Multivesicular compartments proliferate in susceptible and resistant MLA12-barley leaves in response to infection by the biotrophic powdery mildew fungus

There is growing evidence that multivesicular bodies and cell wall-associated paramural bodies participate in the enhanced vesicle trafficking induced by pathogen attack. Here, we performed transmission electron microscopy in combination with cytochemical localization of H2O2 to investigate multivesicular compartments during establishment of compatible interaction in susceptible barley (Hordeum vulgare) and during hypersensitive response in resistant MLA12-barley infected by the barley powdery mildew fungus (Blumeria graminis f. sp. hordei). Multivesicular bodies, intravacuolar vesicle aggregates and paramural bodies proliferated in the penetrated epidermal cell during development of the fungal haustorium. These vesicular structures also proliferated at the periphery of intact cells, which were adjacent to the hypersensitive dying cells and deposited cell wall appositions associated with H2O2 accumulation. All plasmodesmata between intact cells and hypersensitive cells were constricted or blocked by cell wall appositions. These results suggest that multivesicular compartments participate in secretion of building blocks for cell wall appositions not only to arrest fungal penetration but also to contain hypersensitive cell death through blocking plasmodesmata. They may also participate in internalization of damaged membranes, deleterious materials, nutrients, elicitors and elicitor receptors.     

Paramural bodies of Albugo candida

The paramural bodies of Albugo candida were formed solely by elaboration of the plasmalemma. Two major forms were recognized: one consisting of plasmalemmal invaginations projecting into the cytoplasm; the other appearing like a pocket containing a number of vesicles and tubules. It is suggested that the first is the basic form of paramural body. In sporangia the paramural bodies break away from the plasmalemma and undergo autodigestion while in vegetative hyphae their tubules and lamellae break up into vesicles that are finally sequestered into the wall.     

On the ultrastructure of differentiating secondary xylem in willow

Summary Studies of differentiating xylem inSalix fragilis L. show the immediate cambial derivatives to be ultrastructurally similar. The Golgi apparatus is important at all stages of wall synthesis, possibly producing (amongst other substances) hemicellulose material which is carried to the wall in vesicles or multivesicular bodies. The endoplasmic reticulum also contributes one or more components to the developing wall: at some stages during differentiation the endoplasmic reticulum produces electron opaque bodies which appear to be guided towards the wall by microtubules. Compact structures formed from concentric membranes (myelin-like bodies) have been found joined to rough endoplasmic reticulum, but their presence is not explained.Two types of plasmalemma elaboration occur: invagination of the plasmalemma itself to form vesicles which may contain cytoplasmic material; and vesicles between the plasmalemma and cell wall which are the result of single vesicles or multivesicular bodies traversing the plasmalemma. Both systems provide a means for transporting cytoplasmic material across the plasmalemma.Microtubules have been seen associated with all vesicles derived from the cytoplasm which appear to be moving towards the wall. The presence of microtubules may generally be explained in terms of orientation of vesicles, even if they also happen coincidentally to parallel the supposed orientation of microfibrils in the wall itself. It is possible to resolve connections between the microtubules and the plasmalemma.     

Vesicular structures in the dendrites of an insect olfactory receptor

The receptor dendrites of the antennal plaque organs of two species of fulgoroid insects contain 600 Å vesicles. These are concentrated in the distal vesicular region but also occur in the extreme tips of fine dendritic branches at the olfactory surface. At these points pore filaments run from the cuticular pores and are closely apposed to the dendritic membrane. Similar vesicles are found associated with Golgi bodies in the cell body and are also found more distally in multivesicular bodies. The multivesicular bodies appear to rupture and release vesicles. Vesicles also occur in the core of the dendrite cilium and it is assumed that they pass through the core and accumulate in the vesicular region. Similar vesicles appear to be a common feature in many, if not all, insect olfactory receptors.     

浙贝母鳞片细胞休眠解除前后超微结构的变化

浙贝母休眠解除后,鳞片近轴面表皮附近的几层细胞首先降解。本文观察了这些细胞在休眠解除前后超微结构的变化。与休眠状态时的细胞相比,解除休眠后的细胞中颗粒和丝状物的数量明显增加,一些细胞中出现了复杂的膜结构。线粒体的数目增加,常聚集在细胞核和细胞壁旁。胞间连丝的直径略有增加。休眠解除前后的细胞中,常可见到各类囊泡、多泡体与壁旁体,一些小囊泡正在进入细胞中。表明两种时期的细胞间都发生着物质和信息的交换     

水稻胚乳细胞质膜内陷的超微结构和磷酸酶的细胞化学定位

对生长分化期水稻胚乳细胞的质膜内陷进行了超微结构和磷酸酶的细胞化学研究。结果表明 ,胚乳细胞内的小泡、内质网常与胞间连丝相连 ;质膜形态多变 ,功能活跃 ,由局部起伏的波纹状发展成明显内陷 ,深浅不一 ,多呈袋状 ,袋中包含着大小不一的泡状物 ;有些内陷脱离质膜成为胞质中的囊泡 ,表现出活跃的内吞现象。除细胞间隙中含有圆球状的内含物外 ,在质膜内陷和囊泡中常含有大量的内含物。H ATP酶定位结果显示 ,质膜及其邻近的泡状物周围有酶的分布 ;而酸性磷酸酶定位在液泡、胞间隙和其中的泡状内含物周围 ;在质膜及其内陷形成的囊泡中有G6P酶的分布。这些结果表明胞间隙和质膜内陷在物质的运输中可能起着重要作用     

毛竹茎秆纤维细胞发育过程中ATP酶的超微细胞化学定位研究

采用磷酸铅沉淀技术,对毛竹茎秆纤维细胞发育过程中的ATP酶进行了超微细胞化学定位研究.在初生壁形成时期,大量的ATP酶的活性产物沉积在质膜、质膜内陷、运输小泡、胞间连丝等膜体系以及细胞核和各种细胞器上;在次生壁形成的初期,ATP酶在多泡小体和裂解的液泡膜上出现,凝聚并边缘化的染色质上仍然具有ATP酶活性;随着次生壁的逐渐加厚,在前四年中持续存在具有ATP酶活性的质膜内陷结构,以后消失;而在六年生纤维细胞的质膜、运输小泡、纹孔、胞间连丝和凝聚化的染色质上仍然发现有明显的ATP酶分布,并发现在染色质上ATP酶活性会随着凝聚程度的加深而增强.结果表明,ATP酶在毛竹茎秆纤维细胞壁的整个形成过程中发挥重要作用,而纤维细胞的次生壁形成过程是一个由核基因控制的主动的PCD过程;并证实毛竹茎秆纤维细胞的发育有别于其它木本植物纤维细胞的发育过程,这种纤维细胞是一种典型的长寿细胞.     

Multivesicular bodies participate in a cell wall-associated defence response in barley leaves attacked by the pathogenic powdery mildew fungus

Localized cell wall modification and accumulation of antimicrobial compounds beneath sites of fungal attack are common mechanisms for plant resistance to fungal penetration. In barley (Hordeum vulgare) leaves, light-microscopically visible vesicle-like bodies (VLBs) containing H(2)O(2) or phenolics frequently accumulate around cell wall appositions (syn. papillae), in which the penetration attempt of the biotrophic powdery mildew fungus Blumeria graminis f. sp. hordei (Bgh) is halted. By ultrastructural analyses, we demonstrated that the Bgh-induced VLBs represent different structures. VLBs intensively stained by H(2)O(2)-reactive dyes were actually small papillae instead of cytoplasmic vesicles. Other VLBs were identified as osmiophilic bodies or multivesicular compartments, designated paramural bodies (PMBs) and multivesicular bodies (MVBs). MVBs seemingly followed two distinct pathways: either they were engulfed by the tonoplast for degradation in the vacuole or they fused with the plasma membrane to release their internal vesicles into the paramural space and hence could be the origin of PMBs. MVBs and PMBs appeared to be multicomponent kits possibly containing building blocks to be readily assembled into papilla and antimicrobial compounds to be discharged against fungal penetration. Finally, we propose that released paramural vesicles might be similar to exosomes in animal cells.     

Application of Electrical Wave Transmission in Basic Research of Grafting

Electrical wave (EW) transmission from scion to stock across the grafting interface was related to the histological changes during the development of graft union. Variation wave (VW) could not be transmitted to stock from scion before isolation layer broken and callus interdigitation. As plasmodesmata formed secondarily at the interface between stock and scion where the isolation layer had disappeared, VW could be transmitted from scion to stock, but its velocity was not rapid until the vascular bridges form between two partners of graft union. Hence, the authors could deduce the degree of graft union formation by measuring whether VW could be transmitted from scion to stock or not and its velocity. EW trans mission method was a new tool for quickly detecting the formation of graft union.     

Immunogold localisation of endogenous immunoglobulin-G in ultrathin frozen sections of the human placenta

Summary Endogenous immunoglobulin-G was localised in ultrathin frozen sections of human term placenta by use of an indirect immuno electron-histochemical methodology. Immunoreactivity of endogenous IgG to rabbit anti-human immunoglobulin-G antibody was visualised by use of protein-A — colloidal gold complex. Gold marked the syncytiotrophoblast in both coated and uncoated regions of the apical plasmalemma, in vesicles and multivesicular bodies, and in vesicles near the basal plasmalemma. Immunoreactivity was also seen in the interstitial space between the trophoblast and the fetal endothelial layer as well as in various types of vesicles within the endothelial cells. No immunoreactivity was seen in the intercellular clefts of the endothelium. The pattern of localisation observed is consistent with receptor-mediated uptake of immunoglobulin-G into the syncytiotrophoblast of the human placenta followed by release into the interstitial space and then vesciular transport through the endothelium.     

电波传递在嫁接基本理论研究中的应用

研究发现电波从接穗跃过嫁接面向砧木传递与嫁接的组织学变化相关。砧木和接穗愈伤组织细胞突破隔离层互相嵌合之前,接穗中产生的电波不能传到砧木。砧木和接穗细胞间产生次生胞间连丝后,电波即可沿接穗传至砧木,但此时传递速度慢。当砧木和接穗间产生维管束桥时,电波下传的速度加快。由此可见,通过检测电波跃过嫁接面与否及下传的速度,就可推断嫁接接合部组织学变化状况。电波传递法是一种快速鉴定嫁接植株发育过程的新方法。     

Immunocytochemical and labelled tracer approaches to uptake and intracellular routing of Immunoglobulin-G (IgG) in the human placenta

Summary Isolated lobules of freshly delivered human term placenta were (a) subjected to an indirect immunoelectron ultracryo method in which the immunoreactivity of endogenous Immunoglobulin-G (IgG) to rabbit anti-human IgG antibody was localized with protein-A-colloidal gold and (b) extracorporeally perfused and human IgG molecules complexed to horseradish peroxidase (HRP) added to the maternal perfusate and the uptake of IgG-HRP over different perfusion durations visualized ultrastructurally by using diaminobenzidine cytochemistry. Immunoreactivity to anti-human IgG antibody was localized all along the apical plasmalemma, in apical coated and uncoated vesicles, in apical and juxtanuclear multivesicular bodies, and in basal vesicles of the syncytiotrophoblast layer of the placenta. The stroma separating the syncytiotrophoblast from the foetal endothelium as well as vesicles within the endothelium were immunoreactive. No immunoreactivity was localized in paracellular clefts of endothelia. A similar distribution of exogenous IgG-HRP was observed for the perfused placentae. When bovine IgG-HRP or HRP alone were used as control tracers no uptake was seen for the former whilst the latter was observed only in early endosomal vesicles of the syncytiotrophoblast. The pattern of localization visualized in both studies is consistent with receptor-mediated uptake of IgG by the syncytiotrophoblast and a vesicular transport of IgG across the foetal endothelium.     

Fine Structure of Cryptococcus neoformans Grown In Vitro as Observed by Freeze-Etching

Cryptococcus neoformans grown on culture media was observed by the freeze-etching technique. In the capsule, short fibrils were seen when freezeetched. This organism was unique in the appearance of the cell wall, which showed two strata. The outer one was dense with particles of about 20 nm in diameter, whereas the inner one was sparse in particles. The appearance of the cell membrane of this organism differed distinctly depending on the culture media. When grown on glycerol medium, the cell membrane possessed, as do other yeasts, clear but somewhat longer and curved invaginations. The membrane of cells grown on nonglycerol medium exhibited, however, only a few invaginations of irregular shape. Instead, characteristically of this organism, the cell membrane showed round depressions of 40 to 200 nm in diameter which were the surface view of the paramural bodies. In cross-fractured cells, both types of paramural bodies were found. Some of them contained a single vesicle of about 50 nm in diameter. These seem to play a role in secreting the cytoplasmic vesicles. Data suggesting the existence of multivesicular bodies in the cytoplasm and multivesicular lomasomes were also obtained. Some of the baglike paramural bodies showed multilayered membrane. These are thought to be plasmalemmasomes. This organism was similar to other yeasts reported in other respects.     

The small GTPase Rab5a is essential for intracellular transport of proglutelin from the Golgi apparatus to the protein storage vacuole and endosomal membrane organization in developing rice endosperm

Rice (Oryza sativa) glutelins are synthesized on the endoplasmic reticulum as larger precursors, which are then transported via the Golgi to the protein storage vacuole (PSV), where they are processed into acidic and basic subunits. Three independent glutelin precursor mutant4 (glup4) rice lines, which accumulated elevated levels of proglutelin over the wild type, were identified as loss-of-function mutants of Rab5a, the small GTPase involved in vesicular membrane transport. In addition to the plasma membrane, Rab5a colocalizes with glutelins on the Golgi apparatus, Golgi-derived dense vesicles, and the PSV, suggesting that Rab5a participates in the transport of the proglutelin from the Golgi to the PSV. This spatial distribution pattern was dramatically altered in the glup4 mutants. Numerous smaller protein bodies containing glutelin and α-globulin were evident, and the proteins were secreted extracellularly. Moreover, all three independent glup4 allelic lines displayed the novel appearance of a large dilated, structurally complex paramural body containing proglutelins, α-globulins, membrane biomarkers for the Golgi apparatus, prevacuolar compartment, PSV, and the endoplasmic reticulum luminal chaperones BiP and protein disulfide isomerase as well as β-glucan. These results indicate that the formation of the paramural bodies in glup4 endosperm was due to a significant disruption of endocytosis and membrane vesicular transport by Rab5a loss of function. Overall, Rab5a is required not only for the intracellular transport of proglutelins from the Golgi to the PSV in rice endosperm but also in the maintenance of the general structural organization of the endomembrane system in developing rice seeds.     

发育过程中苹果果皮和果肉细胞的超微结构

用透射电镜对发育过程中苹果（Ｍａｌｕｓ ｄｏｍｅｓｔｉｃａ Ｂｏｒｋｈ ｃｖ．Ｒｅｄ Ｆｕｊｉ）果皮和果肉细胞的超微结构进行了观察。结果表明,不同发育期果皮细胞的超微结构发生了变化,其中最引人注目的是,内质网自始至终密布于整个细胞质中,而且大多为槽库膨大的、合成功能旺盛的管状粗面内质网,并分泌出大量的具运输功能的小泡;观察到这些小泡与液泡融合的景象;细胞中也存在活跃的高尔基体。超微结构上的这些现象     

银耳（Tremella fuciformis）原基分化前期双核菌丝细胞和分生孢子的边缘体与质膜体

本文报道银耳（Ｔｒｅｍｅｌｌａｆｕｃｉｆｏｒｍｉｓ）原基分化前期．在双核菌丝的幼细胞、成熟细胞和分生孢子中,与质膜相关联的两类膜结构──边缘体和质膜体的形成与功能。根据相似结构的存在．支持小泡或多泡体排出质膜之外附在细胞壁上成为边缘体和参于细胞壁合成的假定。银耳原基分化前期．双核菌丝迅速分裂的幼细胞．其质膜内陷产生泡状质膜体,内含数个小泡,或产生膜状质膜体;在成熟细胞中．质膜内陷通常形成回旋的膜结构──膜状质膜体．内含１—２个电子致密小泡．当这两类质膜体脱离质膜进入细胞质后,有的膜层和小泡局部被消化．因此,推断质膜体具有内吞和输送养料的作用。另外．在桶孔隔膜闭塞一侧电子致密度高的细胞质中．还观察到一种罕见的只有单个膜层的质膜体．其内充满３个电子致密小泡．估计它的形成与功能同膜状质膜体相似。作者认为．桶孔闭塞和质膜体的出现是与银耳原基细胞分化有关联的两个重要特征。最后,在成熟细胞中,尚可以观察到质膜体的膜层能够散开形成内质网．因此．内质网也可以来源于质膜体。     

Fine Structure of Cryptococcus neoformans Grown In Vivo as Observed by Freeze-Etching

Cryptococcus neoformans grown in the parasitic state was observed by the freeze-etching technique and was compared with that grown on culture media. Unlike other yeasts, this organism grown in vivo is very often devoid of the "ordinary" invaginations. The membrane of the cell grown in vivo was almost free from concavity and convexity except for many round depressions which represent the surface view of paramural bodies. Some of the paramural bodies were found to be multivesicular systems. Most were spherical invaginations containing a single vesicle or its ghost remaining after secretion of the vesicles. In clear contrast to the cell grown in vitro, the in vivo cell contained a great number of vesicles in the cytoplasm. These seemed to show high-secretion activity in C. neoformans grown in the parasitic state. On transfer from in vitro to in vivo, this organism enlarged the cell wall, capsule, and cell body. The appearance of a large vacuole, accumulation of storage organelles, and the existence of rodlike structures, seemingly lipid deposits, were also noted in the cytoplasm of the cell grown in vivo. the meaning of these results as well as the mode of capsular production are discussed.     

Multivesicular bodies in the transitional epithelium of the neonatal mouse urinary border.

The origin of late endosomes - multivesicular bodies (MVBs) in the superficial cells of 16 and 17 embryonic old transitional epithelium of mouse urinary bladder was studied by electron microscopy, lectin labelling and HRP tracing. Analysis of hexagonally structured membrane particles, WGA, and RCA I binding sites revealed structural similarity between plasmalemma, fusiform vesicles and multivesicular bodies. Early endosomes are lined by symmetric unit membrane as well as by asymmetric thickened membrane regions. Multivesicular bodies and fusiform vesicles have asymmetric unit membranes. MVBs may be derived from primary endosomes as well as from fusiform vesicles in the cytoplasm.     

FUNCTIONS OF COATED VESICLES DURING PROTEIN ABSORPTION IN THE RAT VAS DEFERENS

The role of coated vesicles during the absorption of horseradish peroxidase was investigated in the epithelium of the rat vas deferens by electron microscopy and cytochemistry. Peroxidase was introduced into the vas lumen in vivo. Tissue was excised at selected intervals, fixed in formaldehyde-glutaraldehyde, sectioned without freezing, incubated in Karnovsky's medium, postfixed in OsO₄, and processed for electron microscopy. Some controls and peroxidase-perfused specimens were incubated with TPP,¹ GP, and CMP. Attention was focused on the Golgi complex, apical multivesicular bodies, and two populations of coated vesicles; large (> 1000 A) ones concentrated in the apical cytoplasm and small (<750 A) ones found primarily in the Golgi region. 10 min after peroxidase injection, the tracer is found adhering to the surface plasmalemma, concentrated in bristle-coated invaginations, and within large coated vesicles. After 20–45 min, it is present in large smooth vesicles, apical multivesicular bodies, and dense bodies. Peroxidase is not seen in small coated vesicles at any interval. Counts of small coated vesicles reveal that during peroxidase absorption they first increase in number in the Golgi region and later, in the apical cytoplasm. In both control and peroxidase-perfused specimens incubated with TPP, reaction product is seen in several Golgi cisternae and in small coated vesicles in the Golgi region. With GP, reaction product is seen in one to two Golgi cisternae, multivesicular bodies, dense bodies, and small coated vesicles present in the Golgi region or near multivesicular bodies. The results demonstrate that (a) this epithelium functions in the absorption of protein from the duct lumen, (b) large coated vesicles serve as heterophagosomes to transport absorbed protein to lysosomes, and (c) some small coated vesicles serve as primary lysosomes to transport hydrolytic enzymes from the Golgi complex to multivesicular bodies.     

多泡体形成过程的细胞化学研究

本实验用酶细胞化学和示踪细胞化学方法观察了睾丸间质细胞中多泡体的形成过程及其与溶酶体的关系。实验结果表明,睾丸间质细胞中多泡体的形成可分三个阶段:首先,一些含内吞物质的泡状结构进入高尔基体区域,与那里的小泡融合,形成内含少量小泡的前多泡体;然后,前多泡体互相融合,形成体积较大、基质电子密度低、内含小泡排列稀疏的低电子密度多泡体;最后,低电子密度多泡体通过表面长出微绒毛样结构并不断断裂的方式去除多余的界膜,形成体积较小、基质电子密度高、内含小泡排列紧密的高电子密度多泡体。因此,多泡体的形成既与内吞活动有关,又与高尔基体区域小泡有关。前多泡体和低电子密度多泡体不含溶酶体酶。在多泡体形成过程中,只有到高电子密度多泡体阶段,才与溶酶体发生关系,从溶酶体中获取溶酶体酶。多泡体形成后,常与自体吞噬泡靠近,可能参与睾丸间质细胞的自体吞噬活动。