木姜子油细胞发育的超微结构研究

利用超薄切片法和透射电镜研究了木姜子（Litsea pungens Hemsl.）油细胞的发育过程。油细胞3层细胞壁的发育可分为4个阶段,阶段1：油细胞仅有初生纤维素壁层,又可分为原始细胞和细胞 泡化两个时期。此阶段质体具透明小泡和黑色嗜锇物质,并与液泡融合。阶段2：木栓质化壁层的形成,片层状木栓质不断叠加在初生纤维素壁内侧,其细胞结构与前期相似,阶段3：内纤维素壁层的形成,较厚而松散的内纤维素壁层叠加在木栓质化壁层的内侧,在内纤维素壁层中可见黑色嗜锇物质,胞间连丝成为被阻塞的特化结构,此时大液泡被嗜锇油脂充满,成为油囊。阶段4：油细胞成熟及细胞质解体,杯形构造由内纤维素壁层向细胞腔内突起形成,油囊由液泡膜包被连接到杯形构造上,油呈浅灰色嗜锇状态,其细胞质和细胞器解体,变得电子不透明或呈杂乱状态。     

Ultrastructural Studies on the Development of Oil Cells in Litsea pungens

The developmental process of oil cells in the shoot of Litsea pungens Hemsl. has been studied with transmission electron microscopy. According to the development of the three layers of cell wall, the developmental process could be divided into 4 stages. In stage 1, the cell wall consisted only of a primary (the outmost) cellulose layer, which might further be divided into two substages, the oil cell initial, and the vacuolizing oil cell. During this stage, there were some small electron translucent vesicles and dark osmiophilic droplets of variant sizes in the different-shaped plastids. It was observed that some dark and gray osmiophilic materials coalesced to vacuoles in the cytoplasm. In stage 2, a lamellated suberin layer accumulated inside the primary cellulose layer. In stage 3, a thicker and looser inner cellulose wall layer was formed gradually inside the suberin layer. Some dark osmiophilic droplets have been observed in this loose inner cellulose wall layer. The plasmodesmata were blocked up and became a special structure. Then, the big vacuole, which is the oil sac, was full of osmiophilic oil. In stage 4, the oil cell became matured and the cytoplasm disintegrated. The oil sac enveloped from plasmalemma was attached to the cupule, which was formed by the protuberance of the inner cellulose wall layer into the lumen. After the maturity of oil cell, the ground cytoplasm began to disintegrate and became electron opaque or exhibited in a disordered state, and the osmiophilic oil appeared light gray.     

Pollen Development in Actinidia deliciosa var. deliciosa: Histochemistry of the Microspore Mother Cell Walls

The development of the microspore mother cell walls in Actinidiadeliciosa (kiwifruit) has been studied using light and electronmicroscopy. The microspore mother cell wall is similar, histochemically,and structurally in anthers from both functionally staminateand functionally pistillate flowers. Deposition, which beginsduring early prophase I, produces an electron-dense multilaminatedwall layer (layer a) and by the end of meiosis I a thick electron-lucentlayer (layer b) to the inside of this multilayered wall. Thereasons for histochemical differences and similarities betweenthese layers are discussed. The original primary wall persistsuntil the late uninucleate microspore stage. Layer (b), whichis probably mainly callose, dissolves at the late tetrad/earlymicrospore stage while layer (a), which probably also containsother polysaccharides, persists and dissolves concurrently withthe primary wall. Actinidia deliciosa, kiwifruit, microspore mother cell wall, callose, histochemistry, light microscopy, electron microscopy, male sterility     

Structural arrangement of polymers within the wall of Streptococcus faecalis.

The structure of the cell wall of Streptococcus faecalis was studied in thin sections and freeze fractures of whole cells and partially purified wall fractions. Also, the structures of wall preparations treated with hot trichloroacetic acid to remove non-peptidoglycan wall polymers were compared with wall preparations that possess a full complement of accessory polymers. The appearance of the wall varied with the degree of hydration of preparations and physical removal of the cell membrane from the wall before study. Seen in freeze fractures of whole cells, the fully hydrated wall seemed to be a thick, largely amorphic layer. Breaking cells with beads caused the cell membrane to separate from the wall and transformed the wall from a predominantly amorphic layer to a structure seemingly made up of two rows of "cobblestones" enclosing a central channel of lower density. Dehydration of walls seemingly caused the cobblestones to be transformed into two bands which continued to be separated by a channel. This channel was also observed in isolated wall preparations treated with hot trichloroacetic acid to remove non-peptidoglycan polymers. These observations are consistent with the interpretation that both peptidogylcan and non-peptidoglycan polymers are concentrated at the outer and inner surfaces of cell walls. These observations are discussed in relation to possible models of wall structure and assembly.     

Relationship of a Wall-Associated Enzyme with Specific Layers of the Cell Wall of a Gram-Negative Bacterium

In untreated cells of the marine pseudomonad studied here, alkaline phosphatase was found to be located in the periplasmic space, at the cell surface, and in the medium into which it had been shed during growth. Washing in 0.5 M NaCl, which removed the loosely bound outer layer, caused a shift of periplasmic enzyme to the outer aspect of the double-track layer and released some of the cell surface-associated enzyme. When the double-track layer of the cell wall was partially deranged, large amounts of this cell wall-associated enzyme were released, and, when the double-track was removed from the cells to produce mureinoplasts, alkaline phosphatase was released into the menstruum. There was no significant association of the enzyme with the peptidoglycan layer of the cell wall, which is the outermost structure of the mureinoplast, and no association of the enzyme with the cytoplasmic membrane of these modified cells. This study has shown that alkaline phosphatase is specifically associated with the outer layers of the cell walls of cells of this organism and is retained within the cell wall by virtue of this association.     

Enzyme layers on glass as a new model for the quantitative study of capture reactions in cytochemistry,with special attention to acid phosphatase

Summary A model system is described for the study of capture reactions for diffusable compounds in enzyme cytochemistry. The model, which allows the investigation of the influence of the composition of the cytochemical medium, the enzymatic activity, and the dimensions of the enzymatic site on the capture reaction, consists of very thin homogeneous layers of enzyme (0.01–0.1 m thick) on glass, which are incubated in the cytochemical medium. The fraction of the total amount of liberated product precipitated in the enzyme layer is dependent not only on the trapping efficiency of the cytochemical medium but also on the concentration of the primary reaction product that can be built up in the enzyme layer. Calculations were performed to determine the steady-state concentration of the primary reaction product that can be built up in the enzyme layer. Acid phosphatase was used as enzyme. The problems associated with the model and its applicability to other types of cytochemical reactions are discussed.     

银杏套细胞发育的解剖学研究

银杏套细胞起源于近雄配子体表达的颈卵器母细胞周围细胞,中央细胞形成时,套细胞呈明显的一层,围绕颈卵器紧密排列,随着颈卵器的发育,套细胞体积逐渐地大,细胞质变浓厚,细胞质中脂滴增多,套细胞和中央细胞的接触壁开始出现局部加厚,在颈卵器发育的泡沫化阶段,套细胞和中央细胞的接触壁不均匀加厚较为显著,在较薄的区域可见胞间连丝,受精前,套细胞中液泡增多,脂滴迅速减少,造淀粉体增大,套细胞与卵细胞的接触壁的不均匀加厚非常明显,精核进入颈卵器以后,卵细胞与套细胞的接触壁和卵细胞的质膜之间形成一个薄厚不均的间隔层,受精卵分裂时,受精卵细胞与套细胞接触壁的凹陷处可见许多小泡和内质网,游离核期时,套细胞内出现大量小液泡,细胞内含物迅速消失,套细胞外形变长,胚胎长出颈卵器后,套细胞逐渐解体消失。     

Characterization of two “Metabacterium” sp. from the gut of rodents

A new giant Gram-negative non-cultivatable symbiotic endospore-forming bacterium was found in the gut of the European hamster. This “Metabacterium” sp., provisionally named “Metabacterium criceti”, sp. n., has a length of approximately 20 μm and thickness of 4 μm. It forms 1 to 2 cylindrical endospores, approximately 9 μm long and 1.4 μm thick. TEM-micrographs show a cell wall structure characteristic of Gram-negative bacteria. Vegetative cells are filled with granules 0.3 μm in diameter which resemble starch granules. The reproduction occurs with binary fission and by formation of two endospores. Of thirteen biochemical components sought, four,i.e. glycogen, triacylglyceroles, peroxidase and alkaline phosphatase, were not found. Starch, acid mucosubstances, DNA, RNA, lipids, proteins, adenosine triphosphatase and acid phosphatase were found in different patterns, depending on the developmental stage of the bacterium. In the vegetative cell stage all these components, with the exception of starch, were found. In the endospore-bearing cell stage, only the starch-like cell component granulose could be detected. In free endospores only DNA, RNA and acid phosphatase were found. Some of the components,i.e. DNA, lipids, starch-like granulose, were linked to certain cell substructures, the distribution of others,viz. polysaccharides, RNA, adenosine triphosphatase and proteins was diffuse. The lipids, found only in vegetative cells, were associated with the cell wall. Presented in part during the19th Congress of the Czechoslovak Society for Microbiology, Košice (Slovakia) September 14–17, 1992. An erratum to this article is available at .     

Microsporogenesis and exine substructure in Uraria crinita (Fabaceae)

This paper intends to elucidate the anther wall development, pollen wall development and exine substructure of Uraria crinita (L.) Desv. ex DC. (Fabaceae). The undifferentiated anther is ovoid-shaped and tetrasporangiated. The anther wall development is basic type, which is comprised of an epidermis, an endothecium layer, two middle layers and a tapetum. Anther-tapetum is glandular type and the cells are uniseriate and uninucleate. Pollen grains are tricolporate and 2-celled at the time of shedding. Before protectum development begins, a glycocalyx layer is inserted against the callose, and the plasma membrane is invaginated, exclusive of the future apertures. Subsequently, the probacula are elongate under the protectum and arise basally from the plasma membrane. The foot layer and endexine formation are concomitant with the callosic wall dissolution. The foot layer is thin and interrupted, but the endexine is thick and continuous. The intine is initially in the vacuoled stage. The substructure in the tectum, bacula and endexine is the same as a rod-shaped in side view. It composed of the loop like striate elements.     

Studies on Mature Seeds of Cuscuta pedicellata and C. campestris by Electron Microscopy

The seeds of Cuscuta pedicellata have been investigated by transmissionand scanning electron microscopy. Additional observations havebeen made on seeds of C. campestris by SEM only. The seed coatconsists of an outer single epidermis, two different palisadelayers, and an inner multiparenchyma layer. The outer epidermalwall in C. pedicellata has a thick cuticle and zones rich inpectic substances. The thicker ‘U-shaped’ cell wallsin the outer palisade layer are strengthened by a wall layerof hemicellulose. The inner palisade layer has thick walledcells with a ‘light line’. The inner cell wall ofthe compressed multiparenchyma layer has a thin cuticle. A fairlythick cuticle is positioned directly on the endosperm surface.The aleurone cell walls are different from the remaining endospermwalls. The latter are thick and believed to be of galactomannans.There is a ‘clear’ zone between the plasmalemmaand the cell wall in the aleurone cells. The embryo cells arepacked with lipids and proteins. In Cuscuta campestris mostendosperm has been absorbed during the seed development. Theembryo apex has two minute leaf primordia. The features of theCuscuta seeds are discussed in relation to functional and environmentalconditions. Cuscuta pedicellata, Cuscuta campestris, seed, seed coat, cuticle, cell walls, endosperm, aleurone cells, galactomannan, embryo, TEM, SEM     

Exine ontogeny in Borago officinalis pollen

The primexine matrix is finely granulo-fibrillar up to callose digestion; it becomes distinctly fibrillar at the free microspore stage. The columellae and the tectum are initiated at the middle tetrad stage, the foot layer and the endexine are initiated when the callose wall digestion begins. The columellae are initiated by the deposition of spiral elements around a clear central zone. This hollow aspect of columella disappears when thickening. The foot layer and the endexine are built by the expansion of plasmalemma derived components. The foot layer appears first at the poles, then at the interapertural levels and at last at the apertures while the endexine appears first at the mesoapertures, then it spreads laterally towards the interapertural levels and, at last, at the poles. The gemmae are formed at the free microspore stage over all the tectum. The thickening of the exine takes place essentially during the free microspore stage and continues during the vacuolate microspore one. Apertures are entirely formed before the complete digestion of the callose wall. The ectoapertures are determined by the lacking of the columellae; the sites of the pericolpal cavities and the mesoapertures result from the plasmalemma retraction even before the setting up of the foot layer and the endexine by which they will be delimited respectively afterwards. The endoapertures are determined by the lacking of compact endexine at their level, and merge into a continuous equatorial belt.     

Wall texture in the spore of a vesicular-arbuscular mycorrhizal fungus

Summary InGlomus epigaeum Daniels and Trappe, a vesicular-arbuscular mycorrhizal fungus, the mature spore has a complex multi-layered wall containing a regular pattern of wall subunits.The outer wall (2–4 m thick) consists of a simple layer of parallel microfibrils. The inner wall (5–6 m thick) is built from two layers possessing different organization. The innermost layer, near the plasmalemma has a texture of apparently dispersed fibrils, whereas the second layer is regularly organized with an arced texture. Ten to twelve bundles of fibrils connected by apparently bow-shaped fibrils are consistently observed. The appearance of this arced organization depends on the section plane and on the angle of observation in the electron microscope as confirmed by tilting experiments. Wall subunits are evident as straight electron transparent fibrils; particularly well-defined in negatively stained frozen sections: their diameter is about 3.5nm.The regular pattern of wall subunits in this fungal cell wall is compared with the textures shown by cellulose fibrils in algae or higher plants and by chitin fibrils in arthropod cuticle.Research work supported by CNR, Italy. Special grant I.P.R.A.—Sub-project 1. Paper No. 55.     

凤仙花花药发育特征

凤仙花花药发育比较特殊: 在造孢细胞时期,花药横切面中央是体积较大、细胞内含物较多的细胞团、包括造孢细胞和绒毡层细胞。花药药壁细胞的细胞质较稀少,与中部细胞界限明晰。花粉母细胞时期的花药药壁由约6层细胞组成,但细胞的界限不明显;绒毡层细胞显示变形流入药室中。到四分体时期,绒毡层细胞进一步退化。开花时,成熟花药的药壁细胞由一层表皮细胞、两层药室内壁细胞和一层中层细胞组成。对凤仙花花药绒毡层的特殊性质进行了讨论。     

Enzyme cytochemistry of the alveolar sacs and golgian-like cisternae in the ciliate Ichthyophthirius multifiliis

In the cell cortex of the parasitic ciliate Ichthyophthirius multifiliis different kinds of cisternae were observed: the alveolar sacs, thick membrane cisternae and the endoplasmic reticulum. The thick membrane cisternae possess coated dilated rims and sometimes could be observed close to the endoplasmic reticulum. Using cytochemical techniques acid phosphatase, thiamine pyrophosphatase and nucleoside diphosphatase activities were detected in the thick membrane cisternae and in the alveolar sacs of trophozoites. In the endoplasmic reticulum acid phosphatase activity was not detected and only very small amounts of thiamine pyrophosphatase and nucleoside diphosphatase reaction product were observed. After exit from the host, a reduction in acid phosphatase activity was evident in the alveolar sacs. At theront stage acid phosphatase activity is absent from these structures. However, high thiamine pyrophosphatase and nucleoside diphosphatase activities remain in the alveolar sacs during the whole life cycle. On the other hand, acid phosphatase, thiamine pyrophosphatase and nucleoside diphosphatase activities were detected in thick membrane cisternae of theronts. Based on the morphological aspects and enzymatic content the thick membrane cisternae of the cell cortex are designated as golgian-like cisternae. The cytochemical results point out a relationship between the alveolar sacs and the Golgi complex.     

Deposition of glucuronoxylans on the secondary cell wall of Japanese beech as observed by immuno-scanning electron microscopy

Summary Glucuronoxylans (GXs), the main hemicellulosic component of hardwoods, are localized exclusively in the secondary wall of Japanese beech and gradually increase during the course of fiber differentiation. To reveal where GXs deposit within secondary wall and how they affect cell wall ultrastructure, immuno-scanning electron microscopy using anti-GXs antiserum was applied in this study. In fibers forming the outer layer of the secondary wall (S₁), cellulose fibrils were small in diameter and deposited sparsely on the inner surface of the cell wall. Fine fibrils with approximately 5 nm width aggregated and formed thick fibrils with 12 nm width. Some of these thick fibrils further aggregated to form bundles which labelled positively for GXs. In fibers forming the middle layer of the secondary wall (S₂), fibrils were thicker than those found in S₁ forming fibers and were densely deposited. The S₂ layer labelled intensely for GXs with no preferential distribution recognized. Compared with newly formed secondary walls, previously formed secondary walls were composed of thick and highly packed microfibrils. Labels against GXs were much more prevalent on mature secondary walls than on newly deposited secondary walls. This result implies that the deposition of GXs into the cell wall may occur continuously after cellulose microfibril deposition and may be responsible for the increase in diameter of the microfibrils.Abbreviations GXs glucuronoxylans - PBS phosphate-buffered saline - RFDE rapid-freeze and deep-etching technique - FE-SEM field emission scanning electron microscope - TEM transmission electron microscope     

Ultrastructure of the ascospores of the new yeast genus Sporopachydermia Rodrigues de Miranda

Development of the ascospores of Sporopachydermia lactativora and S. cereana was studied in ultrathin sections. The spores have a very thick wall consisting of a thin dark outer layer and a double light inner layer the outer part of which is very wide and often irregular. During germination, this part disappears, the outer dark layer breaks up and the inner part of the light layer remains around the protoplast during development to a vegetative cell.This investigation was supported by the Netherlands Organization for the Advancement of Pure Research (Z.W.O.).     

柿树炭疽菌侵染寄主的细胞学研究*

超微结构研究表明,柿树炭疽菌(Colletotrichum gloeosporioides)侵染后在寄主细胞中形成初生菌丝和次生菌丝,寄主细胞膜外沉积了一层厚的电子不透明物质,初生菌丝与具有沉积物的寄主原生质膜之间有一层界面基质(interfacial matrix)。当初生菌丝扩张并侵染相邻细胞时, 围绕着初生菌丝层的界面基质消失,具有沉积物的原生质膜被逐步降解。初生菌丝在穿透寄主细胞壁过程中形成一个漏斗状的菌丝锥,然后穿透寄主细胞壁并迅速膨大, 然后形成厚壁的初生菌丝。初生菌丝在寄主细胞壁中收缩狭窄处产生一个隔膜,隔膜两边菌丝中细胞质的电子密度明显不同,菌丝锥中有浓密的电子密度。死体营养的次生菌丝在死的细胞中繁殖和扩展,并产生分枝。次生菌丝可直接穿透较薄的寄主细胞壁,无缢缩或任何变形现象,菌丝顶端部分未见隔膜产生;在穿透较厚的细胞壁时,靠近顶端处产生隔膜,顶端细胞膨大,使寄主细胞壁撕裂。接种90h后分生孢子盘在枝条表面形成。柿树炭疽菌其侵染过程有两个阶段,即初生菌丝的活体营养阶段和次生菌丝的死体营养阶段。     

Ultracytochemical localization of acid phosphatase in Humicola lutea conidia and mycelia

Electron microscopic cytochemical procedures were used to determine the cellular location of acid phosphatase in the fungus Humicola lutea grown in casein-containing medium lacking in mineral orthophosphates. In our investigations acid phosphatase in nongerminating conidia was localized on the outer side of the cell wall, in the cell wall, and on the exterior surface of the plasma membrane. The reaction product of acid phosphatase in germinating conidia was seen in the outer wall layer while in young mycelium on the cell surface and in the exocellular space. The relationship between phosphatase activities localized in the cell wall and their role in the enzymatic degradation of the phosphoprotein casein providing available phosphates for cell growth is discussed.     

The fine structure of secretion in Hyalophysa chattoni: formation of the attachment peduncle and the chitinous phoretic cyst wall.

The settling tomite stage of the apostome Hyalophysa chattoni secretes a phoretic cyst wall composed of chitin, mucopolysaccharides, and protein. Within 1 1/2 h after settling, an electron-dense proteinaceous cyst layer (the outer layer) is formed from secretions originating at the base of the kineties and from the thick pellicular layer between the kineties. The inner cyst layer, composed primarily of chitin (acidic and neutral polysaccharides are also present), is secreted across the entire cell surface. Cyst wall formation is completed within 6 h. The fine structure of endocyst secretion resembles stages in the secretion of chitin by fungi, yeasts, and arthropods. A proteinaceous attachment peduncle is secreted to anchor the cell to a shrimp host and is formed by the release of electrondense dense secretory bodies from the cell's ventral surface.     

Localization of alkaline phosphatase in three gram-negative rumen bacteria

Of the three species (Bacteroides ruminicola, B. succinogenes, and Megasphaera elsdenii) of anaerobic gram-negative rumen bacteria studied, only B. ruminicola produced significant amounts of alkaline phosphatase. This enzyme, which is constitutive, showed a greater affinity for p-nitrophenylphosphate than for sodium-beta-glycerophosphate and was shown to be located exclusively in the periplasmic space of log-phase cells. Small amounts of this enzyme were released from these cells in stationary-phase cultures, but washing in 0.01 M MgCl(2) and the production of spheroplasts by using lysozyme in 0.01 M MgCl(2) did not release significant amounts of the enzyme. Exposure to 0.2 M MgCl(2) did not release significant amounts of the periplasmic alkaline phosphatase of the cell, and when these cells were spheroplasted with lysozyme in 0.2 M MgCl(2) only 25% of the enzyme was released. Spheroplasts were formed spontaneously in aging cultures of B. ruminicola, but even these cells retained most of their periplasmic alkaline phosphatase. It was concluded that the alkaline phosphatase of B. ruminicola is firmly bound to a structural component within the periplasmic area of the cell wall and that the enzyme is released in large amounts only when the cells break down. The behavior of alkaline phosphatase in this bacterium contrasts with that of conventional periplasmic enzymes of aerobic bacteria, which are released upon conversion into spheroplasts by lysozyme and ethylenediaminetetraacetic acid and by other types of cell wall damage. All three species of bacteria studied here, as well as bacteria found in mixed populations in the rumen, have thick, complex layers external to the double-track layer of their cell walls. In addition, B. ruminicola produces a loose extracellular material.