Fibrous and tubular inclusions in four xylariaceous fungi

Summary Intranuclear and cytoplasmic fibrous inclusions and cytoplasmic tubular inclusions have been studied using electron microscope techniques. The fibrous inclusions are composed of closely packed, rod-like structures. Each rod has an outside diameter of ± 24 nm, a hollow centre and lateral projections along its length. The tubular inclusions occur as densely packed bundles or loose arrays of 10–13 nm diameter tubules which are composed of subunits arranged in a double helical structure. The distribution, origin and possible function of these and similar inclusions is discussed.     

Syncytial-type cell plates: a novel kind of cell plate involved in endosperm cellularization of Arabidopsis

Cell wall formation in the syncytial endosperm of Arabidopsis was studied by using high-pressure-frozen/freeze-substituted developing seeds and immunocytochemical techniques. The endosperm cellularization process begins at the late globular embryo stage with the synchronous organization of small clusters of oppositely oriented microtubules ( approximately 10 microtubules in each set) into phragmoplast-like structures termed mini-phragmoplasts between both sister and nonsister nuclei. These mini-phragmoplasts produce a novel kind of cell plate, the syncytial-type cell plate, from Golgi-derived vesicles approximately 63 nm in diameter, which fuse by way of hourglass-shaped intermediates into wide ( approximately 45 nm in diameter) tubules. These wide tubules quickly become coated and surrounded by a ribosome-excluding matrix; as they grow, they branch and fuse with each other to form wide tubular networks. The mini-phragmoplasts formed between a given pair of nuclei produce aligned tubular networks that grow centrifugally until they merge into a coherent wide tubular network with the mini-phragmoplasts positioned along the network margins. The individual wide tubular networks expand laterally until they meet and eventually fuse with each other at the sites of the future cell corners. Transformation of the wide tubular networks into noncoated, thin ( approximately 27 nm in diameter) tubular networks begins at multiple sites and coincides with the appearance of clathrin-coated budding structures. After fusion with the syncytial cell wall, the thin tubular networks are converted into fenestrated sheets and cell walls. Immunolabeling experiments show that the cell plates and cell walls of the endosperm differ from those of the embryo and maternal tissue in two features: their xyloglucans lack terminal fucose residues on the side chain, and callose persists in the cell walls after the cell plates fuse with the parental plasma membrane. The lack of terminal fucose residues on xyloglucans suggests that these cell wall matrix molecules serve both structural and storage functions.     

Chloroplast microtubules in young leaves ofSedum rotundifolium

Well-defined tubular inclusions were detected in mesophyll chloroplasts of youngSedum rotundifolium leaves. The size and distinctly uniform arrangement of tubular inclusions were the most noticeable features. The chloroplast usually contained a large inclusion, sometimes extending almost as long as the chloroplast. Such inclusions were built up from microtubules exhibiting aggregates of either large plate-like or paracrystalline structures depending on the section angles. These inclusions were often quite huge, measuring as much as 7.1 μm in length and 2.6 μm in width. The diameter of the microtubule was approximately 25 nm. The microtubule aggregates were non-membrane bounded structures enclosed partly by the thylakoids. The microtubules in the aggregate were all displaying the same definite orientation. Cross-sectional views clearly demonstrated the characteristic hexagonal arrangement within paracrystalline structures. Longitudinal sections of the chloroplast microtubules showed that they were in perpendicular orientation to the chloroplast envelope. They were not connected to these membranes in any case. In general, one microtubular aggregate was seen in each chloroplast section. The role of tubular inclusions in the chloroplasts related to the photosynthetic mode was discussed.     

Tubular inclusions within polyploid nuclei ofGerris najas do not react with a monoclonal anti-β-tubulin antibody

Summary Variable aggregates, composed of tubules with a mean diameter of 19 nm, were found exclusively within polyploid nuclei of the midgut, Malpighian tubes, cyst cells, testis epithelium, and trophocytes ofGerris najas. The nuclear inclusions are always in direct contact with the nucleoplasm, and no other structures are associated with them. They appear most abundant within degenerating nuclei of the midgut surface epithelium, where they form paracrystalline bodies or spindle-shaped inclusions with tapered ends. Smaller fusiform inclusions occur in younger epithelial nuclei but not in the diploid nuclei of regenerative cells. In other tissues, mainly spindle-shaped inclusions can be observed, the longest (4.5 m) in cyst cell nuclei. The mean diameter of the tubules determined from transverse sections, resembles that of cytoplasmic microtubules and was verified statistically. The inclusions within trophocyte nuclei failed to react with monoclonal anti--tubulin antibody, although the antibodies could penetrate the nuclei after extensive lysis of the cells.     

A Comparative electron microscopic study of cytoplasmic microtubules and axial unit tubules in a spermatozoon and a protozoan

Cytoplasmic microtubules and axial unit tubules were studied in both sectioned and negatively-stained material. Walls of microtubules of frog lung-fluke (Haematoloechus medioplexus) spermatozoa have a helical substructure, while those of the flagellate, Trypanosoma lewisi, are composed of ten longitudinally-oriented filaments. Cross-bridges occur between some filaments of trypanosome microtubules. Doublet tubules of axial units in both cell types are structurally similar to the trypanosome microtubules, which may indicate similarity of function. Microtubules of fluke spermatozoa appear to be somewhat rigid, are resistant to sonication, and are considered to be mainly supportive. Circular profiles of wall subunits are seen in transverse sections of microtubules of both cell types and in doublet tubules of the trypanosome. Comparisons are made between sectioned and negatively-stained material; while negative-staining better reveals the fundamental substructure of microtubular elements, some distortion appears to occur. In connection with this research, a brief preliminary article demonstrated the presence of subunits in the walls of cytoplasmic microtubules of fluke spermatozoa (Burton, '66). Also, it was shown that the wall of these tubular elements possesses a helical structure, and a diagrammatic representation of the wall structure was set forth.     

The intranuclear mitosis of the ciliates Paracineta limbata and Ichthyophtirius multifiliis

Electron microscope studies on the premetaphase stages of micronuclear divisions of Paracineta limbata and Ichthyophtirius multifiliis showed that spindle material also exists during interphase. In the case of I. multifiliis scattered microtubule fragments persist in the nuclear space; in P. limbata the micronuclei contain a small paracrystalloid which is suggested to be microtubular protein. Wide microtubules, varying in diameter from 300 to 400 Å develop during intranuclear prophase near the nuclear envelope in both cases. There are good reasons to assume that they function as a kind of stem body during the enlargement of the surface area of the nuclear envelope. Later micronuclear prophase stages of both species show a some-what different development. In I. multifiliis, there are scattered groups of short microtubular segments, partly in parallel array, whereas in P. limbata the wide tubules are transformed into normal microtubules of 180–200 Å diameter. The nuclei of both species are similar at late prophase and prometaphase stages. Bundles of interpolar microtubules run between the chromosomes, and single microtubules, presumably induced by the chromosomes, cross them at different angles. The chromosome-induced microtubules appear a short time after the interpolars. At prometaphase stage all microtubules show a highly parallel arrangement and therefore it is suggested that chromosomal tubules reach their final polar orientation by the action of cross-bridges.     

Spermiogenesis and sperm structure in the crabUca tangeri (Crustacea,Brachyura), with special reference to the acrosome differentiation

Summary Early spermatids of the crabUca tangeri consists of the nucleus of granular chromatin and the cytoplasm, which contains a proacrosomal vesicle in close association with membrane lamellae. In the mid spermatids an invagination of the acrosomal vesicle membrane gives rise to the formation of the perforatorium, a spindle-shaped tubule which encloses tubular membranous structures. The pair of centrioles located at the base of the acrosome is not directly involved in perforatorial differentiation. The acrosomal vesicle shows a heterogeneous content composed of the operculum, the thickened ring, and three layers of different materials concentrically arranged around the perforatorium. During the late spermatid stage the nuclear profile differentiates numerous slender arms and the chromatin arranges into fibers. Membranous tubules from the cytoplasm become incorporated into the tubular structures of the perforatorium. The mature spermatozoon has the typical structure of the branchyuran sperm, with a complex acrosome, cupped by the nucleus, and a thin cytoplasmic band intervening between the former main elements. The centrioles are degenerate. The nuclear arms are unusually numerous (more than 20) and lack microtubules or microtubular derivatives.     

ULTRASTRUCTURE OF THE GREEN ALGA DICHOTOMOSIPHON TUBEROSUS WITH SPECIAL REFERENCE TO THE OCCURRENCE OF STRIATED TUBULES IN THE CHLOROPLAST1

The ultrastructure of the siphonous green alga Dichotomosiphon tuberosus (A. Br.) Ernst is compared with that of other siphonous plants. There is a characteristic association between the Golgi bodies and endoplasmic reticulum, but. the mitochondria are not involved in the association as they are in Vaucheria and the phycomycete Saprolegnia. An unusual structure and arrangement of the chloroplasts is described as well as a previously unreported type of “striated tubule” which occurs in most if not all chloroplasts, and amyloplasts. The structure of these tubules is compared with that of other tubules recently found in green algae and higher plants. In addition, cytoplasmic microtubules arranged in the longitudinal direction of the siphon suggest a function in cytoplasmic streaming.     

Configurational changes in helical microtubule frameworks in feeding tentacles of the suctorian ciliateTokophyra

Microtubules at the tip of a resting (non-feeding) tentacle are arranged helically in two concentric tube-shaped arrays. The pitches of the helical paths followed by tubules in the two arrays differ. At the start of feeding these microtubules bend along their longitudinal axes and splay outwards and downwards away from the tentacle tip as it ‘everts’. Tubules in the two arrays slideacross each other as this occurs. Comparison of the fine structure of the tips of feeding and resting tentacles with a dynamic model of the microtubular framework indicates that movement of the tubules is not brought about by active sliding of the tubules against each other or by the action of contractile elements attached along the lengths of tubules. The tips of microtubules forming the inner tube may be pulled downwards by contractile elements in the tentacular pellicle; these tubules apparently push those in. the outer tube to their new positions. The pattern of configurational changes in a tentacle tip at the start of feeding appears to be largely defined by the elastic resistance of the microtubules to bending, and the ways in which tubules are packed and linked together and attached to the pellicle.     

Ultrastructural observation of paracrystalline aggregates of microtubules in anterior pituitary cells of the chinchilla (Chinchilla laniger)

Summary Unusual paracrystalline aggregates of microtubules which have not been described in any other mammalian species were observed in cultured anterior pituitary cells of normal chinchillas as well as in situ in the pituitary glands of these animals. These aggregates appeared as regularly arranged tubular structures in the longitudinal plane, and as a checkerboard pattern of closely and regularly packed microtubules when examined in transverse section. Supplementation with vinblastine, colcemide or colchicine in the culture medium did not change these structures morphologically. Each unit of tubules consisted of an outer wall or parellelogram profile and an inner wall composed of a single hexagonal doublet or in a figure 8 form. The outer wall of the parallelogram was 35×28 nm in length for both sides, while the diagonal of the inner wall was 18×28 nm. These paracrystalline aggregates of microtubules in the chinchilla pituitary cells are morphologically distinct from the paracrystalline assembly of cytoplasmic microtubules induced by vinblastine or other alkaloids.The function and significance of these paracrystalline aggregates in anterior pituitary cells of the chinchilla are uncertain.Supported by USPHS Grant HD 11826     

Mobile and immobile endoplasmic reticulum in onion bulb epidermis cells: short- and long-term observations with a confocal laser scanning microscope

The endoplasmic reticulum (ER) of onion bulb scale epidermis cells consists of long, tubular strands lying deep in the cytoplasm which move quickly and a less mobile peripheral network of tubules and cisternae that change in position, shape and size but that also have immobile, fixed, sites (IFSs). IFSs occur in junctions, at vertexes and at blind endings of tubules as well as at the edges and the surface of cisternae. They are regularly arranged in helicoidal rows and may be knot- or ring-like in structure. They become enlarged by treatment with oryzalin but not with colchicine. They persist for long times (for more than 30 min); together with pulling forces, the surface tension and other factors, they determine the configuration and motion of the peripheral network. New polygons of the network are mainly formed by the development of new tubules that become joined with other parts of the network. Polygons disappear by contraction and fusion of tubules. The inner, rapidly moving ER tubules remain connected with the peripheral network over longer distances by sliding junctions. Cytochalasin D causes an accumulation of the ER into patches, a fusion of tubules into cisternae and changes in shape, which indicate the loss of pulling forces. In contrast to animal cells (but like the movement of the inner tubular strands), the latter is dependent upon the actomyosin system; microtubules are not involved. Despite the differences in the organizing components, the peripheral ER in onion bulb scale epidermis cells and that of the borders of cultured animal cells are similar in morphology and motility.     

Tubule bundle-inclusions in vacuoles of Tamarix aphylla L. mesophyll cells

Summary Vacuoles of differentiating mesophyll cells of Tamarix aphylla contain an amorphous electron-dense material in which stacks of parallel aligned striations are embedded. Cross-sections of the striations disclosed that they represent profiles of longitudinally sectioned bundles of tubules (tubule outer diameter 9.0 nm, tubule wall thickness 1.8 nm). In advanced mesophyll cell development, the amorphous vacuolar material disappears, whereas the bundles of tubules turn into bundles of double helices (double helix diameter 14.5 nm). Cytochemical treatment of mesophyll cells with the enzymes pepsin and trypsin has revealed that both the bundles of tubules/double helices and the embedding material are constituted of protein. The possible functional role of the vacuolar inclusions is discussed.     

Ultrastructure of P-protein inHevea brasiliensis during sieve-tube development and after wounding

Summary P-protein and the changes it undergoes after wounding of sieve tubes of secondary phloem in one- to two-year old shoots ofHevea brasiliensis has been studied using electron microscopy. The P-protein in the form of tubules with a diameter of 8–9 nm and a lumen of 2–2.5 nm occurred in differentiating sieve elements and appeared as compact bodies which consisted of small aggregates of the tubules. As the sieve elements matured, these P-protein bodies dispersed with a disaggregation of the tubules before they turned into striated fibrils, 10–11 nm in diameter. In wounding experiments, as the mature sieve elements collapsed after cutting, their striated P-protein converted into tubules. These tubules were the same in ultrastructure as the tubules in differentiating sieve elements and they often were arranged in crystalline aggregates.     

与蚕豆萎蔫病毒2号胞间运动和长距离转运相关的寄主细胞超微结构研究

电镜超微结构观察和免疫金标记显示:受蚕豆萎蔫病毒2号(Broad bean wilt virus 2,BBWV 2)中国分离物B935侵染的豌豆(Pisum sativum)和蚕豆(Vicia faba)叶细胞中膜结构增生,形成膜结构增生区,病毒以结晶体和管状体形式存在于细胞质中。在病变早期,叶肉细胞的胞间连丝处连接有小管结构,病毒样颗粒呈纵列排在小管中,穿越胞间连丝的小管能被BBWV 2的金标记抗体特异性标记。维管束组织的薄壁细胞、伴胞及转移细胞内存在膜增生区及病毒管状体,在筛管壁附近存在的病毒样颗粒能被BBWV 2金标记抗体特异性标记。实验结果表明BBWV 2胞间运动形式与豇豆花叶病毒(CPMV)相似,以完整粒子通过在胞间连丝处形成的小管结构穿越胞间连丝;细胞质中存在的直径160nm管状体只是一种病毒聚集体,与胞间运动无直接关系;该病毒在筛管中可能也是以完整粒子形式进行长距离转运的。     

Electron Microscopic Study of Trypanosoma cruzi Periplast in Tissue Cultures. I. Number and Arrangement of the Peripheral Microtubules in the Various Forms of the Parasite's Life Cycle*

SYNOPSIS. In an electron microscopic study, counts of peripheral microtubules were made in spheromastigotes and trypomastigotes in tissue cultures of embryonic heart muscle cells. In interphase spheromastigotes there were, at the level of the nucleus, ～ 114 microtubules; in dividing forms, there were ～ 222. In trypomastigotes, the number of microtubules varied according to the level of the section—there were fewer than 40 tubules in the pointed ends of an organism, while in the central segment the number of these elements ranged from 60 to 115. The highest number of microtubules was found in the region containing the Golgi complex. The distance between the microtubules was constant, equalling 44 nm, even at the pointed ends of a trypanosome. This suggests that the microtubules course parallel to one another. Cross sections and randomly arranged, variable length, longitudinal sections of the tubules were noted around the kinetosomes in dividing organisms.     

Further studies on ultrastructure of plants infected withPetunia ringspot virus

Summary New methods of fixation and embedding have revealed in plants infected withPetunia ringspot some structural features not described before. These include the X-bodies, 80 per cent of which are formed by tubular elements which are responsible for the positive staining specific for Golgi apparatus. The tubular elements are morphologically similar to agranular endoplasmic reticulum as described in some animal cells. The rest of the inclusion is formed by normal cytoplasmic elements in which are embedded long rod-shaped tubules 600 Å wide and more than 7,000 Å long, cross sections of which are formed by 10 subunits. These subunits are arranged in a helical way to form the large tubules. These subunits are probably the actual virus particles, which would be icosahedral and would form tubular crystals. Icosahedral virus particles would also form true crystalline inclusions.It is not known what the role of the agranular endoplasmic reticulum and of some dense osmiophilic bodies found in it may be in the multiplication of the virus. However, these components induced by the virus infection probably result in the manufacture of some proteins or other substances necessary for virus multiplication.     

Mop-top tubules. The ultrastructure of unusual tubular elements associated with two different leaf symptoms of potato mop-top virus infected potato plants

Summary Potato mop-top virus, one of the most commonly occurring viruses in virus tested stocks of seed potatoes in the United Kingdom induces four different haulm symptoms which are both climate and variety dependent. Ultrastructural examination showed that the aucuba symptom recognizable as bright yellow patches on the leaves, and the mop-top symptom characterised by the dwarfed and bunched habit of the plants, both contained in their leaf cells, tufts and clusters of microtubule-like elements, although the other ultrastructural features associated with each symptom were quite different. These mop-top tubules occurred in the cytoplasm, between the cell wall and the plasma membrane, and in the vacuole, and have been demonstrated in every cell type.The mop-top tubules were 18–22 nm wide with a 2.5–3.0 nm thick wall and often branched. No regular substructure could be discerned. Complete virus particles were rarely seen. These mop-top tubules are compared with plant microtubules and P-protein tubules, and the topic of viral inclusions and their relevance to virus classification is discussed.     

The structure and mitotic behaviour of a nuclear inclusion in a fern

Summary A study has been made of the structure and behaviour during mitosis of a crystalline inclusion within cell nuclei of roots of Dryopteris filix-max. The inclusion within the interphase nucleus is an aggregate of randomly oriented crystals. All the crystals are similar, and consist of a cubic array of particles of unit spacing approximately 100 Å. During mitosis, the inclusions are eliminated from the nucleoplasm at prometaphase. The crystals reappear within the nucleus at early interphase by a process of random crystallisation from a preformed mass of amorphous material. The results are discussed in the light of previous work on nuclear inclusions in plants and of current theories of the mode of action of microtubules.     

Kristalloide Einschlüsse im Zytoplasma pflanzlicher Zellen

Summary In healthy as well as dahlia mosaic sick plants ofVerbesina encelioides, Sanvitalia procumbens, Zinnia elegans, Calendula spec. andDahlia hybrids, leaf cell vacuoles are found in the marginal cytoplasm which contain protein crystals. They are single membrane-limited products of the endoplasmatic reticulum. They may be found mainly in the older leaves and especially in those of virus infected plants. The crystalline structures consist of tetragonally arranged tubules of 105 Å in diameter, separated by an interspace about 35 Å wide. There are similar structures in virus infected plants ofFragaria vesca, but not inChenopodium quinoa, where the vacuoles contain no bodies. This cell organelle is compared with crystalline inclusions already described. Its significance and relations to the virus disease are discussed.     

Cytokinesis in microspore mother cells of Impatiens sultani

Summary Cytokinesis in Impatiens sultani microspore mother cells is simultaneous. It starts with the formation of small ingrowths of the surrounding callosic wall. Next, an incomplete cell plate is formed by fusion of small dictyosome vesicles. The cell plate consists of a network of anastomosing tubules and sacs. Aggregates of fusing vesicles are associated with bundles of microtubules, which are oriented perpendicular to the plane of the future cell walls. In the sacculate parts of the cell plate, some callose is deposited, while the associated microtubules disappear. The cell walls ultimately develop by enlargement of the previously formed wall ingrowths, which successively incorporate the elements of the cell plate. The enlargement and thickening of the walls is not accompanied by a further fusion and incorporation of dictyosome vesicles.