烟草花粉母细胞细胞融合过程中细胞骨架的超微结构观察

应用普通电镜和DGD去包埋技术 ,研究了烟草花粉母细胞中的细胞融合现象及细胞融合过程中细胞骨架的变化。观察发现 ,处于凝线期的花粉母细胞 ,其内含物 ,包括细胞器和染色质 ,主要通过胞质通道向相邻细胞发生转移。DGD去包埋观察发现 ,花粉母细胞中核骨架与细胞质内及胞间连丝和胞质通道内胞质骨架连接成一个整体。在整个细胞融合过程中 ,均有核骨架纤维与染色质相连。本文讨论了细胞骨架在细胞融合过程中的作用     

烟草花粉母细胞中果胶酶细胞化学定位及其在次生胞间连丝和胞质通道形成中的作用

对果胶酶在烟草(Nicotiana　tabacum　L.)花粉母细胞减数分裂前期Ⅰ的活性进行了电镜细胞化学定位,以研究其在次生胞间连丝和胞质通道形成中的作用。结果表明:在细线期,酶反应产物主要存在于光面内质网及其衍生小泡内;偶线期,当次生胞间连丝和胞质通道开始形成时,反应产物明显增强。同时果胶酶活性也出现在细胞壁上,尤其是出现在简单或者复杂分枝状的胞间连丝和胞质通道内部或沿着它们附近细胞壁的中胶层分布。胞吐小泡中的酶活性表明果胶酶和纤维素酶一样,也是通过内质网及其衍生小泡由胞吐作用分泌到细胞壁的,并在那里通过特异性降解果胶质而和纤维素酶协同作用导致次生胞间连丝和胞质通道的形成。     

烟草花粉母细胞中果胶酶细胞化学定位及其在次生胞间连丝和胞质通道形成中的作用

对果胶酶在烟草(Nicotiana tabacum L.)花粉母细胞减数分裂前期Ⅰ的活性进行了电镜细胞化学定位,以研究其在次生胞间连丝和胞质通道形成中的作用.结果表明:在细线期,酶反应产物主要存在于光面内质网及其衍生小泡内;偶线期,当次生胞间连丝和胞质通道开始形成时,反应产物明显增强.同时果胶酶活性也出现在细胞壁上,尤其是出现在简单或者复杂分枝状的胞间连丝和胞质通道内部或沿着它们附近细胞壁的中胶层分布.胞吐小泡中的酶活性表明果胶酶和纤维素酶一样,也是通过内质网及其衍生小泡由胞吐作用分泌到细胞壁的,并在那里通过特异性降解果胶质而和纤维素酶协同作用导致次生胞间连丝和胞质通道的形成.     

洋葱花粉母细胞细胞内、细胞间微梁骨架的超微结构观察(英文)

以洋葱(AlliumcepaL.)花粉母细胞为材料,采用DGD包埋去包埋原位技术,对花粉母细胞不同发育时期的细胞内、细胞间微梁骨架的超微结构进行了电镜观察。结果发现,花粉母细胞核内存在粗细不等的微梁骨架,与核仁和染色体紧密相连,随着发育的推移,其均一性发生改变。在核周有核纤层样的结构存在,与细胞核和胞质中的微梁骨架紧密相连,到前期结束时解体。洋葱花粉母细胞内具有发达的胞质微梁骨架,这种结构在减数分裂前期Ⅰ变化不明显。在胞间连接(胞间连丝和胞质通道)内,也有精细的微梁骨架分布,并且与两端细胞中的骨架相连。在凝线期的花粉母细胞中观察到细胞融合现象,有胞质或核内微梁骨架与穿壁转移的胞质小球和核小球内骨架相连。此时细胞核偏向一边,但细胞的其余部位仍充满了胞质微梁骨架。初步探讨了核微梁骨架与核仁和染色体之间的关系,核纤层与细胞核之间的关系,以及细胞内、细胞间微梁骨架与细胞融合之间的关系     

小麦胚乳细胞内,细胞间胞质骨架分布格局的超微结构观察

以小麦（ＴｒｉｔｉｃｕｍａｅｓｔｉｖｕｍＬ．）幼嫩胚乳为材料,经ＴｒｉｔｏｎＸ１００抽提、ＤＧＤ（ｄｉｅｔｈｙｌｅｎｅｇｌｙｃｏｌｄｉｓｔｅａｒａｔｅ）渗透、包埋,制备去包埋剂超薄切片,对细胞内、细胞间胞质骨架的分布格局与特征进行了电镜观察。由所获图像可见,胞质骨架呈主要由微管、微丝组成的三维网络结构;特别值得注意的是,有不少５～７ｎｍ的微丝在多处从网络表层向胞壁界面方向突出,并时而可见其横贯分界壁连接相邻骨架网络而将相邻细胞骨架联成一体。胚乳组织中微丝的跨胞分布以两种形式存在,直径达１００～２００ｎｍ微丝束的跨越和单个微丝的分散贯穿,看来这与该组织中开放态胞间通道与正常胞间连丝同时并存相吻合。初步讨论了微丝参与正常胞间连丝结构的可能性。     

洋葱花粉母细胞细胞内、细胞间微染骨架的超微结构观察

以洋葱（Allium cepa L.）花粉母细胞为材料,采用DGD包埋去包埋原位技术,对花粉母细胞不同发育时期的细胞内、细胞间微染骨架的超微结构进行了电镜观察。结果发现,花粉母细胞核内存在的粗细不等的微染骨架,与核仁和染色体紧密相连,随着发育的推移,其均一性发生改变。在核周有核纤层样的结构存在,与细胞核和胞质中的微染骨架紧密相连,到前期结束时解体。洋葱花粉母细胞内具有发达的胞质微染骨架,这种结构在减数分裂前期Ⅰ变化不明显。在胞间连接（胞间连丝和胞质通道）内,也有精细的微染骨架分布,并且与两端细胞中的骨架相连。在凝线期的花粉母细胞中观察到细胞融合现象,有胞质或核内微梁骨架与穿壁转移的胞质小球和核小球内骨架相连。此时细胞核偏向一边,但细胞的基余部位仍充满了胞质微染骨架,初步探讨了核微染骨架与核仁和染色体之间的关系,核纤层与细胞核之间的关系。以及细胞内、细胞间微染骨架与细胞融合之间的关系。     

陆地棉（Gossypium hirsutum）绒毡层细胞间的胞质通道

陆地棉（Ｇｏｓｓｙｐｉｕｍｈｉｒｓｕｔｕｍ）绒毡层细胞间的胞质通道王毅,娄成后,杨世杰（北京农业大学农业生物学院．北京１０００９４）用透射电镜的系统观察表明,陆地棉纺毡层细胞分化过程中在细胞壁间有胞质通道的形成。小孢子母细胞减数分裂前,绒毡层细胞间由...     

胞松素B对黑麦花粉母细胞发育的影响

采用花药离体培养研究了胞松素B对黑麦花粉母细胞的发育及显微和亚显微结构的影响。细线期和偶线期接种的花药在含有5—40μmol/L胞松素B的White培养基上培养24小时以后,部分花药不能进一步发育,被抑制在偶线期阶段,并且偶线期花药的百分率随胞松素B浓度的增加而增加。在被抑制的偶线期的花药中,多数花药的花粉母细胞被抑制在细胞融合期或细胞融合前时期。电子显微镜观察发现,相邻花粉母细胞间的胞间连丝通道仍然处于开放状态,内部充满细胞质基质和一些细胞器,细胞核停留在胞间连丝通道口附近,通道内部及附近细胞质内的微管系统依然存在。文章讨论了胞松素B对细胞融合的作用。     

植物胞质分裂发生机制

胞质分裂(cytohnesis)是指在同一细胞中在新形成的两个子核之间形成新的间隔,将母细胞一分为二的过程。胞质分裂存在于任何一种生命形式中,从单细胞的细菌到多细胞的真核生物都能进行胞质分裂。近些年由于细胞学方法的改进和研究材料增多等因素,使得对植物胞质分裂发生机制的研究取得了很大的进展。现对植物中不同类型的胞质分裂在细胞学、分子生物学方面的研究进展作一综述。     

光周期对光敏胞质不育小麦花药发育过程中Ca^2＋分布的影响

运用焦锑酸钾沉淀法,研究了不同光照条件下光敏胞质不育小麦（Ｔｒｉｔｉｃｕｍ ａｅｓｔｉｖｕｍ Ｌ．）花药发育过程中Ｃａ＾２＋的分布。短日照条件下,小孢子形成和花粉发育过程中胞内Ｃａ＾２＋在数量、分布上有变化;小孢子表面逐渐积累Ｃａ＾２＋,至成熟花粉表面覆盖一层Ｃａ＾２＋,胞质Ｃａ＾２＋较少;药隔和药壁组织通过抽外体或共质体途径运输Ｃａ＾２＋供给花粉的发育;长日照条件下,花粉败育发生在不同时期,早期     

Ultrastructural Observations on the Distribution of the Intra-and Intercellular Cytoskeleton of the Endosperm Cells in Triticum aestivum

By means of Triton X-l00 extraction and DGD (diethylene glycol distearate) embedment-free section method the distribution pattern and characteristics of intra- and intercellular cytoskeleton of endosperm cells of Triticum aestivum L. were studied with electron microscopy. Threedimensional architecture of the cytoskeleton could be recognized as a meshwork mainly composed of microtubules (MT) and microfilaments (MF). Attention was stressed on the interface of the adjoining cytoskeletal frameworks where an attractive phenomenon observed was that the MF extruding from the surface of the cytoskeleton often traversed the whole wall boundary and connected the neighbouring frameworks into an entity. In the endosperm tissue two types of transcellular MF distribution could be distinguished, the MF in bundles traversing the enlarged intercellular channels and the MF individually penetrating the wall boundary; that seemed to coordinate with the co-presence of normal and modified plasmodesmata in the same wall. The above observations demonstrated the intercellular cytoskeletal continuity within the symplast and confirmed that the MF was the main constituent of the traversing cytoplasmic strands, the possibility of MF being organized as a structural element of the normal plasmodesmata was also discussed.     

Actin-regulated water permeability of two transport channels of plasmodesmata in roots of winter wheat cultivars varying in drought resistance

In roots of 5-6-day old seedlings of three cultivars of the winter wheat, varying in drought-resistance: Bezostaya 1 (low resistant), Mironovskaya 808 (resistant), and Albidum 114 (highly resistant) water permeability of two transport channels of plasmodesmata was studied at the action of cytochalasin B, which is known to inhibit polymerization of cytoskeleton actin filaments, by a pulse method of NMR, on the background of increasing water loss in the seedlings. It has been found that the registered coefficients of water self diffusion, two of which (D2 and D3) depend on the water permeability of different transport channels of plasmodesmata, differ in opposite directions. This may suggest that in roots of drought-resistant plants, after a moderate water loss, a diffusive water flow through the cytoplasmic symplast increases (demonstrated by an increase of D2), while that through the vacuolar symplast decreases (seen by an increase of D3). After a high water loss in seedlings, we noticed an even greater increase in water permeability of the cytoplasmic symplast, and a decrease in water permeability of the vacuolar symplast, however, in the roots of low resistant cultivars these changes were poorly expressed, if at all. Under stress-less conditions cytochalasin B would result in an increased water transport through the cytoplasmic channel of plasmodesmata due apparently to a destruction of their actin-myosin sphincters. Both weak and average degrees of water loss would strengthen the cytochalasin B exerted influence on plasmodesmal water conductance, that may testify to a synergetic action of these two factors. After a significant water loss this action was kept only partially, because the inhibitor, on blocking the cytoplasmic channel, did increase at the same time the effect of water stress, limiting water flows through the vacuolar symplast and, simultaneously, raising the water inflow to the apoplast.     

在三维结构上对百合花粉母细胞actin的免疫定位

传统的切片仅仅能够显示样品的平面结构,不能用于细胞中三维网络结构的研究。笔者在DGD(diethylene glycol distearate)包埋去包埋的基础上,结合电镜免疫胶体金技术对大卫百合花粉母细胞胞间及胞内细胞的骨架系统进行了研究,观察到高反差细胞微梁结构的三维网络,actin这一细胞骨架的主要成员被定位在该微梁结构纤维上。三维结构上的研究表明,actin不但是植物细胞核及细胞质骨架的成员,而且也存在于胞间连接结构(胞质桥和胞间连丝)中,推测它可能与细胞融合有关。实验结果同时表明,三维结构免疫胶体金技术对于细胞骨架和核基质的结构蛋白研究是行之有效的。     

Getting connected: actin-based cell-to-cell channels in plants and animals

It has been known for more than one hundred years that plant cells are interconnected by cytoplasmic channels called plasmodesmata. This supracellularity was generally considered to be an exotic feature of walled plants containing immobile cells that are firmly enclosed within robust walls. Unexpectedly, intercellular channels in mobile animal cells have been discovered recently. These are extremely dynamic and sensitive to mechanical stress, which causes their rapid breakage and retraction. Both plasmodesmata and nanotubular cell-to-cell channels are supported by the actin cytoskeleton and exclude microtubules. In this article, we discuss the relevance of cell-to-cell channels not only for intercellular communication but also for the development and morphogenesis of multicellular organisms. We also suggest possible parallels between the cell-to-cell transport of endosomes and intracellular pathogens.     

Structural changes in the vacuole and cytoskeleton are key to development of the two cytoplasmic domains supporting single-cell C<Subscript>4</Subscript> photosynthesis in <Emphasis Type="Italic">Bienertia sinuspersici</Emphasis>

Bienertia sinuspersici Akhani has an unusual mechanism of C₄ photosynthesis which occurs within individual chlorenchyma cells. To perform C₄, the mature cells have two cytoplasmic compartments consisting of a central (CCC) and a peripheral (PCC) domain containing dimorphic chloroplasts which are interconnected by cytoplasmic channels. Based on leaf development studies, young chlorenchyma cells have not developed the two cytoplasmic compartments and dimorphic chloroplasts. Fluorescent dyes which are targeted to membranes or to specific organelles were used to follow changes in cell structure and organelle distribution during formation of C₄-type chlorenchyma. Chlorenchyma cell development was divided into four stages: 1—the nucleus and chloroplasts occupy much of the cytoplasmic space and only small vacuoles are formed; 2—development of larger vacuoles, formation of a pre-CCC with some scattered chloroplasts; 3—the vacuole expands, cells have directional growth; 4—mature stage, cells have become elongated, with a distinctive CCC and PCC joined by interconnecting cytoplasmic channels. By staining vacuoles with a fluorescent dye and constructing 3D images of chloroplasts, and by microinjecting a fluorescence dye into the vacuole of living cells, it was demonstrated that the mature cell has only one vacuole, which is traversed by cytoplasmic channels connecting the CCC with the PCC. Immunofluorescent studies on isolated chlorenchyma cells treated with cytoskeleton disrupting drugs suspended in different levels of osmoticum showed that both microtubules and actin filaments are important in maintaining the cytoplasmic domains. With prolonged exposure of plants to dim light, the cytoskeleton undergoes changes and there is a dramatic shift of the CCC from the center toward the distal end of the cell.     

Effects of myosin ATPase inhibitor 2,3-butanedione 2-monoxime on distributions of myosins, F-actin, microtubules, and cortical endoplasmic reticulum in maize root apices

2,3-Butanedione 2-monoxime (BDM) is a general inhibitor of myosin ATPases of eukaryotic cells, and its effects on animal and yeast cells are well described. Using immunofluorescence and electron microscopy, we have analyzed the impacts of BDM on distributions of plant myosins, actin filaments (AFs), microtubules (MTs), and cortical endoplasmic reticulum (ER) elements in various cell types of maize root apices. Treatment of growing maize roots with BDM altered the typical distribution patterns of unconventional plant myosin VIII and of putative maize homologue(s) of myosin II. This pharmacological agent also induced a broad range of impacts on AFs and on cortical ER elements associated with plasmodesmata and pit fields. BDM-mediated effects on the actomyosin cytoskeleton were especially pronounced in cells of the root transition zone. Additionally, BDM elicited distinct reactions in the MT cytoskeleton; endoplasmic MTs vanished in all cells of the transition zone and cortical MTs assembled in increased amounts preferentially at plasmodesmata and pit-fields. Our data indicate that AFs and MTs interact together via BDM-sensitive plant myosins, which can be considered as putative integrators of the plant cytoskeleton. Morphometric analysis revealed that cell growth was prominently inhibited in the transition zone and the apical part, but not the central part, of the elongation region. Obviously, myosin-based contractility of the actin cytoskeleton is essential for the developmental progression of root cells through the transition zone.     

Inhibitors of myosin,but not actin,alter transport through <Emphasis Type="Italic">Tradescantia</Emphasis> plasmodesmata

Actin and myosin are components of plasmodesmata, the cytoplasmic channels between plant cells, but their role in regulating these channels is unclear. Here, we investigated the role of myosin in regulating plasmodesmata in a well-studied, simple system comprising single filaments of cells which form stamen hairs in Tradescantia virginiana flowers. Effects of myosin inhibitors were assessed by analysing cell-to-cell movement of fluorescent tracers microinjected into treated cells. Incubation in the myosin inhibitor, 2,3-butanedione monoxime (BDM) or injection of anti-myosin antibodies increased cell–cell transport of fluorescent dextrans, while treatment with the myosin inhibitor N-ethylmaleimide (NEM) decreased cell–cell transport. Pretreatment with the callose synthesis inhibitor, deoxy-d-glucose (DDG), enhanced transport induced by BDM treatment or injection of myosin antibodies but did not relieve NEM-induced reduction in transport. In contrast to the myosin inhibitors, cell-to-cell transport was unaffected by treatment with the actin polymerisation inhibitor, latrunculin B, after controlling for callose synthesis with DDG. Transport was increased following azide treatment, and reduced after injection of ATP, as in previous studies. We propose that myosin detachment from actin, induced by BDM, opens T. virginiana plasmodesmata whereas the firm attachment of myosin to actin, promoted by NEM, closes them.     

Cytochemical localization of cellulase activity in pollen mother cells of david lily during meiotic prophase I and its relation to secondary formation of plasmodesmata

Summary Cellulase activity was localized at the ultrastructural level in pollen mother cells (PMCs) of David lily [Lilium davidii var.willmottiae (Wilson) Roffill] at different stages of meiotic prophase I. The enzyme was observed to appear at the early leptotene stage and reached its highest level at the subsequent zygotene stage, and its subcellular distribution revealed by the presence of electron-dense deposits of reaction product was found to be restricted exclusively to the endoplasmic reticulum (ER), the vesicles derived from that, and the cell wall, especially at the sites of secondary plasmodesmata and cytoplasmic channels where the wall was being digested. Other cytoplasmic organelles, such as dictyosomes and Golgi vesicles, lacked such deposits of reaction product. After zygotene the enzyme activity decreased abruptly, and at the pachytene stage only very few deposits could be observed in the cell wall. Our results indicate that cellulase is synthesized on rough ER and secreted directly via the smooth ER and ER-derived vesicles into the cell wall by exocytosis, where it brings about local wall breakdown, leading to the secondary formation of plasmodesmata and cytoplasmic channels.