Inhibition and Recovery of Cell Wall Formation in Root Hairs of Ceratopteris thalictroides

The processes occurring in root hairs of Ceratopteris thalictroidesduring recovery after 10 min incubation in cellulose crystallizationinhibitors (Congo Red and Calcofluor White) and in cellulosesynthesis inhibitors (Coumarin and Dichlorobenzonitrile) werestudied using light and electron microscopy. All these drugscause reversible cessation of growth of the root hairs. AfterCR and CW treatment the nuclei proceed in their normal directionof movement, whereas after Cm and DCB treatment the directionof movement is reversed immediately. After CR and CW treatment the organization of the cytoplasmin the root hair tip is temporarily altered. Cell wall synthesiscontinues, although the resulting wall has a different appearance.After Cm and DCB treatment, the cytoplasm in the root hair tiphas disintegrated and cell wall synthesis is stopped. Recoveryof cell wall synthesis results in a new cell wall between thedisintegrated cytoplasm and the remaining cytoplasm. The effects of the drugs on cytoplasmic organization are discussedin relation to their specificity in inhibiting cell wall synthesis. Key words: Ceratopteris thalictroides, root hairs, cellulose synthesis, inhibition of cell wall formation     

伸展素在黄瓜胞壁中的转化及其与羟脯氨酸和异联酪氨酸的关系

用~(14)C-Pro和~3H_2-Tyr离体暗培养黄瓜子叶,发现细胞质、SECW和RCW中的Hyp/Pro和Idt/Tyr都随时间呈线性增加。这两种比值后两者高于前者,而两种比值增加速度之比在胞质部分最大、RCW中最小。表明在胞质和胞壁中都有Pro羟化和Tyr异联化过程,但羟化作用主要发生在胞质中,异联化主要在RCW中。 理化处理(高渗溶液和CHM)和放射性示踪证明胞质中存在HRGP库;它被分泌到胞壁后,先以离子键与壁结合,后转变为共价键与壁结合的伸展素。     

Structural aspects of bulge formation during root hair initiation

Using light and electron microscopy, the early stages of root hair initiation were investigated under control conditions and in a situation where F-actin polymerization was effectively inhibited by latrunculin B. Trichoblasts in their early stage of bulge formation possessed large vacuole traversed by cytoplasmic strands and enclosed within a narrow peripheral layer of cytoplasm. The nucleus was settled at the inner periclinal cell wall, typically opposite the site of bulge formation. Within the bulging area, dense cytoplasm and numerous ER elements, and other organelles were accumulated, together with pleiomorphic membrane-bound structures, the identity and nature of which will require further studies. These unusual structures, which were associated with the outer cell wall, contained material similar to that of the cell wall. Similar cell wall-like bodies were observed also in the cytoplasm and sometimes within vacuoles. The possible role of these novel organelles of plant cells in cell wall thinning/degradation or in the turgor pressure maintenance are discussed. Latrunculin B treatment allowed bulge formation but prevented the switch from the slow and diffuse expansion of bulge into the rapid tip-growth characteristic of the emerging root hair. Moreover, the cytoplasm of the bulging domain became extensively vacuolated and lacked abundant ER elements and other organelles including the membrane-bound structures. These results indicate important roles of F-actin in the switch from diffuse to highly polarized tip growth.     

Dermonecrotic activity of a cell wall preparation from Fusobacterium necrophorum

A cell wall preparation of Fusobacterium necrophorum induced haemorrhagic necrosis in the skins of guinea pigs and rabbits. Effects in mice and rats were weak or absent. The toxic activity of the cell wall preparation was not reduced by heat treatment. A dermonecrotic toxin was isolated from the cell wall preparation with sodium dodecylsulphate and concentrated by precipitation with ethanol. A preparation of the bacterial cytoplasm from Fus. necrophorum induced mainly erythema.     

Studies of Ultrastructure and ATPase Localization of Mature Embryo Sac in Vicia faba L.

Studies of ultrastructure and ATPase localization of the mature embryo sac in Vicia faba L. show that the egg cell has no cell wall at thechalazal end, it has a chalazally located nucleus and a large micropylar vacuole. There are many nuclear pores in the nuclear membrane. The cytoplasm is restricted around the nucleus. Dictyosome and mitochondria are few. There are some starch grains and lipid grains in the egg cytoplasm. There are no obvious differences between two synergids. No cell wall is seen at the chalazal end either, but there are some vesicles which project to vacuole of the central cell and fuse with its vacuolar membrane. Plasmodesmata connections occur within the synergid wall where it is adjacent to the central cell. The synergid has a micropylarly located nucleus and a chalazal vacuole, the nucleus is irregularly shaped. The synergid cytoplasm is rich in organelles. The filiform aparatus is of relatively heterogeneous structure. The central cell is occupied by a large vacuole and its cytoplasm is confined to a thin layer along the empryo sac wall, but is rich in various organelles, starch grains and lipid bodies. Nucleolar vacuoles are often present two polar nuclei. The nuclear membranes of two polar nuclei have partly fused. ATPase reactive product was located obviously at the endoplasmic reticulum in cytoplasm of the egg cell and central cell. The embryo sac wall consists of different density of osmiophilic layer. There are some wall ingrowths in chalazal region of the embryo sac. The long-shaped and cuneate cells of chalazal region are peculiar. Special tracks of ATPase reactive products are visible at their intercellular space which may be related to transportation of nutrients.     

The Development and Structure of the Generative Cell Wall in Polygonatum simizui

The developmental structure and components of the generative cell wall in Polygonatum sirnizui Kitag were studied by means of cytochemical and electron microscope observation. The early generative cell wall separating the generative and vegetative cytoplasm contains callose and cellulose. From the time when the generative cell detaches from the intine untill it is freely suspended in the cytoplasm of the vegetative cell, the wall becomes progressively thinner and does not show the specific fluorescence when stained with aniline blue and cai- cofluor white although it remains PAS positive. At later developmental stage when the generative cell moves into the pollen tube but before its initiation of mitosis, an envelope with weak PAS positive reaction appears on the surface of the cell. Its morphological nature is similar to that of the sperm cell discribed as the "periplasm”. This study proves that a cell wall is present in the generative cell of Polygonatum simizui throughout the developmental process, althrough changes in structure and components of the wall may occur. The properties of the generative cell wall at different stages, its significance in differentiation between generative and vegetative cytoplasm and translocation of nutrient materials, and the possible mechanism of the detachment of the generative cell from the intine are the subjects to discussion.     

大葱卵器及受精后助细胞的超微结构

章丘大葱（Ａllium fistulosum L.cv.Zhangqiu)的卵器由１个卵细胞及２个助细胞组成,观察到不少卵器没有卵细胞,只有２个助细胞。卵细胞的核及大部分细胞质位于细胞的合点端,１个大液泡占据了细胞其他部位。卵细胞含有很多的核糖体及多聚核糖体、嵴明显的线粒体、粗面内质网、高尔基体具小泡,卵细胞似是一个活跃的细胞。细胞外被细胞壁,其合点端及侧方与助细胞共同壁不连续,助细胞有一较大的核,位于细胞膨大的部位,众多的小液泡遍布细胞中。核糖体及聚合核糖体、线粒体,粗面内质网及风心圆环状粗面内质丰富,高尔基体及小泡常见,反映了其活跃的代谢作用。助细胞合点端及侧方与卵细胞、中央细胞的共同壁不连续,与卵细胞共同壁含胞间连丝,壁不连续处,有不状多层膜结构伸入卵细胞质,显示助细胞可能对卵细胞提供营养,伟粉后,一个助细胞退化,宿存助细胞至随胚胚期尚存在,它经历了一个缓慢的退化过程,出现质壁分离,细胞质变稀,液泡扩大,细胞器逐渐减少,在椭形胚期,宿存助细胞核内的染色质及核仁消失,有细胞质侵入核内,因宿存助细胞壁变厚,细胞质出现现脂滴,宿存助细胞可能仍有合成功能,宿存助细胞壁出现若干无壁部位,细胞内的营养物质可能通过无壁部位向胚乳转运,供游离核胚乳及胚乳细胞化初期的发育。     

THE HOST-PARASITE INTERFACE OF ALBUGO CANDIDA ON RAPHANUS SATIVUS

The fine structure of the intercellular hyphae of the obligate parasite Albugo candida infecting radish does not differ markedly from that described previously for cells of Peronospora manshurica. The stalked, capitate haustoria do not contain nuclei and are packed with mitochondria and lomasomes. The fungal plasma membrane and cell wall are continuous from the intercellular hypha throughout the haustorium except that there is no evidence of fungal cell wall around a portion of the haustorial stalk proximal to the haustorial head. Within the vacuolate host mesophyll cell, the haustorium is always surrounded by host plasma membrane and with at least a thin layer of host cytoplasm. The host cell wall invaginates at the point of haustorial penetration to form a short sheath around the region of penetration, but normally there is no host cell wall around the balance of the haustorium. About 1% of the haustoria observed were necrotic, and these were invariably walled-off completely from host cytoplasm by host cell wall. An amorphous, moderately electron-dense encapsulation lies between the haustorium proper and the host plasma membrane and extends into the penetration region between the sheath and the fungal cell wall. Invaded host cells contain more ribosomal-rich ground cytoplasm than uninfected cells. Glandular-like systems of tubules and connecting vesicles are often numerous in host cytoplasm in the vicinity of haustorial heads. These tubules open into the encapsulation, their limiting unit membranes being continuous with the host plasma membrane. We suggest that these represent a secretory mechanism of the host specifically induced by the parasite.     

Ultrastructure of Male Gametophyte in wheat I. The Formation of Generative and Vegetative Cells

The present study of the formation of the generative and vegetative cells in wheat has demonstrated some cytological details at the ultrastructural level. The phragmoplast formed in telophase of the first microsporic mitosis extended centrifugally until it connected with the intine of the pollen grain. A new cell wall was then formed to separate the generative and the vegetative cells. By unequal cytokinesis the former is small and the latter large. In early developmental stage of male gametophyte, the organelles in the cytoplasm of the generaVive cell and the vegetative cells are similar, including mitochondria, dictyosomes, rough endoplasmic retieulum, free and clustered ribosomes and plastids, but microtubules were observed only in the early cytokinesis stage. In the further developmental stage of the male gemetophyte, the generative cell gradually detached from the intine of pollen grain and grew inward to the cytoplasm of the vegetation cell. When the generative cell became round and free in the cytoplasm of the vegetative cell, the wall materials between plasma membranes of the cytoplasm of the generative and the vegetative cells disappeared completely, so that it was a naked cell with a double-layer membrane at this time. The heterogeneity between both cells was then very conspiceous. The organelles in the cytoplasm of the generative cell have hardly any changed besides the degeneration of plastids, but in vegetative cytoplasm the mitochondria and plastids increased dramatically both in number and size. The rapid deposition of starch in the plastids of the cytoplasm of the vegetative cell made the most conspicuous feature of the vegetative cell in mature pollen grain. The significance of the presence of a temporary cell wall in generative cell and heterogeneity between generative and vegetative cells are discussed.     

Wall-associated kinases are expressed throughout plant development and are required for cell expansion

The mechanism by which events in the angiosperm cell wall are communicated to the cytoplasm is not well characterized. A family of five Arabidopsis wall-associated kinases (WAKs) have the potential to provide a physical and signaling continuum between the cell wall and the cytoplasm. The WAKs have an active cytoplasmic protein kinase domain, span the plasma membrane, and contain an N terminus that binds the cell wall. We show here that WAKs are expressed at organ junctions, in shoot and root apical meristems, in expanding leaves, and in response to wall disturbances. Leaves expressing an antisense WAK gene have reduced WAK protein levels and exhibit a loss of cell expansion. WAKs are covalently bound to pectin in the cell wall, providing evidence that the binding of a structural carbohydrate by a receptor-like kinase may have significance in the control of cell expansion.     

The ultrastructure of the salt gland of Spartina foliosa

Summary The salt gland in Spartina foliosa is composed of two cells, a large basal cell and a smaller, dome-shaped cap cell which is located on a neck-like protrusion of the basal cell. There is no cuticular layer separating the salt gland from the mesophyll tissue. The basal cell has dense cytoplasm which contains numerous mitochondria, rod-like wall protuberances, and infoldings of the plasmalemma which extend into the basal cell and partition the basal cell cytoplasm. The protuberances originate on the wall between the basal and the cap cells and are isolated from the basal-cell cytoplasm by the infoldings of the plasmalemma. While the cap cell has no partitioning membrane system or wall protuberances, it resembles the basal cell by having dense cytoplasm and numerous mitochondria.The basal cell seems to be designed for efficient movement of ions toward the cap cell. The long, dead-end extracellular channels in the basal cell of Spartina appear comparable to surface specializations seen in the secreting epithelium of animal cells which carry out solute-linked water transport. The number of mitochondria and their close association with the plasmalemma extensions suggest that they have an important role in the transfer of ions through the basal cell.The accumulated ions would move into the extracellular spaces along an osomotic gradient where the accompanying passive flow of water would move the ions into the cap-cell wall and from there the solution would pass out through the pores in the cuticle.     

扁豆成熟胚囊的超微结构

本文对扁豆（Dolichos lablab)成熟胚囊的超微结构进行了研究,在成熟胚囊中,卵细胞和助细胞仅在珠孔端1／3有细胞壁,靠近合点端,卵细胞一助细胞,卵细胞－中央细胞,助细胞－中央细胞之间没有细胞壁存在,相邻细胞的质膜靠在一起,在卵细胞和中央细胞的质膜间,有些地方存在中等电子密度的物质,卵细胞的细胞质中含有很多的线粒体和质体,内质网和高尔基体较少,助细胞的珠孔端有一复杂的丝状器,靠近珠孔端的细胞质中有很多管状的内质网,表明助细胞可能具有分泌功能,在助细胞的合点端,含有丰富的粗糙内质网,助细胞和卵细胞的质膜之间有很多囊泡状的结构,中央细胞内含有丰富的线粒体,高尔基体和内质网,中央细胞的壁向内形成突起,在周缘细胞质中含有丰富的脂滴。     

植物叶片原生质体分离的可能机制

分析了植物叶片在分离液环境中形成原生质体的过程,文中提出,分离液配方中的酸性物质使植物叶片处于酸性环境中并导致植物正常细胞首先发生细胞壁酸性降解,随后出现原生质体脱离细胞壁进入分离液,继而又进一步发生质膜的酸性降解,使细胞核和细胞器进入分离液中,最终分离液中的细胞器以细胞核为中心进行细胞器重组,最后产生外貌形态一致的新的原生质体。植物细胞壁和质膜是植物细胞的包被系统。植物细胞包被系统的酸性降解使植物细胞器重组并产生新的原生质体成为可能。     

Effect of penicillin on the morphology of a Neisseria meningitidis strain liberating free endotoxin. An electron microscope study

An endotoxin-liberating strain of Neisseria meningitidis plasmolysed extensively after 2 h of exposure to 100 times MIC values of benzypenicillin. The peptidoglycan layer could be demonstrated after 2 h of treatment in places where the cytoplasm still was close to the cell wall. After 20 h, however, this layer was complete undetectable. In untreated cells the peptidoglycan layer could more easily be found in older cultures than in very young cultures. An increased adhesiveness and aggregation to other bacterial cells and to cell wall material could be observed after 2 h of penicillin treatment, and more pronouncedly after 20 h. A high yield of free cell wall material could be demonstrated after 2 h of penicillin treatment. This corresponded well to an increased content of free endotoxin in the filtrates from the cell cultures treated with penicillin, compared to untreated controls. After 20 h of treatment, free cell wall material had formed large aggregates or was adherent to the cell walls of ghost cells. The corresponding endotoxin analysis showed a reduced content of filtrable endotoxin. Possible implications of the structural changes in relation to penicillin treatment of patients are discussed.     

High resolution shadowing of Mycobacterium leprae.

Metal shadow casting techniques for transmission electron microscopic examination was used to determine the morphological characteristics of Mycobacterium leprae in untreated and treated patients. This technique is used to visualize bacterial surface structures by thermal evaporation of platinum alloys under moderate vacuum. This method gives a high contrast image at relatively low resolution and is useful for correlating micro-morphology quantitatively to early therapeutic effects of anti-leprosy drugs. Using these techniques in untreated cases, the surface structures of M. leprae were uniformly filled with relatively homogenous protoplasm surrounded by a cell wall. Most of the bacilli had thick cell walls with prominent banded and fibrous structures on the surface of the cell body. The cell wall was not detached in any of the solid bacilli in untreated cases. The bacilli varied in size and some of them were swollen in their mid-portion. Some bacilli were very short and completely filled with cytoplasm; therefore, these short bacilli were counted as solid bacilli in electron microscopic morphological index (EM-MI) determination. During treatment, mainly the cytoplasms of the bacilli were affected, and degeneration was observed. Ultrastructurally, the cytoplasm was shrunken and detached from the cell wall indicating mild degeneration. After moderate degeneration, the cytoplasm appeared fragmented. In advanced degeneration, all structures except the cell walls collapsed completely and no fibrous or band structures were visible on the surfaces of the cell walls. Therefore, these bacilli were counted as non-solid bacilli for EM-MI determination. This study shows that transmission electron shadowing gives more accurate counts than standard light microscopy of intact M. leprae bacilli in patient specimens.     

High resolution shadowing of Mycobacterium leprae

Metal shadow casting techniques for transmission electron microscopic examination was used to determine the morphological characteristics of Mycobacterium leprae in untreated and treated patients. This technique is used to visualize bacterial surface structures by thermal evaporation of platinum alloys under moderate vacuum. This method gives a high contrast image at relatively low resolution and is useful for correlating micro-morphology quantitatively to early therapeutic effects of anti-leprosy drugs. Using these techniques in untreated cases, the surface structures of M. leprae were uniformly filled with relatively homogenous protoplasm surrounded by a cell wall. Most of the bacilli had thick cell walls with prominent banded and fibrous structures on the surface of the cell body. The cell wall was not detached in any of the solid bacilli in untreated cases. The bacilli varied in size and some of them were swollen in their mid-portion. Some bacilli were very short and completely filled with cytoplasm; therefore, these short bacilli were counted as solid bacilli in electron microscopic morphological index (EM-MI) determination. During treatment, mainly the cytoplasms of the bacilli were affected, and degeneration was observed. Ultrastructurally, the cytoplasm was shrunken and detached from the cell wall indicating mild degeneration. After moderate degeneration, the cytoplasm appeared fragmented. In advanced degeneration, all structures except the cell walls collapsed completely and no fibrous or band structures were visible on the surfaces of the cell walls. Therefore, these bacilli were counted as non-solid bacilli for EM-MI determination. This study shows that transmission electron shadowing gives more accurate counts than standard light microscopy of intact M. leprae bacilli in patient specimens.     

Effect of hexachlorobenzene and acetone on algal growth: physiology and ultrastructure

The effect of hexachlorobenzene (HCB) and acetone on growth and ultrastructure of the freshwater alga Chlorella pyrenoidosa was studied. The algal cells were grown for 76 h under continuous light in 10 ppm HCB with 0.33% acetone or in 0.33% acetone alone; the control cells were grown in nutrient solution only. As was deduced from determinations of dry matter, carbohydrates, chlorophyll content and total nitrogen, 0.33% acetone in nutrient solution slightly decreased the growth of the cells without having any influence on their ultrastructure while 3.33% acetone affected the ultrastructure of the cells severely. An incubation of Chlorella with 10 ppm HCB in nutrient sultion containing 0.33% acetone led to a drastic decrease of all growth parameters studied, total nitrogen and chlorophyll content being affected most strongly. These latter observations were in accord with the changes in ultrastructure showing damage to the cell membranes, disintegrated cytoplasm and sometimes even break down of cell organells leaving only starch grains, the pyrenoid and some endomembranes. In addition to these cells with severe lesions, quite normal cells were found.     

Encystment ofBodo caudatus

Summary Before formation of the cyst wall, the food vacuoles are lost, the cell rounds up and the flagella lie close against the body in a flagellar groove. At this early stage, the contractile vacuole is very active, the Golgi apparatus is prominent and the basophilic cytoplasm is composed of closely packed ribosomes. As the cyst wall is secreted, layer by layer, the large Golgi apparatus is replaced by several smaller membrane stacks and mitochondrial changes occur involving local loss and modification of the cristae. Some parts of the mitochondrion undergo degenerative changes and may become surrounded by bacilliform bodies. These same bodies are also associated with small particles of sequestered cytoplasm which are present throughout the encystment process and are believed to be autophagic vacuoles. As the cyst wall thickens, cell shrinkage is manifest as a number of membrane invaginations. The final cyst wall is of uneven thickness and possesses a single operculum which is visible only by electron microscopy. Probable cyst wall precursor is found in small vesicles scattered throughout the cytoplasm.     

薏苡胚乳细胞化的超微结构观察

采用透射电镜对薏苡早期的胚乳细胞化进行了研究,在胚乳游离核时期,胚乳游离核及细胞质绕中央细胞分布,游离核间没有发现胚囊壁内突、成膜体等结构。胚乳细胞化过程中初始垂周壁形成过程如下：（１）胚乳细胞质中出现液泡,使细胞质和核向中央液泡推进：（２）一对相邻细胞核间液泡成对存在,且呈垂周分布,而且两液泡间的细胞质很狭窄;（３）在这狭窄的细胞质中出现成行排列的小泡;（４）小泡融合形成细胞板,细胞板悬于两液泡