离体细胞共培养中科间细胞共质体的形成

离体培养下选出的绿色胡萝卜（Ｄａｕｃｕｓ ｃａｒｏｔａ）细胞系和白色普通烟草（Ｎｉｃｏｔｉａｎａｔａｂａｃｕｍ ）细胞系,各自具有独特的细胞结构标志,在愈伤组织、光镜和电镜３ 个水平上均可区分。对两个细胞系进行分散、混合、Ｋ＋ 液低渗处理后在固体培养基上共培养,１０—１５ ｄ 后可观察到两种细胞的镶嵌生长。光镜和电镜下均观察到烟草细胞和胡萝卜细胞之间隔离层的存在与消失。在隔离层消失的区域可见到异种细胞间次生胞间连丝的形成,从而将独立的两个共质体连成一个统一的共质体。对科间细胞共质联系的建立过程进行了讨论,认为细胞接触后首先非特异粘连——以隔离层形成并适度加厚为标志,然后特异的细胞识别在隔离层中启动,从而导致隔离层或消失而重新建立共质联系或加厚、木质化、木栓化     

Intercellular Symplastic Connection and Isolation of the Unloading Zone in Flesh of the Developing Grape Berry

The uhrastructure and intercellular connection of the sugar unloading zone (i. e. the phloem in the dorsal vascular bundle and the phloem-surrounding the assimilate sink-cells) of grape ( Vitis vinifera x V. labrusca cv. Jingchao) berry was observed via transmission electron microscopy. The results showed that during the early developmental stages of grape berry, numerous plasmodesmata were found in the phloem between sieve element (SE) and companion cell (CC), between SE/CC complexes, between SE/CC complex and phloem parenchyma cell and in between phloem parenchyma cells, which made the phloem a symplastic integration, facilitating sugar unloading from sieve elements into both companion cells and phloem parenchyma cells via a symplastic pathway. On the contrary, there was almost no plasmodesma between phloem and its surrounding flesh photoassimilate sink-cells, neither in between the flesh photoassimilate sink-cells giving rise to a symplastic isolation both between phloem and its surrounding flesh photoassimilate sink-cells, as well as among the flesh photoassimilate sink-cells. This indicated that both the sugar unloading from phloem and pestphloem transport of sugars should be mainly via an apoplastic pathway. Dining the ripening stage, most of the plasmodesmata between SE/CC complex and the surrounding phloem parenchyma cells were shown to be blocked by the electron-opaque globules, and a phenomenon of plasmolysis was found in a number of companion cells, indicating a symplastic isolation between SE/CC complex and its surrounding parenchynm cells during this phase. The symplastic isolation between the whole phloem and its surrounding photoassimilate sink-cells during the early developmental stages shifted to a symplastic isolation within the phloem during the ripening phase, and thus the symplastic pathway of sugar unloading from SE/CC complex during the early development stages should be replaced by a dominant apoplastic unloading pathway from SE/CC complex in concordance.     

Filament Disruption in Funaria Protonemata: Occlusion of Plasmodesmata

Plasmodesmata are occluded when Funaria chloronemata are fragmented by the development of tmema cells (TCs). The TC deposits a new wall layer along the cross wall toward the neighbouring non-sister cell (NC). This wall layer cuts off the plasmodesmata and its connection with the cross wall is soon lost. The plasmodesmata become isolated when the NC forms a new wall layer along the former cross wall. At the end of TC development, before its disintegration, the sister cell (SC) also deposits a new wall layer along the cross wall toward the TC, cutting off the plasmodesmata. For some time the plasmalemma of the plasmodesmata remains connected to the NC or the TC, whereas the desmotubule soon disappears. Relicts of the plasmalemma remain even after the isolation of the plasmodesmata and the disintegration of the TC. During the decay of the plasmodesmata, a cylinder of electron-dense material is frequently formed along the border of the plasmodesmatal channel. This may extend over the surface of the cell wall. Eventually, the plasmodesmatal channel is filled with wall material. Callose is only observed around functional plasmodesmata and does not seem to play a role in their occlusion.     

灵武长枣果实维管束韧皮部及周围细胞的超微结构特征

为了探讨灵武长枣果实光合同化物韧皮部卸载和运输的途径,该研究采用透射电镜技术,对不同发育时期灵武长枣果实维管束韧皮部及其周围薄壁细胞的超微结构特征进行了分析.结果表明:筛管/伴胞复合体及其周围韧皮薄壁细胞间在果实膨大前期富含胞间连丝,而韧皮薄壁细胞与周围库细胞以及相邻库细胞间几乎不存在胞间连丝,形成共质体隔离;筛管/伴...     

Studies on graft unions

Summary De novo formation of cytoplasmic cell connections are studied at the graft interface of 5 day old in vitro heterografts ofVicia faba onHelianthus annuus. Continuous and half plasmodesmata, both branched and unbranched, are described at various stages of development in non-division walls between unlike and like dedifferentiated callus cells. In apical portions of protruding callus cells and in the contact zone between opposing cells extremely thin wall parts with a striking ER/plasmalemma contact are observed. During subsequent thickening of the modified wall parts cytoplasmic strands enclosing constricted ER cisternae are entrapped within the newly deposited wall material. These cytoplasmic strands represent half plasmodesmata which—in case of fusion with corresponding structures of adjoining cells across the loosened wall matrix — form continuous cell connections. Golgi vesicles secreting wall material are involved in the process of forming half and continuous plasmodesmata, thus following the same mechanism of plasmodesmata development as described for isolated protoplasts in cell cultures. The findings suggest the existence of a unifying mechanism of secondary formation of plasmodesmata showing far-reaching similarities with the establishment of primary cell connections.     

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。     

Are there symplastic connections between the endosperm and embryo in some angiosperms?—a lesson from the Crassulaceae family

It is believed that there is symplastic isolation between the embryo (new sporophyte) and the endosperm (maternal-parental origin tissue, which nourishes the embryo) in angiosperms. However, in embryological literature there are rare examples in which plasmodesmata between the embryo suspensor and endosperm cells have been recorded (three species from Fabaceae). This study was undertaken in order to test the hypothesis that plasmodesmata between the embryo suspensor and the endosperm are not so rare but also occur in other angiosperm families; in order to check this, we used the Crassulaceae family because embryogenesis in Crassulaceae has been studied extensively at an ultrastructure level recently and also we tread members of this family as model for suspensor physiology and function studies. These plasmodesmata even occurred between the basal cell of the two-celled proembryo and endosperm cells. The plasmodesmata were simple at this stage of development. During the development of the embryo proper and the suspensor, the structure of plasmodesmata changes. They were branched and connected with electron-dense material. Our results suggest that in Crassulaceae with plasmodesmata between the endosperm and suspensor, symplastic connectivity at this cell-cell boundary is still reduced or blocked at a very early stage of embryo development (before the globular stage). The occurrence of plasmodesmata between the embryo suspensor and endosperm cells suggests possible symplastic transport between these different organs, at least at a very early stage of embryo development. However, whether this transport actually occurs needs to be proven experimentally. A broader analysis of plants from various families would show whether the occurrence of plasmodesmata between the embryo suspensor and the endosperm are typical embryological characteristics and if this is useful in discussions about angiosperm systematic and evolution.     

Rapid detection of plasmodesmata in purified cell walls

Summary Plasmodesmata are complex channels within the plant cell wall, which create plasma membrane and symplastic continuity between neighbouring cells. To detect plasmodesmata in cell wall preparations fromNicotiana cle elandii, we have used 3,3-dihexyl-oxacarbocyanine iodide (DiOC₆), a cationic amphiphilic fluorescent probe, widely employed for general studies of membrane structure and dynamics. Punctate fluorescent staining was readily seen in pit fields, small depressions within the cell wall known to be rich in plasmodesmata. Scanning electron microscopy was used to demonstrate that the punctate staining corresponded to plasmodesmata. Treatment of cell wall fragments with chloroform-methanol to remove lipids did not alter the staining of plasmodesmata. In contrast, pronase E-sodium dodecyl sulfate treatment completely abolished staining, indicating that the DiOC₆ labelling of plasmodesmata may be protein rather than lipid specific. Although not membrane mediated, DiOC₆ staining of plasmodesmata is a simple, rapid, and specific tool for the detection of plasmodesmata in isolated cell walls and will prove useful for studies of plasmodesmal location, structure, and composition.     

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.     

Transport of assimilates in the developing caryopsis of rice (Oryza sativa L.)

Assimilates entering the developing rice caryopsis traverse a short-distance pathway between the terminal sieve elements of the pericarp vascular bundle and the aleurone layer. The ultrastructure of this pathway has been studied. Sieve elements in the pericarp vascular bundle are smaller than their companion cells.The sieve elements show few connections with surrounding vascular parenchyma elements but are connected to companion cells by compound plasmodesmata. Companion cells, in turn, are connected to vascular parenchyma elements by numerous compound plasmodesmata present in wall thickenings. Assimilates leaving the sieve element — companion cell complex must laterally traverse cells of the pigment strand before they come into contact with the aleurone layer. The pigment strand cells have modified inner walls made up of a suberin-like material. This material may act as a permeability barrier isolating the apoplast from the symplast of the pigment strand. The walls of the pigment strand cells are traversed by numerous plasmodesmata. Water may be conducted to the endosperm through the isolated cell-wall system of the pigment strand while assimilates possibly move via plasmodesmata. High frequencies of plasmodesmata occur at the junction between the pigment strand and the nucellus and also between adjacent cells of the nucellus. By contrast, plasmodesmata are absent between the nucellus and the aleurone layer and also between the nucellus and the seed coat. A predominantly circumferential and symplastic transport pathway is likely between the pigment strand and nucellus. In view of the total absence of plasmodesmata between the nucellus and the aleurone layer assimilates entering the endosperm may have to cross the plasmalemma of the nucellus. It is possible that constraints to the flow of assimilates may occur in the short-distance pathway between the terminal sieve element — companion cell complexes and the endosperm, and this is discussed.     

荔枝雄花性别决定过程中细胞超微结构的变化

荔枝雄花雌蕊原基在大孢子母细胞减数分裂后开始衰退.内质网历经增生扩展,穿壁相连,同心缠绕,多条平行弯曲,不规则堆叠.内质网和高尔基体产生许多囊泡,囊泡在细胞内含物的降解和运输过程中起着重要的作用.线粒体在雌蕊原基细胞衰败的前、中期数量增加,后期分批降解.过氧化物酶体在雌蕊原基细胞衰败的中期紧挨核短暂出现.细胞核的染色质凝集断裂;核周腔扩大,形成胀泡;染色质趋边,外泄.细胞原生质表现出有序的、在膜包裹下的降解,首先是核糖体,而后依次是:过氧化物酶体、内质网、高尔基体、线粒体、核.雌蕊原基的衰败历程可能是一种程序性细胞死亡的过程.     

灵武长枣果实ATPase超微细胞化学定位和功能研究

采用ATPase超微细胞化学定位技术,研究灵武长枣果实不同发育阶段韧皮部和果肉库薄壁细胞ATPase分布特征,以明确灵武长枣果实ATPase超微细胞化学定位特征和功能。结果显示:(1)第一次快速生长期SE/CC复合体与周围的薄壁细胞有丰富的胞间连丝,形成共质体连续,韧皮部薄壁细胞之间有丰富的胞间连丝,ATPase反应物在韧皮部各细胞分布较少。(2)缓慢生长期ATPase反应物在韧皮部各细胞分布逐渐增加。(3)第二次快速生长期SE/CC复合体与周围的薄壁细胞缺乏胞间连丝,形成共质体隔离,韧皮薄壁细胞及果肉库薄壁细胞的胞间连丝较少,囊泡和膜泡在筛管、韧皮薄壁细胞和库薄壁细胞中很丰富,质膜、液泡膜、囊泡膜、细胞壁和胞间隙的ATPase活性较高。研究表明,果实在第一次快速生长期同化物从筛分子的卸出主要采取共质体途径,缓慢生长期同化物卸出时可能为共质体和质外体途径共存,第二次快速生长期则主要以质外体途径为主,证明果实不同发育阶段韧皮部同化物卸出路径存在差异。     

Paramural Bodies in Callus Beside the Isolation Layer of the Graft Union

The isolation layer of the graft union is a changeable component. It is formed and thickened during the early stage (Fig. 6) and disrupted, thinned and even disappeared durmg tile later stage of development of the graft (Fig. 1, 2, 14). A number of vesicles, paramarat bodies (Fig. 3, 5, 8, 9, 11, 13, 19), multivesicular bodies (Fig. 4, 15, 16, 18, 20) and concentric membrane bodies (Fig. 7) are observed in callus 'beside the isolation layer during both stages of development based on transmission electron microscopy. The paramural body comprises invagination of plasmalemma containing numerous vesicles and/or tubules situated between the cell wall and the plasmalemma. The multivesicular body is a organelle about 0.5-μm in diameter with a single membrane surrounding several smaller single vesicles. It is possible that multivesicular body and/or single iesicles transverse the plasmalemma to produce paramural body. The figures show that the paramural bodies appear always beside the isolalion layer at the different stages of development of the graft union and the multivesicular bodies appear mostly near the region where plasmodesmata are secondarily formed between the stock and the scion (Fig. 15, 17, 20). This may reflect that paramural body and multivesi- cular body, as well as single vesicles, are capable of performing vesicular transport. The deposition and reabsorption of material of the isolation layer occur due to vesicular transport. All the above facts seem to indicate that paramural body, multivesicular body and single vesicles can be both endocytotic and exocytotic. The present study supports the theory of vesicular transport, and authors suggest that transcellular cytosis occurs not only through plasmodesmata but also through plasmalemma in oapoplast. The single vesicles, paramural bodies and multivesicular bodies take an active part in the transport process of symplast-apoplast-symplast.     

Symplastic and apoplastic sugar contents in gall tissues and callus of the sumac (Rhus chinensis MILL.)

To elucidate the role of cell wall in interaction with gall-inducing organisms, symplastic and apoplastic sugar contents in different shapes of gall tissue of the sumac (Rhus chinensis Mill.) were compared with those of the callus. The gall tissues with vascular cylinders, intercellular spaces and callus were fractionated into symplastic [methanol (MeOH), hot water (HW), and starch] fractions and apoplastic [pectin, hemicellulose, trifluoroacetic acid (TFA)-soluble, and cellulose] fractions. Symplastic sugar content of gall tissues was higher than that of callus. In apoplastic (cell wall) fractions, the cellulose content of gall tissues was lower than that of callus, due to large amount of pectin with high ratio of uronic acid (UA) and hemicellulose with low ratio of UA. Analysis of neutral sugar component of the hemicellulosic, TFA-soluble fraction showed that arabinose (side chain) and galactose (backbone) of arabinogalactan were rich in gall tissues and callus. The gall tissues had higher glucose and lower xylose contents than the callus. These results suggest that the structure of cell wall polysaccharides of gall changed during its development with an increase in symplastic sugar contents. The feeding activities occuring in gall by the gall-inducers were discussed.     

Studies on graft unions

Summary The occurrence of plasmodesmata in the graft interfaces of two heteroplastic grafts (Impatiens walleriana onImpatiens olivieri andHelianthus annum onVicia faba) has been studied. For both systems two types of intercellular strand are described: 1. Continuous plasmodesmata interconnecting the cells of stock and scion and 2. half plasmodesmata traversing the wall part of one partner cell without connection to the abutting cell. Single strands or branched forms occur in both types of plasmodesma. In the case of half plasmodesmata, branchings with extended median nodules predominate. The distribution of half and continuous plasmodesmata varies with the different areas of a graft interface: in the region of bridging vascular tissues most cell connections are continuous. In areas where cortex or pith-derived callus cells and those of misaligned tissues (cortex/vascular tissue; cortex/pith; pith/vascular tissue) match, discontinuous strands predominate.Branched half plasmodesmata also occur in presumably fused walls between related callus cells; they are typical structures secondarily formed in non-division walls.The results are discussed with regard to compatibility/incompatibility phenomena in heterografts and the development and function of interspecific cell bridges.     

Temporal and spatial regulation of symplastic trafficking during development in Arabidopsis thaliana apices

Plasmodesmata provide symplastic continuity linking individual plant cells. However, specialized cells may be isolated, either by the absence of plasmodesmata or by down regulation of the cytoplasmic flux through these channels, resulting in the formation of symplastic domains. Maintenance of these domains may be essential for the co-ordination of growth and development. While cells in the center of the meristem divide slowly and remain undifferentiated, cells on the meristem periphery divide more frequently and respond to signals determining organ fate. Such symplastic domains were visualized within shoot apices of Arabidopsis, by monitoring fluorescent symplastic tracers (HPTS: 8-hydroxypyrene 1,3,6 trisulfonic acid and CF: carboxy fluorescein). Tracers were loaded through cut leaves and distributed throughout the whole plant. Confocal laser scanning microscopy on living Arabidopsis plants indicates that HPTS moves via the vascular tissue from leaves to the apex where the tracer exits the phloem and moves symplastically into surrounding cells. The distribution of HPTS was monitored in vegetative apices, and just prior to, during, and after the switch to production of flowers. The apices of vegetative plants loaded with HPTS had detectable amounts of tracer in the tunica layer of the meristem and in very young primordia, whereas the corpus of the meristem excluded tracer uptake. Fluorescence signal intensity decreased prior to the onset of flowering. Moreover, at approximately the time the plants were committed to flowering, HPTS was undetectable in the inflorescence meristem or young primordia. Later in development, after several secondary inflorescences and mature siliques appeared, inflorescence apices again showed tracer loading at levels comparable to that of vegetative apices. Thus, analysis of fluorescent tracer movement via plasmodesmata reveals there is distinct temporal and spatial regulation of symplastic domains at the apex, dependent on the developmental stage of the plant.     

胞间连丝的次生形成和次生变化

本文介绍了胞间连丝次生形成和次生变化的研究进展。用统计特定细胞壁区段上胞间连丝数量与密度的变化,电镜观察嫁接组合中接穗与砧水间细胞壁上胞间连丝的形成等方法,证明了在植物生长发育过程中,存在着胞间连丝的次生形成。在某些特定部位,某一发育阶段,已形成的胞间连丝常会发生可逆的次生变化,这种变化和植物发育过程中的共质体隔离以及物质运输的调节有关。     

胞间连丝的次生形成和次生变化

本文介绍了胞间连丝次生形成和次生变化的研究进展,用统计特定细胞壁区段上胞间连丝数量与密度的变化,电镜观察嫁接组合中接穗与砧水间细胞壁上胞间连丝的形成等方法,证明了在植物生长发育过程中,存在着胞间连丝的次生形成。在某些特定部位,某一发育阶段,已形成的胞间连丝常会发生可逆的次生变化,这种变化和植物发育过程中的共质体隔离以及物质运输的调节有关。     

Changes in dye coupling of stomatal cells of Allium and Commelina demonstrated by microinjection of Lucifer yellow

Lucifer yellow has been microinjected into stomatal cells of Allium cepa L. epidermal slices and Commelina communis L. epidermal peels and the symplastic spread of dye to neighboring cells monitored by fluorescence microscopy. Dye does not move out of injected mature guard cells, nor does it spread into the guard cells when adjacent epidermal or subsidiary cells are injected. Dye does spread from injected subsidiary cells to other subsidiary cells. These results are consistent with the reported absence of plasmodesmata in the walls of mature guard cells. Microinjection was also used to ascertain when dye coupling ceases during stomatal differentiation in Allium. Dye rapidly moves into and out of guard mother cells and young guard cells. Hovewer, dye movement ceases midway through development as the guard cells begin to swell but well before a pore first opens. Since plasmodesmata are still present at this stage, the loss of symplastic transport may result from changes in these structures well in advance of their actual disappearance from the guard cell wall.Abbreviations DIC differential interference contrast - GMC guard mother cell - LY Lucifer yellow - Pd plasmodesmata You can observe a lot by watching Lawrence Berra, as quoted in Sports Illustrated, vol. 60 (No. 14), p. 94, 2 April 1984     

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.