Seasonal changes in the ultrastructure of the vascular cambium of Robinia pseudoacacia

 The ultrastructure of the vascular cambium of Robinia pseudoacacia L. was examined in trunk tissues collected over a 2 ¹/₂ year period. During dormancy, fusiform cells are densely cytoplasmic with many small vacuoles and centrally located nuclei. Mitochondria are round to oval in sectional view. The plastids are variable in shape, have few internal membranes, and generally lack starch grains. The plasmalemma is smooth in outline. Proteinaceous material occurs in the vacuoles and many lipid droplets are scattered throughout the ground substance. Smooth tubular ER, often highly dilated, predominates, but short segments of rough ER are also present. Abundant free ribosomes are evenly distributed throughout the ground substance and the dictyosomes are inactive. Microtubules are parietal and have various orientations. During reactivation, the plasmalemma becomes irregular in outline and begins to form invaginations. Concurrently, the proteinaceous material disappears, the vacuoles begin to fuse, polysomes appear, and the dictyosomes begin to produce vesicles. During the period of cambial activity, fusiform cells are highly vacuolate, and the nuclei are centrally located. The mitochondria are round, oval, or elongate. Now the plastids contain phytoferritin, starch grains, or both. Many large invaginations of the plasmalemma intrude into the vacuole, pushing the tonoplast inward and pinching off into the vacuole, which lacks proteinaceous material. Lipid droplets are scarce. Most ER is rough, and ribosomes are generally aggregated as polysomes. Dictyosomes are actively producing vesicles. During the transition to dormancy, the fusiform cells gradually assume the appearance typical of the dormant cambium.     

Ultrastructure of dormant buds of Salix sp. in early winter

In the apex of dormant buds of Salk a histological zonation comparable to that found in growing buds was observed. However, significant changes in relative volumes of cell components between dormant and growing buds were noted; the dormant buds had a lower volume density of vacuoles and higher relative volumes of mitochondria, plastids, lipid bodies, and starch grains than the growing buds. In the leaf primordia the relative volume of nuclei decreased with age, while the relative volume of plastids and mitochondria increased. The large central vacuole found in cells of e.g. the pro-cambium and rib meristem in growing buds is split into many smaller ones during the winter. A high content of tannin and calcium oxalate crystals was noted in dormant buds. They also accumulate lipids and starch. Phytoferritin may appear in plastids. Stacked ER and concentric sheaths of ER around lipid bodies appear, probably as a consequence of either anaerobic conditions or water stress. Several indications of metabolic activities in the seemingly dormant buds were found; plasmatubules at the plasmalemma particularly in the procambium, sheaths of smooth ER around the plastids, electron opaque globules (probably calcium-binding sites), and vacuoles that seemed to be autophagic.     

Histogenesis of the transmitting tract in Strelitzia reginae

Organisation and development of the stigmatic, stylar and ovarian parts of the transmitting tract in Strelitzia reginae were evaluated. They were characterised by 1) cell shape, 2) appearance of distal cell wall, 3) type of plastid, 4) and vacuolar system. The long stigmatic trichomes have a secondary irregular wall layer separated from the primary wall. Cell structures include pleomorphic plastids with vesiculated thylakoids and frequently a crystal or a lipidic globule. In early stages of bud development the extensive endoplasmic reticulum (ER) is smooth, whereas it is mainly rough in older buds. Coated vesicles are frequent, as are dictyosomes. Prominent invaginations along the plasma membrane contain floccular deposits in the older flower buds. These deposits are similar in structural appearance to material in the large vacuoles. The basal part of the stigma has wedge-shaped cells with wall ingrowths. Three stages of stigmatic secretion during the development were characterised. The stylar canal is initially narrow but widens subsequently. The cuticle is detached and the apical cell walls show a fringed surface; from this wall inbuddings develop shortly before anthesis. At this stage sheets of rough ER are evident in the cell cortex. The plastids have a few vesiculated thylakoids, proteinaceous crystals and starch grains. The epithelial cells of young buds have numerous vacuoles, the volume of which decreases in more mature cells. The stylar canal is filled with a secretion at all stages of bud development. The face of the ovarian transmitting tract, lining the placenta, is smooth in young buds but lobed in older ones due to the division pattern of the epithelial cells. These cells are large, elongated and culumnar as young but narrow and wedge-shaped when more mature. Cell wall inbuddings are formed late during bud development.     

STRUCTURE AND DEVELOPMENT OF THE SIEVE ELEMENTS IN PSILOTUM NUDUM

Shoot tissue of Psilotum nudum (L.) Griseb. was fixed in glutaraldehyde and postfixed in osmium tetroxide for electron microscopy. Young sieve elements can be distinguished from contiguous parenchyma cells by their distinctive plastids, the presence of refractive spherules, and the overall dense appearance of their protoplast. The refractive spherules apparently originate in the intracisternal spaces of the endoplasmic reticulum (ER). With increasing age the sieve-element wall undergoes a marked increase in thickness. Concomitantly, a marked increase occurs in the production of dictyosome vesicles, many of which can be seen in varying degrees of fusion with the plasmalemma. Other fibril- and vesicle-containing vacuoles also are found in the cytoplasm. In many instances the delimiting membrane of these vacuoles was continuous with the plasmalemma. Vesicles and fibrillar materials similar to those of the vacuoles were found in the younger portions of the wall. At maturity the plasmalemma-lined sieve element contains a parietal network of ER, plastids, mitochondria, and remnants of nuclei. The protoplasts of contiguous sieve elements are connected by solitary pores on lateral walls and pores aggregated into sieve areas on end walls. All pores are lined by the plasmalemma and filled with numerous ER membranes which arise selectively at developing pore sites, independently of the ER elsewhere in the cell. P-protein and callose are lacking at all stages of development.     

An ultrastructural study of early endosperm development and synergid changes in unfertilized cotton ovules

Excised, unfertilized cotton (Gossypium hirsutum L.) ovules were cultured for 1–5 days postanthesis and embryo-sac development was studied with the electron microscope. In some ovules the two polar nuclei fuse and the diploid endosperm nucleus goes through a limited number of free nuclear divisions after 2–3 days in culture. Each nucleus has two nucleoli, in contrast to nuclei of fertilized triploid endosperm which have three nucleoli. Precocious cell walls form between the endosperm nuclei on the 3rd day in culture. The morphology of the plastids, mitochondria, rough endoplasmic reticulum (RER), dictyosomes and microbodies, and the amount of starch and lipid in the diploid cellular endosperm are similar to those of the central cell. A few large helical polysomes appear close to plastids and mitochondria. After 2 days in culture, one of the two synergids in the unfertilized cultured ovules shows degenerative changes which in fertilized ovules are associated with the presence of the pollen tube, i.e., increase in electron density, collapse of vacuoles, irregular darkening and thickening of mitochondrial and plastid membranes, disappearance of the plasmalemma and the membranes of the plasmalemma and the membranes of the RER. The second synergid remains unchanged in appearance. The egg cell does not shrink or divide or show structural changes characteristic of the cotton zygote. Embryo-sac development is arrested on the 4th and 5th days in culture. The nucellus continues growth and at 14 days crushes the degenerate embryo sac.     

Differentiation and Ultrastructure of the Protophloem Sieve Elements of Minor Veins in the Maize Leaves: Changes in the Protoplast

Protophloem sieve element differentiation in the minor veins of the maize ( Zea mays L. ) leaves was first evidenced as an increase of the wall thickness, which began in the comers of the cell and then extended to other parts of the wall, and the appearance of long rough endoplasmic reticulum cistemae distributed throughout the cytoplasm, and then the presence of characteristic crystalloid inclusions within the plastids. As differentiation progressed, long cisternae of rough endoplasmic reticulum appeared to transform into shorter forms and eventually aggregated into small stacks, losing their ribosomes during the process. The nuclei degenerated, although frequently persisted until very late in differentiation the stages of maturation, as darkly stained amorphous aggregates surrounded by double nuclear envelope or only inner membrane of nuclear envelope. Subsequently, the nuclear envelope collapsed and became discontinuous. At the beginning of nuclear degeneration the perinuclear spaces were partly dilated and sometimes the outer nuclear envelope in the dilated portions then ruptured, and was accompanied by the disappearance of the cytoplasmic portion near it. During the peried of nuclear degeneration, in addition to the endoplasmic reticulum, plastids and mitochondria underwent structural modification, while components such as ribosomes, cytoplasmic ground substances, vacuoles and dictyosomes disintegrated and disappeared. At maturity, the surviving protoplasmic components, including plasmalemma, mitochondria, small stacked smooth endoplasmic reticulum and P-type plastids with crystalloids, became parietal in position. As differentiation of adjacent metaphloem sieve elements proceeded, the protoplasmic components of the mature protophloem sieve elements progresively degenerated and finally obliterated.     

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.     

ULTRASTRUCTURE OF THE SHOOT APEX OF CHENOPODIUM ALBUM AND CERTAIN OTHER SEED PLANTS

The ultrastructure of cells of the vegetative shoot apices is described for Chenopodium album, Kalanchoë blossfeldiana and K. laxiflora, Bryophyllum daigremontianum, Nicotiana rustica, and N. tabacum (Maryland Mammoth), and Ginkgo biloba. A less intensive study was made of the last three listed. The structures and organelles usually associated with meristematic cells were observed: dictyosomes, plastids (in various stages of development), mitochondria, endoplasmic reticulum (ER), vacuoles, lipid droplets, and plasmalemma. In addition, spherosome-like structures were observed in all zones of the shoot apices. Also, multivesicular bodies were observed in C. album and B. daigremontianum. Ribosome density is greater in cells of the flank meristem. Proplastids, plastids with prolamellar bodies, or grana have a differential distribution in the apex, characteristic for a particular species. Confirmation could not be given to the concept that vacuoles arise as a series of local dilations in long extensions of the so called "smooth ER." The tonoplast and ER are distinguishable at the time of inception of a vacuole, although the tonoplast may arise from the ER. Rapid growth of a vacuole and/or fusion with other vacuoles may result in irregularly shaped prevacuoles. No vacuoles were observed to originate from cisternae of dictyosomes in the species studied.     

STUDIES ON THE ULTRASTRUCTURE OF THE GUARD CELLS OF OPUNTIA

The guard cells of Opuntia contain numerous mitochondria, elements of endoplasmic reticulum, dictyosomes, and microbodies. A complex array of small to large vacuoles which contain small, membrane-bounded vesicles occur in each guard cell. The variety of cytoplasmic constituents and vacuoles suggest that the guard cells are complex in function. A highly reduced grana-fretwork system within the plastids indicates that the photosynthetic capacity of the guard cells is probably rather low. No plasmodesmata occur in the walls between the guard cells and the subsidiary cells while there are numerous invaginations of the guard cell plasmalemmas. Many of the variations in the plasmalemma probably indicate that the plasmalemma is a highly active interface.     

Ultrastructural localization of cytoplasmic phosphatases in preimplantation mouse embryos.

The appearance and localization of the cytoplasmic phosphatases [acid phosphatase (AcPase) as a marker of lysosomes, TPPase as a marker of the Golgi apparatus, and NDPase (IDPase) as enzymatic marker of the endoplasmic reticulum (ER)] were cytochemically studied on the ultrastructural level in secondary oocytes and in preimplantation mouse embryos. The detectable AcPase activity, located on the inner surface of the membrane delimiting some cytoplasmic vacuoles (lysosomes and autophagic vacuoles), appears at the eight-cell stage and grows pregressively stronger up to the blastocyst stage. Golgi-associated reaction for TPPase was detectable in oocytes, dropped in one-cell embryos and became negative in the two-cell embryos. The reaction for TPPase and IDPase was present in plasma membranes of oocytes and early embryos and appeared in the delimiting membrane of some cytoplasmic vesicles in eight-cell embryos. Some activity of IDPase was found in small segments of the ER at the morula and blastocyst stage. The observed results suggest that the lysosomes are the first organelles in early embryos showing activity of the marker enzymes of the phosphatase type, while the activity of other marker enzymes is mainly concentrated in the plasma membrane of blastomeres. It cannot be excluded, however, that positive reaction for TPPase and IDPase in the plasma membrane results from nonspecific action of other phosphatases.     

THE LATERAL MERISTEM AND ITS DERIVATIVES IN THE CORM OF ISOETES MURICATA

Corm tissue of Isoetes muricata Dur. was fixed in glutaraldehyde and postfixed in osmium tetroxide for electron microscopy. Very young secondary sieve elements can be distinguished from contiguous cambial cells by their distinctive plastids and by the presence of crystalline and/or fibrillar proteinaceous material in dilated cisternae of rough endoplasmic reticulum (ER). At maturity, the sieve elements are lined by the plasmalemma and a parietal, anastomosing network of smooth ER. Degenerate nuclei persist in all mature sieve elements. In addition, mature sieve elments contain plastids and mitochondria. Sieve-area pores are present in all walls. The lateral meristem of I. muricata consists of 2–3 layers of cells year-round. Judging from numerous collections made between October 1972 and July 1975, new sieve-element differentiation precedes cambial activity by about a month. Early in May, 1–2 cells immediately adjacent to already mature sieve elements differentiate directly into sieve elements without prior division. In early June, at about the time sieve-element differentiation is completed, cambial division begins. Division is sporadic, not uniform throughout the meristem. Dormancy callose accumulates in the secondary sieve elements in late October, and is removed in early May, at about the same time new sieve-element differentiation begins. Cells of the dormant cambium are characterized by the presence of numerous small vacuoles and large quantities of storage materials, including lipid droplets, starch grains, and tannin. By contrast, active cambial cells contain few large vacuoles with little or no tannin, and they have little storage material.     

Development and ultrastructure of the primary phloem in the shoot of Ephedra viridis (Ephedraceae)

Sieve cell differentiation in the primary phloem of Ephedra viridis is first indicated by an increase in thickness of the wall, which begins in the corners of the cell, and next by the proliferation of smooth tubular endoplasmic reticulum (ER). As differentiation proceeds, cisternae of rough ER form stacks along the wall, losing their ribosomes in the process. Concomitantly, all of the mitochondria, plastids, and ER become parietal in distribution, and the vacuoles collapse. Nuclear degeneration is pycnotic and accompanied by the formation of tubular invaginations of the nuclear envelope into the peripheral chromatin. At maturity, an anastomosing network of smooth ER borders the plasmalemma, interconnecting aggregates of smooth tubular ER located primarily opposite the sieve areas. In addition to ER, the mature sieve cell contains mitochondria, plastids, and remnants of the degenerate nucleus, all of which are parietal in distribution. P-protein is lacking at all stages of sieve cell development. Sieve pore and compound median cavity development is similar to that reported for the sieve cells of conifers. Albuminous cells are associated with the sieve cells of the metaphloem throughout the shoot but with sieve cells of the protophloem only in the node. Among their cytoplasmic components are broad bundles of microfilaments spatially associated with a complex system of rough and smooth ER.     

Ultrastructural localization of acid phosphatase in cultured cells ofDaucus carota

Summary Acid phosphatase localization has been studied, using the lead salt method, in suspension-cultured cells of the wild carrot. Enzyme activity in most of the cells was restricted to the walls and vacuoles. However, in some senescent cells activity was also seen in the nucleus, at one face of the dictyosomes, and in nearby dictyosome-derived vesciles.The activity in the walls was closely associated with the central portion of the wall which ultimately disintegrates in auxin-containing media. However, the large vesicles which accumulate in this portion of the wall as it breaks down never showed acid phosphatase activity, nor did the multivesicular bodies which appear to transfer vesicular material into the wall space. Although multivesicular bodies in plant cells resemble the multivesicular lysosomes of animal cells, no evidence could be obtained in this study for the presence in such bodies of hydrolytic enzymes.     

Ulfrastructure of the histological zones in growing vegetative buds of Salix spp.

Ultrastructural differentiation in the shoot apex of growing vegetative buds of Salix was studied, and some micrographs analysed morphometrically. The distribution of inorganic phospahte (P;) was analysed cytochemically. A distinct histological zona–tion was observed in the apex. The relative volumes of nuclei and plastids were significantly higher in the central tunica zone than in the peripheral one. The corpus differed from the central tunica zone by significantly lower volume density of nuclei and higher of vacuoles and mitochondria. During differentiation of the rib meristem vacuole volume increased significantly, while the relative volumes of nuclei, mitochondria, nucleoli, and heterochromatin decreased. It was not possible to decide whether the vacuoles originate from ER or GERL. Morphogenesis of chloroplasts with large starch grains and grana from proplastids was evident in the rib meristem; dedifferentiation to S–plastids was found in the protophloem. Prolamellar bodies were observed in the procambium plastids. The protophloem was characterized by P–protein and spiny vesicles. P_i was found in the nucleoli of most epidermis cells, several procambium cells, and a few chlorencyma cells, but never in the tunica of the growing apical and developing lateral buds. P_i also occurred in some plasmalemma–somes and occasionally in the walls in connection with plasmodesmata.     

Localization of Phloem Oxidases

Among oxidases, cytochrome oxidase has been localized in mitochondria of all phloem cells, catalase has been visualized in parenchyma peroxisomes and peroxidase has been localized in cell walls and in several cell organelles. In angiosperms, peroxidase is present in all phloem cell walls; it is sensitive to cyanide inhibition excepted in sieve areas and around plasmodesmata between sieve tubes and companion cells. In some species, this cyanide resistant oxidasic activity can be localized without exogenous H₂O₂. Peroxidase is localized on ribosomes, inside vacuoles, on the tonoplast and often on the plasmalemma in companion cells and differentiating sieve elements. In young sieve cells some dictyosomes can exhibit a strong peroxidasic activity. In mature parenchyma cells peroxidase can be associated with ER cisternae but not with vacuoles.     

Studies of the Mature Pollen of Spinacia oleracea after Freeze Substitution and Observed with Confocal Laser Scanning Fluorescence Microscopy

The three-dimensional configuration of the nuclei of the trinucleate pollen grain of Spinacia oleracea L. has been examined by means of confocal laser scanning fluorescence microscopy (CLSM). It shows the presence of a male germ unit (MGU) in which all three nuclei are usually positioned in the periphery of the pollen grain. After freeze fixation and freeze substitution, the ultrastructure is better preserved than with standard chemical fixation. It shows the presence inside the pollen grain of mitochondria, dictyosomes, large starch-containing plastids, endoplasmic reticulum (ER), vacuoles and the MGU. In the sperm cells mitochondria, vesicles, dictyosomes and ER are also found. No microtubules were found in the grain and only very few inside the sperm cells. This is in contrast with earlier published results where fluorescent-labeled antibodies were used.     

Seasonal variation in oxidase activity in the procambium of Salix buds

Seasonal changes in peroxidase activity in the procambium of Salix buds were elucidated by cytochemical methods combined with transmission electron microscopy. Three different substates were used, viz. diaminobenzidine (DAB), tetramethylbenzidine (TMB), and p-phenylenediamine (PPD) + pyrocathecol (PC). The specificity of the reactions was tested with the aid of several inhibitors: sodium diethyldithiocarbamate (DDC), KCN, and 3-amino-1,2,4-triazole (AT, inhibitor of catalase), and by exclusion of H₂O₂. Peroxidase activity was found mainly in the middle lamella region of the walls, at the plasmalemma, in the ER, nuclear envelopes, and many small vesicles (possibly lysosomes). The results indicate the presence of many isoenzymes with various pH optima and sensitivity to inhibitors. The enzyme activity was highest during dormancy (autumn and winter), perhaps a component of the hardening process. It decreased considerably in the spring simultaneously with dormancy breaking.     

STRUCTURE AND DEVELOPMENT OF THE SIEVE ELEMENT IN THE STEM OF LYCOPODIUM LUCIDULUM

Stem tissue of Lycopodium lucidulum Michx. was fixed in glutaraldehyde and postfixed in osmium tetroxide for electron microscopy. Although their protoplasts contain similar components, immature sieve elements can be distinguished from parenchymatous elements of the phloem at an early stage by their thick walls and correspondingly high population of dictyosomes and dictyosome vesicles. Late in maturation the sieve-element walls undergo a reduction in thickness, apparently due to an “erosion” or hydrolysis of wall material. At maturity, the plasmalemma-lined sieve elements contain plastids with a system of much convoluted inner membranes, mitochondria, and remnants of nuclei. Although the endoplasmic reticulum (ER) in most mature sieve elements was vesiculate, in the better preserved ones the ER formed a tubular network closely appressed to the plasmalemma. The sieve elements lack refractive spherules and P-protein. The protoplasts of contiguous sieve elements are connected with one another by pores of variable diameter, aggregated in sieve areas. As there is no consistent difference between pore size in end and lateral walls these elements are considered as sieve cells.     

玉米叶片原形成层细胞衰退过程的超微结构研究

在玉米（ＺｅａｍａｙｓＬ．）叶片原形成层发育的不同阶段常出现细胞衰退的现象,其衰退方式可分为自溶型和凝缩型。自溶型衰退的特点是细胞发生一系列降解活动最后导致细胞自溶死亡。衰退早期的细胞出现明显的自体吞噬活动,伴随核糖体和细胞质电子密度下降。衰退最后阶段的细胞自溶导致所有原生质组分的解体和消失,其中细胞核和质膜是最后消失的组分。凝缩型衰退的起动则伴随细胞质密度的增大。随着衰退的进行,原生质组分以一定顺序逐步退化解体：首先是高尔基体和内质网,接着是质体、细胞核和部分线粒体。但到衰退后期当细胞被挤压塌陷时仍有质膜、核糖体和线粒体存在。     

Ultrastructure of endopolyploid antipodals inAconitum vulparia Rchb.

Summary The ultrastructure of antipodals ofAconitum vulparia Rchb. was studied at two stages of development: at the earlier stage the endosperm has several nuclei, at the later one the endosperm is multinucleate. Over the investigated period the antipodal size enlarges distinctly. The wall ingrowths increase in size and number. Finally, they occur throughout the antipodal walls except for a small area in the extreme chalazal part, sunk deep into the nucellar podium. There are no plasmodesmata in the antipodal cell walls. The cytoplasm is dense and rich in ribosomes; it shows weak vacuolation. The rough endoplasmic reticulum is well developed. At the later stage dilated cisternae of endoplasmic reticulum are formed. Mitochondria, plastids and active dictyosomes are abundant. At the later stage some giant mitochondria are present; their matrix contains a large clear area with fine fibrils and an aggregation of fibrillar material. At this stage of development plastids have two types of inclusions: electron-transparent vacuoles and aggregations of electron-dense granules. The giant endopolyploid nuclei are considerably larger than those at the mature embryo sac stage; they are lobed on all sides.During the studied periodA. vulparia antipodals seem to be at their most active state.