scholarly journals STRUCTURE OF ISOLATED PLANT GOLGI APPARATUS REVEALED BY NEGATIVE STAINING

1966 ◽  
Vol 28 (2) ◽  
pp. 169-179 ◽  
Author(s):  
William P. Cunningham ◽  
D. James Morré ◽  
H. H. Mollenhauer

Sucrose-gradient-purified dictyosomes of plant Golgi apparatus appear, after glutaraldehyde stabilization, as stacks of highly fenestrate and tubate cisternae when negatively stained with phosphotungstic acid, shadowed with heavy metal, or OsO4-stained in thin section. The tubular proliferations (diameter 200 to 400 A) extend for several microns from the central region and are united at intervals into an anastomosing network. Associated with the tubules are two kinds of vesicles which are distinguishable on the basis of texture, size, shape, and staining characteristics. One vesicle type is rough-surfaced, nearly spherical, and of uniform dimensions (diameter approximately 600 A). Metal shadowing shows that these vesicles remain spherical after drying. The other vesicle type is smooth-surfaced and varies in both size and shape. Intercisternal elements are revealed, by negative staining, on the surface of internal cisternae after fragmentation of the dictyosome. The progressive differentiation of cisternae from the forming face to the maturing face is observed in thin sections of these isolated preparations. The morphological characteristics observed in negatively stained dictyosomes indicate regions of functional specialization within the dictyosome cisternae and reveal a dictyosome structure more extensive than that envisioned from sections.

1978 ◽  
Vol 30 (1) ◽  
pp. 151-174
Author(s):  
J.G. Robertson ◽  
M.P. Warburton ◽  
P. Lyttleton ◽  
A.M. Fordyce ◽  
S. Bullivant

Peribacteroid membranes and bacteroid envelope inner membranes have been isolated from developing lupin nodules. Isolation of the peribacteroid membranes was achieved by first preparing membrane-enclosed bacteroids free from other plant organelles or membranes. The peribacteroid membranes were then released by osmotic shock and purified by centrifugation to equilibrium on sucrose gradients. The bacteroids were broken in a pressure cell and the bacteroid envelope inner membranes were isolated using sucrose gradient fractionation of the bacteroid total envelope preparation. The density of the peribacteroid membranes decreased during the period of development of N2-fixation in lupin nodules from 1.148 g/ml for nodules from 12-day plants to 1.137 g/ml for nodules from 18-day plants. The density of the bacteroid envelope inner membranes from nodules from 18-day plants was 1–153 g/ml. The identity and homogeneity of the isolated membranes was established, by comparison with membranes in intact nodules, using phosphotungstic acid and silver staining of thin sections and particle densitites on faces of freeze-fracture replicas of the membranes. Analyses for NADH oxidase and succinate dehydrogenase, spectral analyses and gel-electrophoretic analysis of proteins were also used to characterize the membrane and soluble protein fractions from the nodules. The ratio of lipid to protein was 6.1 for the peribacteroid membranes and 2.5 for the bacteroid envelope inner membranes. Leghaemoglobin was localized in the plant cytoplasm in lupin nodules and not in the peribacteroid space.


1974 ◽  
Vol 62 (2) ◽  
pp. 491-504 ◽  
Author(s):  
William P. Cunningham ◽  
L. Andrew Staehelin ◽  
Robert W. Rubin ◽  
Ross Wilkins ◽  
Mary Bonneville

Isolated Golgi complexes can be recognized in phosphotungstate (PTA) negative stain as stacks of membranous plates surrounded by a complex anastomosing network of tubules and vesicles. The extent of this tubular network is, however, much greater than can be observed in thin sections of whole cells. To determine which of the steps leading to the final negatively stained image may produce the observed changes, we have monitored each of the steps by other electron microscope and biochemical methods. The first damage to the membranes seems to occur during the initial isolation procedure as judged by the appearance of smooth patches on the freeze-fractured membrane faces that are normally covered with particles. Subsequent suspension of the Golgi fraction in water, to dilute the sucrose for negative staining, leads to the disappearnce of the stacking, to some tubulation and some vesiculation of the membranes as judged by thin section and freeze-cleave microscopy. The latter technique also reveals an increase in smooth-cleaving membrane faces. Application of the negative stain to the water-washed Golgi fraction, finally, produces extensive tubular arrays and a simultaneous decrease in the remaining large membranous vesicles. The freeze-cleaved tubular membranes appear essentially smooth except for small patches of aggregated particles. Parallel gel electrophoresis studies of the membranes and of the water and negative stain wash extracts indicate that protein extraction is involved in these morphological changes. PTA seems to be a particularly effective solvent for certain membrane proteins that are not removed by the water wash. These observations suggest that removal of membrane proteins alters structural restraints on the membrane lipids so that they behave semiautonomously like myelinics and form new artificial structures. This does not eliminate the possibility, however, that some tubules also exist in the Golgi apparatus in vivo.


Author(s):  
E. N. Albert

Silver tetraphenylporphine sulfonate (Ag-TPPS) was synthesized in this laboratory and used as an electron dense stain for elastic tissue (Fig 1). The procedures for the synthesis of tetraphenylporphine sulfonate and the staining method for mature elastic tissue have been described previously.The fine structure of developing elastic tissue was observed in fetal and new born rat aorta using tetraphenylporphine sulfonate, phosphotungstic acid, uranyl acetate and lead citrate. The newly forming elastica consisted of two morphologically distinct components. These were a central amorphous and a peripheral fibrous. The ratio of the central amorphous and the peripheral fibrillar portion changed in favor of the former with increasing age.It was also observed that the staining properties of the two components were entirely different. The peripheral fibrous component stained with uranyl acetate and/or lead citrate while the central amorphous portion demonstrated no affinity for these stains. On the other hand, the central amorphous portion of developing elastic fibers stained vigorously with silver tetraphenylporphine sulfonate, while the fibrillar part did not (compare figs 2, 3, 4). Based upon the above observations it is proposed that developing elastica consists of two components that are morphologically and chemically different.


Author(s):  
J. Quatacker ◽  
W. De Potter

Mucopolysaccharides have been demonstrated biochemically in catecholamine-containing subcellular particles in different rat, cat and ox tissues. As catecholamine-containing granules seem to arise from the Golgi apparatus and some also from the axoplasmic reticulum we examined wether carbohydrate macromolecules could be detected in the small and large dense core vesicles and in structures related to them. To this purpose superior cervical ganglia and irises from rabbit and cat and coeliac ganglia and their axons from dog were subjected to the chromaffin reaction to show the distribution of catecholamine-containing granules. Some material was also embedded in glycolmethacrylate (GMA) and stained with phosphotungstic acid (PTA) at low pH for the detection of carbohydrate macromolecules.The chromaffin reaction in the perikarya reveals mainly large dense core vesicles, but in the axon hillock, the axons and the terminals, the small dense core vesicles are more prominent. In the axons the small granules are sometimes seen inside a reticular network (fig. 1).


Author(s):  
S.R. Allegra

The respective roles of the ribo somes, endoplasmic reticulum, Golgi apparatus and perhaps nucleus in the synthesis and maturation of melanosomes is still the subject of some controversy. While the early melanosomes (premelanosomes) have been frequently demonstrated to originate as Golgi vesicles, it is undeniable that these structures can be formed in cells in which Golgi system is not found. This report was prompted by the findings in an essentially amelanotic human cellular blue nevus (melanocytoma) of two distinct lines of melanocytes one of which was devoid of any trace of Golgi apparatus while the other had normal complement of this organelle.


Author(s):  
I. R. Khuzina ◽  
V. N. Komarov

The paper considers a point of view, based on the conception of the broad understanding of taxons. According to this point of view, rhyncholites of the subgenus Dentatobeccus and Microbeccus are accepted to be synonymous with the genus Rhynchoteuthis, and subgenus Romanovichella is considered to be synonymous with the genus Palaeoteuthis. The criteria, exercising influence on the different approaches to the classification of rhyncholites, have been analyzed (such as age and individual variability, sexual dimorphism, pathological and teratological features, degree of disintegration of material), underestimation of which can lead to inaccuracy. Divestment of the subgenuses Dentatobeccus, Microbeccus and Romanovichella, possessing very bright morphological characteristics, to have an independent status and denomination to their synonyms, has been noted to be unjustified. An artificial system (any suggested variant) with all its minuses is a single probable system for rhyncholites. The main criteria, minimizing its negative sides and proving the separation of the new taxon, is an available mass-scale material. The narrow understanding of the genus, used in sensible limits, has been underlined to simplify the problem of the passing the view about the genus to the other investigators and recognition of rhyncholites for the practical tasks.


1974 ◽  
Vol 60 (1) ◽  
pp. 92-127 ◽  
Author(s):  
Melvyn Weinstock ◽  
C. P. Leblond

The elaboration of dentin collagen precursors by the odontoblasts in the incisor teeth of 30–40-g rats was investigated by electron microscopy, histochemistry, and radioautography after intravenous injection of tritium-labeled proline. At 2 min after injection, when the labeling of blood proline was high, radioactivity was restricted to the rough endoplasmic reticulum, indicating that it is the site of synthesis of the polypeptide precursors of collagen, the pro-alpha chains. At 10 min, when the labeling of blood proline had already declined, radioactivity was observed in spherical portions of Golgi saccules containing entangled threads, and, at 20 min, radioactivity appeared in cylindrical portions containing aggregates of parallel threads. The parallel threads measured 280–350 nm in length and stained with the low pH-phosphotungstic acid technique for carbohydrate and with the silver methenamine technique for aldehydes (as did extracellular collagen fibrils). The passage of label from spherical to cylindrical Golgi portions is associated with the reorganization of entangled into parallel threads, which is interpreted as the packing of procollagen molecules. Between 20 and 30 min, prosecretory and secretory granules respectively became labeled. These results indicate that the cylindrical portions of Golgi saccules transform into prosecretory and subsequently into secretory granules. Within these granules, the parallel threads, believed to be procollagen molecules, are transported to the odontoblast process. At 90 min and 4 h after injection, label was present in predentin, indicating that the labeled content of secretory granules had been released into predentin. This occurred by exocytosis as evidenced by the presence of secretory granules in fusion with the plasmalemma of the odontoblast process. It is proposed that pro-alpha chains give rise to procollagen molecules which assemble into parallel aggregates in the Golgi apparatus. Procollagen molecules are then transported within secretory granules to the odontoblast process and released by exocytosis. In predentin procollagen molecules would give rise to tropocollagen molecules, which would then polymerize into collagen fibrils.


1964 ◽  
Vol 20 (2) ◽  
pp. 217-233 ◽  
Author(s):  
G. W. Claus ◽  
L. E. Roth

The morphological features of the cell wall, plasma membrane, protoplasmic constituents, and flagella of Acetobacter suboxydans (ATCC 621) were studied by thin sectioning and negative staining. Thin sections of the cell wall demonstrate an outer membrane and an inner, more homogeneous layer. These observations are consistent with those of isolated, gram-negative cell-wall ghosts and the chemical analyses of gram-negative cell walls. Certain functional attributes of the cell-wall inner layer and the structural comparisons of gram-negative and gram-positive cell walls are considered. The plasma membrane is similar in appearance to the membrane of the cell wall and is occasionally found to be folded into the cytoplasm. Certain features of the protoplasm are described and discussed, including the diffuse states of the chromatinic material that appear to be correlated with the length of the cell and a polar differentiation in the area of expected flagellar attachment. Although the flagella appear hollow in thin sections, negative staining of isolated flagella does not substantiate this finding. Severe physical treatment occasionally produces a localized penetration into the central region of the flagellum, the diameter of which is much smaller then that expected from sections. A possible explanation of this apparent discrepancy is discussed.


Phytotaxa ◽  
2016 ◽  
Vol 270 (2) ◽  
pp. 71 ◽  
Author(s):  
SLAVOMÍR ADAMČÍK ◽  
MIROSLAV CABOŇ ◽  
URSULA EBERHARDT ◽  
MALKA SABA ◽  
FELIX HAMPE ◽  
...  

The current generally accepted concept of Russula maculata defines the species by yellow-brownish spots on the basidiomata, an acrid taste, a yellow spore print and a red pileus. This concept was tested using collections originating from various geographical areas mainly in Europe. Analyses of the ITS region suggested that there were three species within this broad concept. One of them, R. maculata, was identified based on the sequence from the epitype. Two other species, R. nympharum and R. sp., are described here as newly identified species. The European species R. maculata and R. nympharum grow in deciduous forests, are similar in their field aspect and are distinctly different in micro-morphological characteristics of spores, pleurocystidia and pileipellis. An Asian species, R. sp., is associated with pine and has smaller basidiomata and spores. These three species form the R. maculata complex and represent the sister clade to the R. globispora complex. This clade consists of species also characterized by a yellow-brownish context discolouration but with a different type of spore ornamentation. All of the other tested species had an acrid taste and yellow spore print but did not have a conspicuous yellow-brownish context discolouration and were placed in various unrelated clades.


Author(s):  
Wei-hong Wang ◽  
Xue-gang Luo ◽  
Zhe Wang ◽  
Yu Zeng ◽  
Feng-qiang Wu ◽  
...  

To investigate the heavy metal and metalloid contamination of soil around a Huanan uranium tailings pond, abandoned in 1998, we defined a study area of 41.25 km2 by a natural boundary and targeted 5 elements’ (U, Mn, As, Pb, Cr) single contamination and comprehensive pollution as the assessment contents. First, we collected 205 samples and evaluated them with the contamination factor (CF) method aiming at judging whether the single target element concentration exceeded the local background value and environmental quality standard. We obtained CF1 (the background value of a certain target element as the baseline value) and CF2 (the environmental quality standard for soils as the baseline value). Second, we evaluated the ecological risk of the key pollutant U with the risk assessment code (RAC) method, taking the 27 samples whose CF2 > 1 as examples and concluded that the environmental risk of U was relatively high and should arouse concern. Third, we selected comprehensive pollution index (CPI) to assess the compound pollution degree of five target elements. Fourth, we constructed the U contamination and CPI’s continuous distribution maps with spatial interpolation, from which we worked out the sizes and positions of slightly, moderately and strongly polluted zones. Finally, we analyzed the spatial variability of U and CPI with the aid of a geostatistical variogram. We deduced that the spatial variation of uranium was in close relationship with local topography, and probably precipitation was the driving force of U contamination diffusion, whereas CPI exhibited weak spatial dependence with random characteristics. The above work showed that 3.14 km2 soil near the pond was fairly seriously polluted, and the other 4 elements’ single contaminations were less serious, but the 5 target elements’ cumulative pollution could not be ignored; there were other potential pollution sources besides the uranium tailings pond. Some emergency measures should be taken to treat U pollution, and bioremediation is recommended, taking account into U’s high bioavailability. Further, special alerts should be implemented to identify the other pollution sources.


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