The Ligule of Isoetes lacustris: Ultrastructure, Mucilage Composition, and a Possible Pathway of Secretion

Mature ligules of Isoetes lacustris can be divided anatomicallyinto three ultrastructurally different regions. First, the basalregion contains large numbers of two types of protein bodies.Second, the cells of the sub-marginal region are characterizedby a well developed Golgi apparatus closely associated withthe tubular compartments of the endoplasmic reticulum. Third,the peripheral region consists of one to three layers of cellsshowing an extremely well developed rough asternal and smoothtubular endoplasmic reticulum (ER). The tubular ER compartmentsare frequently observed in close attachment to the plasmalemma.The outermost peripheral cells are covered with a mucilaginouslayer. The dry matter in the mucilage consists of 49 per centpolysaccharides and 22 per cent proteins. The polysaccharidefraction, analysed by ion exchange chromato-graphy, consistsmainly of glucose, arabinose, galactose and uronic acids. Theprotein fraction was analysed by SDS gel dectrophoresis andby high performance liquid-chromatographic separation of theamino acids. The analysis shows a protein pattern very similarto that of the peripheral ligule tissue. It is suggested, therefore,that the material of the external mucilage is secreted by theperipheral ligule cells. The secretional mechanism appears tobe a direct release of polysaccharides and proteins by the tubularcomponents of the ER. There is no indication of secretion viathe Golgi apparatus. Because of its high activity in proteinsynthesis and secretion, it is suggested that the ligule isa vestigial structure, which, in extinct genera, might havefunctioned as a digestive organ. Isoetes lacustris, endoplasmic reticulum, ligule, ultrastructure, Polysaccharide secretion, protein secretion     

Structure and Development of Sieve Elements in the Root of Isoetes muricata Dur.

Root tissues of Isoetes muricata Dur. were fixed in glutaraldehydeand postfixed in osmium tetroxide for electron microscopy. Veryyoung root sieve elements can be distinguished from contiguousparenchyma cells by the presence of crystalline and/or fibrillarproteinaceous material in dilated cisternae of rough endoplasmicreticulum (ER). Similar crystalline-fibrillar material accumulatesin the perinuclear space. During differentiation, the portionsof ER enclosing this proteinaceous substance become smooth surfacedand migrate to the cell wall. Along the way many of them formmultivesicular bodies which fuse with the plasmalemma, dischargingtheir contents toward the wall. Nuclear degeneration is pycnotic.At maturity, the sieve element contains a degenerate, filiformnucleus, plastids, and mitochondria. In addition, the wall ofthe mature sieve element is lined by a plasmalemma and a parietalnetwork of smooth ER. Sieve-area pores are present in both endand lateral walls of mature sieve elements. Whereas a singlecluster of pores occurs in each end wall, the pores of the lateralwalls are solitary and few in number.     

Activities of Carboxylation Enzymes in Freshwater Macrophytes

Fifteen species of freshwater macrophytes, mainly from cool,temperate waters, were assayed for ribulose bisphosphate carboxylase-oxygenase(RuBPCase) and phosphoenolpyruvate carboxylase (PEPCase) activities.In extracts from all the species RuBPCase was the most activecarboxylation enzyme, and the RuBPCase/PEPCase ratio was atleast 2·0, even for the submersed species Isoetes lacustrisL. and Littorella unifiora (L.) Aschers. which have been reportedto show Crassulacean Acid Metabolism (CAM) activity. The PEPCaseactivity in I.lacustris was lower than that found in some non-CAM-likespecies. In this respect, I.lacustris and L unifiora differfrom most terrestrial CAM plants. However, these two species,along with Potamogeton praelongus Wulf. and Juncus bulbosusvar.fluitans L., had the lowest RuBPCASE/PEPCase ratios, lowerthan found in terrestrial C₃ species; suggesting that the potentialfor substantial photosynthetic metabolism of C₄ acids existsin some temperate, submersed plants. In the three amphibiousspecies (Potamogeton polygonifolius Pourr., Mentha aquaticaL., and Hippuris vulgaris L.) examined, the aerial leaves exhibitedhigher RuBPCase activities than the submersed leaves. Key words: Ribulose bisphosphate carboxylase-oxygenase, phosphoenolpruvate carboxylase, freshwater macrophytes     

Localization of Phenolic Compounds in Tannin-secreting Cells from Sambucus racemosa L. Shoots

The localization of phenolic compounds was investigated in twokinds of idioblastic tannin cells: (1) mononucleate tannin cellsclose to the promeristem in which they are initiated, and comparedwith the surrounding tissue, and (2) the very long coenocytes(i.e. almost as long as the whole internode of Sambucus) andproducing large amounts of phenolics. One likely pathway forthe appearance of these compounds in the central vacuole ispostulated from their localization: phenolic precursors occuroutside the ER cisternae and enter the interior of parts ofthe ER cisternae which undergo fragmentation and dilatation.Many small vacuoles so formed fuse and lead to the formationof a large central vacuole. Phenolic compounds, tannin cells, idioblasts, Sambucus racemosa L., shoots     

Structure and Function in the Grass Ligule: the Newly-initiated Ligule of Lolium temulentum L., a Light and Electron Microscope Study

Initiation and early development of the membranous ligule ofLolium temulentum L. was studied by light and transmission electronmicroscopy. The ligule appeared to be derived solely from theleaf adaxial epidermis and at this stage had a structure andultrastructure typical of a meristematic tissue. Cells of thefuture collar region of the leaf appear to be initiated at thesame time as the ligule. The siting of the ligule upon the leafis briefly discussed. Darnel, ligule (initiation), Lolium temulentum L., Poaceae, ultrastructure     

Dilated Endoplasmic Reticulum Cisternae in Differentiating Xylem of Minor Veins of Mimosa pudica L. Leaf

The differentiating tracheary elements in the xylem of minorveins in Mimosa pudica L. contain, as is usual, complete nucleateprotoplasts. Within the latter, dictyosomes with associatedvesicles and endoplasmic reticulum (ER) are prominent components.The ER cisternae show an uncommon feature of developing voluminousdilations accumulating finely fibrous material. Similar dilatedER cisternae occur in parenchyma cells associated with the trachearyelements. Most of the dilated cisternae are elongated, taperingat both ends, and almost circular in transections. They varyin size. The largest measured was 10 µm long and 3 µmwide. In the tracheary elements the cisternae break down asthe protoplast disintegrates. For a time, mature cells containfibrous material, apparently the product of the dilated cisternae,at least in part. In the parenchyma cells the dilated cisternaeare released into the vacuoles after the associated trachearyelements reach maturity. They become structurally modified anddisintegrate. The timing in the appearance and disintegrationof the dilated ER cisternae suggests that these structures havesome function with reference to the differentiation of trachearyelements in the xylem.     

Effects of brefeldin A on the Golgi apparatus, the nuclear envelope, and the endoplasmic reticulum in a green alga,Scenedesmus acutus

Summary The effects of brefeldin A (BFA) on the structure of the Golgi apparatus, the nuclear envelope, and the endoplasmic reticulum (ER), and on the thiamine pyrophosphatase (TPPase) activity in these organelles were examined in a green alga,Scenedesmus acutus, to obtain evidence for the existence of a retrograde transport from the Golgi apparatus to the ER via the nuclear envelope. InScenedesmus, Golgi bodies are situated close to the nuclear envelope throughout the cell cycle and receive the transition vesicles not directly from the ER, but from the nuclear envelope. BFA induced the disassembly of Golgi bodies and an increase in the ER cisternae at the trans-side of decomposed Golgi bodies in interphase cells and multinuclear cells before septum formation. The accumulated ER cisternae connected to the nuclear envelope at one part. TPPase activity was detected in all cisternae of Golgi bodies, but not in the nuclear envelope or the ER in nontreated cells. On the contrary, in BFA-treated cells, TPPase activity was detected in the nuclear envelope and the ER in addition to the decomposed Golgi bodies. When septum-forming cells were treated with BFA, the disassembly of Golgi bodies was less than that in interphase cells, and TPPase activity was detected in the Golgi cisternae but not in the nuclear envelope or the ER. These results suggest mat BFA blocks the anterograde transport from the nuclear envelope to the Golgi bodies but does not block the retrograde transport from the Golgi bodies to the nuclear envelope in interphase and multinuclear cells.Abbreviations BFA brefeldin A - ER endoplasmic reticulum - TPPase thiamine pyrophosphatase     

Some observations on coleoptile cell ultrastructure in ungerminated grains of rice (Oryza sativa L)

Summary The ultrastructure of coleoptile cells of ungerminated rice grains has been examined following fixation in glutaraldehyde and osmium tetroxide. In many respects the cell structure resembles that reported for other dormant seed tissues: the cells contain protein bodies and lipid droplets, mitochondria and plastids show little internal structure but cytoplasm invaginates into many plastids; golgi cisternae cannot be discerned. Rough ER is present as cisternae surrounding protein bodies, as occasional regions of parallel layers, and in concentric whorls where it alternates with smooth paired membranes of an unknown nature. The ribosomes on the ER are at least partly arranged into regular rows. Various crystalline, presumably proteinaceous, inclusions lie in the groundplasm, plastids and nuclei.     

STRUCTURE AND DEVELOPMENT OF SIEVE ELEMENTS IN THE LEAF OF ISOETES MURICATA

Leaf tissue of Isoetes muricata Dur. was fixed in glutaraldehyde and postfixed in osmium tetroxide for electron microscopy. The very young sieve elements can be distinguished from contiguous parenchyma cells by their distinctive plastids and the presence of crystalline and fibrillar proteinaceous material in dilated cisternae of the rough ER. During differentiation, the portions of ER enclosing this proteinaceous substance become smooth surfaced and migrate to the cell wall. Along the way they apparently form multivesicular bodies which then fuse with the plasmalemma, discharging their contents to the outside. At maturity, the sieve element contains an elongate nucleus, which consists of dense chromatin material, and remnants of the nuclear envelope. In addition, the mature sieve element is lined by a plasmalemma and a parietal, anastomosing network of smooth ER. Both plastids and mitochondria are present. P-protein is lacking at all stages of development. Tonoplasts are. not discernible in mature sieve elements. The end walls of mature sieve elements contain either plasmodesmata or sieve pores or both, but only plasmodesmata occur in the lateral walls.     

Cytochemical Localization of Phenolic Compounds in Columella Cells of the Root Cap in Seeds of Brassica napus--Changes in the Localization of Phenolic Compounds during Germination

Changes in the localization of phenolic compounds were investigatedin columella cells of embryonic roots in 1-year-old Brassicanapus seeds during 48 h of imbibition and germination. In dry,dormant seeds, phenolic compounds were located in the apoplasticcompartment between the cell wall and plasmalemma in the outermostlayer of the columella. These apoplastic phenolic deposits disappearedduring the activation processes associated with imbibition andgermination, but new deposits appeared successively in the nucleus,ER cisternae, protein bodies and on the outer surface of theroot cap. A large number of phenolic deposits were observedin the outermost part of the columella, becoming less frequenttowards the initial centre. Their appearance coincided withrestoration of the ER and immediately preceded cytological activation.After the primary root had emerged from the seed coat, depositsof phenolic compounds disappeared from the cytoplasm, but simultaneouslyappeared in the vacuoles. Copyright 1999 Annals of Botany Company Brassica napus var. oleifera, root columella, germination, phenolic compounds localization.     

Root Cap Structure in Isoetes macrospora Dur

Root meristem cells of Isoetes macrosporausually have one plastidwhich is associated with the prominent nucleus, numerous ribosomesand mitochondria, and small vacuoles. During mitosis each plastidappears to replicate so that each daughter cell contains oneplastid. The cell walls of the meristem cells are traversedby numerous plasmodesmata. Central cells of the root cap lackdistally displaced plastids but have one or more amyloplastsassociated with the nucleus. These cells also contain largeprotein deposits. Peripheral root cap cells are characterizedby being vacuolated, and by possessing a few dictyosomes andprotein deposits. They appear to be sloughed infrequently. Isoetes macrospora Dur, root cap, protein bodies, ultrastructure     

Electron microscopic and cytochemical studies of the mouse subcommissural organ

Summary In mice most of the ependymal cells of the subcommissural organ (SCO cells) are densely packed with dilated cisternae of the endoplasmic reticulum (ER) containing either finely granular or flocculent materials. The well developed supra-nuclear Golgi apparatus consists of stacks of flattened saccules and small vesicles; the two or three outer Golgi saccules are moderately dilated and exhibit numerous fenestrations; occasional profiles suggesting the budding of coated vesicles and formation of membrane-bound dense bodies from the ends of the innermost Golgi saccules are seen. A few coated vesicles and membrane-bound dense bodies of various sizes and shapes are also found in the Golgi region.The contents of the dilated ER cisternae are stained with periodic acid-silver methenamine techniques. In the Golgi complex the two or three inner saccules are stained as deeply as the dense bodies, and the outer saccules are only slightly stained. The stained contents of ER cisternae are more electron opaque than those of the outer but less opaque than those of the inner Golgi saccules and the dense bodies.Acid phosphatase activities are localized in the dense bodies, some of the coated vesicles in the Golgi region, and in the one or two inner Golgi saccules.On the basis of these results the following conclusions have been reached: (1) In mouse SCO cells the finely granular and the flocculent materials in the lumen of ER cisternae contain a complex carbohydrate(s) which is secreted into the ventricle to form Reissner's fiber; (2) the secretory substance is assumed to be synthesized by the ER and stored in its cisternae, and the Golgi apparatus might play only a minor role, if any, in the elaboration of the secretory material; (3) most of the dense bodies in the mouse SCO cells are lysosomal in nature instead of being so-called dark secretory granules.Sponsored by the National Science Council, Republic of China.     

Studies of Diurnal Acid Fluctuations in British Isoetid-type Submerged Aquatic Macrophytes

British isoetid species are examined for the presence of diurnalfluctuations in tritratable acidity (to pH 6·4), in plantscollected directly from a small lake and in plants grown inconstant conditions in the laboratory. Wide diurnal fluctuationsare present in Isoetes lacustris and in both submerged and terrestrialpopulations of Littorella uniflora. They are absent in Lobeliadortmanna, Subularia aquatica, Eriocaulon septangulare, Ranunculusflammula and Pilularia globulifera. The significance of submerged CAM is discussed in relation toother carbon accumulating mechanisms in isoetids and in considerationof their general ecology. Crassulacean acid metabolism, photosynthesis, isoetid, oligotrophic lakes     

Juvenile Hormone Induction of the Insect Yolk Protein Precursor

In Leucophaea maderae the female specific protein, or vitellogenin,is being synthesized and secreted exclusively by the adult femalefat bodies. This specific protein, which is induced by thejuvenilehormones makes up approximately 90% of the total yolk proteins.It is a lipophosphoprotein of low phosphorus (0.14) content.The female specific protein is synthesized on polysomes boundto membranes of the ergastoplasmic reticulum (ER). Microsomevesicles obtained from active tissues are heavily studded withribosomes and are considerably more dense than those from "inactive"tissues. The nascent polypeptide chains of the vitellogeninare secreted into the cisternae of the ER and can be releasedby Na deoxycholate digestion of the membranes. Similarly, thenon-specific serum proteins are also secreted into the cisternaeof the ER. All evidence points to the fact that microsomes maycarry mixed populations of polysomes, those associated withspecific and those associated with non-sex-specific proteinsynthesis. The significance of the polysomal association withmembranes for the synthesis of exportable sex-specific proteinis discussed.     

Fine Structure of Nucellar Cells During Development of the Embryo Sac in Oenothera biennis L.

Developing nucellar cells in Oenothera biennis L. present distinctpatterns of differentiation at the chalaza and around the embryosac. The cytoplasm of nucellar cells surrounding the tetradof megaspores displays cytolysomes, lipid bodies and membranesof smooth ER enveloping different cytoplasmic components. Concomitantto the differentiation of the embryo sac the nucellar cellsconstituting these ‘parietal layers’ undergo cytoplasmicdegeneration with shrinkage and flattening. In addition to theregular nucellar cells the chalaza at this stage presents threeother types: One with pycnotic nuclei, paramural bodies andcytoplasm filled with polymorphic vacuoles containing membranes,granular or flocculent material and multivesicular bodies. Asecond cell type shows swollen perinuclear cisternae aroundtheir pycnotic nuclei and large cytoplasmic vacuoles accumulatingtannins. The third type of cells is characterized by large numbersof starch grains and advanced disorganization of cytoplasmicorganelles; these cells probably become reservoirs after death. Oenothera biennis L., evening primrose, embryo sac, nucellus, cytolysomes, ultrastructure     

Disturbance of the secretory pathway inMicrasterias denticulata by tunicamycin and cyclopiazonic acid

Summary Both tunicamycin, an inhibitor of N-linked glycosylation of proteins, and cyclopiazonic acid, which inhibits the Ca²⁺-dependent ATPase in the ER, influence the secretory pathway at the ER level and lead to a cessation of cell growth inMicrasterias. Electron microscopical investigations reveal that the mode of action of the two inhibitors differs. While tunicamycin treatment results in a disintegration of the Golgi bodies into small vesicles, cyclopiazonic acid prevents products being supplied from the ER, resulting in the dilatation of ER cisternae and a reduction in the number of Golgi cisternae, combined with a loss of dictyosomal activity. The disturbed cell wall formation under tunicamycin indicates that N-linked glycosylation of proteins is required for normal cell growth inMicrasterias. Moreover, our studies reveal that changes in cytoplasmic free calcium concentration, as a consequence of ATPase inhibition in the ER by cyclopiazonic acid, may inhibit wall material secretion by interrupting the normal ER-dictyosome association.Abbreviations CPA cyclopiazonic acid - ER endoplasmic reticulum - TM tunicamycin     

The Ultrastructure of Tetrasporogenesis in the Marine Red Alga Chondria tenuissima (Good, et Woodw.) (Ceramiales, Rhodomelaceae)

Tetraspore development has been studied in Chondria tenuissimausing light and electron microscopy. The transformation of tetrasporangialmother cells into mature tetrasporangia involves a series ofstructural changes, especially of dictyosomes and of the nucleus.The youngest stage of tetrasporogenesis consists of a uninucleatetetraspore mother cell with synaptonemal complexes present duringearly prophase of meiosis I. Mitochondria are aggregated aroundthe nucleus, dictyosome activity is low, and proplastids occurin the peripheral cytoplasm. The cleavage furrows are initiatedalmost concomitantly with commencement of meiosis. When thecleavage furrows are initiated, spherical bodies bounded bytwo membranes are found within the cytoplasm; they develop intovacuoles with fibrillar contents (fv₁), which increase in sizeduring tetraspore development by fusing with each other andwith Golgi vesicles. The Golgi vesicles and the vacuoles withfibrillar contents (fv₁) contribute material to the developingtetraspore wall. During the middle stage of tetraspore formationthe vacuoles with fibrillar contents (fv₁) are dominant, dictyosomeactivity increases, as well as the number of plastids and mitochondria;starch formation also increases. Stacked cisternae of the endoplasmicreticulum are found within the peripheral part of the nucleus.The same nuclear structures are also observed in tetrasporangiaof the marine red alga Gastroclonium clavalum. The final stageis characterized by the disappearance of vacuoles with fibrillarcontents (fv₁) and of the stacked ER within the nucleus, presenceof straight, large dictyosomes which produce cored vesicles,an abundance of starch grains and by the formation of fullydeveloped chlorqplasts. The cored vesicles contain Thiéry-positivematerial and contribute to the formation of vacuoles with fibrouscontents (fv₂) as they are dominant in the tetraspores beforetheir liberation. Rhodophlyla, Chondria, tetrasporogenesis, ultrastructure, Golgi apparatus     

Changes in the Endoplasmic Reticulum During Sieve Element Differentiation in Triticum aestivum

The changes in structure of the endoplasmic reticulum (ER) andits associations with other cell components have been studiedin differentiating protophloem sieve elements of root tips ofTriticum aestivum. In the young sieve elements single ER cisternaebearing ribosomes are dispersed in the cytoplasm. As differentiationprogresses ER increases in amount while a small proportion ofit aggregates into stacks or becomes associated with the nuclearenvelope and the mitochondria. These modifications occur inthe last two sieve elements containing ribosomes and coincidewith most dramatic changes in the degenerating nucleus. Stacksconsist of relatively few ER cisternae and may be encounteredfree in the cytoplasm or applied to the nuclear envelope. Electron-densematerial accumulates between the contiguous cisternae of thestacks. ER-attached ribosomes persist even in nearly maturesieve elements, but their pattern of arrangement becomes changed.The structural evidence indicates that only a few highly degradedER elements are retained in fully mature sieve elements. Triticum aestivum, root protophloem, sieve elements, endoplasmic reticulum, differentiation     

Acid phosphatase activity during spore differentiation of the red algaeGigartina teedii andChondria tenuissima

The acid phosphatase activity during carposporogenesis inGigartina and tetrasporogenesis inChondria was studied using the Gomori technique. During the first steps of gonimoblast maturation ofGigartina, portions of cytoplasm are ensheathed by ER cisternae with acid phosphatase activity, giving rise to autolysosomal concentric membrane bodies. In a similar way large mucilage sacs are severed. They extrude their contents in a kind of exocytosis. Multivesicular bodies, concentrically arranged cisternae and extracytoplasmic compartments, each with acid phosphatase activity, remain in young carpospores for some time, probably as remnants of the autophagocytotic and exocytotic events. The Golgi apparatus is poorly developed in gonimoblast cells and young carpospores. It becomes a prominent cell component in maturing carpospores and then participates in cell wall formation. Only some of the dictyosomal cisternae contain acid phosphatase; these are irregularly distributed in the dictyosome. — In pre- and postmeiotic tetraspore mother cells ofChondria massive lead deposits are found in the dictyosomes and in adjacent Golgi vesicles. Finer lead precipitates occur in ER cisternae, especially in those which are sequestering starch-grain-containing portions of the cytoplasm to give rise to autolysosomes. During cell cleavage, the dictyosomes aggregate. They become devoid of acid phosphatase activity with the exception of vesicles at the trans face. Later, Golgi stacks associate and have common, Gomori positively reacting, narrow cisternae at the cis face. The Golgi apparatus derived cored vesicles do not contain lead precipitates whereas the Golgi cisternae in the final stage of tetrasporogenesis show acid phosphatase activity. Variations in acid phosphatase distribution are explained in the light of current models of membrane flow.Dedicated to Univ.-Prof. DrO. Härtel on the occasion of his 80th birthday.