ULTRASTRUCTURAL FEATURES OF DEVELOPING SIEVE ELEMENTS IN LEMNA MINOR L.—SIEVE PLATE AND LATERAL SIEVE AREAS

Both intact and cut duckweed plants were prepared for electron microscopy. Plants which are prepared intact do not exhibit callose formation during development of sieve-plate pores. Future pore sites can be recognized by the presence of median cavities that are unassociated with callose platelets. These cavities are first seen in the region of the compound middle lamella and are lined by a plasmalemma. As end walls thicken, the cavities increase in size until open pores of uniform width are formed. Mature sieve plates of intact-prepared plants are also devoid of callose. Fully opened pores are lined by a plasmalemma and are only traversed by an occasional tubule of endoplasmic reticulum. Plants which have been cut prior to fixation possess mature sieve plates containing callose. The pores of developing sieve plates in cut plants exhibit small amounts of callose. Except for the lack of callose, lateral wall connections between sieve elements and contiguous cells are similar in development and mature state to those reported for other species.     

Sieve-plate pores in leaf veins of Hordeum vulgare

Summary The sieve-plate pores of sieve elements in leaf veins of Hordeum vulgare, fixed in glutaraldehyde with postfixation in osmium tetroxide, were lined by the plasmalemma and variable amounts of callose. All pores were filled with endoplasmic reticulum, which was continuous from cell to cell. Mature sieve elements lacked P-protein.     

P-protein distribution in mature sieve elements of Cucurbita maxima

Summary Portions of the hypocotyls of 16-day-old Cucurbita maxima plants, from which the cotyledons and first foliage leaves had been removed 2 days earlier, were fixed in glutaraldehyde and postfixed in osmium tetroxide for electron microscopy. In well over 90% of the mature sieve elements examined the P-protein was entirely parietal in distribution in both the lumina and sieve-plate pores. In addition to the parietal P-protein, the unoccluded sieve-plate pores were lined by narrow callose cylinders and the plasmalemma. Segments of endoplasmic reticulum also occurred along the margins of the pores.     

Sieve-plate pores in tobacco and bean

Summary Tobacco and bean plants were wilted and then fixed as whole plants with formaldehyde-glutaraldehyde for electron microscopy. In some tobacco plants the sieve-plate pores were large, with little callose. Light slime plugs were present, but there was no compaction of P-protein in the pores. Some pores in wilted bean plants were also unplugged. In other plants of both tobacco and bean the sieve-plate pores were plugged. The pores in unwilted control plants of both tobacco and bean were invariably plugged. Tobacco plants were also cut into thin slices and then immediately fixed. In specimens prepared in this way there was little callose in the pores, and many of the pores were not plugged with P-protein. These observations provide additional evidence that sieve-plate pores may be unplugged in vivo.     

Aspects of sieve element ultrastructure in Primula obconica

Summary At maturity, the enucleate sieve element of Primula obconica is lined with a parietal layer of cytoplasm consisting of plasmalemma, one or more cisterna-like layers of endoplasmic reticulum, numerous mitochondria and plastids, and a membrane which apparently separates these cytoplasmic components from a large central cavity. The central cavity contains numerous longitudinally oriented slime tubules. We believe these tubules normally form strands which run the length of the cell and traverse consecutive cells through the sieve-plate pores. Developmental aspects are discussed.This research has been supported by NSF Grant GB 3193.     

Ultrastructural changes in the cell wall,nucleus and cytoplasm of pollen mother cells during meiotic prophase I inLycopersicon esculentum (Mill.)

Summary Throughout the premeiotic to late prophase I stages of meiosis in the anthers of tomato (Lycopersicon esculentum) extensive changes occurred in the ultrastructure of pollen mother cells (PMCs). During early prophase, the wall of each PMC developed a layered appearance and was broadened both by the widening of the middle lamella as well as by intensive deposition of microfibrils in the wall. By late prophase, however, the microfibrils adjacent to the plasmalemma dissipated. At the same time, callose was deposited between the wall and the plasmalemma. The nucleus of the PMCs also underwent changes. During early prophase, the nucleolus consisted of a linear series of three segments, with a separation of the granular and fibrillar portions. By late prophase, the nucleoli were less distinct as the nucleus was highly vacuolate. Mitochondria were initially simple with lightly stained matrix and few cristae but, during the course of prophase, they acquired a more densely-stained matrix with dilated cristae. Plastids remained relatively undifferentiated and, at late prophase, many were convoluted in appearance and constricted at intervals indicating their division. Cytoplasmic connections between adjacent PMCs were broad enough to permit the passage of organelles and were retained through to metaphase I. These cytological and wall changes appear to be a prerequisite for the subsequent development of microspores.Abbreviations PMC pollen mother cell - NOR nucleolus organizing region     

LAUDERIA ANNULATA—A MARINE CENTRIC DIATOM WITH AN ELONGATE BILOBED NUCLEUS1

Lauderia annulata Cleve is probably unique among marine centric diatoms in possessing an elongate dumbbell-shaped nucleus. A lobe at, each end of the nucleus lies adjacent to each valve during interphase and each lobe resembles a typical eucaryotic nucleus. The central portion of the thin strand, which passes through the vacuole connecting the two lobes is Feulgen positive and is nuclear membrane bounded. A group of micro tubules occurs in this strand clustered excentrically between, the nuclear membrane and the tonoplast. Evidence for the coordinated functioning of both nuclear lobes is suggested, by the aggregation of chloroplasts around, both lobes when shade-adapted cells are exposed, to intense white or blue (400–500 nm) light.     

P PROTEIN IN THE PHLOEM OF CUCURBITA : II. The P Protein of Mature Sieve Elements

During maturation of sieve elements in Cucurbita maxima Duchesne, the P-protein bodies (slime bodies) usually disperse in the tonoplast-free cell. In some sieve elements the P-protein bodies fail to disperse. The occurrence of dispersal or nondispersal of P-protein bodies can be related to the position of the sieve elements in the stem or petiole. In the sieve elements within the vascular bundle the bodies normally disperse; in the extrafascicular sieve elements the bodies often fail to disperse. Extrafascicular sieve elements showing partial dispersal also occur. The appearance of the sieve plate in fixed material is related to the degree of dispersal or nondispersal of the P-protein bodies. In sieve elements in which complete dispersal occurs the sieve plate usually has a substantial deposit of callose, and the sieve-plate pores are filled with P protein. In sieve elements containing nondispersing P-protein bodies the sieve plate bears little or no callose, and its pores usually are essentially "open." The dispersed P-protein components may aggregate into loosely organized "strands," which sometimes extend vertically through the cell and continue through the sieve-plate pores; but they may be oriented otherwise in the cell, even transversely.     

SLIME SUBSTANCE AND STRANDS IN SIEVE ELEMENTS

Studies of the secondary phloem of 6 species of woody dicotyledons revealed that slime is not normally dispersed throughout the vacuole of mature sieve elements, but occurs in the form of discrete strands that traverse the cell and run from cell to cell through the sieve-plate pores. As many as 5 fine strands, each measuring less than 0.5μ in diameter, were observed in a single pore. Less than 30% of pore area was occupied by strands. Thus, the pores are mostly open, and intervacuolar continuity exists between cells. These structural characteristics of pores offer strong support for the concept of mass flow.     

Sieve Element of Impatiens sultanii: 1. Wound Reaction

A Study of wound reaction of the metaphloem of the stem by white-light,fluorescence, and electron microscopy provides evidence forthe structure of mature sieve elements in the intact plant.Starch grains usually are retained in plastids which are locatedagainst the lateral walls of sieve elements and concentratedat both ends of each cell. Slime plugs and dense connectingstrands in the sieve plates seem to result from reactions tocutting or penetration of the killing agent; after appropriatemethods of killing, the contents of a connecting strand maybe only slightly denser, if any, than the milieu on either sideof the sieve plate. A strange accumulation of slime, consistingof streamers directed toward the wound surface from each sieveplate, occurred in tissue boiled immediately after incisionof the phloem. Callose is present in sieve elements of intactplants when the tissue is killed within 4 seconds after injury.Callose is accumulated in response to wounding in added amounts,but only after 5 minutes or more and only within about 15 sieveelements from the wound. Quantities of callose sufficient forplugging the sieve plate accumulate after 30 minutes or more.Sieve-plate callose and deposits on the nearby lateral wallsproduce a cup-shaped mass which is called a cup deposit.     

Analysis of the reaction products from incubation of sugarcane vacuoles with uridine-diphosphate-glucose: no evidence for the group translocator

Isolated sugarcane (Saccharum spp. hybrid H50-7209) vacuoles incorporate radioactivity during incubation with labeled UDP-glucose by a mechanism which was postulated to be responsible for sucrose storage in the vacuoles (UDP-glucose group translocator). Analysis of the reaction products in the medium revealed that several enzymic processes are going on during incubation with UDP-glucose such as production of hexose phosphates, UMP, and sugars, all of which seem unrelated to the incorporation of radioactivity into vacuoles. The incorporated radioactivity was identified mainly as (1→3)-β-glucan (callose) of polymerization grades up to more than 20. Callose occurs as a contaminant at the surface of isolated vacuoles coming from the plasmalemma. The properties of UDP-glucose incorporation into the vacuolar preparation compared favorably with known properties of callose synthase. The low mol wt glucans that are found are probably degradation products of labeled callose due to hydrolases, which are liberated by centrifugation of vacuoles. The labeled disaccharide, which chromatographically had been formerly identified as sucrose, is laminaribiose. No sucrose (or sucrose phosphate) could be identified in the vacuole preparation after incubation with UDP-glucose. Thus, the mechanism of sucrose storage in sugarcane vacuoles is still open.     

Transmission Electron Microscopy of Meront Development of Eimeria vermiformis Ernst,Chobotar and Hammond, 1971 (Apicomplexa,Eucoccidiorida) in the Mouse,Mus musculus1

First and second generation meronts of Eimeria vermiformis developed in epithelial cells of the crypts of Lieberkühn. They were usually between the host cell nucleus and the basement membrane. Sporozoite organelles dedifferentiated with the first generation meront's development except for the refractile body and the apical complex, which persisted. After several nuclear divisions, the apical complex dedifferentiated further until only micronemes remained attached by a duct system to the plasmalemma. The form of the apical complex was highly variable. Sometimes the duct system was absent and the micronemes were attached directly to the plasmalemma or a dense material on it. Crescent body-like material was often present in the parasitophorous vacuole next to the microneme structure. The microneme structure was not present in second generation meronts but evaginations of the plasmalemma, cytoplasmic outpocketings, and cytoplasmic vesicles were associated with the round granular bodies in the parasitophorous vacuoles. During first generation merogenesis, invaginations from the parasitophorous vacuole formed channels into the meront along which merozoites budded. Micropores were often at the ends of these invaginations. These and other micropores of the meront had a dense U-shaped band for a collar while those of the merozoites had a collar with a double band of dense material that connected to the inner membrane. First generation merozoites budded randomly from the meront, resulting in a residual body that was usually in the middle of the parasitophorous vacuole. Second generation merozoites budded in one direction, resulting in a peripheral residual body and merozoites that were parallel in an oblong parasitophorous vacuole.     

LIGHT MICROSCOPE INVESTIGATION OF SIEVE-ELEMENT ONTOGENY AND STRUCTURE IN ULMUS AMERICANA

At maturity the sieve elements of Ulmus americana L. contain a parietal network of very fine strands of slime which is continuous from one sieve element to the next through the sieve-plate pores. Upon injury this parietal network, which is derived from the slime bodies of immature sieve elements, sometimes becomes distorted into longitudinally oriented strands. Some of these strands frequently extend the length of the cells and often are continuous from one sieve element to the next through the sieve-plate pores. At times past such strands have erroneously been interpreted as normal constituents of the mature sieve-element protoplast. Many mature sieve elements of U. americana contain nuclei, which apparently persist for the life of the sieve elements. In addition, some evidence has been found in mature sieve elements for the presence of a membrane which delimits the parietal layer of cytoplasm, including its network of slime strands, from the vacuolar region of the cell.     

The orientation of nucleus, nucleus-associated body and protruding nucleolus in aggregating Dictyostelium discoideum

Dictyostelium discoideum growing or developing on cellulose dialysis membranes were fixed with acrolein vapour for electron microscopy. In interphase amoebae, nucleoli began to protrude from the nuclei. The percentage of cells with protruding nucleoli increased during aggregation by a value approximately twice as high in aggregation streams as in centers. Cells in pseudoplasmodia showed only a low percentage and protrusions disappeared at early culmination stage. The protrusions did not reappear when cells from dissociated pseudoplasmodia migrated toward cAMP. Thus the formation of the protrusions did not depend solely on chemotaxis; rather, it was specific to the aggregation stage. In aggregation streams, the nucleus was anterior in the cell, with the protrusion at its anterior periphery. In contrast, the nucleus associated body (NAB) was evident at the cell's mid-point. This orientation of nucleus and NAB in the aggregating slime mould amoeba is contrary to that seen in human neutrophils or cultured mouse 3T3 cells.     

Pericentriolar processes of photoreceptor cell basal bodies in the mammalian retina

Pericentriolar processes (arm-like fibers) of the migrating centrioles (diplosome) in differentiating retinal photoreceptor cells were examined in six mammalian species (hamster, vole, rat, rabbit, ferret, cat). These processes emanate in a radial fashion from one end of the centrioles comprising the photoreceptor diplosome. The pericentriolar processes of the basal body are first observed as the diplosome migrates toward the apical plasmalemma, suggesting that centrioles are committed early-on to developing such processes. One pericentriolar process arises from each set of microtubular triplets comprising the centriole and are apicoexternally oriented at an angle of between 30 and 60 degrees with the centriolar axis. Prior to the arrival of one of the centrioles at the apical plasmalemma these processes connect with an electron-dense portion of a centriole-associated vacuole. The diplosome migrates to the apical plasmalemma where one centriole (the presumptive basal body) orients perpendicularly to the apical plasmalemma. The centriole-associated vacuole appears to fuse with the plasmalemma. The pericentriolar processes appear to attach to this fusion site on the plasmalemma which is a region of the membrane characterized by increased electron density (the basilar plate). Invagination of the apical membrane, which occurs at this same site, is accompanied by a lengthening of the microtubules forming a cilium and is observed as an outpouching of the plasmalemma within the aforementioned invagination. The associated vacuole apparently becomes continuous with the apical plasmalemma. These pericentriolar processes appear to be functionally involved in ciliogenesis and offer structural stability between the basal body, the plasmalemma and indirectly the cilium.     

DEVELOPMENT AND STRUCTURE OF PHLOEM IN THE PETIOLE OF LUFFA CYLINDRICA

Differentiation of external phloem is earlier than that of internal phloem in the young petiole of Luffa cylindrical. For a single sieve-tube element, one to six companion cells are present. The young sieve element shows many globular slime bodies which fuse longitudinally and disperse into the cytoplasm. Simultaneously the nucleus loses its stainable contents and later disorganizes. The contents of the sieve element are in the form of plugs, strands or a granular mass. Undispersed slime in the form of discrete bodies along the lateral walls is also observed. During one stage, at least, the dispersed slime and other contents of a mature sieve element lie at the periphery around a central cavity. A special type of phloem-parenchyma cell shows disorganizing chloroplasts, an extruded nucleolus, and callose on primary pit fields.     

Another View of the Ultrastructure of Cucurbita Phloem

The primary phloem of young internodes of Cucurbita maxima wasstudied with the electron microscope. Phloem parenchyma cellsare highly vacuolated and contain nuclei, endoplasmic reticulum,ribosomes, mitochondria, chloro-plasts, and occasional dictyosomes.As compared with parenchyma cells, the most distinctive featuresof companion cells are their extremely dense cytoplasm, lowdegree of vacuolation, lack of chloroplasts, and numerous sieve-elementconnexions. Companion cells contain plastids with few internalmembranes. At maturity the enucleate sieve element is linedby a plasmalemma, one or more cistema-like layers of endoplasmicreticulum, and a membrane which apparently delimits the parietallayer of cytoplasm from a large central cavity. In OsO_4–-andglutaraldehyde-fixed elements, the central cavity is traversedby numerous strands, which run from cell to cell through thepores of sieve plates and lateral sieve areas, and which arederived ontogenetically from the slime bodies of immature cells.Numerous normal-appearing mitochondria are present in the parietallayer of cytoplasm. The pores of sieve plates and lateral sieveareas are lined with cytoplasm. The ultrastructural detailsof young sieve elements differ little from those of other youngnucleate cells. During sieve-element development, the sieveelement increases in vacuolation. At the same time, slime bodiesdevelop in the cytoplasm. With glutaraldehyde fixation, thesebodies often exhibit a double-layered limiting membrane. Asthe sieve element continues to differentiate, the slime bodiesincrease in size and the parietal layer of cytoplasm becomesvery narrow. Presently, the slime bodies begin to disperse andtheir contents fuse. This phenomenon occurs in the parietallayer of cytoplasm, while the latter is still delimited fromthe large central vacuole by a distinct tonoplast. The initiationof slime-body dispersal more or less coincides with perforationof the pore sites, and many pores are traversed by slime earlyin their development. Before slime-body dispersal, all dictyosomesand associated vesicles disappear from the cytoplasm. Eventually,the tonoplast diappears and the slime becomes distributed throughoutthe central cavity in the form of strands. Nuclei and ribosomesdisappear before breakdown of the tonoplast. Sieve elementsare connected with companion cells and parenchyma cells by plasmodesmata.     

Ricerche Sull'infrastruttura del Coleottile di Avena

Abstract

Researches on ultrastructure of Avena coleoptile. 3. The sieve elements. — A study on the ultrastructural organization of the mature sieve elements of Avena coleoptile has been carried out. Data suggest that functional phloem tubes are alive and remain alive until they are working. Judging on morphological basis, the metabolic activity of sieve elements should be of peculiar type and low in comparison to that of the companion cells. In fact the cytoplasm is located in a narrow parietal strand, mitochondria, Golgi apparatus and endoplasmic reticulum are present, but they appear very modified; plastids and nucleus are absent. The cytoplasm is bounded externally by a normal plasmalemma, whilst the vacuole has no visible limits: a tonoplast is, therefore not identifiable.

The strands connecting the superimposed sieve elements with one another through the sieve plate result to be made by a double membrane system very similar to the endoplasmic reticulum, which we believe to realize cytoplasmic continuity between phloem tubes.

The data reported are more favorable to the existence in the sieve tubes of an active mechanism of translocation of organic solutes than a passive mass-flow.

The collaboration of companion cells in the translocation mechanism has been discussed.     

SOME ASPECTS OF SIEVE CELL ULTRASTRUCTURE IN PINUS STROBUS

The immature sieve cell of Pinus strobus contains all of the protoplasmic components commonly encountered in young cell types. In addition, it contains slime bodies with distinct double-layered limiting membranes. The mature sieve cell is lined by a narrow layer of cytoplasm consisting of a plasmalemma, one or more layers of anastomosing tubules of endoplasmic reticulum, numerous mitochondria, starch granules and crystal-like bodies. Each mature cell contains a necrotic nucleus. Ribosomes and dictyosomes are lacking. Strands derived ontogenetically from the slime bodies of the immature cell traverse the central cavity and are continuous with those of neighboring sieve cells through the plasmalemma-lined pores of the sieve areas. Sieve-area pores are also traversed by numerous endoplasmic membranes. A membrane was not found separating the parietal layer of cytoplasm from the large central cavity.     

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.