores closely associated with the bodies. Wall pores were also observed in four species lacking tri-lamellar bodies. The possible role of these structures is discussed.     

Karyotype analysis ofAscaris lumbricoides var.suum

Twelve synaptonemal complexes are present in both oocyte and spermatocyte pachytene nuclei ofAscaris lumbricoides var.suum, as determined by 3-D reconstruction of the nuclear contents from electron microscopy of serial sections and therefore, n=12 in the strain ofAscaris described here. In the female the heterochromatic end of each synaptonemal complex is attached to the nuclear envelope and the other end is free in the nucleoplasm. In the male neither end ot the synaptonemal complex is attached, but there is a heterochromatic knob at one end of each complex. Five additional large heterochromatic masses are present in the spermatocyte nucleus and these may be the sex chromosomes described by earlier workers.     

The plasmalemma surface area exposed to the soil solution is markedly reduced by maturation of the exodermis and death of the epidermis in onion roots

The dimorphic exodermis of the root of onion (Allium cepa L.) consists of long and short cells, both of which have Casparian bands. The long cells and some of the short cells also have suberin lamellae. The proportion of short cells with lamellae increases with distance from the root tip and with plant age, but is not influenced by drought stress. In young regions of onion roots, characterized by a mature endodermis and an immature exodermis, the plasmalemma surface area that can be contacted by the soil solution is 90·9 mm² per mm length of root, i.e. the sum of the plasmalemma surface areas of the epidermis, immature exodermis, cortical parenchyma and endodermis external to the Casparian band. This is reduced to 14·5–14·7 mm² by the development of a Casparian band in the exodermis, which cuts off access to the cortical parenchyma, and by the development of suberin lamellae, which cut off access to the plasmalemmae of the long and some of the short cells of the exodermis. Death of all the epidermal cells, a consequence of drought, further reduces this area to 0·205–0·0183 mm², i.e. the area of the outer tangential plasmalemmae of the short cells without suberin lamellae. In this condition, the root's capacity for ion uptake should be reduced but its capacity to resist water loss to the soil should be increased.     

Zoosporulation in Labyrinthula sp.; an Electron Microscope Study1

SYNOPSIS. Zoosporulation in Labyrinthula sp. in monoxenic culture was initiated by aggregation of spindle cells into reticulate sori. The spindle cells then changed into rounded or oval cells and formed, de novo, 2 pairs of centrioles at opposite sides of each nucleus. A pair of granular aggregates (protocentrioles) ～ 240 mμ in diameter served as precursor bodies during centriole formation. Spindle microtubules around the prophase nucleus connected the pairs of centrioles but were not found in the nucleoplasm until nuclear envelope fragmentation occurred. Prophase nuclei of uninucleated sporangia contained synaptinemal complexes; therefore, meiosis is presumed to occur. The envelope fragments moved toward the centrioles and regrouped to form the nuclear membranes of the daughter cells. Alternating nuclear and cytoplasmic divisions subdivided the preparation into 8 cells which differentiated into laterally biflagellated zoospores. Flagellar development involved growth of the kinetosome microtubules into a bud which formed over the kinetosome tangential to the cell surface. Kinetosomes were derived directly from centrioles with little differentiation other than addition of an electron-dense core to the lumen of the centriole. Zoospore ultrastructure included a stigma comprised of a row of electron-dense granules located slightly under the plasmalemma and posterior to the pair of kinetosomes. A single row of 17–21 microtubules lay parallel to the stigma granules, one or more being connected to the anterior kinetosome. A striated fiber apparatus similar to that found in some phytoflagellates connected the midregions of the kinetosomes. Fibers 1.0–1.2 μ long were attached to the plasmalemma around the base of the anterior flagellum. Zoospores settled on the substrate and differentiated directly into spindle cells. Since synaptinemal complexes were observed the planonts are probably haploid zoospores and probably not gametes since planogametic copulation was not observed.     

STUDIES ON THE ULTRASTRUCTURE OF THE GUARD CELLS OF OPUNTIA

The guard cells of Opuntia contain numerous mitochondria, elements of endoplasmic reticulum, dictyosomes, and microbodies. A complex array of small to large vacuoles which contain small, membrane-bounded vesicles occur in each guard cell. The variety of cytoplasmic constituents and vacuoles suggest that the guard cells are complex in function. A highly reduced grana-fretwork system within the plastids indicates that the photosynthetic capacity of the guard cells is probably rather low. No plasmodesmata occur in the walls between the guard cells and the subsidiary cells while there are numerous invaginations of the guard cell plasmalemmas. Many of the variations in the plasmalemma probably indicate that the plasmalemma is a highly active interface.     

CULTURALLY INDUCED VARIATION IN FOUR MORPHOLOGICALLY DIVERSE BLUEGREEN ALGAE

Characters used in taxonomic placement at levels from family to species appeared, disappeared, or varied widely, while others remained constant, in cultures of four disparate species of blue-green algae (Cyanophyta) under varying conditions of culture. Species of Anabaena and Gloeocapsa displayed fairly stable characteristics. Species of Calothrix and Lyngbya exhibited much greater variability, especially in twisting of filaments, presence or absence of constrictions at the cross walls, abundance of heterocysts, pseudovacuoles, fragmentation of filaments, and change of color. Color change was associated with abrupt pH shift in Lyngbya and could be reversed by adding nitrate. In the putatively nitrogen-fixing Anabaena, similar change in color with age was not associated with sharp pH shift, nor could it be reversed with nitrate. In general, growth on agar in turbulent conditions and with decreased CO₂–O₂ exchange was less than that obtainable under other conditions, and media with acid pH were partially or totally inhibitory to growth.     

Diurnal changes in the vertical distribution of phytoplankton in hypertrophic Lake Kasumigaura,Japan

The daily vertical migration of five species;Microcystis aeruginosa (Kütz.) Trevis,Anabaena spiroides Klebahn f.crassa (L.) Elenkin,Aphanizomenon flos-aquae (L.) Ralfs,Melosira granulata (E). Ralfs, andCoscinodiscus lacustris Grun. was studied using a close-interval water sampler on a calm summer day in Lake Kasumigaura. Many colonies ofMicrocystis were observed at the middle of the water column (approx. 1.5 m depth) in the afternoon, and at the surface in the early morning.Anabaena occurred mostly in the upper layer whileAphanizomenon tended to be uniformly distributed. The difference in migration patterns suggests thatMicrocystis is superior toAnabaena andAphanizomenon in obtaining both light and nutrients from this lake. Among diatoms,Melosira remained at the bottom of the water column throughout day and night, but Coscinodiscus was uniformly distributed.     

MORPHOLOGICAL AND PHYLOGENETIC STUDIES ON UNICELLULAR DIAZOTROPHIC CYANOBACTERIA (CYANOPHYTES) ISOLATED FROM THE COASTAL WATERS AROUND SINGAPORE1

Six unicellular diazotrophic cyanobacteria were isolated from the coast around Singapore. The isolates grew under both light:dark (L:D) cycles and continuous illumination (CL) in media without combined nitrogen and exhibited an ability to fix nitrogen (as measured by acetylene reduction) under aerobic conditions. The cells of all isolates were surrounded by a thick fibrous outer wall layer, and they divided by transverse binary fission. The arrangement of photosynthetic thylakoids was of the dispersed type. Three isolates were identified as form‐genus Gloeothece as cells were divided in a single plane, and the other three isolates were identified as form‐genus Gloeocapsa as cells were divided in multiple planes. Phylogenetic analyses based on the DNA sequences of the genes encoding 16S rRNA and dinitrogenase reductase (nifH) revealed the following: (i) Our six isolates formed a monophyletic cluster. (ii) The monophyletic cluster was subdivided into two phylogenetic groups, which taxonomically corresponded with the form‐genera Gloeothece and Gloeocapsa. However, (iii) a diazotrophic strain of form‐genus Gloeothece, Gloeothece membranacea (Rabenh.) Bornet PCC6501, was not closely related to our isolates, and (iv) some, but not all, diazotrophic unicellular strains of form‐genus Cyanothece were observed to be in a close relationship with our isolates.     

The heat shock response and acquired thermotolerance in three strains of cyanobacteria

The heat shock response of three cyanobacterial strains,Anabaena sp. Strain PCC (paris Culture Collection) 7120,Plectonema boryanum Strain PCC 6306, andSynechococcus sp. Strain PCC 7942, was characterized by polyacrylamide gel electrophoresis.Anabaena produced 33 heat shock proteins,P. boryanum 35 proteins, andSynechoccus 19 proteins. The rapid response to heat shock was consistent for all three strains, although the number of time-dependent proteins varied. All strains developed thermotolerance when first pretreated with a sublethal heat shock and then challenged with a previously lethal temperature. A 30-min 30°C incubation was required between the heat shock and challenge forSynechococcus, but not forAnabaena andP. boryanum. Synechococcus cells required a higher challenge temperature (51° vs. 49°C) than the other two strains to destroy control cells that were not pretreated with a heat shock.     

SIEVE-ELEMENT ULTRASTRUCTURE IN PLATYCERIUM BIFURCATUM AND SOME OTHER POLYPODIACEOUS FERNS: THE NACREOUS WALL THICKENING AND MATURATION OF THE PROTOPLAST

Sieve elements of various ages were examined in petioles and midribs of Platycerium bifurcatum (Cav.) C. Chr. and Phlebodium aureum (L.) J. Sm., only older ones in similar parts of leaves of Polypodium schraderi Mett. and Microgramma lycopodioides (L.) Copel. Nacreous walls apparently are formed by most, if not all, protophloem and metaphloem sieve elements in all four species. In Platycerium and Phlebodium nacreous wall formation is closely correlated with the appearance of numerous membranes or vesicles in the region of the wall. These extracytoplasmic membranes apparently are derived from protrusions of the plasmalemma. After the nacreous layer is fully thickened, many endoplasmic reticulum (ER) membranes apparently end up outside the plasmalemma of Platycerium, where they degenerate and gradually intergrade in appearance with the fibrillar material comprising the nacreous thickening. In Phlebodium, Polypodium, and Microgramma the ER forms multivesicular bodies. As the cells approach maturity, the membranes delimiting the multivesicular bodies fuse with the plasmalemma and their vesicular contents, which are not discharged into the region of the wall, disappear. Gradually, the nacreous layer decreases in thickness and disappears. At maturity the enucleate sieve-element protoplasts of all four species are essentially similar. They are lined by a plasmalemma and a parietal, anastomosing network of ER and contain both plastids and mitochondria. The plastids in Polypodium and Microgramma are chloroplasts, but those in Platycerium and Phlebodium lack grana and intergrana lamellae.     

CYTOLOGY OF BLUE-GREEN ALGAE. I. THE CELLS OF SYMPLOCA MUSCORUM

Pankratz , H. S., and C. C. Bowen . (Iowa State U., Ames.) Cytology of blue-green algae. I. The cells of Symploca muscorum . Amer. Jour. Bot. 50(4): 387–399. Illus. 1963.—The cellular morphology of Symploca muscorum is described, based upon electron micrographs utilizing improved techniques of specimen preparation. Except for a limiting plasma membrane, ribosomes, and Feulgen-positive chromatin, the cells have little resemblance to those of higher organisms. The longitudinal components of the cellular envelope consist of a 200–300 mμ fibrous sheath and a complex inner investment about 35 mμ thick which includes at least 3 distinctly layered wall elements in addition to the typical 7-mμ unit membrane forming the plasma membrane. A row of very small elongate “pores” pierce the inner investment on each side of, and immediately adjacent to, the junction of the longitudinal walls and the crosswalls. Crosswalls vary in thickness from 3 to 20 mμ, depending upon their age, and arise as elaborations of the inner layers of the longitudinal inner investment. The photosynthetic lamellar component of the cytoplasm consists of flattened sacs formed from unit membranes. The lamellae are concentrated in the peripheral region of the cell and usually are parallel to the longitudinal wall. These often extend from one crosswall to the next but, except for a few cases, are not continuous with the plasma membrane at either end. The Feulgen-positive nucleoplasm appears as an anastomosing system of lightstaining regions containing fibrils 2–5 mμ in diameter. The morphology and interrelationship of a number of other cellular elements are described: (1) structured granules range up to 0.5μ in diameter and occur near crosswalls; (2) polyhedral bodies, 0.2–0.5μ in diameter, are closely associated with the nucleoplasm; (3) “cylindrical bodies” characteristically consist of 2 concentric cylinders, are about 13 mμ in diameter and up to lμ in length; (4) “α granules” are spherical or somewhat elongate elements about 30 mμ in diameter and characteristically associated with the photosynthetic lamellae and structured granules; (5) “β granules” are spherical, highly osmiophilic granules which range from 30 to 90 mμ in diameter; (6) ribosomes, 10–15 mμ, in diameter, are most numerous near the nucleoplasm; (7) plasmodesms penetrate the crosswalls between adjacent cells. The cells of this organism can best be described as being in a “steady state” of division, and there is no evidence of any kind of organized distribution of the nucleoplasm to daughter cells during the constant progress of cytokinesis.     

Androecial development and systematics inFlacourtiaceae s. l.

Androecial development of 13 species belonging to six tribes ofFlacourtiaceae has been investigated. While inScolopieae andFlacourtieae the stamens develop centrifugally, inErythrospermeae, Oncobeae andPangieae they are initiated in a centripetal sequence or a sequence that is neither distinctly centripetal nor centrifugal. The distribution of these developmental patterns coincides with the distribution of other characters (e.g. cyanogenic compounds, salicoid leaf teeth) and therefore supports a split of the family intoFlacourtiaceae s. str. (containing theScolopieae, Homalieae, Prockieae, Flacourtieae, Casearieae andBembicieae) andKiggelariaceae (withErythrospermeae, Oncobeae andPangieae) and is in accordance with results of recentrbcL studies.