CYTOLOGY OF DIFFERENTIATING TRACHEARY ELEMENTS I. ORGANELLES AND MEMBRANE SYSTEMS

The distribution of organelles, membrane systems, and ribosomes is not at any time obviously related to the pattern of secondary wall in helically thickened tracheary elements in leaves of Beta vulgaris L. (sugar beet) and Cucurbita maxima Duchesne, fixed with potassium permanganate and osmium tetroxide. During the differentiation of the secondary wall, cisternae of the endoplasmic reticulum and dictyosomes are particularly conspicuous, and the dictyosomes are associated with numerous vesicles. Similar vesicles appear to be in various stages of fusion with the secondary wall thickenings. The tracheary elements contain plastids which may include starch granules. Ribosomes occur free in the cytoplasm and in association with endoplasmic membranes.     

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.     

Characterization of fibrous compound of Golgi vesicles in tapetal cells ofDelphinium

Summary It was shown that Golgi structures abundantly appearing in tapetal cells ofDelphinium Ajacis L. developing anthers, prior to meiocytes meiosis, show a fine fibrous material within their vesicles. At the time of the formation of tapetal cell wall this fibrous component, released by an exocytotic process, is incorporated into the cell wall. The membrane of dictyosomes derived vesicles participates in the development of plasma membrane. Fibrous material appears to be morphologically similar to the fibrils of tapetal cell wall; this cell wall gives a positive reaction for cellulose and pectins, as visible in the light microscope. Moreover, the fibrous and pectinase resistant compound of dictyosomes derived vesicles and the fibrils of cell wall disappear partly after cellulase digestion which proves their cellulosic character. On the other hand pectinase treatment as well as ruthenium red staining suggest associated with cellulose pectins within Golgi vesicles.     

CELL SHAPE AND WALL PATTERN IN RELATION TO CYTOPLASMIC ORGANIZATION IN PEDIASTRUM SIMPLEX

P. simplex is a single-pronged, fenestrated species of Pediastrum. Comparison is made in regard to cell differentiation and structure with P. boryanum, a 2-pronged, unfenestrated species, with emphasis on the origin of cell wall pattern and the regulation of cell shape. The characteristic wall pattern is initiated with the deposition of plaques of wall material of the outer wall layer when zoospores have assembled in the colony. The pattern is postulated to be templated in the plasma membrane. The inner, thicker wall layer is fibrillar and deposited from vesicles derived from the golgi apparatus. In P. simplex 2–4 dictyosomes are present in contrast to the single dictyosome of P. boryanum. The dictyosomes lie at the concave inner face of the nucleus. Blebs of its ribosome-free outer membrane are contributed to the forming face of the golgi apparatus. Parallel microtubules underlie the plasma membrane in the aggregating zoospores and disappear after the initiation of wall formation. The possible role of microtubules and other organelles in the determination of cell shape in Pediastrum is discussed.     

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.     

Numerical and structural changes in dictyosomes during zygospore germination ofClosterium ehrenbergii

Summary Numerical and structural changes in dictyosomes during the germination of zygospores inClosterium ehrenbergii were examined by electron microscopy. In the dormant mature zygospores, two parallel cisternac were seen which were derived from the disorganization of dictyosomes during the maturation of zygospores. After the induction of germination, the two parallel cisternae developed into dictyosomes with ten or eleven cisternae. The dictyosomes doubled in number by division every day for four days and reached, at the time of germination, a density of distribution similar to that found in the youngest zygospore. On the 4th day after the induction of germination, dictyosomes produced two kinds of vesicles which appear to be involved in the formation of new cell wall layers. The germination of the zygospore was effected by the escape of the cell covered with the new cell wall layers through the broken old cell wall layers.     

CYTOCHALASIN B INFLUENCES DICTYOSOMAL VESICLE PRODUCTION AND MORPHOGENESIS IN THE DESMID EUASTRUM1

Cytochalasin B (CB) applied to young developing cells of the desmid Euastrum oblongum Ralfs ex Ralfs, at concentrations that do not entirely inhibit cytoplasmic streaming, retarded cell growth and caused malformations of cell shape. While the basic symmetry of the cell was maintained, only the first indentations were formed and the cell body appeared to be swollen. Electron microscopic investigations revealed that vesicle production at the dictyosomes was disturbed by cytochalasin. In contrast to untreated control cells, where vesicles with electron-dense contents (“dark vesicles”) were formed during primary wall formation, vesicles pinched off by the dictyosomes during CB treatment exhibited an “empty” appearance. These vesicles, which correspond to the “dark vesicles” in size, were accumulated around the dictyosomes without being transported to the plasma membrane and were frequently connected to the trans-cisternae of the Golgi bodies. We speculate that CB may influence the transfer of products from the endoplasmic reticulum (ER) to the dictyosomes via transition vesicles, which results in a disturbed vesicle production at the Golgi bodies. CB also causes a shift in ER and dictyosome distribution. Moreover, a cortical actin system appears to be involved in the cell shaping of Euastrum. The arrangement of microtubules around the nucleus is not affected by the drug.     

THE ULTRASTRUCTURE OF CARPOSPOROGENESIS IN THE MARINE HEMIPARASITIC RED ALGA ERYTHROCYSTIS SACCATA1

The ultrastructure of carposporogenesis for Erythrocystis saccata is described. The fusion and gonimoblast cells contain few organelles, and chloroplasts are in a proplastid state, with pit plugs between gonimoblast cells dissolving early in development. Carpospore development may be separated into 3 stages, the first stage being characterized by the appearance of straight-profiled dictyosomes, fibrous vesicles, and an increase of discoid thylakoids within the chloroplasts. During the second, stage the dictyosomes assume a curved profile and striped vesicles are formed by the endoplasmic reticulum. The third stage is initiated by the disappearance of striped vesicles and the appearance of straight-profiled dictyosomes secreting vesicles with cores. Mature carpospores consist of many cored vesicles, fibrous vesicles, and floridean starch grains. A single wall layer surrounds each carpospore since the carposporangial wall becomes incorporated into a mucilaginous matrix surrounding the spores.     

EFFECT OF LATRUNCULIN B AND BREFELDIN A ON CYTOKINESIS IN THE BROWN ALGA SCYTOSIPHON LOMENTARIA (SCYTOSIPHONALES,PHAEOPHYCEAE)1

In zygotes of the brown alga Scytosiphon lomentaria (Lyngb.) Link, cytokinesis proceeds by growth of membranous sacs, which are formed by fusion of Golgi vesicles and flat cisternae accumulated at the future cytokinetic plane. It has been reported that depolymerization of actin filaments by latrunculin B does not inhibit mitosis. However, this molecule prevents the formation of the actin plate, which appears at the region of intermingled microtubules from each centrosome just before and during cytokinesis. In this study, zygotes treated with latrunculin B were observed using EM. Remarkably, this reagent inhibited the formation of flat cisternae. Golgi vesicles gathered around the midzone between the two daughter nuclei and fused with the plasma membrane there. As a result, the plasma membrane invaginated, in a complicated manner, into the cytoplasm. However, these invaginations of the plasma membrane never produced a continuous partition membrane. The ultrastructure of zygotes treated with brefeldin A, which prevents Golgi‐mediated secretion, was also examined. Flat cisternae appeared at the future cytokinetic plane, and a new cell partition membrane was formed. However, the partition membrane became thick, because it was filled with amorphous material rather than the normal rigid fibrous material. These results suggested that actin is involved in the formation of flat cisternae, where it is necessary for completion of the new cell partition membrane, and that Golgi vesicles may play an important role in the deposition of cell wall material.     

THE ULTRASTRUCTURE OF CARPOSPOROGENESIS IN CALOGLOSSA LEPRIEURII (DELESSERIACEAE,CERAMIALES)

Carposporogenesis in Caloglossa leprieurii is divided into three cytological stages. At stage I, the young spores have few plastids and little starch. Abundant dictyosomes secrete a gelatinous wall layer in scale-like units. At stage II, dictyosomes produce a second fibrillar wall component in addition to the gelatinous constituent. Large fibrillar vesicles accumulate in the cytoplasm. Production of gelatinous material decreases in this stage. By stage III, starch grains and fully developed plastids are abundant. Rough endoplasmic reticulum occupies much of the peripheral cytoplasm. A dense, granular proteinaceous component appears in the wall in association with the fibrillar layer. Arrays of randomly oriented tubules are scattered in the cytoplasm. The mature carpospore is surrounded by an outer gelatinous wall layer and an inner fibrillar layer. Few dictyosomes persist in the mature spore. Carposporogenesis in Caloglossa is compared with that in other red algae.     

Ultrastructure of the differentiating zygotospores of Porphyra leucosticta (Rhodophyta)

The ultrastructure of zygotosporogenesis is described for the red alga Porphyra leucosticta Thuret. Packets of eight zygotosporangia, each packet derived from a single carpogonium are interspersed among vegetative cells. Zygotospore differentiation in Porphyra can be separated into three developmental stages. (i) Young zygotospores exhibit a nucleus and a large centrally located, lobed plastid with pyrenoid. Mucilage is produced within concentric membrane structures during their dilation, thus resulting in the formation of mucilage sacs. Subsequently, these sacs release their contents, initiating the zygotospore wall formation. Straight‐profiled dictyosomes produce vesicles that also provide wall material. During the later stages of young zygotospores, starch polymerization commences, (ii) Medium‐aged zygotospores are characterized by the presence of fibrous vacuoles. These are formed from the ‘fibrous vacuole associated organelles’. The fibrous vacuoles finally discharge their contents. (iii) Mature zygotospores are recognized by the presence of numerous cored vesicles produced by dictyosomes. Cored vesicles either discharge their contents or are incorporated into the fibrous vacuoles. There is a gradual reduction of starch granules during zygotospore differentiation. Mature zygotospores are surrounded by a fibrous wall, have a large chloroplast with pyrenoid and well‐depicted phycobilisomes but are devoid of starch granules.     

THE FINE STRUCTURE OF DIFFERENTIATING XYLEM ELEMENTS

Differentiating xylem elements of Avena coleoptiles have been examined by light and electron microscopy. Fixation in 2 per cent phosphate-buffered osmium tetroxide and in 6 per cent glutaraldehyde, followed by 2 per cent osmium tetroxide, revealed details of the cell wall and cytoplasmic fine structure. The localized secondary wall thickening identified the xylem elements and indicated their state of differentiation. These differentiating xylem elements have dense cytoplasmic contents in which the dictyosomes and elements of rough endoplasmic reticulum are especially numerous. Vesicles are associated with the dictyosomes and are found throughout the cytoplasm. In many cases, these vesicles have electron-opaque contents. "Microtubules" are abundant in the peripheral cytoplasm and are always associated with the secondary wall thickenings. These microtubules are oriented in a direction parallel to the microfibrillar direction of the thickenings. Other tubules are frequently found between the cell wall and the plasma membrane. Our results support the view that the morphological association of the "microtubules" with developing cell wall thickenings may have a functional significance, especially with respect to the orientation of the microfibrils. Dictyosomes and endoplasmic reticulum may have a function in some way connected with the synthetic mechanism of cell wall deposition.     

AN ULTRASTRUCTURAL BASIS FOR HYPHAL TIP GROWTH IN PYTHIUM ULTIMUM

Hyphae of the fungus Pythium ultimum extend by tip growth. The use of surface markers demonstrates that cell expansion is limited to the curved portion of the hyphal apex. Growing and non-growing regions are reflected in internal organization as detected by light and electron microscopy. The young hypha consists of three regions: an apical zone, a subapical zone and a zone of vacuolation. The apical zone is characterized by an accumulation of cytoplasmic vesicles, often to the exclusion of other organelles and ribosomes. Vesicle membranes are occasionally continuous with plasma membrane. The subapical zone is non-vacuolate and rich in a variety of protoplasmic components. Dictyosomes are positioned adjacent to endoplasmic reticulum or nuclear envelope, and vesicles occur at the peripheries of dictyosomes. A pattern of secretory vesicle formation by dictyosomes is described which accounts for the formation of hyphal tip vesicles. Farther from the hyphal apex the subapical zone merges into the zone of vacuolation. As hyphae age vacuolation increases, lipid accumulations appear, and the proportional volume of cytoplasm is reduced accordingly. The findings are integrated into a general hypothesis to explain the genesis and participation of cell components involved directly in hyphal tip growth: Membrane material from the endoplasmic reticulum is transferred to dictyosome cisternae by blebbing; cisternal membranes are transformed from ER-like to plasma membrane-like during cisternal maturation; secretory vesicles released from dictyosomes migrate to the hyphal apex, fuse with the plasma membrane, and liberate their contents into the wall region. This allows a plasma membrane increase at the hyphal apex equal to the membrane surface of the incorporated vesicles as well as a contribution of the vesicle contents to surface expansion.     

ULTRASTRUCTURE OF SPERMATIAL DEVELOPMENT IN THE PARASITIC RED ALGAE LEVRINGIELLA GARDNERI AND ERYTHROCYSTIS SACCATA12

Morphologically, the development of spermatia in Levringiella gardneri and Erythrocystis saccata is identical, although cytologically several differences are evident. Mature spermatia contain 1 or 2 large spermatial vesicles that contain fibrous material, several small mitochondria, some proplastids, and are surrounded by a wall, either single-layered as in Erythrocystis or triple-layered as in Levringiella. Spermatial vesicles are formed by aggregations of endoplasmic reticulum in Levringiella, whereas concentric membrane bodies and dictyosomes may be involved in Erythrocystis. In addition to being fibrillar, the contents of the vesicle assume a convoluted appearance in Levringiella. Several spermatia are formed per mother cell and are connected by small pit connections which rupture to allow spermatial release from the spermatangial branch.     

ANALYSIS OF MUCILAGE SECRETION AND EXCRETION IN MICRASTERIAS (CHLOROPHYTA) BY MEANS OF IMMUNOELECTRON MICROSCOPY AND DIGITAL TIME LAPSE VIDEO MICROSCOPY1

Two different, independent, and alternative modes of mucilage excretion were found in the unicellular green alga Micrasterias denticulata Bréb. under constant culture conditions. The cells were capable of either excreting mucilage over all their cell surface or they extruded mucilage from one of their polar ends, which enabled directed movement such as photoorientation or escape from unfavorable environmental conditions. By means of a polyclonal antibody raised against Micrasterias mucilage, the secretory pathway of Golgi derived mucilage vesicles from their origin to their discharge was analyzed by means of conventional and energy filtering TEM. Depending on the stage of the cell cycle, mucilage vesicles were subjected to maturation processes. This may occur either after they have been pinched off from the dictyosomes (e.g. during cell growth) or when still connected to trans‐Golgi cisternae, as in the case of interphase cells. Only fully grown mature vesicles contained mucilage in its final composition as indicated by antibody labeling. After fusion of mucilage vesicles with vacuoles, no immunolabeling was found in vacuoles, indicating that the vesicle content was digested. Mucilage vesicles fused with the plasma membrane in areas of cell wall pores but were also able to excrete mucilage at any site directly through the respective cell wall layer. This result disproves earlier assumptions that the pore apparatus in desmids are the only mucilage excreting areas at the cell surface. Both mechanisms, excretion through the pores and through the cell wall, lead to formation of mucilage envelopes covering the entire cell surface.     

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 CARPOSPOROPHYTE DEVELOPMENT IN THE RED ALGA GLOIOSIPHONIA VERTICILLARIS (CRYPTONEMIALES,GLOIOSIPHONIACEAE)

The ultrastructure of carposporophyte development is described for the red alga Gloiosiphonia verticillaris Farl. The auxiliary cell produces gonimoblast initials, which divide to produce two types of gonimoblast cells—the nondividing vacuolate cells and terminal generative gonimoblast cells. The generative gonimoblast cells form clusters of carpospore initials, which eventually differentiate into carpospores. After gonimoblast filaments are formed, the auxiliary cell undergoes autolysis, causing degeneration of septal plugs between the auxiliary cell and adjacent cells, thus forming a fusion cell. Since this cell lacks starch and appears degenerate throughout carposporophyte development, a nutritive function cannot be ascribed to the fusion cell. Carpospore differentiation is simple and proceeds through three developmental stages. Young carpospores structurally resemble gonimoblast cells, because they contain undeveloped plastids, large quantities of floridean starch, and are surrounded by extensive mucilage instead of a distinct wall. In addition, dictyosomes form and begin to produce vesicles with fibrous contents representing carpospore wall material. During the intermediate stage, dictyosomes continue to produce vesicles that contribute additional carpospore wall material, thereby compressing the mucilage and creating a darker-staining layer outside the carpospore wall. Plastids form internal thylakoids by invaginations of the inner membrane of the peripheral thylakoid. The endoplasmic reticulum forms large granular vacuoles that appear to be degraded during subsequent stages of development. Mature carpospores form cored vesicles. They also contain mature chloroplasts, large amounts of floridean starch, and occasionally granular vacuoles. During this stage, interconnecting carpospore-carpospore and carpospore-gonimoblast cell septal plugs begin to undergo degeneration. This process may be mediated by tubular structures.     

THE FINE STRUCTURE OF SIEVE ELEMENTS OF NEREOCYSTIS LÜTKEANA

The sieve elements of Nereocystis from the base of phylloids contain numerous small vesicles, cytoplasm, ribosomes, and the usual organelles and membrane systems, including nuclei, plastids, mitochondria, dictyosomes, and endoplasmic reticulum. They have a thick secondary wall layer which is deposited along the longitudinal walls and at the sieve plate excluding the sieve pores. The sieve pores range in diameter from 100 to 400 nm and are lined by plasmalemma. The sieve elements from the hollow basal parts of the pneumatocyst show essentially the same features but have larger and fewer vesicles, relatively little cytoplasm, larger sieve pores, 400–900 nm in diameter, and may lack a nucleus. In old sieve elements there are large deposits of callose on the sieve plate and along the longitudinal wall; the vesicles seem to break down, and the protoplast appears necrotic. It is concluded that the trumpet hyphae and sieve tubes are basically the same type of cell, and that the trumpet-shape of the sieve elements is due to their passive stretching during extension growth of the organ in which they occur. There are minor but significant differences among the sieve elements from different regions of the thallus which may reflect possible levels of structural specialization of the sieve elements within the same plant.     

Etude des premiers stades de développement de la Coccidie Coelotropha durchoni

Summary This study considers the earlier growth stages of Coccidium Coelotropha durchoni in its host, Nereis diversicolor. Before evolving into free trophozoites and gamontes in coeliac fluid, the parasites remain in muscular and coeliac cells in microscopic intracellular form. Electron microscope reveals that these stages show an intermediary fine structure between that of a sporozoite — from which they keep some typical characteristics such as the conoid, the fibers and the involuted tubuli — and that of the future free trophozoites. The wall consisting in two clear membrans is provided with one or several micropores. The classical cytoplasmic organites clearly stand out: dictyosomes show constant relationship with ergastoplasm, the mitochondria contain short inner tubuli. Besides the paraglycogen granules and lipoid vacuoles, at least three types of vacuoles may be observed. Peculiar topographic relationship connects mitochondria and paraglycogen granules probably in formation. In the nucleus with classical membrane and heterogeneous structure, a rather voluminous nucleolus may be seen.     

FINE STRUCTURE AND COMPOSITION OF THE EGG APPARATUS BEFORE AND AFTER FERTILIZATION IN QUERCUS GAMBELII: THE FUNCTIONAL OVULE

A study of the egg apparatus of Quercus gambelii was made at both the light and the electron microscope levels. This investigation was concerned primarily with the changes that occur in these cells before and after the process of fertilization and what role, if any, is played by the synergids in this phenomenon. The synergids before fertilization are, on the basis of ultrastructure, healthy, intact, functional cells. They have numerous mitochondria, dictyosomes, endoplasmic reticulum, ribosomes, and a typical nucleus. A prominent filiform apparatus is present, but the cell wall only extends a short distance around the micropylar end of the cells. Just before fertilization, one of the synergids degenerates. This is the synergid that receives the pollen tube and its discharge, including both male gametes. Dictyosomes increase in number and activity in the other synergid (persistent synergid) after fertilization. Eventually a complete cell wall forms around both of the synergids. No plasmodesmata are present in these walls. The egg has numerous mitochondria, dictyosomes, endoplasmic reticulum, and ribosomes, both free in the cytoplasm and attached to the endoplasmic reticulum. Lipid bodies are characteristic of this cell. A cell wall is present only around the micropylar end of the egg. After fertilization, little change occurs in the zygote. The number and activity of the dictyosomes increase, apparently in correlation with cell wall formation. The number of lipid bodies increases. The zygote is approximately the same size as the egg. Plastids are scarce, and starch grains are typically absent from all cells of the egg apparatus. It is suggested that the synergids function in the secretion of chemotropic substances that guide the growth of the pollen tube. Comparisons are made between the egg apparatus of Quercus gambelii and that of the other plants studied thus far.