Sporoderm development in Liriodendron chinense (Magnoliaceae): a probable role of the endoplasmic reticulum

The developmental events in the sporoderm and the cytoplasm of Liriodendron chinense microspore from the early tetrad stage until late free microspore stage were observed. Various forms of the endoplasmic reticulum (ER) and surprising unusual aggregates of ER seen during microspore development attract special attention. Being scanty at early and middle tetrad stage, while primexine matrix (glycocalyx) acquires well-defined form, the ER becomes distinct in unusual forms at the late tetrad stage. Thin long tubules with an osmiophilic contents, which cannot be compared with the tubular smooth endoplasmic reticulum (SER), undulate through the cytoplasm. Towards the end of the tetrad period when callose begins to disintegrate, and a distinct tectate-columellate pattern of the ectexine becomes evident, two new forms of the SER occur in the cytoplasm. Instead of single tubules observed previously, 3–tubuled aggregates meander through the cytoplasm, the middle tubule contains an osmiophilic substance. The second form of the SER looks like an ordinary tubular SER, but has ampoule-like dilations with dark granular contents. Later on the tubules undergo major changes: multi-tubuled aggregates of parallel tubules overcrowd the cytoplasm, the outer tubules of each aggregate carrying ribosomes. These aggregates undulate through the cytoplasm, branch, and are associated with lipid globules. The tips of many aggregates are pressed to the plasmalemma. The ontogenetic period of time of the presence of these ER aggregates, their structure and localization in the microspore cytoplasm allow me to assume that these ER aggregates synthesize sporopollenin precursors.     

ULTRASTRUCTURE OF TETRASPOROGENESIS IN THE PARASITIC RED ALGA LEVRINGIELLA GARDNERI (SETCHELL) KYLIN12

Ultrastructural studies on tetraspore formation in Levringiella gardneri revealed that 3 stages may be recognized during their formation. The youngest stage consists of a uninucleate tetraspore mother cell with synaptonemal complexes present during early prophase of meiosis I. Mitochondria are aggregated around the nucleus, dictyosome activity is low, and chloroplasts occur in the peripheral cytoplasm. A 4-nucleate tetraspore mother cell is formed prior to tetrahedral cell cleavage, and an increase in the number of chloroplasts and mitochondria occurs. Small straight-profiled dictyosomes secrete vesicles into larger fibrous vesicles or contribute material to the developing tetraspore wall. During the second stage of tetraspore formation, striated vesicles form within endoplasmic reticulum, semicircular profiled dictyosomes secrete vesicles for fibrous vesicles or wall material, and starch formation increases. The final stage is characterized by the disappearance of striated vesicles, presence of straight, large dictyosomes which secrete cored vesicles, and an abundance of starch grains. Cleavage is usually complete at this stage and the tetraspore wall consists of a narrow outer layer of fibrillar material and an inner, electron transparent layer. These spores are surrounded by a tetrasporangial wall which was the original wall surrounding the tetraspore mother cell.     

Pre-fertilization degeneration of both synergids in Brassica campestris ovules

Summary In Brassica campestris, both synergids of the ovule degenerate before the arrival of the pollen tube. Synergid degeneration does not depend on pollination. At the non-degenerated stage, the synergids are completely filled with a complexly organized cytoplasm containing numerous mitochondria with many cristae, a large number of dictyosomes with many associated vesicles, and a very extensive rough endoplasmic reticulum. The degenerative changes that occur in the cytoplasm of the synergids are characterized by a loss of visibility of the membranes of the endoplasmic reticulum and the simultaneous formation of dense deposits on the surrounding membranes of the mitochondria. Locally, the plasma membranes of the synergids disappear, and some ground plasma of the synergids penetrates into the space between the plasma membranes of the egg cell and the central cell.     

Formation of protein storage bodies during embryogenesis in cotyledons of Sinapis alba L.

An electron microscopic investigation of fine structural changes in post-meristematic cotyledon mesophyll cells during the period of storage protein accumulation (16–32 d after pollination) showed that the rough ER, the Golgi apparatus and the developing vacuome are intimately involved in the formation of storage protein bodies (aleurone bodies). At the onset of storage protein accumulation (16–18 d after pollination) storage protein-like material appears within Golgi vesicles and preformed vacuoles. At a later stage (24 d after pollination) similar material can also be detected within vesicles formed directly by the rough endoplasmic reticulum (ER). It is concluded that there are two routes for storage protein transport from its site of synthesis at the ER to its site of accumulation in the vacuome. The first route involves the participation of dictyosomes while the second route bypasses the Golgi apparatus. It appears that the normal pathways of membrane flow in the development of central vacuoles in post-meristematic cells are used to deposit the storage protein within the protein bodies. Thus, the protein body can be regarded as a transient stage in the process of vacuome development of these storage cells.Abbreviation ER endoplasmic reticulum     

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     

Cytochemical localization of cellulase activity in pollen mother cells of david lily during meiotic prophase I and its relation to secondary formation of plasmodesmata

Summary Cellulase activity was localized at the ultrastructural level in pollen mother cells (PMCs) of David lily [Lilium davidii var.willmottiae (Wilson) Roffill] at different stages of meiotic prophase I. The enzyme was observed to appear at the early leptotene stage and reached its highest level at the subsequent zygotene stage, and its subcellular distribution revealed by the presence of electron-dense deposits of reaction product was found to be restricted exclusively to the endoplasmic reticulum (ER), the vesicles derived from that, and the cell wall, especially at the sites of secondary plasmodesmata and cytoplasmic channels where the wall was being digested. Other cytoplasmic organelles, such as dictyosomes and Golgi vesicles, lacked such deposits of reaction product. After zygotene the enzyme activity decreased abruptly, and at the pachytene stage only very few deposits could be observed in the cell wall. Our results indicate that cellulase is synthesized on rough ER and secreted directly via the smooth ER and ER-derived vesicles into the cell wall by exocytosis, where it brings about local wall breakdown, leading to the secondary formation of plasmodesmata and cytoplasmic channels.     

Apomixis in plants: structural and functional aspects of diplospory inPoa nemoralis andP. palustris

Summary Data on structural and functional aspects of mitotic diplospory and later stages of apomictic seed formation are reported forPoa palustris andP. nemoralis. In this study, the plant material of two Russian populations ofP. nemoralis andP. palustris were used for transmission electron microscope observations. Seed formation was investigated by phase contrast microscopy in two populations ofP. nemoralis collected in The Netherlands. The processes of transformation of the megasporocytes to the megaspore mother cells of diplosporous embryo sacs, and thereafter to one- and two-nucleate diplosporous embryo sacs (Antennaria type) were characterized by an increase of cell size, structural and functional reorganization of the nucleus, nucleolus, and cytoplasm, and cell isolation as a result of thickening of the cell wall. These were accompanied by an increase in the cell metabolic activity inferred from visual evidence of the activation of nucleus, nucleolus, endoplasmic reticulum, dictyosomes, mitochondria, and from the appearance of a dense population of ribosomes and polysomes. The diplosporous embryo sac of the Antennaria type was characteristic for bothP. nemoralis andP. palustris. No signs of the presence of synaptonemal complexes were observed during the process of diplosporous-embryo-sac megaspore mother cell differentiation and division. About 90–95% of the diploid egg cells of diplosporous embryo sacs were able to produce apomictic embryos. These embryos developed before anthesis. However, many of them degenerated at the globular stage because of lack of endosperm. The ultrastructural events occurring during the process of diplospory of apomictic species, and meiosis and megagametogenesis of sexually reproduced plants are discussed.Abbreviations DMC megaspore mother cell of diplosporous embryo sac - TEM transmission electron microscopy - ER endoplasmic reticulum     

The endomembrane system and mechanism of membrane flow in the green alga,Gloeomonas kupfferi (Volvocales,Chlorophyta) I. An ultrastructural analysis

Summary The endomembrane system of the chlamydomonad flagellate,Gloeomonas kupfferi (Chlorophyta), is complex. It consists of a proliferating ER network, a perinuclear complex of 14–18 dictyosomes and 8–12 vacuoles and an anterior contractile vacuole complex. The ER network extends from the nuclear envelope outwards, ensheafhs a dictyosome, extends out through a lobe of the chloroplast and terminates in the thin zone of peripheral cytoplasm between the chloroplast and plasmamembrane. The individual dictyosome is polar with distinct cis- and trans-faces. The cis-face is closely associated with transition vesicles emerging from the adjacent ER. Large vesicles emerge from peripheral swellings of terminal cisternae. The dictyosome-associated ER is connected to the peripheral vacuolar system. During cell division and cytokinesis, changes in the endomembrane system occur. Dictyosomes divide and quickly separate to form perinuclear complexes around the daughter nuclei. Each dictyosome undergoes morphological changes during this wall precursor-producing stage. ER lines the furrow zone and is closely associated with phycoplast microtubules. A discussion of the endomembrane system in membrane flow mechanics is provided.Abbreviations ER endoplasmic reticulum - OsFeCN Osmium ferricyanide     

Histochemical and ultrastructural changes in the haustorium of date (Phoenix dactylifera L.)

Summary During imbibition ofPhoenix dactylifera embryos, all cotyledon cells show the same changes: protein and lipid bodies degrade, smooth endoplasmic reticulum (ER) increases in amount, and dictyosomes appear. At germination, the distal portion of the cotyledon expands to form the haustorium. At this time, epithelial cells have a dense cytoplasm with many extremely small vacuoles. Many ribosomes are present along with ER, dictyosomes, and mitochondria. The parenchyma cells have large vacuoles and a small amount of peripheral cytoplasm. Between 2 and 6 weeks after germination, epithelial cells still retain the dense cytoplasm and many organelles appear: glyoxysomes, large lipid bodies, amyloplasts, large osmiophilic bodies, and abundant rough and smooth ER which appear to merge into the plasmalemma. A thin electron-transparent inner wall layer with many small internal projections is added to the cell walls. Starch grains appear first in the subsurface and internal parenchyma and subsequently in the epithelium. Lipid bodies, glyoxysomes, protein, and osmiophilic bodies occur in the epithelial and subepithelial cell layers but not in the internal parenchyma. At 8 weeks after germination, the cytoplasm becomes electron transparent, vacuolation occurs, lipid bodies and osmiophilic bodies degrade, and the endomembranes disassemble. After 10 weeks, the cells are empty. These data support the hypothesis that the major functions of the haustorium are absorption and storage.     

Developmental changes in the inner epidermis of the bean seed coat

Summary The inner epidermis of the bean seed coat shows remarkable structural changes during seed development. At the globular stage of development, a moderately electron-dense substance begins to accumulate in the outer tangential and radial walls of the cells. The staining and fluorescence characteristics, together with the localization of peroxidase in the wall, suggest that this electron-dense material is a phenolic substance. At the same stage of embryo development, structural specialization can be detected in the cytoplasm of the epidermal cells with an increase in the abundance of organelles, especially the endoplasmic reticulum, mitochondria, and dictyosomes. These structural features are similar to those in the underlying branched parenchyma cells. As the seed rapidly expands during the maturation stage of embryo development, the epidermal cells and the inner layers of the branched parenchyma cells begin to degenerate. Small ruptures can be detected in the epidermis, exposing the branched parenchyma cells. These structural changes are discussed in relation to their possible functions during embryo development.     

Evolution of the cytoplasmic organelles during female meiosis in Pisum sativum L.

In this paper we have traced the evolution of the cytoplasmic organelles in the female germinal cell of Pisum sativum L., from the beginning of meiosis to the early stages of the maturing megaspore, in order to correlate the morphological changes with the physiological aspects of megasporogenesis.A process of intense cytoplasmic vacuolation takes place in the megaspore mother cell (MMC) during prophase I, probably proceeding from the smooth endoplasmic reticulum and dictyosomes; it results in the formation of big vacuoles, which play a role in MMC polarization. By means of this polarization most plastids and mitochondria are incorporated into the functional megaspore at the end of meiosis.There are plastid and mitochondria cycles which consist of dedifferentiation followed by redifferentiation, During these cycles a transient morphology appears, called a cup-shaped form, which we interpret as an expression of low organelle activity.The wall of the MMC thickens throughout megasporogenesis and loses its plasmodesmata during middle prophase I. The ribosome population is reduced during prophase I and then restored during the early stages of the megaspore maturing process, as shown by the quantitative study that we have carried out. The nucleolar cytoplasmic bodies play a part in this restoring process. These bodies have a special morphology and appear to be originated from the activity of the nucleolar organizing region (NOR) during nucleolar disorganization in prophase I.We think that this cytoplasmic evolution is a response to nuclear genic recombination, in order to provide the most adequate expression of the zygote genome.Abbreviations EDTA ethylene-diamine-tetracetic acid - ER endoplasmic reticulum (SER: smooth ER) - MMC megaspore mother cell - NOR nucleolar organizing region - RNP ribonucleoproteins This work has been partially supported by the Comisión Asesora para la investigación Cientifica by Técnica Projects n^o 613/02 and 613/10     

Functional anatomy of the mechanoreceptor cells in tendrils of Bryonia dioica Jacq.

The ultrastructure of epidermal cells of tendrils of Bryonia dioica Jacq. (Cucurbitaceae) was determined using microscopic, histochemical and immunochemical techniques with focus on the tactile blep, the mechano-receptor of these cells. Tactile bleps resemble bordered pits in structure and probably in formation. They contain cytoplasm rich in endoplasmic reticulum, dictyosomes, mitochondria and microbodies. The cytoplasm is highly vesiculated and usually contains lipid-body-like structures. Cytoskeletal elements (microtubules, actin filaments) are uniquely arranged in the tactile blep, and chlorotetracycline-fluorescence analysis shows large amounts of membrane-associated calcium within the tactile blep. We propose a physically interconnected cytoskeleton-membrane device as the immediate force sensor and transducer which creates a primary intracellular signal, for which calcium is a likely candidate.Abbreviations CTC chlorotetracycline - ER endoplasmic reticulum - MT microtubule This work was funded by grants of the Deutsche Forschungsgemeinschaft to E.W.W. and G.W. (SFB 1462) and by the Fonds der Chemischen Industrie (literature provision).     

Inhibition of ion uptake to barley roots by cycloheximide

In Petunia pollen tubes growing in the style there appear to be two ways of callose deposition. The first one is callose deposition outside the plasma membrane as a distinct layer closely appressed to the cell wall. The second one is callose deposition within the cytoplasm as distinct callose grains, leading to the formation of callose plugs. This second way is accompanied by a characteristic ultrastructure of the cytoplasm, namely strong electron-density of the plasma matrix, partial absence of the plasma membrane and the absence of plastids and dictyosomes. For both ways of callose deposition a mechanism is proposed and the function of callose plugs is discussed.Abbreviation RER rough endoplasmic reticulum     

Ultrastructure of Nectaries of Vinca rosea L., Vinca major L. and Citrus sinensis Osbeck cv. Valencia and its Relation to the Mechanism of Nectar Secretion

The ultrastructural changes in nectar-secreting cells of Vincarosea, Vinca major, and Citrus sinensis during their ontogeneticdevelopment are described. The most pronounced changes occurin the amount and morphology of the endoplasmic reticulum (ER).The amount of ER elements increases gradually and reaches amaximum at the stage of secretion. At this stage the ER is thedominant element in the cytoplasm. A process of swelling ofa lamellar ER, followed by formation of vesicles, was notedin the secretory cells during the stage of secretion. Many vesicularelements appeared to be in a close association with the plasmalemma.It is suggested that sugar is secreted as a solution by meansof vesicles derived from the ER.     

Membranes in the spindle of Iris pollen mother cells during the second division of meiosis

Summary Spindle membrane in the second division of meiosis inIris comprises endoplasmic reticulum (ER) and small spherical vesicles (40–80 nm diameter). The vesicles are in all respects similar to those of dictyosomes and it is proposed that they originate in the Golgi system. ER on the other hand is generated from the nuclear envelope (NE). Invaginations and evaginations of the inner and outer membranes of the NE are present at interphase and they enlarge during prophase II and break to form spindle ER elements at the beginning of prometaphase II. The area enclosed by this ER continues to increase until the entire spindle is filled with irregular profiles at mid-prometaphase II. During late-prometaphase II the quantity of ER decreases and, concurrent with an increase in the number of microtubules (MTs), the cisternae align alongside MT bundles and accumulate at the poles. It is suggested that ER increases in quantity by growth of NE membranes. Furthermore, the alignment of ER along the interpolar axis and its transport polewardsprior to anaphase suggest the action of a force on each ER element.     

朱顶红花粉萌发过程中营养细胞质的超微结构变化

本文应用透射电镜对朱顶红成熟花粉水合、活化和萌发的动态过程中营养细胞质的结构和组成变化进行了观察。成熟花粉具质体、线粒体、内质网、高尔基体。微丝束以聚集体的形式存在。花粉活化后,细胞器的数目和结构发生显著变化：质体和线粒体的片层明显增加,内质网片层狭窄,高尔基体活跃产生小泡,脂体降解及微丝聚集体散开。花粉萌发后,细胞质中出现周质微管和被刺小泡,此期细胞器的变化不明显。微丝以纤丝状遍布整个花粉管中。     

Brefeldin A induces reversible dissociation of the Golgi apparatus in the green algaMicrasterias

Summary The fungal metabolite brefeldin A (BFA) causes inhibition of cell growth inMicrasterias denticulata after 2 h incubation, combined with slight malformation of the cell shape. The BFA effects on cell development are accompanied by a gradual decrease in the number of Golgi cisternae and severe structural and morphological changes of the dictyosomes which are already visible after only 10 min exposure. When the treatment is prolonged the number of dictyosomes is markedly reduced, leading to almost complete loss of Golgi bodies, particularly in the young semicell. Groups of primary wall material-containing vesicles accumulated in areas of former dictyosomes, and previously unknown vesicular bodies are found. Restitution of almost normal dictyosomes occurs within 5 h when the cells are allowed to recover from BFA treatment.Micrasterias cells incubated in BFA at concentrations below 15 M maintain their ability to divide over several generations. Our results indicate that, of the various inhibitors of the secretory pathway tested against growingMicrasterias cells, BFA is the only drug which induces complete and reversible dissociation of dictyosomes in the growing semicell. This allows deductions about the function of the processes targeted by BFA during cell development inMicrasterias.Abbreviations BFA brefeldin A - CPA cyclopiazonic acid - ER endoplasmic reticulum - TM tunicamycin     

ULTRASTRUCTURE OF DEVELOPING SIEVE ELEMENTS IN THLASPI ARVENSE L. II. MATURATION

Developing sieve elements of pennycress (Thlaspi arvense L.) were studied with the electron microscope. The maturation of sieve elements involved loss of ribosomes from cytoplasm; degeneration of nulcei; modification of endoplasmic reticulum (ER); loss of tonoplast; and disappearance of dictyosomes and dictyosomes vesicles, coated vesicles, microtubules, and microbodies. Such changes produce a mature, presumably conducting cell that contains no nucleus or central vacuole but which retains a thin layer of peripheral cytoplasm with plastids, mitochondria, and smooth ER. Some similar changes have been described in a variety of developing sieve elements of angiosperms, but coated vesicles and microbodies previously have not been followed through sieve-element maturation. Likewise, few developmental studies have been made of plant sieve elements that exhibit two types of P-protein, the tubular type and the granular P-protein body.     

Ultrastructure and cytochemistry of the pearl gland in Piper regnellii (Piperaceae)

Pearl glands are scattered throughout the lamina of developing leaves and rarely found on adult leaves of Piper regnellii (Piperaceae). The pearl gland is a bicellular secretory trichome composed of a short broad basal cell and a spatula-like, semiglobular apical cell. Four different stages of the pearl gland were determined during its ontogenesis: origin, pre-secretory, secretory and post-secretory. During the pre-secretory stage, mitochondria, ribosomes, dictyosomes, rough endoplasmic reticulum, and plastids with electron dense inclusions were present in the cytoplasm of the apical cell. During the secretory stage, the most remarkable characteristics of the apical cell are the proliferation of dictyosomes and their vesicles, rough endoplasmic reticulum, and modified plastids. At this stage, electron-dense oil drops occur in the plastids as well as scattered within the cytoplasm, proteins and polysaccharides are seen in the plastids, vesicles, and vacuoles. Only polysaccharides are present in the periplasmic space, wall cavities, and on the surface of the apical cell. The polysaccharides are one of the main components of the mucilagenous exudate that covers the developing leaf structures. The apical cell of the senescing trichomes undergoes a progressive degeneration of its cellular components, the plastids being the first organelles to undergo lysis.     

Ultrastructural changes in the MP3 neuron of the mollusk Lymnaea stagnalis after cryopreservation of the isolated brain

Investigation of a possibility of long-term storage of frozen (-196 degrees C) viable neurons and nervous tissue is one of the central present day problems. In this study ultrastructural changes in neurons of frozen-thawed snail brain were examined as a function of time. We studied the influence of cryopreservation, cryoprotectant (Me2SO), cooling to 4-6 degrees C, and a prolonged incubation in physiological solution at 4-6 degrees C on dictyosomes of Golgi apparatus, endoplasmic reticulum (ER) cisternae and mitochondria. It has been found that responses of these intracellular structures of cryopreserved neurons to the above influences are similar: dissociation of Golgi dictyosomes, swelling of endoplasmic reticulum cisternae and mitochondrial cristae. Both freezing-thawing and cryoprotectant were seen to cause an increase in the number of lysosomes, liposomes, myelin-like structures, and to form large vacuoles. The structural changes in molluscan neurons caused by cryopreservation with Me2SO (2 M) were reversible.