Cadmium adsorption by the non-living biomass of microalgae grown in axenic mass culture

The freeze-dried (extracted and non-extracted) biomass of 15 microalgal species grown in axenic mass culture and belonging to the Cyanobacteria, Chloro-, Eustigmato-, Phaeo-, Rhodo- and Tribophyceae were investigated for their ability to adsorb cadmium (Cd) ions from aqueous solutions. For comparison, other standard adsorbing materials (activated carbon, silica gel, siliceous earth) were included in the studies. The biomass of 11 microalgae exhibited a higher Cd adsorption than the standard materials. Extraction of the algal biomass increased the Cd adsorption capability of some, but not all microalgae. High Cd adsorption was found inAnabaena lutea, Nodularia harveyana, andNostoc commune (Cyanobacteria),Chlamydomonas sp. (Chlorophyceae),Bumilleriopsis filiformis (Tribophyceae), and inEctocarpus siliculosus, Halopteris scoparia andSpermatochnus paradoxus (Phaeophyceae). The specific surface (m² cm^–3) of the various microalgae was determined by means of laser diffractometry.Anabaena inaequalis andA. lutea (Cyanobacteria) and the Phaeophyceae had especially high Cd adsorption per surface unit. Most of the Cd adsorbed to these various materials could be desorbed subsequently with diluted mineral acid (pH 2).     

Phototactic responses in the gametes of the brown alga,Ectocarpus siliculosus

The action spectrum of phototaxis was determined and the photoreceptive mechanism was studied in Ectocarpus gametes (Ectocarpales, Phaeophyceae) using a computerized cell-tracking system. The fine structures of the stigma and the flagellar swelling were analyzed, and the reflective function of the stigma was demonstrated for the first time. Under monochromatic light stimulation, Ectocarpus gametes show mainly positive phototaxis between 370 nm and 520 nm. The action spectrum has a minor peak near 380 nm, and two major peaks at 430 nm and 450 nm or 460 nm and a shoulder at 470 nm adjoining a remarkable depression near 440 nm. Under unilateral stroboscopic illumination with more than four pulses per second, the gametes show clear phototaxis. However, the response is disturbed at lower frequencies. Addition of methyl cellulose, which increases the viscosity of the medium and slows down gamete rotation, decreases the threshold frequency. These results indicate that rotation of the gamete plays an essential role in the photoreceptive mechanism. Under equal intensities of bilateral illumination at an angle of 90°, most of the gametes swim on the resultant between the two light beams. This response is disturbed when the angle of the two light beams is as large as 120°. Observations by transmission electron microscopy show that the flagellar swelling fits precisely into a concave depression of the chloroplast at the central region of the stigma. Electron-dense material is present in that sector of the flagellar swelling which faces away from the stigma. Epifluorescence microscopy without a barrier filter and epipolarization microscopy reveal that stigmata reflect blue light. A hypothesis is formulated which discusses the possibility that the reflected light is focused onto the flagellar swelling.We are grateful to Jochen Schäfer and Elke Reinecke for their technical assistance and Dr. G. Konerman for access to epipolarization microscopy. We are also grateful to the Alexander von Humboldt Foundation for a Research Fellowship to H.K. and the Deutsche Forschungsgemeinschaft (University of Freiburg, Freiburg, FRG) for financial aid to D.-P.H.To whom correspondence should be addressed.     

Observation of Woronin bodies in Arthrinium aureum by scanning electron microscopy

Woronin bodies are cytoplasmic organelles of filamentous fungi that can be observed on one, or both sides of each septum. The goal of this paper is to illustrate the presence of them in hyphae of Arthrinium aureum by means of scanning electron microscopy and to show that they act as a safety plug to close septa pores in hypha. Results show that Woronin bodies as an immediate response to prevent a cytoplasm loss. Results support hypothesis proposed previously in literature. This revised version was published online in June 2006 with corrections to the Cover Date.     

Observation of Protoplasts from the Conidia of Arthrinium Aureum by scanning electron microscopy

Protoplast formation from conidia of Arthrinium aureum was achieved with Lysing Enzyme L-2265 (Sigma Chemical) from Trichoderma harzianum. Scanning electron microscopy of conidia protoplasts showed the acquired spherical shape.     

Morphology,morphometry and electron microscopy of HeLa cells infected with bovine Mycoplasma

Summary The host-parasite relationship of HeLa M cells artificially infected with a bovine species of Mycoplasma was studied by light microscopy, transmission electron microscopy and scanning electron microscopy. The use of morphometry to quantitate some of the findings was explored. The parasites were seen in locations extracellular to the cell surface. The detection of small numbers of organisms by light microscopy was well demonstrated by use of the fluorescent antibody technique. Scanning electron microscopy proved to be an excellent method for revealing the surface details of cell-parasite morphology. Ultra-thin sections showed that the parasites are aligned mostly parallel to the plasma membrane of the host cell but separated by a gap of 10 nm. Morphometry indicated an average of 69 organisms per cell surface occupying 1.7% of the surface area. An increase of 26% in diameter of the HeLa cells, possibly as a result of infection, was observed.The authors wish to thank Christiana Ulness and Andrea Erickson for expert technical assistance and Arnold Schmidt for the operation of the scanning electron microscope. This work was supported by grants from the U.S.P.H.S.: AI 09586, AI 10743, and AI 06720     

Scanning electron microscopy of the conidia produced by the mycelial form of Paracoccidioides brasiliensis

The conidia produced by the mycelial form of Paracoccidioides brasiliensis were examined by scanning electron microscopy for the first time. Several different conidial types were characterized. These included intercalary arthroconidia, several types of septate conidia that are formed from other conidia, pedunculate conidia, and terminal hyphal conidia. In addition, the ultrastructure of the supporting pedestal of the pedunculate conidium was found to be separated from the mother conidium by a septum in some instances, and at other times it was not.     

施氮对镉胁迫下山杨幼苗叶片氮磷钾吸收及镉积累量的影响

为探究山杨在重金属镉胁迫与增施外源氮复合处理条件下的生长及元素积累差异,该研究以山杨幼苗为材料,通过盆栽实验研究在重金属镉(Cd)胁迫下增施外源氮(N)对其形态、叶绿素、淀粉和可溶性糖含量、叶片中全氮(N)、全磷(P)、全钾(K)以及Cd含量的影响。结果表明：(1)单独Cd处理显著抑制山杨幼苗的叶片宽度和茎粗;施加外源N能够缓解Cd胁迫对植株的毒害,其叶长、叶宽和茎粗较单独Cd处理显著增加。(2)单独Cd处理后山杨叶片的叶绿素含量和淀粉含量均比对照显著下降,而其可溶性糖含量显著增加;与单独Cd处理相比,Cd+N复合处理后山杨的叶绿素含量显著增加,淀粉含量却显著下降,而可溶性糖含量略有降低。(3)与对照相比,单独Cd处理后,山杨叶片全N含量显著下降,全P含量显著增加;单独N和Cd+N复合处理后叶片全N含量均显著增加,全P含量均显著下降;全K含量在各处理下均无显著差异。(4)在Cd胁迫下山杨幼苗叶片的Cd含量比对照极显著增加,且Cd+N复合处理后叶片的Cd含量约是单独Cd处理的2倍。研究认为,增施氮素处理可显著提高山杨幼苗对镉胁迫环境的适应能力以及叶片对镉的富集能力。     

Intracellular localisation of phytochrome and ubiquitin in red-light-irradiated oat coleoptiles by electron microscopy

The intracellular localisation of phytochrome and ubiquitin in irradiated oat coleoptiles was analysed by electron microscopy. We applied indirect immunolabeling with polyclonal antibodies against phytochrome from etiolated oat seedlings or polyclonal antibodies against ubiquitin from rabbit reticulocytes, together with a goldcoupled second antibody, on serial ultrathin sections of resin-embedded material. Immediately after a 5-min pulse of red light-converting phytochrome from the red-absorbing (Pr) to the far-redabsorbing (Pfr) form-the label for phytochrome was found to be sequestered in electron-dense areas. For up to 2 h after irradiation, the size of these areas increased with increasing dark periods. The ubiquitin label was found in the same electrondense areas only after a dark period of 30 min. A 5 min pulse of far-red light, which reverts Pfr to Pr, given immediately after the red light did not cause the electron-dense structures to disappear; moreover, they contained the phytochrome label immediately after the far-red pulse. In contrast, after the reverting far-red light pulse, ubiquitin could only be visualised in the electron-dense areas after prolonged dark periods (i.e. 60 min). The relevance of these data to light-induced phytochrome pelletability and to the destruction of both Pr and Pfr is discussed.Abbreviations FR far-red light; Pfr - Pr far-red-absorbing and red-absorbing forms of phytochrome, respectively - R red light     

Low and high voltage electron microscopy of mitosis and cytokinesis in maize roots

The structure and distribution of cytoplasmic membranes during mitosis and cytokinesis in maize root tip meristematic cells was investigated by low and high voltage electron microscopy. The electron opacity of the nuclear envelope and endoplasmic reticulum (ER) was enhanced by staining the tissue in a mixture of zinc iodide and osmium tetroxide. Thin sections show the nuclear envelope to disassemble at prophase and become indistinguishable from the surrounding ER and polar aggregations of ER. In thick sections under the high voltage electron microscope the spindle is seen to be surrounded by a mass of tubular (TER) and cisternal (CER) endoplasmic reticulum derived from both the nuclear envelope and ER, which persists through metaphase and anaphase. At anaphase strands of TER traverse the spindle between the arms of the chromosomes. The octagonal nuclear pore complexes disappear by metaphase, but irregular-shaped pores persist in the membranes during mitosis. It is suggested that these form a template for pore-complex reformation during telophase. Phragmoplast formation is preceded by an aggregation of TER across the spindle at anaphase. Evidence is presented to suggest that the formation of the desmotubule of a plasmodesma is by the squeezing of a strand of endoplasmic reticulum between the vesicles of the cell plate.Abbreviations CER cisternal endoplasmic reticulum - ER endoplasmic reticulum - HVEM high voltage electron microscope - TER tubular endoplasmic reticulum - ZIO zinc iodide/osmium tetroxide     

Scanning electron microscopy of the bullfrog's retina and pigment epithelium

Summary The retina and pigment epithelium of the bullfrog (Rana catesbiana) were studied with the scanning electron microscope. Fixed-dehydrated tissues were critical point dried with CO₂, then cracked in the plane of the long axis of the photoreceptors. The cellular layers of the retina and the lateral surfaces of pigment epithelial cells were visualized. The four major types of frog photoreceptor were identified: red rod, green rod, single cone, and double cone. Cone myoids were observed to be contracted in light-adapted retinas and elongated in more dark adapted retinas.This work was supported by a career development award EY-18,083 to the author and research grant EY 00468 to Dr. Kenneth T. Brown.The author gratefully acknowledges the skillful technical assistance of Ms. Maria T. Maglio.     

Membrane turnover in crab photoreceptors studied by high-resolution scanning electron microscopy and by a new technique of thick-section transmission electron microscopy

Summary Crab photoreceptors were examined after treatment by the osmium-DMSO-osmium method for high-resolution scanning electron microscopy. This technique of specimen preparation was also adapted for transmission electron microscopy, enabling sections up to 1 urn thick to be viewed in a conventional microscope at 75 kV. With appropriate pretreatment, some cytoskeletal elements can be visualised by both techniques. The methods were then used to investigate some of the daily changes known to occur in photoreceptor cell structure. Striking differences were found in the structure of Golgi bodies present in retinula cells during the synthesis and breakdown phases of the daily cycle of photoreceptor membrane turnover. Cyclic changes were also noticed in the mitochondria of retinula cells, and additional evidence was found for a previously proposed model of rhabdomeral microvillus formation.     

Localization of mannan at the surface of yeast protoplasts by scanning electron microscopy

The (13)glucanase of Basidiomycete QM 806 was used to prepare Saccharomyces cerevisiae and Candida utilis protoplasts. Plasma membranes isolated from S. cerevisiae contained a small amount of mannose and traces of glucose and ribose. Randomly distributed -mannan was detected by scanning electron microscopy at the surface of prefixed protoplasts using colloidal gold labelled with Concanavalin A as a marker. C. utilis protoplasts were also marked with anti-mannan antibodies. Again the distribution of mannan was random. This experiment indicated also that plasma membrane mannan has the same immunochemical determinants as cell wall mannan. It is hypothesized that mannan is mainly located in the outer layer of plasma membranes.     

A transmission electron microscopy investigation: the membrane complex in spermatogenesis of <Emphasis Type="Italic">Fenneropenaeus chinensis</Emphasis>

The transforming characteristics of the membrane complex in spermatogenesis of Fenneropenaeus chinensis have been studied by using transmission electron microscopy. Two types of membrane complex have been investigated based on their sources: one originating from nucleus and the other from cytoplasm. The first one, consisted of annular structures, monolayer membrane blebs, and double or multi-lamellar membrane vesicles, emerges in the primary spermatocyte, then diffuses with the nuclear membrane and finally enters the cytoplasm. This type of membrane complex seems to play an important role in the materials transfusion from nucleus to cytoplasm, and it mainly exists inside the primary spermatocyte with some inside the secondary spermatocyte. The latter, originated from cytoplasm, is formed during the anaphase of spermiogenesis. It also exists in mature sperm, locating at both sides of the nucleus under the acrosomal cap. This type of membrane complex mainly comprises rings of convoluted membrane pouches, together with mitochondria, annular lamina bodies, fragments of endoplasmic reticulum, nuclear membrane and some nuclear particles. It releases vesicles and particles into the acrosomal area during the formation of the perforatorium, suggesting a combined function of the endoplasmic reticulum, mitochondria and Golgi’s mechanism.     

Settlement of the diazotrophic,phytoeffective bacterial strain Pantoea agglomerans on and within winter wheat: An investigation using ELISA and transmission electron microscopy

The aim of this study was to investigate the ability of Pantoea agglomerans, a plant growth-promoting bacterium, to colonize various regions and tissues of the wheat plant (Triticum aestivum L.) by using different inoculation methods and inoculum concentrations. In addition, the enzyme-linked immunosorbent assay (ELISA) and transmission electron microscopy (TEM) were used to determine: (a) the ability of the bacterial cells to grow and survive both on the surface and within internal tissue of the plant and (b) the response of the plant to bacterial infection. After inoculation, cells of the diazotrophic bacterial strain P. agglomerans were found to be located in roots, stems and leaves. Colony development of bacterial cells was only detected within intercellular spaces of the root and on the root surface. However, single bacterial cells were observed in leaves and stems on the surface of the epidermis, in the vicinity to stomatal cells, within intercellular spaces of the mesophyll and within xylem vessels. Inoculated bacterial cells were found to be able to enter host tissues, to multiply in the plant and to maintain a delicate relationship between endophyte and host. The density of bacterial settlement in the plant in all experiments was about 10⁶ to 10⁷ cells per mL root or shoot sap. Establishment was confirmed by a low coefficient of variation of ELISA means at these concentrations.     

Cell types of the endocrine pancreas in the shark Scyliorhinus stellaris as revealed by correlative light and electron microscopy

Summary In the pancreas of Scyliorhinus stellaris large islets are usually found around small ducts, the inner surface of which is covered by elongated epithelial cells; thus the endocrine cells are never exposed directly to the lumen of the duct. Sometimes, single islet cells or small groups of endocrine elements are also incorporated into acini. Using correlative light and electron microscopy, eight islet cell types were identified:Only B-cells (type I) display a positive reaction with pseudoisocyanin and aldehyde-fuchsin staining. This cell type contains numerous small secretory granules (Ø280 nm). Type II- and III-cells possess large granules stainable with orange G and azocarmine and show strong luminescence with dark-field microscopy. Type II-cells have spherical (Ø700 nm), type III-cells spherical to elongated granules (Ø450 × 750 nm). Type II-cells are possibly analogous to A-cells, while type III-cells resemble mammalian enterochromaffin cells. Type IV- cells contain granules (Ø540 nm) of high electron density showing a positive reaction to the Hellman-Hellerström silver impregnation and a negative reaction to Grimelius' silver impregnation; they are most probably analogous to D-cells of other species. Type VI-cells exhibit smaller granules (Ø250 × 500 nm), oval to elongated in shape. Type VI-cells contain small spherical granules (Ø310 nm). Type VII-cells possess two kinds of large granules interspersed in the cytoplasm; one type is spherical and electron dense (Ø650 nm), the other spherical and less electron dense (Ø900 nm). Type VIII-cells have small granules curved in shape and show moderate electron density (Ø100 nm). Grimelius-positive secretory granules were not only found in cell types II and III, but also in types V, VI, and VII. B-cells (type I) and the cell types II to IV were the most frequent cells; types V to VII occurred occasionally, whereas type VIII-cells were very rare.This work was supported by a fellowship from the Ministry of Education of Japan and the Deutsche Forschungsgemeinschaft, Bonn-Bad Godesberg (La 229/8)     

Contrasting pattern of somatic and zygotic embryo development in alfalfa (Medicago sativa L.) as revealed by scanning electron microscopy

Scanning electron microscopy has been used to investigate the morphological changes occurring during the development of alfalfa somatic embryos. Embryos were initiated from callus, transferred to suspension culture and matured on solid agar medium. This developmental pattern was compared to that of zygotic embryos developing in ovulo. Somatic embryos begin as distinct pro-embryos within the callus tissue pieces placed in suspension culture. They become globular and heart-shaped while on solid agar medium and then undergo cotyledon elongation and maturation. Somatic embryos develop comparatively slower at early stages of development and faster at the later stages than zygotic embryos. They lack a well-defined suspensor and have a very rough, poorly-differentiated epidermis, the first layer of which is lost after pro-embryo formation. The cotyledons of somatic embryos are multiple and poorlydeveloped; there appears to be a correlation between the amount of surface roughness of the developing embryo and the extent to which polycotyledony occurs.     

Scanning electron microscopy of photoreceptor cells in the light- and dark-adapted retina of Haplochromis burtoni (Cichlidae,Teleostei)

Summary The photoreceptor layer in the retina of Haplochromis burtoni (Cichlidae, Teleostei) was studied by scanning electron microscopy. Three types of receptors were identified: rods, single-cones and double-cones. The three-dimensional arrangement of these photoreceptors is described in the light- and dark-adapted retina. The surface of the inner segment of the photoreceptor cells displays fine vertical fissures which give rise to slender processes. These so called calycal processes which are of different lengths in rods and cones, surround the beginning of the smooth-surfaced outer segment. The myoid, the contractile part of the receptor, which is located beneath the ellipsoid, was examined in the single-cones of the dark-adapted retina. It is a slender structure with surface infoldings. The myoid, studied by transmission electron microscopy, contains bundles of parallel myofilaments, which are thought to be contractile.This investigation was supported by grants of the Deutsche Forschungsgemeinschaft (Sonderforschungsbereich 51-E/10)     

Scanning electron microscopy of the wall of the third ventricle in the brain of Rana temporaria. Part IV.

Summary The surface specializations of the wall of the third cerebral ventricle of Rana temporaria were investigated with the scanning electron microscope. These specializations can be divided into three types: cilia, large bulbous protrusions, and microvillus-like protrusions.Most parts of the ventricular surface are densely ciliated. In contrast, other regions are either scantily ciliated or devoid of cilia. Four areas of the ventricular surface are studded with numerous large bulbous protrusions. These large protrusions can be divided into two types: One type consists of intraventricular end bulbs of dendrites of secretory neurons. The other type is represented by large cytoplasmic extensions of ependymal cells.In the third ventricle of Rana, microvillus-like surface specializations of ependymal cells are ubiquitous structures. Generally, filiform protrusions of varying length are the predominant type. The microvillus-like specializations are transient structures, the number of which varies according to different physiological states of the ependymal cells.This investigation was supported by a grant from the Belgian Nationaal Fonds voor Geneeskundig Wetenschappelijk Onderzoek     

Deposition of glucuronoxylans on the secondary cell wall of Japanese beech as observed by immuno-scanning electron microscopy

Summary Glucuronoxylans (GXs), the main hemicellulosic component of hardwoods, are localized exclusively in the secondary wall of Japanese beech and gradually increase during the course of fiber differentiation. To reveal where GXs deposit within secondary wall and how they affect cell wall ultrastructure, immuno-scanning electron microscopy using anti-GXs antiserum was applied in this study. In fibers forming the outer layer of the secondary wall (S₁), cellulose fibrils were small in diameter and deposited sparsely on the inner surface of the cell wall. Fine fibrils with approximately 5 nm width aggregated and formed thick fibrils with 12 nm width. Some of these thick fibrils further aggregated to form bundles which labelled positively for GXs. In fibers forming the middle layer of the secondary wall (S₂), fibrils were thicker than those found in S₁ forming fibers and were densely deposited. The S₂ layer labelled intensely for GXs with no preferential distribution recognized. Compared with newly formed secondary walls, previously formed secondary walls were composed of thick and highly packed microfibrils. Labels against GXs were much more prevalent on mature secondary walls than on newly deposited secondary walls. This result implies that the deposition of GXs into the cell wall may occur continuously after cellulose microfibril deposition and may be responsible for the increase in diameter of the microfibrils.Abbreviations GXs glucuronoxylans - PBS phosphate-buffered saline - RFDE rapid-freeze and deep-etching technique - FE-SEM field emission scanning electron microscope - TEM transmission electron microscope     

New insights into the ultrastructure, permeability, and integrity of conodont apatite determined by transmission electron microscopy

New crystalline structures have been observed in argon ion‐milled conodont elements from a diverse suite of Ordovician taxa (‘Cordylodus robustus’, Drepanoistodus suberectus, Panderodus gracilis, Plectodina? sp., Aphelognathus sp., Periodon aculeatus), using transmission electron microscopy (TEM). Electron diffraction patterns of albid tissue reveal that the component crystals are extraordinarily large, in the order of hundred(s) of microns. These large albid crystals show typical cancellate porosity, although a distinctly lamellar structure has also been observed within a large albid crystal positioned between hyaline lamellar and cancellate albid tissues. There is a distinct absence of ‘interlamellar space’ within all hyaline tissues examined, which are characterized by a polycrystalline matrix of micron‐scale elongate crystals that are both strongly aligned and tightly bound within a broader lamellar structure. Optical opacity, caused by light scattering within large (≥ 0.5 µm) pores, is also a feature of both albid and polycrystalline lamellar crown tissues. Accordingly, conodont hard tissues are differentiated by crystal size and shape, as well as inter‐ and intracrystalline porosity. These new observations highlight the structural complexities of conodont histologies and the need for more comprehensive investigations particularly of transitional crown tissues, which are not well defined by terms typically used in the literature. Their histological structures are interpreted to be a product of in vivo crystallization and thus provide new insights into the relative porosity, permeability, and inherent integrity of the tissues as well as their growth relationships. Accordingly, these data not only have implications for earlier histological and palaeobiological interpretations of conodont hard tissues but are also fundamental in determining their chemical integrity, which is crucial for characterizing palaeoseawater composition and palaeoenvironmental change. The potential for conodont apatite to retain primary chemical information depends on crystal size and permeability, so the large albid crystal domains are consistent with parallel geochemical studies that suggest that cancellate albid crown is more resistant to diagenetic modification.