Cytochemistry and Ultrastructure of the Mucilage Secreting Trichomes of Nymphoides peltata (Menyanthaceae)

The young developing leaves in the buds of Nymphoides peltataare covered by a hyaline mucilage. The mucilage contains freesugars, polysaccharides and proteins. The most abundant monosaccharidesof the polysaccharide fraction are arabinose and galactose.Therefore, the major component of the mucilage is probably anarabinogalactan or arabinogalactan protein. The mucilage issecreted by glandular trichomes. It is suggested that both thepolysaccharide and the protein fraction of the mucilage aretransported to the plasmamembrane by vesicles of the Golgi apparatus(granulocrine secretion). Secretory proteins are probably synthesizedin the rough endoplasmic reticulum and transported to the Golgiapparatus via transition vesicles. Polysaccharides were localizedin Golgi vesicles by ultracytochemistry. After exocytosis thesecretion is accumulated between the cell wall and the cuticle;this leads to the formation of protrusions on the outer wallsof the glandular cells. Finally, the cuticle is ruptured andthe secretion is released. The biological function of the mucilageis not known. Possibly the mucilage is a lubricant or a protectionfrom desiccation. Nymphoides peltata (S.G. Gmel.) O. Ktz., trichomes, mucilage secretion, cytochemistry, ultrastructure     

Pistil Exudate Production and Pollen Tube Growth in Lilium longiflorum Thunb.

Exudate production in the pistil of Lilium longiflorum was studiedin relation to pollen tube growth, using scanning electron microscopy(SEM), transmission electron microscopy and light microscopy.In contrast with conventional fixation for SEM, during whichthe exudate of L. longiflorum largely washes away, the exudateremains present through freezing in case of cryo-SEM. Usingthe latter method we observed that exudate production on thestigma and in the style started before anthesis. Just underneaththe stigma the exudate was first accumulated at the top of eachsecretory cell, followed by a merging of those accumulationsas exudate production proceeded. Exudate is also produced bythe placenta. It was however not possible to determine whetherany of this fluid originated from the micropyle. Apart fromthe cell shape and the cuticle present in between the secretorycells, the ultrastructure of the secretory cells covering theplacenta was comparable to those of the stylar canal. The transferwall of the secretory cells of the placenta originated fromfusing Golgi vesicles but the endoplasmic reticulum seemed tohave an important role as well. After pollination the pollen tubes grew across the stigma andentered the style through one of the slits in the three stigmalobes. The pollen tubes grew straight downward through the styleand were covered by exudate. As the pollen tubes approachedthe ovary their growth was restricted to the areas with secretorycells. In the cavity the pollen tubes formed a bundle and theybent from this bundle in between the ovules towards the micropylarside. There they bent again to stay close to the secretory cells.After bud pollination the pollen tube growth was retarded. Laterarriving pollen tubes had a tendency to grow close to the secretorycells of the style, which resulted in a growth between thesecells and preceding pollen tubes. If there was still a littleexudate produced, it resulted in a lifting up of the pollentubes, out of the exudate. The relationship between exudateproduction and pollen tube growth is discussed. Both the speedand the guidance of the pollen tube seemed determined by theproperties of the exudate.Copyright 1994, 1999 Academic Press Cryo-scanning electron microscopy, exudate, Lilium longiflorum, lily, ovary, pollination, pollen tube growth, secretory cell, stigma, style     

Cytochemical and Morphological Evidence for the Involvement of the Plasma Membrane and Plastids in Mucilage Secretion in Aloe arborescens

Most of the volume of Aloe arborescens leaves (volumetric density= 0.68±0.08) is occupied by a mucilage tissue. The mucilagesof Aloe species are glucomannans of great medical and pharmaceuticalimportance. An electron microscopical and histochemical studywas carried out, following the development of the tissue andsecretion processes. In contrast to other reports on polysaccharide secretion inplants, no observable activity was found in the Golgi apparatus.In the young stages of leaf development, positive histochemicalstaining for polysaccharides and structural changes were foundmainly in the plastids. In the mature leaves the mucilage cellsremain alive and intact and secretory activity appears to becentered in the plasma membrane. Aloe arborescens, cytochemistry, mucilage, plasma membrane, plastids, secretion     

Developmental Aspects of Ultrastructure, Histochemistry and Receptivity of the Stigma of Nicotiana sylvestris

The bilobed papillate stigma of Nicotiana sylvestris Speg. andComes, is covered at maturity with a copious exudate containinglipid, protein and carbohydrate. The stigma is receptive fromthe very early stage of development and it also stains positivelyfor esterase activity. The stigma has three distinct zones:an epidermis with papillae; a subepidermal secretory zone; anda parenchymatous ground tissue. The behaviour of the cells ofthese three zones has been followed from 6 d before anthesisto one day after anthesis and pollination. The cells of theepidermis and the secretory zone stain intensively for lipids,proteins and carbohydrates in the initial stages. The secretoryzone develops large intercellular spaces containing heterogenoussecretory products which also stain positively for the aforesaidthree compounds. At maturity the secretory products are releasedto the surface through gaps formed in the epidermis by cellseparation. The main secretion of the stigma is produced bythe cells of the secretory zone. Less secretion is derived fromthe stigmatic papillae. Some amount of secretion is also releasedfrom the stylar transmitting tissue adjoining the stigma. Theglandular cells of the stigma contain numerous plastids, mitochondria,ribosomes, ER, cytoplasmic lipid droplets and some dictyosomes.The plastids and the vacuoles in the secretory cells of thestigma have a lot of electron dense (osmiophilic) inclusionsrespectively in the initial and later stages of development.The former are probably involved in the production of thesematerials. It is suggested that the proteins are directly secretedby rough ER compartments whereas smooth ER is involved in thesynthesis of lipidic materials. The carbohydrate moiety of theexudate is released by the eccrine mode (sugar mono- and dimers)with some addition of polymers by disintegration of the middlelamellae. The means by which the lipidic and osmiophilic materialis extruded remains unclear. Nicotiana sylvestris, stigma receptivity, organization, stigmatic secretory system, stigmatic exudate     

The Epidermal Cell Structure of the Secondary Pollen Presenter in Vangueria infausta (Rubiaceae: Vanguerieae) Suggests a Functional Association with Protruding Onci in Pollen Grains

Secondary pollen presentation is a well-known phenomenon in the Rubiaceae with particularly conspicuous pollen presenters occurring in the tribe Vanguerieae. These knob-like structures are formed by a modification of the upper portion of the style and stigma, together known as the stylar head complex. In the flower bud and shortly before anthesis, the anthers surrounding the stylar head complex dehisce and release pollen grains which adhere to the pollen presenter. The epidermal cells of the pollen presenter facing the anthers are radially elongated with a characteristic wall thickening encircling the anticlinal walls of each cell towards the distal end. These cells were studied in the pollen presenter of Vangueria infausta using electron and light microscopy in conjunction with histochemical tests and immunohistochemical methods. Other prominent thickenings of the cell wall were also observed on the distal and proximal walls. All these thickenings were found to be rich in pectin and possibly xyloglucan. The terms “thickenings of Igersheim” and “bands of Igersheim” are proposed to refer, respectively, to these wall structures in general and those encircling the anticlinal walls of each cell near the distal end. The epidermal cells have an intricate ultrastructure with an abundance of organelles, including smooth and rough endoplasmic reticulum, Golgi apparatus, mitochondria and secretory vesicles. This indicates that these cells are likely to have an active physiological role. The pollen grains possess prominent protruding onci and observations were made on their structure and development. Walls of the protruding onci are also rich in pectin. Pectins are hydrophilic and known to be involved in the dehydration and rehydration of pollen grains. We hypothesise that the thickenings of Igersheim, as well as the protruding onci of the pollen grains, are functionally associated and part of the adaptive syndrome of secondary pollen presentation, at least in the Vanguerieae.     

ULTRASTRUCTURAL EVIDENCE FOR SECRETORY PHASES IN THE LIFE HISTORY OF STIGMATIC HAIRS OF COTTON: A DRY TYPE STIGMA

Stigmatic hairs of the cotton flower were studied through their developmental stages up to anthesis. Stigmatic hairs invariably develop from a densely straining band of epidermal cells opposite the transmitting tissue cells. At anthesis, these are single cell structures measuring up to 300 μm long. At the 5-mm stage of stylar length (7–10 days before anthesis), some stigmatic hair cells begin to accumulate an osmiophilic substance between the plasmalemma and the cell wall, possibly synthesized in the endoplasmic reticulum. This material is apparently never secreted outside the cell wall. Immediately following this secretory phase in some stigmatic hair cells a second secretory phase starts. A dense osmiophilic substance, different in appearance from the previous phase, accumulates in the vacuoles of each hair cell. Concomitantly, dimorphism develops in the cytoplasmic densities of stigmatic hair. Some stigmatic hair cytoplasm appears very dense and shows signs of degeneration while other cytoplasm appears normal. A third secretory phase, which begins at anthesis, occurs in the normal hair cells. This phase is characterized by enhanced activity in the cytoplasm of the endoplasmic reticulum and Golgi apparatus. Large vesicles containing granular material are seen fusing with the plasmalemma. Coincident with this activity there is dissolution of the middle layers of the cell wall and the cuticle is ruptured at various points. The dense osmiophilic substance that had accumulated in the vacuole breaks down into fine granular material. Significance of these changes is discussed in relation to the pollen germination mechanism on the dry type stigma of cotton.     

SECRETIONS FROM THE PISTIL OF LILIUM LONGIFLORUM

The pistil of Lilium longiflorum secretes two forms of exudate, one from the stigma surface and the other from the canal cavity. Electrophoretic studies of these exudates have revealed quantitative and qualitative differences in protein profiles. The exudatic components which are transferred to the cell wall by endoplasmic reticulum and Golgi vesicles, are stored within the cell wall of the secretive tissues and secreted from the cell walls directly. The cell wall structure of these secretive tissues differs. The canal cell wall has thick characteristic ingrowths that are supplied mainly from Golgi vesicles, while the papilla cell wall of the stigma is thinner, lacks ingrowths, and is supplied from ER vesicles.     

Pollen--stigma Interactions: Development and Cytochemistry of Stigma Papillae and their Secretions in Annona squamosa L. (Annonaceae)

The development and cytochemical features of the stigma andstyle have been investigated in Sugar apple, Annona squamosaL., using light and electron microscopy. The pistil is a syncarpwith an open stylar canal. Papillae of epidermal origin lineboth the surface of the stigma and the inner face of the stylarcanal. The papillae contain organelles characteristic of secretorycells with a highly thickened cellulosic wall. The wall is multi-layered,the zones differing in their microfibrillar stacking and orientation.The stigma is of the ‘wet’ type and the surfaceexudate is heterogeneous in microscopic appearance and reactscytochemically for proteins, carbohydrates and lipids. The surfacecuticle undergoes dissolution prior to anthesis. A secretionalso appears in the thickened middle lamella of the sub-epidermalcell layer which reacts cytochemically for pectinaceous acidicpolysaccharides. Esterase activity of papillae is indicative of the receptiveareas, and it is also related to the onset of receptivity. Acidphosphatase activity is intense in the sub-epidermal cell layerswhich probably reflects their secretory activity. Pollinationtriggers a copious flow of secretion onto the stigma surfacewhich engulfs the pollen grains. It appears that most of theacidic polysaccharides of this secretion come from the middlelamella of the sub-epidermal cell layer. Compatible pollen tubes have no apparent barriers to overcomeon their route to the embryo sac and the inherent protogynousdichogamy seems to control the acceptance or rejection of compatiblepollen. Annona squamosa L., sugar apple, stigma, style, secretions     

Immunocytochemical Localization of Phenylalanine Ammonia-Lyase and Cinnamyl Alcohol Dehydrogenase in Differentiating Tracheary Elements Derived from Zinnia Mesophyll Cells

Secondary wall thickening is the most characteristic morphologicalfeature of the differentiation of tracheary elements. Isolatedmesophyll cells of Zinnia elegans L. cv. Canary Bird in differentiationmedium are converted to tracheary elements, which develop lignifiedsecondary wall thickenings. Using this system, we investigatedthe distribution of two enzymes, phenylalanine ammonia-Iyase(PAL) (EC 4.3.1.5 [EC] ) and cinnamyl alcohol dehydrogenase (CAD)(EC 1.1.1.195 [EC] ), by both biochemical and immunological methods.Both PAL and CAD appear to be key enzymes in the biosynthesisof lignin precursors, and they have been shown to be associatedwith the differentiation of tracheary elements. Cultured cellswere collected after various times in culture. The culture mediumwas separated from cells by centrifugation and designated fraction(1), the extracellular fraction. The collected cells were homogenizedand separated into four fractions: (2) cytosol; (3) microsomes;(4) cell walls (loosely bound material); and (5) cell walls(tightly bound material). PAL activity was detected in eachfraction. The extracellular fraction consistently had the greatestPAL activity. Moreover, PAL activity in the cytosolic fractionincreased rapidly prior to lignification, as it did in boththe microsomal and the cell wall (tightly bound) fractions duringlignification. Antisera against PAL and against CAD detectedthe proteins with molecular masses that corresponded to thoseof PAL and CAD in Zinnia. Immuno-electron microscopy revealedthat, in differentiating tracheary elements, PAL was dispersedin the cytoplasmic matrix and was located on Golgi-derived vesiclesand on the secondary wall thickenings. "Cell-free" immuno-lightmicroscopy supported the putative distribution of PAL on lignifyingsecondary walls. The pattern of distribution of CAD was similarto that of PAL. Thus, both PAL and CAD seemed to be localizedin secondary wall thickenings. From the results of both biochemicalassays and immunocytochemical staining, it appeared that atleast two types of PAL and CAD are present in differentiatingcells. One type of each enzyme is distributed in the cytosol,while the other is secreted from the Golgi apparatus and transportedby Golgi-derived vesicles to the secondary wall thickenings. (Received April 19, 1996; Accepted November 18, 1996)     

The Structure and Cytochemistry of the Pistil of Sternbergia lutea (Amaryllidaceae)

Studies were carried out on structural and cytochemical aspectsof the pistil of Sternbergia lutea (L.) KerGawl. The stigmais of the wet papillate type; the papillae are unicellular andare arranged densely around the rim of a funnel-shaped stigma.The stigma exudate is limited and is confined to the bases ofthe papillae and the inner lining of the stigma. The papillaeare smooth in the distal part and are covered with intact cuticle-pelliclelining. The cuticle is disrupted at places towards the baseof the papillae releasing the exudate. The exudate is rich inpectins and other polysaccharides but poor in proteins and lipids.The papillae show dense cytoplasmic profiles with extensiveendoplasmic reticulum (ER), abundant mitochondria, polyribosomesand active dictyosomes. The style is hollow. The stylar cavityis surrounded by two to four layers of glandular cells. In theyoung pistil the canal is lined with a continuous cuticle, butin the mature pistil the cuticle becomes disrupted and the canalis filled with the secretion produced by the cells of the surroundingglandular tissue. Ultrastructurally, the cells of the glandulartissue are very similar to the stigmatic papillae. The innertangential wall of the cells bordering the canal is uniformlythicker than other walls. The secretion in the stylar canal,as well as the intercellular spaces of the glandular tissue,stain intensely for pectins and polysaccharides but poorly forproteins and lipids. Pollen tubes grow through the stylar canal.Structural and cytochemical details of the pistil of Sternbergiaare compared with other hollow-styled systems. Pistil, Sternbergia lutea (L.) Ker-Gawl., stigma and style, structure and cytochemistry     

Effect of brefeldin A on the structure of the Golgi apparatus and on the synthesis and secretion of proteins and polysaccharides in sycamore maple (Acer pseudoplatanus) suspension-cultured cells.

Brefeldin A (BFA), a specific inhibitor of Golgi-mediated secretion in animal cells, has been used to study the organization of the secretory pathway and the function of the Golgi apparatus in plant cells. To this end, we have employed a combination of electron microscopical, immunocytochemical, and biochemical techniques to investigate the effects of this drug on the architecture of the Golgi apparatus as well as on the secretion of proteins and complex cell wall polysaccharides in sycamore maple (Acer pseudoplatanus) suspension-cultured cells. We have used 2.5 and 7.5 micrograms/mL of BFA, which is comparable to the 1 to 10 micrograms/mL used in experiments with animal cells. Electron micrographs of high-pressure frozen and freeze-substituted cells show that although BFA causes swelling of the endoplasmic reticulum cisternae, unlike in animal cells, it does not induce the disassembly of sycamore maple Golgi stacks. Instead, BFA induces the formation of large clusters of Golgi stacks, an increase in the number of trans-like Golgi cisternae, and the accumulation in the cytoplasm of very dense vesicles that appear to be derived from trans Golgi cisternae. These vesicles contain large amounts of xyloglucan (XG), the major hemicellulosic cell wall polysaccharide, as shown by immunocytochemical labeling with anti-XG antibodies. All of these structural changes disappear within 120 min after removal of the drug. In vivo labeling experiments using [3H]leucine demonstrate that protein secretion into the culture medium, but not protein synthesis, is inhibited by approximately 80% in the presence of BFA. In contrast, the incorporation of [3H]fucose into N-linked glycoproteins, which occurs in trans-Golgi cisternae, appears to be affected to a greater extent than the incorporation of [3H]xylose, which has been localized to medial Golgi cisternae. BFA also affects secretion of complex polysaccharides as evidenced by the approximate 50% drop in incorporation of [3H]xylose and [3H]fucose into cell wall hemicelluloses. Taken together, these findings suggest that at concentrations of 2.5 to 7.5 mu g/mL BFA causes the following major changes in the secretory pathway of sycamore maple cells: (a) it inhibits the transport of secretory proteins to the cell surface by about 80% and of hemicelluloses by about 50%; (b) it changes the patterns of glycosylation of N-linked glycoproteins and hemicelluloses; (c) it reduces traffic between trans Golgi cisternae and secretory vesicles; (d) it produces a major block in the transport of XG-containing, dense secretory vesicles to the cell surface; and (e) it induces the formation of large aggregates of Golgi apparatus of plant and animal cels share many functional and structural characteristics, the plant Golgi apparatus possesses properties that make its response to BFA unique.     

Structure of the style and wet non-papillate stigma of Colophospermum mopane, Caesalpinioideae: Detarieae

The capitate stigma of Colophospermum mopane (Kirk ex Benth.) Kirk ex J. Leonard is an intensely folded bilobed structure. The epidermal layer of the stigma consists of non-papillate cells. Before anthesis the epidermis is covered with a cuticle and thin proteinaceous layer. Elongated subepidermal cells constitute the secretory zone. Cell disintegration in the central region of each stigma lobe leads to cavities that become connected to the central cavity in the style. During early anthesis it appears as if the receptive surface of the stigma is confined to the depressions of the stigma surface and to the cleft between the two stigma lobes as the secretory product and pollen grains are mainly confined to these areas. The secretory products of the stigma and style are released during five different stages from prior to anthesis to late anthesis. The stigmatic exudate appears complex and consists of carbohydrates, proteins and lipids. The style has a hollow, lysigenous, fluid-filled canal that is not lined with an epidermal layer or cuticle. The stylar canal is continuous with the opening between the two stigma lobes and provides an open route for the passage of exudate. The stylar exudate is PAS-positive. The dorsal and ventral bundles that supply the style branch in such a way as to almost form a cylinder around the central transmitting tissue and stylar canal. New sieve elements proliferate before anthesis.  © 2002 The Linnean Society of London, Botanical Journal of the Linnean Society , 2002, 139 , 295–304.     

The secretory stigma inLycopersicum peruvianum Mill.: Ontogenesis and glandular activity

Summary The aim of the present study has been to elucidate the cytology of the glandular stigma inLycopersicum peruvianum Mill. and the mechanism of the secretory process during the stigma development.The glandular stigma (papillae and superficial stigmatic tissue) has been studied by light and electron microscopy (S.E.M. and T.E.M.).At anthesis, the longitudinal intercellular spaces are filled with exudate in the form of heterogeneous droplets and form an intercommunicating system which allows transmitting tissue to communicate with superficial papillae.The presence of cytoplasmic droplets similar in appearance to the exudate has been noted in the developing stigmatic tissue. Cytoplasmic events which may be related to their production include transitory vacuolar accumulations, modifications of the morphology of plastids and development of smooth endoplasmic reticulum. At anthesis, observations possibly related to the origin of these droplets include their contiguity with endoplasmic reticulum cisternae and their relationship to the Golgi apparatus.Some observations suggest that cytoplasmic droplets are extruded by a process of exocytotic secretion. In addition, the cytoplasmic transport of secretory products in a non-visible form may explain the involvement of several cell compartments and the heterogeneity of the secretory products.Research partially supported by C.N.R.S. R.C.P. 429 and by C.N.R. programme Biology of reproduction.     

Development of Golgi Apparatus in the Root Cap Cells of Maize (Zea mays L.) as Affected by Compacted Soil

Effects of soil mechanical impedance on the development of Golgiapparatus in the root cap cells of maize were studied undercontrolled soil-water conditions Heavily compacted soil (bulkdensity = 1.50 g cm^–2) had 3.3 to 3.4 times greater mechanicalimpedance than control soil (bulk density = 1.33 g cm^–3),but their oxygen diffusion rates were not significantly differentThe number of dictyosomes and the number and area of secretoryvesicles per unit area of tangentially sub-peripheral root capcells in the heavily compacted soil increased compared to thosein the control These results suggest that secretory activityof the root cap cells is promoted by soil mechanical impedance Dictyosome, Golgi apparatus, maize, mucilage, root cap, secretory activity, secretory vesicles, soil mechanical impedance, Zea mays L     

Changes in the secretory activity of the Golgi apparatus during the cell cycle in root tips of maize (Zea Mays L.)

Epidermal cells of maize roots were studied to determine the distribution of Golgi apparatus-derived secretory vesicles in various stages of cell division. The following conclusions were reached: 1) The pattern of Golgi apparatus secretion varies with the cell cycle. 2) Large numbers of secretory vesicles are incorporated into the cell plate. 3) Secretory vesicles from the Golgi apparatus are incorporated primarily in walls undergoing expansion. 4) Secretory vesicles are smaller during mitosis and the first part of cytokinesis than they are during interphase. 5) Secretory vesicles account for at least 12–23% of cell-plate plasma membrane and an estimated 25% of cell-plate volume.     

Lipoprotein secretion by rat liver Golgi apparatus. Lipoprotein particles and lipase activity.

Lipoprotein particles of the size range of very low density lipoproteins in smooth endoplasmic reticulum, peripheral elements of the Golgi apparatus, and secretory vesicles of the immature Golgi apparatus face are 55 to 80 nm in diameter. Particles in mature secretory vesicles are smaller (45 nm). Concomitant with the change in particle size, the lumina of mature vesicles increase in electron density. A technique to fractionate immature and mature secretory vesicles was based on precipitation of a cupric-ferrocyanide complex (Hatchett's brown) through the action of a NADH-ferricyanide oxido-reductase resistant to glutaraldehyde which is characteristic of the membranes of mature secretory vesicles and of the plasma membrane of liver. Mature secretory vesicle fractions so isolated were enriched in cholesterol and depleted in triglycerides relative to immature vesicles on a phospholipid basis. Lipase activity was present in secretory vesicle fractions of the Golgi apparatus as shown by biochemical analysis and by cytochemistry. Cytochemical studies showed lipase to be present in both mature and immature vesicles but most evident in immature vesicles. The findings suggest that some very low density lipoprotein particles are converted to particles of smaller diameter during transit through Golgi apparatus. A lipase-mediated hydrolysis of triglycerides may relate to the transformation.     

The Stimulation of Acid Phosphatase Activity in the Stalked Gland of Drosera rotundifolia

Localization of acid phosphatases (phosphomonoesterases II EC3.1.3.2 [EC] ) was studied in the secretory cells of stalked glandtissue of Drosera rotundifolia L. using a modified Gomori procedurewith p-nitrophenol phosphate (pNP) as substrate. In unstimulatedand 24 h stimulated tissue, some acid phosphatase activity waslocalized in vacuoles, cell wall regions and cuticular poresof only a few cells. Following stimulation for either 48, 72or 96 h, acid phosphatase activity was additionally observedin most gland cells within the nuclear envelope, endoplasmicreticulum and dictyosome cisternae and their associated vesicles,suggesting a de novo synthesis of acid phosphatases. Acid phosphatase, cytochemistry, Drosera rotundifolia, secretory cells     

Developmental Features of Cell Wall Formation in Sieve Elements of the Grass Aegilops comosa var. thessalica

In differentiating sieve elements of Aegilops comosa var. thessalicadictyosomes are abundant and they produce numerous smooth vesicles.Coated vesicles seem to bud from smooth ones. Since both kindsof vesicles appear both in the cytoplasm and in associationwith the plasmalemma, it is proposed that they move to and fusewith the plasmalemma transferring products for cell wall synthesis.During differentiation sub-plasmalemmal microtubules are initiallyscarce and randomly oriented but soon afterwards they becomenumerous and transversely oriented to the long axis. Cellulosemicrofibrils in the cell wall appear to run parallel to themicrotubules and the latter may regulate microfibril orientation. Root protophloem sieve elements develop wave-like wall thickenings,which are, during development, overlaid by microtubules perpendicularto the long axis. Just after maturation these thickenings progressivelybecome smooth and finally the walls appear uniform in thickness.The wave-like wall thickenings may function as stored wall material,utilized in later stages of development when wall material willbe needed and its synthesis will be impossible because of theabsence of a synthesizing mechanism in the highly degraded protoplastsof mature sieve elements. It is suggested that in this way thethickenings may enable root protophloem sieve elements to growand keep pace with the active clongation of the surroundingcells. Aegilops comosa var. thessalica, sieve elements. cell wall, microtubules, dictyosomes, coated vesicles, wave-like thickenings     

Dictyosome-like structures from guinea-pig testes lack galactosyltransferase,a Golgi apparatus marker

Summary More than twenty different enzyme activities of fractions containing dictyosome-like structures (DLS) as a dominant cell component were monitored. Plasma membrane vesicles were a major contaminant of the DLS fractions, which, presumably as a consequence, were enriched somewhat in plasma membrane markers. The lysosomal enzymes arylsulfatase and latent acid phosphatase were present in the DLS fractions as were the Golgi apparatus activities thiamine pyrophosphatase and nucleoside diphosphatase. The presence of the latter two enzymes in DLS, plus NADH-ferricyanide reductase, has been verified from cytochemistry. On the other hand, the Golgi apparatus marker, galactosyltransferase, was not enriched in DLS fractions and appeared to be absent. This latter finding, verified from cytochemistry with isolated DLS fractions and, in situ, from [³H]galactose incorporation by testis tubules with analysis by autoradiography, provides the first clear biochemical characteristic that serves unequivocally to distinguish DLS from conventional Golgi apparatus.Work supported in part by a grant from the National Institutes of Health HD 11508     

Post Golgi apparatus structures and membrane removal in plants

Summary In nongrowing secretory cells of plants, large quantities of membrane are transferred from the Golgi apparatus to the plasma membrane without a corresponding increase in cell surface area or accumulation of internal membranes. Movement and/or redistribution of membrane occurs also in trans Golgi apparatus cisternae which disappear after being sloughed from the dictyosome, and in secretory vesicles which lose much of their membrane in transit to the cell surface. These processes have been visualized in freeze-substituted corn rootcap cells and a structural basis for membrane loss during trafficking is seen. It involves three forms of coated membranes associated with the trans parts of the Golgi apparatus, with cisternae and secretory vesicles, and with plasma membranes. The coated regions of the plasma membrane were predominantly located at sites of recent fusion of secretory vesicles suggesting a vesicular mechanism of membrane removal. The two other forms of coated vesicles were associated with the trans cisternae, with secretory vesicles, and with a post Golgi apparatus tubular/vesicular network not unlike the TGN of animal cells. However, the trans Golgi network in plants, unlike that in animals, appears to derive directly from the trans cisternae and then vesiculate. The magnitude of the coated membrane-mediated contribution of the endocytic pathway to the formation of the TGN in rootcap cells is unknown. Continued formation of new Golgi apparatus cisternae would be required to maintain the relatively constant form of the Golgi apparatus and TGN, as is observed during periods of active secretion.