Phospholipid-synthesizing Enzymes Associated with Golgi Dictyosomes from Pea Tissue

Golgi dictyosomal membranes isolated from pea (Pisum sativum) stem tissue, using a combination of rate zonal and isopycnic sucrose density centrifugation, were shown to bear cytidine diphosphate-choline:diglyceride phosphorylcholinetransferase, CDP-ethanolamine:diglyceride phosphorylethanolaminetransferase, and CTP:phosphorylcholine cytidyltransferase activities. Although the majority of the activity of the phospholipid-synthesizing enzymes was associated with the endoplasmic reticulum, the activity found in the Golgi system was about 25% of the total activity. These results suggest that Golgi dictyosomes probably synthesize at least part of the membrane phospholipids that they may need for their secretory function and for dictyosomal proliferation during cell growth, rather than importing this material entirely from the endoplasmic reticulum.     

ULTRASTRUCTURE AND ACID PHOSPHATASE IN PEDICEL ABSCISSION OF HIBISCUS

Pedicel abscission in Hibiscus rosa-sinensis was investigated by light and electron microscopy. During the pre-abscission period endoplasmic reticulum declined somewhat, dictyosomes increased in number and apparent activity, and mitochondria maintained their numbers. The observations suggested that dictyosomal vesicles were migrating to and fusing with the plasma membrane. The enzyme acid phosphatase was associated with dictyosomes and dictyosomal saccules, with small vacuoles and invaginations of the plasma membrane, and in the paramural region between the plasma membrane and the cell wall. Our interpretation is that acid phosphatase, (and probably also the enzymes involved in cell wall dissolution) are transported via an endoplasmic reticulum-dictyosome-vesicle carrier system to the paramural regions of the cell. In more general terms, our observations support the view that the enzymes involved in the cell wall hydrolysis of abscission are synthesized within a compartmentalized, lysosomal system prior to their release and action.     

ROLE OF DICTYOSOMES IN WALL FORMATION DURING CELL DIVISION OF CHLORELLA VULGARIS

The behavior of dictyosomes in wall formation during cell division of Chlorella vulgaris follows a definite pattern. During formation of the partition membrane they migrate into the equatorial plane and pair. There is a close spatial relationship between the dictyosomes and the partition membrane which, itself, may be derived from the fusion of dictyosomal vesicles. Dictyosomes also may participate significantly in the deposition of new wall material.     

Cellular compartments in mitotic cells: ultrahistochemical identification of Golgi elements in PtK-1 cells

This investigation focuses on the identification, distribution, and transport of intracellular membrane systems during mitosis. The membranes of the Golgi apparatus can be identified cytochemically by staining for acid phosphatase (acPase) and thiamine pyrophosphatase (TPPase) activity. Using this approach we are able to study the disintegration of the Golgi apparatus during mitosis and to follow the dislocation as well as the organized reappearance of Golgi elements after the completion of mitosis. We are able to demonstrate that during mitosis the activity of both enzymes is strong enough to react with the substrate applied during the staining procedure. Furthermore, we observe a characteristic pattern of membrane distribution in mitotic cells. During interphase the TPPase reaction is characteristically limited to one or two cisternae of a dictyosomal stack. The acPase reaction stains the membranes of the total stack, of the GERL, of some vesicles and cisternae near the dictyosomes and lysosomes. After the mitotic breakdown of the dictyosomal stacks the forming vesicles still stain positively and are distributed over the entire cytoplasm. At late anaphase and early telophase the enzyme activity occurs not only in the reconstituting dictyosomes but also in the nuclear envelope and in some ER cisternae. The extended spectrum of membrane structures indicating Golgi enzyme activity becomes obvious. This phenomenon favors the idea that at least some functions of the Golgi apparatus persist during mitosis.     

Number of dictyosomes in a single cell of the green algaMicrasterias

The numbers of dictyosomes in cells ofM. crux-melitensis andM. pinnatifida were counted at various stages in the cell cycle. Dictyosomes synchronously doubled in number by dividing at the premitotic stage and then were separated into two groups by the septum, thus reducing the dictyosomal number to the ordinal number in each new cell. The number remained the same throughout the cell cycle until the next premitotic stage.     

Seasonal Variation in the Ultrastructure of the Cambium in New Zealand Grown Pinus radiata D. Don

Examination of Pinus radiata cambium from trees growing in thecentral North Island of New Zealand has revealed that many ofthe structural changes occurring in other tree species at theonset or cessation of cambial activity are not found in thisspecies. The winter cambium bears a closer resemblance to thesummer cambium than it does to the winter cambium of any otherangiosperm or gymnosperm described in the literature. In particularthere is little change in vacuolar structure, endoplasmic reticulumform, and dictyosomal activity during the year. The only changeswhich take place involve a slight increase in vacuole volume,storage of starch in vacuoles, and a decrease in numbers ofspherosomes during the summer. These observations confirm that,while the degree of cambial activity is reduced in the winter,complete dormancy is absent.     

Regions of lead uptake inLemna minor plants and localization of this metal within selected parts of the root

Investigations were carried out to determine the sites of lead uptake within the frond and the root ofLemna minor. With the sodium rhodizonate four regions favoured in lead uptake were distinguished: the frond region between the base and the node, the basal part of the root, and the regions at the proximal and distal ends of the root cap. For analysis in electron microscope only the root regions were chosen. The highest rate of lead uptake was found in the basal part of the root. Lead was present in the apoplast of this region after 5 min of exposure and was observed in the stelar cells after 30 min of incubation. Lead deposits were detected mostly in the cell walls adjacent to the plasma membrane and in the lumen of several endomembrane compartments - the endoplasmic reticulum (ER), dictyosomal vesicles, nuclear envelope and the vacuoles. Lead induced changes of cell ultrastructure; an increase in the number of membraneous structures, swelling of ER cisternae and distortion of the dictyosomal cisternae were observed after 2 to 6 h of exposure. We wish to thank Mrs. G. Winiecka for her technical assistance in preparing the photographs.     

Disturbance of the secretory pathway inMicrasterias denticulata by tunicamycin and cyclopiazonic acid

Summary Both tunicamycin, an inhibitor of N-linked glycosylation of proteins, and cyclopiazonic acid, which inhibits the Ca²⁺-dependent ATPase in the ER, influence the secretory pathway at the ER level and lead to a cessation of cell growth inMicrasterias. Electron microscopical investigations reveal that the mode of action of the two inhibitors differs. While tunicamycin treatment results in a disintegration of the Golgi bodies into small vesicles, cyclopiazonic acid prevents products being supplied from the ER, resulting in the dilatation of ER cisternae and a reduction in the number of Golgi cisternae, combined with a loss of dictyosomal activity. The disturbed cell wall formation under tunicamycin indicates that N-linked glycosylation of proteins is required for normal cell growth inMicrasterias. Moreover, our studies reveal that changes in cytoplasmic free calcium concentration, as a consequence of ATPase inhibition in the ER by cyclopiazonic acid, may inhibit wall material secretion by interrupting the normal ER-dictyosome association.Abbreviations CPA cyclopiazonic acid - ER endoplasmic reticulum - TM tunicamycin     

Acid phosphatase activity during spore differentiation of the red algaeGigartina teedii andChondria tenuissima

The acid phosphatase activity during carposporogenesis inGigartina and tetrasporogenesis inChondria was studied using the Gomori technique. During the first steps of gonimoblast maturation ofGigartina, portions of cytoplasm are ensheathed by ER cisternae with acid phosphatase activity, giving rise to autolysosomal concentric membrane bodies. In a similar way large mucilage sacs are severed. They extrude their contents in a kind of exocytosis. Multivesicular bodies, concentrically arranged cisternae and extracytoplasmic compartments, each with acid phosphatase activity, remain in young carpospores for some time, probably as remnants of the autophagocytotic and exocytotic events. The Golgi apparatus is poorly developed in gonimoblast cells and young carpospores. It becomes a prominent cell component in maturing carpospores and then participates in cell wall formation. Only some of the dictyosomal cisternae contain acid phosphatase; these are irregularly distributed in the dictyosome. — In pre- and postmeiotic tetraspore mother cells ofChondria massive lead deposits are found in the dictyosomes and in adjacent Golgi vesicles. Finer lead precipitates occur in ER cisternae, especially in those which are sequestering starch-grain-containing portions of the cytoplasm to give rise to autolysosomes. During cell cleavage, the dictyosomes aggregate. They become devoid of acid phosphatase activity with the exception of vesicles at the trans face. Later, Golgi stacks associate and have common, Gomori positively reacting, narrow cisternae at the cis face. The Golgi apparatus derived cored vesicles do not contain lead precipitates whereas the Golgi cisternae in the final stage of tetrasporogenesis show acid phosphatase activity. Variations in acid phosphatase distribution are explained in the light of current models of membrane flow.Dedicated to Univ.-Prof. DrO. Härtel on the occasion of his 80th birthday.     

Three-Dimensional Reconstruction of a Dictyosome using Serial Sections

A single dictyosome from a suspension cultured sycamore cellhas been reconstructed on an enlarged scale using electron micrographsobtained from a series of 12 thin sections as the basis forthe moulding of profile slices. Slices were hardened and stucktogether to form single cisternae, tubules and vesicles. Thesestructural elements were joined, true-to-scale, to give a three-dimensionalfigure. The model thus obtained is more realistic than otherwell-known dictyosomal models previously presented in the literature.In contrast to these our model exhibits occasional membranecontinuities between adjacent cisternae of the dictyosomal stackand entertains the possibility of similar connections betweencisternae of the endoplasmic reticulum and the Golgi apparatus. Key words: Acer pseudoplatanus, Dictyosome reconstruction     

Relationship between microbial diversity and nitrogenase activity of Stipagrostis pennata rhizosheath

Nitrogen is the key factor for plant survival and growth, especially in the desert. Stipagrostis pennata, a sand born drought-resistant plant, could colonize pioneerly in Gurbantunggut Desert during revegetation. One strategy for their environment adaptation was the rhizosheath formatted by root-hair, mucilaginous exudates, microbial components, and soil particles, for which not only provides a favorable living microenvironment but also supplies essential nutrients. To understand the relationship between microorganisms living in rhizosheaths and the nitrogen nutrition supply, the microbial diversity and nitrogenase activity was estimated during the growth of S. pennata. Five samples of the rhizosheath, which based on the development periods of the plant, regreen, flowering, filling, seed maturating, and withering period, were collected. The nitrogenase activity was estimated by acetylene reduction and the microbial diversity was analyzed by 16S rRNA high-throughput sequencing. The results showed that the nitrogenase activity was increased slowly during regreen to flowering, while reached a peak rapidly at filling sample and then decreased gradually. A total of 274 operational taxonomic units (OTUs) were identified and significant differences in community structure and composition at each growth period. Among them, the main phyla included Actinobacteria and Proteus, which were the most abundant phyla in all periods. In addition, the microbial diversity in the grain filling period was higher than other periods in view of the analysis of alpha diversity and beta diversity. Furthermore, principal component analysis (PCA) analysis showed that the microbial communities in the filling period was low in similarity with other periods. Most importantly, the OTUs associated with nitrogen fixation is the most during the filling period, involving Phagecidae and Fucoraceae. Overall, the study not only revealed the differences in nitrogenase activity among different developmental periods in S. pennata, but also explored the potential bridges between it and community structure and diversity of bacteria.     

Pigment biogenesis in freshwater shrimp ventral nerve chord chromatophores

Summary The possible biogenesis of two pigment granule types present in the monochromatic, brown chromatosomes enveloping the ventral nerve chord of the freshwater palaemonid shrimps Macrobrachium acanthurus, M. heterochirus and M. olfersii is examined by transmission electron microscopy in thin section and freeze fracture replicas. Prominent, membrane limited granules are suggested to have their origin in a complex, juxtanuclear, smooth endoplasmic reticulum labyrinth, continuous with the nuclear envelope. Amembranous, lipocarotenoid granules possibly derive from the external surface of the smooth endoplasmic reticulum. Nuclear envelope and SER membranes contain numerous 11 nm diameter intramembranous particles while pigment granule membranes exhibit fewer particles. A dictyosomal origin for the lipocarotenoid granules is discounted. Granulogenesis is suggested to be a continuous process in crustacean chromatophores.     

Recurrent seasonal changes in bacterial growth efficiency,metabolism and community composition in coastal waters

The microbial response to environmental changes in coastal waters of the eastern Cantabrian Sea was explored for four years by analysing a broad set of environmental variables along with bacterial community metabolism and composition. A recurrent seasonal cycle emerged, consisting of two stable periods, characterized by low bacterial metabolic activity (winter) from October to March, and high bacterial metabolic activity (summer) from May to August. These two contrasting periods were linked by short transition periods in April (T_A) and September (T_S). The phylogenetic groups Alphaproteobacteria and Bacteroidetes were dominant during winter and summer respectively, and their recurrent alternation was mainly driven by the bloom of eukaryotic phytoplankton before T_A and the bloom of prokaryotic phytoplankton before T_S. Bacterial growth efficiency remained high and stable during the winter and summer periods but dropped during the two short transition periods. Our results suggest that bacterial growth efficiency should be considered a very resilient property that reflects different stages in the adaptation of the bacterial community composition to the environmental changes occurring throughout the seasonal cycle in this coastal ecosystem.     

Protease activity inAgaricus bisporus during periodic fruiting (flushing) and sporophore development

Protease activity from sporophores and mycelium of the mushroomAgaricus bisporus was assayed during periodic cropping (flushing) and from sporophores during maturation. When the sporophores were harvested at the same developmental stages (pins or buttons) during cropping, proteolytic activity of the sporophores was found to oscillate with the same periodicity as the flushing cycle. For pin mushrooms (an early stage of development), peaks of activity occurred during the interflush periods, whereas for button mushrooms (a later stage of development) peak proteolytic activity coincided with the periods of maximum production. The proteolytic activity in the mycelium remained low and varied little with time. Of the tissues within the sporophore, gill tissue had a higher activity than the stipe or pileus. The changes in activity during sporophore development or maturation depended on the period in the flushing cycle when the sporophore was initiated. The results are discussed in relation to the possible role and regulation of flush co-ordinated proteases.     

Diurnal Variations in the Activity of Phosphoenolpyruvate Carboxylase and NADP-Malic Enzyme During the Early Steps of Interaction between Glycine max and Bradyrhizobium japonicum

Two important enzyme in organic acid metabolism, phosphoenolpyruvate carboxylase (PEPC) and NADP-malic enzyme (NADP-ME), show marked diurnal rhythms in their activities during the establishment of the soybean - B. japonicum symbiosis. The pH of the nutrient solution changes in parallel with NADP-ME activity, being maximal during the night periods, whereas activity of PEPC was highest during the day periods. The results from the experiments with stem girdled plants indicated that the activity of root PEPC is modulated to a great extent by the supply of photosynthates from the shoots. It was also established that succinate application in the nutrient solution during inoculation altered significantly the pattern of assayed enzyme activities. Although our experiments did not reveal the precise mechanism of the involvement of root PEPC and NADP-ME in soybean response to inoculation with B. japonicum, they indicated the pattern of their activity during the first 72 h postinoculation which are critical for establishment and functioning of the symbiosis. This revised version was published online in July 2006 with corrections to the Cover Date.     

The golgi-mediated scale production in the marine haptophycean alga Ochrosphaera neapolitana Schussnig

Abstract

Some ultrastructural features of cells of the marine haptophycean alga, Ochrosphaera neapolitana Schussnig in the palmelloid stage were examined. Chloroplasts which are contained in a compartment isolated from the cytoplasm by ER profiles and nuclear envelope, display trilamellated thylakoids running along the major axis. The stalked pyrenoid with inner bilamellated thylakoids, protrudes in a large membrane-bounded vacuole. Other structures, as the haptonematic and flagellar bases, autophagic vacuoles and mitochondria, are typical of the chrysophycean and haptophycean genera so far investigated.

The Golgi apparatus is represented by a single dictyosome composed of stacked cisternae fonctioning in a way that they form organic scales which constitute the main part of the cell covering. The scales, build up of microfibrils disposed parallel each to other, lie in cisternal lumina of the dictyosomal maturing face; scaly cisternae are numerous in the peripheral cytoplasm and are observed merging in the plasma membrane and discharging the content outside the protoplast.

Dictyosomal activity is evidenced morphologically by massive vesicle production. Three kinds of membrane-bounded vesicles were identified in the present material: i) inner-granulated vesicles, arising from the maturation face; ii) coated vesicles, scattered in the cytoplasm or at the periphery of the golgi body, and iii) dense-cored vesicles, present in the proximity of the maturation face. The possible functional relationships related to scale production and assembly outside the protoplast, and between the nucleus and dictyosome are discussed.     

Genetic changes in mating activity in laboratory strains of Drosophila subobscura

Eleven populations of Drosophila subobscura that had been maintained in laboratory conditions during different periods of time were examined for evidence of genetic divergence in mating activity. The results indicate that mating activity increases with the time of maintenance under laboratory conditions.     

Some aspects of the ecology of the desert snail Sphincterochila prophetarum in relation to energy and water flow

Summary Between 1976–1978, we studied the ecology of the Sphincterochila prophetarum, the most common snail, on stony slopes in Northern Negev. Energy and water flow determinations were made at the individual and population levels on a field observation basis and metabolic data were derived from laboratory simulations.The snails are active during 5–7% of the time during the year, while the remaining 95% of the time, the snails are in aestivation under stones. The energy and water inputs during the short feeding periods are very fast. Only 8% of the total energy in the eaten food are assimilated. Eighty-nine percent of the total assimilated energy are utilized in respiration during the various periods of activity and dormancy.The water loss during the feeding and activity period is about 37% of the total annual input. The snails lose twice as much the amount of water during the winter dormancy periods than in the summer dormancy period.Dedicated to Prof. Evenari and Dr. Springer     

Partitioning of temporal activity among desert lizards in relation to prey availability and temperature

Partitioning of activity time within ecological communities potentially reduces interspecific competition and increases the number of species that can coexist. We investigated temporal activity in a highly diverse lizard assemblage in the Simpson Desert, central Australia, to determine the degree of partitioning that occurs. Three periods were defined, daytime (sunrise to sunset), early night (sunset to midnight) and late night (midnight to sunrise), and live captures of lizards were tallied for each period during two sampling months (September and November 2007). We also quantified the activity times of potential invertebrate prey and measured ambient temperatures during the different time periods to investigate any associations between these factors and lizard activity. Some 77% of captures of 13 lizard species were made by day, with Ctenotus pantherinus, Egernia inornata (Scincidae) and Nephrurus levis (Gekkonidae) the only species showing extended nocturnal activity. Activity of both species of skink was recorded at temperatures 4°C lower than those for agamid and varanid lizards early in the night, and at temperatures as low as 18–20°C. Surface‐active invertebrates differed in composition between time periods and were less abundant during the late night period in the drier of the two sample months (September), but were distributed equally over time in the other month. Termites were active in subterranean galleries at night in September and mostly by day in November, but available at all times on surface/subsurface baits. We conclude that activity is distributed unevenly within this lizard assemblage, with partitioning facilitated by the ready availability of invertebrate prey and by lizards having relatively broad temperature tolerances that, in some cases, permit opportunistic exploitation of resources beyond usual times of activity.