Preparation of chloroplast DNA from pea plastids isolated in a medium of high ionic strength

A simple, rapid, and inexpensive method for the preparation and purification of chloroplast DNA (cpDNA) from pea has been developed. The crucial step is the isolation of chloroplasts in a medium of high ionic strength (I congruent equal to 1.40 M). CpDNA from pea prepared according to this method has successfully been used for restriction enzyme mapping, Southern transfers, and cloning.     

The major light-harvesting complex of Photosystem II: aspects of its molecular and cell biology

The light-harvesting complex of photosystem II (LHC II) contains one major (LHC IIb) and at least three minor chlorophyll-protein components. The apoproteins of LHC IIb (LHCP) are encoded by nuclear genes and synthesized in the cytoplasm as a higher molecular weight precursor(s) (pLHCP). Several genes coding for pLHCP have been cloned from various higher plant species. The expression of these genes is dependent upon a variety of factors such as light, the developmental stage of the plastids and the plant. After its synthesis in the cytoplasm, pLHCP is imported into plastids, inserted into thylakoids, processed to its mature form, and assembled into LHC IIb. The pathway of assembly of LHC IIb in the thylakoid membranes is currently being investigated in several laboratories. We present a model that gives some details of the steps in the assembly process. Many of the steps involved in the synthesis and assembly are dependent on light and the stage of plastid development.Abbreviations PS Photosystem - LHC II Light-harvesting complex of PS II - LHCP Apoproteins of LHC IIb - pLHCP Precursor of LHCP - PAGE Polyacrylamide gel electrophoresis     

Correction of chlorophyll-defective male-sterile winter oilseed rape (Brassica napus) through organelle exchange: Characterization of the chlorophyll deficiency

Jarl, C. I., Ljungberg, U. K. and Bornman, C. H. 1988. Correction of chlorophyll-defective male-sterile winter oilseed rape ( Brassica napus ) through organelle exchange: Characterization of the chlorophyll deficiency. - Physiol. Plant. 72: 505–510.
As is known, the introduction of male-sterile Raphanus sativus L. cytoplasm into Brassica napus L. results in male-sterile oilseed rape plants, which display a temperature-related chlorophyll defect. The influences of temperature and irradiance on this defect were investigated. Compared to a line of normal (green phenotype) male-fertile oilseed rape, the male-sterile line had reduced chlorophyll content, fewer chloroplasts per cell, an altered ultrastructure of the chloroplasts and reduced activities of both photosystems, although the relative amounts of the photosystems and the chlorophyll a/b ratio were similar. The lower activity of the photosystems is explained by a decreased functional antennae size and a reduced efficiency in the interactions between the nuclear-encoded light-harvesting proteins and the reaction centres coded for by the plastome. Some thylakoid polypeptides differed in proportion between the male-fertile line with green phenotype and the male-sterile line with chlorotic phenotype. Characters, in which the two lines exhibited differences, are ascribed to difficulties in molecular communication between the oilseed rape nucleus and the radish cytoplasm, which are combined in the deficient male-sterile line.     

Temperature-induced modifications in size and pattern of microtubular organelles in a ciliate,Dileptus II. Formation and spatial arrangement of the microtubular skeleton of the oral parts

Summary Microtubular organelles formed during continuous exposure to high or low temperature were studied. Neither heat nor cold prevents formation of the microtubular skeleton in the oral parts of the ciliateDileptus. Elevated temperature causes the formation of short microtubular fibres, while the size of the oral structure is not diminished. This leads to failure in sculpturing of the cytostome. Heat-treatment may also alter the localization of the anchor site of fibres around the circumference of the basal bodies, and the orientation of the fibres. Cold-treatment evokes the formation of small mouthparts containing a lower number of organelles, although these are properly shaped and there are no deviations in the position or orientation of fibres. It seems that low temperature may suppress the rate of formation of microtubular organelles, while elevated temperature affects their patterning.Abbreviations MTOC microtubule organizing centre - T transverse fibres - B basal body - Cy cytostome - K kinetodesma - P postciliary fibre - C compound fibre - L lamina - F filamentous bundle - M monokinetid     

Molecular motors in axonal transport

Neurons require a large amount of intracellular transport. Cytoplasmic polypeptides and membrane-bounded organelles move from the perikaryon, down the length of the axon, and to the synaptic terminals. This movement occurs at distinct rates and is termed axonal transport. Axonal transport is divided into the slow transport of cytoplasmic proteins including glycolytic enzymes and cytoskeletal structures and the fast transport of membrane-bounded organelles along linear arrays of microtubules. The polypeptide compositions of the rate classes of axonal transport have been well characterized, but the underlying molecular mechanisms of this movement are less clear. Progress has been particularly slow toward understanding force-generation in slow transport, but recent developments have provided insight into the molecular motors involved in fast axonal transport. Recent advances in the cellular and molecular biology of one fast axonal transport motor, kinesin, have provided a clearer understanding of organelle movement along microtubules. The availability of cellular and molecular probes for kinesin and other putative axonal transport motors have led to a reevaluation of our understanding of intracellular motility.     

Elektronenmikroskopische Untersuchungen an spezifischen Organellen von Endothelzellen des Frosches (Rana temporaria)

Zusammenfassung Endothelzellen der Wirbeltiere enthalten spezifische Organellen, deren Funktion unbekannt ist. Diese Organellen werden beim Frosch (Rana temporaria) nach unterschiedlichen Fixierungen elektronenmikroskopisch untersucht. Die Organellen sind walzenförmig mit mannigfachen Abweichungen, bis zu 2 lang und 0,1 bis 0,5 dick. Ihre oft unterbrochene Außenmembran ist dicker als zytoplasmatische Membranen. Das Innere der Organellen besteht aus Tubuli, die in eine elektronendichte Matrix eingebettet sind. Die Dichte dieser Matrix zeigt deutliche Abstufungen. Die Tubuli sind möglicherweise aus einer spiralförmigen Molekülkette aufgebaut. Das Verteilungsmuster der Organellen wird mit stereologischen Methoden untersucht. Die größte Volumendichte weisen die Aortae thoracicae mit 8% auf. Die Volumendichte der Organellen im Zytoplasma der Endothelzellen scheint mehr von der Entfernung der betreffenden Gefäßstrecke zum Herzen abzuhängen als von der Gefäßgröße. Es werden Verbindungen der Organellen zu zytoplasmatischen Membransystemen aufgezeigt. Auf Besonderheiten des Endothels, darunter Aggregationen von Ribosomen, wird hingewiesen.

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Electron microscopic studies on specific organelles of endothelial cells in the frog (Rana temporaria)

Summary Endothelial cells of vertebrates contain specific organelles of unknown function. These organelles are studied by electron microscopy with different fixations. The organelles are rod-shaped with many variations, up to 2 in length and 0.1 to 0.5 in thickness. Their outer membrane, which is often discontinuous, is thicker than cytoplasmic membranes. The density of the matrix shows distinct gradations. The organelles contain tubules, possibly built up by a spiral molecular chain. The distribution of the organelles is investigated with stereological methods. Their volume density in endothelial cell cytoplasm appears to depend more on the distance from the heart than on vessel size, the thoracic aortae showing the highest organellae content of 8%. Connections between organelles and cytoplasmic membranes are demonstrated. Particularities of endothelium, among them aggregations of ribosomes, are pointed out.

Herrn Prof. Dr. med. F. Hammersen und Fräulein Dr. med. M. Lewerenz danke ich herzlich für Anregungen, Unterstützung und Kritik. Die Arbeit entstand während eines vom Deutschen Akademischen Austauschdienst dankenswerterweise gewährten Stipendiums; sie wurde unterstützt vom Schweizerischen Nationalfonds zur Förderung der wissenschaftlichen Forschung, Kredit-Nr. 3952.     

Lung alveolar cell cytosomes: A consideration of their significance

Summary An ultrastructural comparison of mammalian, reptilian, and amphibian lung alveolar cells, and avian lung atrial cells reveals that morphologically similar cytoplasmic bodies (cytosomes) occur in these cells. The cytosomes, which appear generally as osmiophilic, lamellae-containing, membrane-bound, round bodies 0.3 to 0.5 in diameter, are also similar to bodies occurring in epithelial cells of both physoclistous and physostomatous swimbladders of fishes. Because the function of both lung alveolar and swimbladder epithelial cells is gas-handling, the possibility is raised that the morphologically similar lamellae-containing bodies of these vertebrate cells are functionally identical. One function, suggested by other investigators, is that, in mammalian lungs, these bodies supply a surface-tension lowering material (surfactant). Because several assumptions concerning this proposed function remain unproved, an alternative proposal is speculatively explored. The suggestion is offered that cytosomes contain an antioxidant needed to protect alveolar and swimbladder cells against the toxic effects of the relatively high concentration of oxygen to which these cells are exposed.Supported by a research grant from the American Cancer Society, Oregon Division, Inc.     

Molecular characterization of cDNAs encoding low-molecular-weight heat shock proteins of soybean

Three cDNA clones (GmHSP23.9, GmHSP22.3, and GmHSP22.5) representing three different members of the low-molecular-weight (LMW) heat shock protein (HSP) gene superfamily were isolated and characterized. A fourth cDNA clone, pFS2033, was partially characterized previously as a full-length genomic clone GmHSP22.0. The deduced amino acid sequences of all four cDNA clones have the conserved carboxyl-terminal LMW HSP domain. Sequence and hydropathy analyses of GmHSP22, GmHSP22.3, and GmHSP22.5, representing HSPs in the 20 to 24 kDa range, indicate they contain amino-terminal signal peptides. The mRNAs from GmHSP22, GmHSP22.3, and GmHSP22.5 were preferentially associated in vivo with endoplasmic reticulum (ER)-bound polysomes. GmHSP22 and GmHSP22.5 encode strikingly similar proteins; they are 78% identical and 90% conserved at the amino acid sequence level, and both possess the C-terminal tetrapeptide KQEL which is similar to the consensus ER retention motif KDEL; the encoded polypeptides can be clearly resolved from each other by two-dimensional gel analysis of their hybrid-arrest translation products. GmHSP22.3 is less closely related to GmHSP22 (48% identical and 70% conserved) and GmHSP22.5 (47% identical and 65% conserved). The fourth cDNA clone, GmHSP23.9, encodes a HSP of ca. 24kDa with an amino terminus that has characteristics of some mitochondrial transit sequences, and in contrast to GmHSP22, GmHSP22.3, and GmHSP22.5, the corresponding mRNA is preferentially associated in vivo with free polysomes. It is proposed that the LMW HSP gene superfamily be expanded to at least six classes to include a mitochondrial class and an additional endomembrane class of LMW HSPs.     

细叶黄芪叶肉原生质体发育早期几种细胞器的变化

在细叶黄芪叶肉原生质体发育早期,细胞器的变化较大。离体培养4h后,线粒体的嵴和基质物质开始增加。培养3—5天后,线粒体的数量增加5倍以上,此时可见大部分线粒体围绕细胞核分布。在培养24h后,高尔基体开始发育,它们主要分布在细胞质周边区域。多糖细胞化学染色表明,高尔基体内沉积着大量嗜银物质。培养1天后,粗面内质网开始发育。培养3天时,部分叶绿体边缘出现一些空隙结构。随着叶绿体内膜结构的消失,淀粉粒增大,叶绿体逐渐转变为造粉质体。     

On Paraptychostomum almae N. G., N. Sp., a Commensal Ciliate from the Digestive Tract of Oligochaetes of the Cameroons, in a New Subclass Hysterocinetia

ABSTRACT. Morphological and ultrastructural studies on a new ciliate, Paraptychostomum almae , from the digestive tract of an oligochaete ( Alma emini ) from the Cameroons are carried out. The flattened cell has a large size; its left lateral face bears an anterior thigmotactic zone that includes seven-nine short kinetal segments. The somatic cortex is composed of flattened alveoli, a thin epiplasm and a microfibrillar ecto-endoplasmic boundary. Kineties are made of monokinetids, each particularly characterized by a long anteriorly directed kinetodesmal fiber, and a hyperdivergent postciliary ribbon. The postero-ventral buccal apparatus consists of a short peristome and a deep longitudinal infundibulum. The paroral organelle is a long stichodyad. The three adoral organelles are of different types: ADI and AD3 are of the membranoid type, respectively with two and one rows of ciliated kinetosomes; AD2 is of the peniculus type with six-seven rows of ciliated kinetosomes. A microfibrillar network with nodes arises from all the buccal kinetosomes and extends under the naked wall. Mitochondria are small and numerous and dispersed throughout the whole cell. The existence of an AD2 with more than two rows of kinetosomes warrants the creation of the new genus Paraptychostomum and a new family, Ptychostomatidae. The presence of a distinct ecto-endoplasmic boundary and of somatic kinetids exclusive without transversal dense tractus, hyperdivergent postciliary ribbons, and dispersed numerous mitochondria, added to particularities of the stomatogenesis, allow us to clearly separate hysterocinetians from the scuticociliates and to set up for them the new subclass Hysterocinetia, within the class Oligohymenophorea, with a single new order Hysterocinetida.