细叶黄芪叶肉原生质体发育早期几种细胞器的变化

在细叶黄芪叶肉原生质体发育早期,细胞器的变化较大。离体培养4h后,线粒体的嵴和基质物质开始增加。培养3—5天后,线粒体的数量增加5倍以上,此时可见大部分线粒体围绕细胞核分布。在培养24h后,高尔基体开始发育,它们主要分布在细胞质周边区域。多糖细胞化学染色表明,高尔基体内沉积着大量嗜银物质。培养1天后,粗面内质网开始发育。培养3天时,部分叶绿体边缘出现一些空隙结构。随着叶绿体内膜结构的消失,淀粉粒增大,叶绿体逐渐转变为造粉质体。     

细叶黄芪叶肉原生质体发育早期几种细胞器的变化

在细叶黄芪叶肉原生质体发育早期,细胞器的变化较大。离体培养４ｈ后,线粒体的嵴和基质物质开始增加。培养３－５天后,线粒体的数量增加５倍以上,此时可见大部分线粒体围绕细胞核分布。,在培养２４ｈ后,高尔基体开始发育,它们主要分布在细胞质周边区域。多糖细胞化学染色表明,高尔基体内沉积着大量嗜银物质。培养１天后,粗面内质网开始发育。培养３天时,部分叶绿体边缘出现一些空隙结构。随着叶绿体内膜结构的消失,淀粉粒     

应用荧光增白剂VBL研究原生质体细胞壁的再生

对新分离和培养的烟草(Nicotiana tabacum)叶肉原生质体用荧光增自剂VBL染色,于波长3600～4400光源的荧光显微镜下观察表明,新分离的烟草叶肉原生质体没有发绿色荧光的细胞壁存在,只见发微弱深红色荧光的叶绿体,而培养4～6天以上的原生质体再生壁发绿色荧光,证明含纤维素成分。这种方法亦可用于观察原生质体的出芽分裂和细胞分裂。     

小麦与叶锈菌互作过程中细胞程序性死亡的细胞学观察

小麦(Triticum aesetivum)品种洛夫林10和叶锈菌(Puccinia recondita f.sp tritici)小种162、165分别组成不亲和组合与亲和组合。透射电镜观察表明,在小麦与叶锈菌的不亲和组合中,接种后12h,侵染点周围叶肉细胞核变形;接种后24h,核内染色质开始凝聚,并趋于细胞核边缘,同时叶绿体膨胀;接种后48h,核内染色质凝聚加剧,叶绿体开始解体;最终在接种后72h,细胞核、叶绿体完全解体,线粒体开始退化。此外,内质网和液泡共同行使溶酶体功能,吞噬各种细胞器残体及原生质降解组分。以上结果表明:在小麦与叶锈菌不亲和组合的互作过程中,寄主细胞呈现细胞程序性死亡的典型特征。而在亲和组合中,叶肉细胞间隙中可见有大量菌丝蔓延,菌丝与寄主细胞壁接触后分化产生吸器母细胞。菌丝的存在对寄主细胞的超微结构产生一定影响。从接种后24h开始,与菌接触的细胞出现质膜下陷,叶绿体稍显膨胀;在接种后48h、72h,大部分叶绿体膨胀,而其它细胞器无明显变化。     

小麦与叶锈菌互作过程中细胞程序性死亡的细胞学观察

小麦(Triticum aesetivum)品种洛夫林10和叶锈菌(Puccinia recondita f.sp tritici)小种162、165分别组成不亲和组合与亲和组合。透射电镜观察表明,在小麦与叶锈菌的不亲和组合中,接种后12h,侵染点周围叶肉细胞核变形;接种后24h,核内染色质开始凝聚,并趋于细胞核边缘,同时叶绿体膨胀;接种后48h,核内染色质凝聚加剧,叶绿体开始解体;最终在接种后72h,细胞核、叶绿体完全解体,线粒体开始退化。此外,内质网和液泡共同行使溶酶体功能,吞噬各种细胞器残体及原生质降解组分。以上结果表明：在小麦与叶锈菌不亲和组合的互作过程中,寄主细胞呈现细胞程序性死亡的典型特征。而在亲和组合中,叶肉细胞间隙中可见有大量菌丝蔓延,菌丝与寄主细胞壁接触后分化产生吸器母细胞。菌丝的存在对寄主细胞的超微结构产生一定影响。从接种后24h开始,与菌接触的细胞出现质膜下陷,叶绿体稍显膨胀;在接种后48h、72h,大部分叶绿体膨胀,而其它细胞器无明显变化。     

缺铁对黄瓜叶内细胞超微结构及某些形态的影响

利用透射电镜,对缺铁黄瓜幼叶肉细胞的超微结构及植株的某些形态学进行了观测,发现缺铁后的幼叶细胞内首先受到影响的细胞器是叶绿体,其次是线粒体,细胞核,内质网结构完好,幼苗长势衰弱,叶色发黄,叶绿素含量下降。     

黄独冻后黑暗培养胚性愈伤组织及其再生植株的电镜观察

本研究以黄独为材料,对其冻后黑暗培养胚性愈伤组织及其再生植株进行电镜观察。结果表明:黄独胚性愈伤组织在冻后没有经过一定时间的黑暗培养直接放于正常的光周期下培养,极易使细胞内容物外泄,且造成部分细胞器和细胞核消失或裂解形成空隙,线粒体和高尔基体等细胞器的结构不完整,出现空隙或内容物溢出或断裂现象;而黄独冻后胚性愈伤组织经过3 d的黑暗培养后再置于光周期下培养,受损的细胞较少,细胞排列较紧密,且细胞器降解较少,很少出现空隙等现象,线粒体、高尔基体等细胞器的结构也较为完整。此外,与黄独带芽茎段常温继代再生试管苗相比,黄独胚性愈伤组织经过冻后黑暗培养后,其再生试管苗的根、茎、叶结构没有出现显著变化,且两者的气孔参数和叶绿体数量也均无显著性差异。本试验结果可为植物种质资源超低温保存后的黑暗培养提供了亚显微结构证据。     

基因工程

<正> 用给体一受体原生质体融合技术研究了叶绿体和线粒体控制的性状在种间的转移。野生烟草或普通烟草白化系(V BW)用作受体原生质体,下一射线照射的花烟草、印度烟草和波状叶烟草原生质体用作细胞器给体。融合细胞经培养后再生成胞质杂种植株。     

缺铁对黄瓜叶肉细胞超微结构及某些形态的影响

利用透射电镜,对缺铁黄瓜幼叶肉细胞的超微结构及植株的某些形态学进行了观测,发现缺铁后的幼叶细胞内首先受到影响的细胞器是叶绿体,其次是线料体,细胞核、内质网结构完好。幼苗缺铁２０天后,大部分叶绿体的基料片层和间质片层受到了严重的破坏。少量线粒体出现空泡化倾向,植株在形态学上表现为幼苗长势衰弱,叶色发黄、叶绿素含量下降     

缺铁对黄瓜叶肉细胞超微结构及某些形态的影响

利用透射电镜,对缺铁黄瓜幼叶肉细胞的超微结构及植株的某些形态学进行了观测,发现缺铁后的幼叶细胞内首先受到影响的细胞器是叶绿体,其次是线粒体,细胞核、内质网结构完好。幼苗缺铁20天后, 大部分叶绿体的基粒片层和间质片层受到了严重的破坏。少量线粒体出现空泡化倾向,植株在形态学上表现为幼苗长势衰弱,叶色发黄、叶绿素含量下降。     

The bloom-forming ciliate Mesodinium rubrum harbours a single permanent endosymbiont

Whether the red tide Mesodinium rubrum contains a permanent cryptophyte symbiont or whether it only sequesters chloroplasts from cryptophyte prey was addressed using electron microscopy and the dynamics of photosynthesis, chloroplasts and nuclei. Mesodinium rubrum contains a branched cryptophyte symbiont consisting of many chloroplasts, mitochondria, nucleomorphs, an endoplasmic reticulum and one nucleus. The volume of the symbiont constitutes 36% of the consortium and it is separated from its host by a single-cell membrane. The chloroplasts of Mesodium are larger and morphologically different from two Teleaulax species that served as prey. The symbiont nucleus is also much larger than Teleaulax nuclei. Although M. rubrum is functionally a phototroph, sustained growth beyond two to four generations requires ingestion of prey, but less than one prey cell per generation suffices for maximum growth. This suggests that either the ciliate or its symbiont needs an essential growth factor for continuous growth.     

Subcellular fractionation of wheat leaf protoplasts by centrifugal elutriation

A new method using centrifugal elutriation for subcellular fractionation of plant cells has been developed. This method takes advantage of the fact that particles sedimenting in a gravitational field can be eluted by flow against the field. A wheat protoplast homogenate was fed into an elutriation rotor spinning at high speed and the flow rate into the rotor was gradually increased. The smaller and less dense materials such as mitochondria, microbodies, endoplasmic reticulum, and cytoplasm were elutriated earlier than the larger and denser nuclei and chloroplasts. The intact chloroplasts, free of mitochondria, microbodies, endoplasmic reticulum, and cytoplasm, could be obtained within 40 min following the rupture of protoplasts. The chlorophyll-free mitochondria could be obtained within 80 min.     

An ultrastructural and cytochemical investigation of the colonial green algaPediastrum tetras during zoospore formation

Summary Ultrastructural changes during zoospore formation and aggregation into motile, aggregating zoospores were examined in the colonial green algaPediastrum tetras. Developing zoospores are characterized by irregularly shaped nuclei, presence of peripheral networks of rough endoplasmic reticulum, chloroplasts with tightly apposed thylakoids and dictyosome cisternae which are compressed and reduced in size. A single membrane bound organelle with a fine granular matrix of moderate electron density of diameter ranging from 0.2 m to 0.6 m and associated with chloroplasts, mitochondria and endoplasmic reticulum was found only in adult cells. Although this organelle has the morphology of a microbody, it did not stain with 3,3-diaminobenzidine (DAB) at pH 9.6 or pH 7.6, whereas mitochondrial membranes stained. No DAB staining was observed along the cell wall or the plasma membrane of zoospores, or associated with endoplasmic reticulum, plastid membranes or dictyosomes.     

The biology ofChlamydomyxa montana: Ultrastructure of the cyst

Summary Cells ofChlamydomyxa montana Lankester photosynthesize within a cyst under bright light and ingest diatoms in an excysted amoeboid form during periods of low light intensity. Cyst cell walls are partially comprised of cellulose and vary in thickness. The cells are multinucleate and intracellular bacteria are closely associated with the nuclei. A pair of centrioles is also present adjacent to individual nuclei. Chloroplasts are numerous and thylakoids are generally organized into bands of three. No endoplasmic reticulum was found surrounding the plastids, nor was a complete girdling lamella ever observed within.C. montana chloroplasts appear to be its own but this protist does not precisely fit into an algal class.     

花生胚乳细胞化的超微结构观察

花生（ＡｒａｃｈｉｓｈｙｐｏｇｅａｅＬ．）心形胚期的胚乳游离核多瓣裂,或具长尾状结构。胚乳细胞质内有大量线粒体、质体、高尔基体、小泡及少量内质网。中央细胞壁有壁内突。球胚及心形胚期常见胚乳瘤。心形胚晚期,胚乳开始细胞化,胚乳细胞壁形成有３种方式,分别存在于不同的胚珠中：（１）从胚囊壁产生自由生长壁形成初始垂周壁,具有明显的电子密度深的中层,其生长主要靠末端的高尔基体小泡及内质网囊泡的融合。两相邻的自由生长壁末端或其分枝末端相连形成胚乳细胞。（２）核有丝分裂后产生细胞板,细胞板向外扩展并可分枝。间期的非姊妹核间也观察到形成了细胞板。小泡与微管参与细胞板的扩展,高尔基体和内质网是小泡的主要来源。细胞板的扩展末端相互连接,形成胚乳细胞的前身。小泡继续加入细胞板的组成,以后形成胚乳细胞壁。（３）胚乳细胞质中,出现一些比较大的不规则形的片段性泡状结构,它们可能来源于高尔基体小泡,这些片段性泡状结构随机相连形成细胞壁,未见微管参与。胚乳细胞外切向壁及经向壁上有壁内突。     

Acyl-CoA dependent acylation of phospholipids in the chloroplast envelope

Acyl-CoAs are substrates for acyl lipid synthesis in the endoplasmic reticulum. In addition, they may also be substrates for lipid acylation in other membranes. In order to assess whether lipid acylation may have a role in plastid lipid metabolism, we have studied the incorporation of radiolabelled fatty acids from acyl-CoAs into lipids in isolated, intact pea chloroplasts. The labelled lipids were phosphatidylcholine (PC), phosphatidylglycerol (PG), phosphatidylinositol and free fatty acids. With oleoyl-CoA, the fatty acid was incorporated preferably into the sn-2 position of PC and the acylation activity mainly occurred in fractions enriched in inner chloroplast envelope. Added lysoPC stimulated the activity. With palmitoyl-CoA, the fatty acid was incorporated primarily into the sn-1 position of PG and the reaction occurred at the surface of the chloroplasts. As chloroplast-synthesized PG generally contains 16C fatty acids in the sn-2 position, we propose that the acylation of PG studied represents activities present in a domain of the endoplasmic reticulum or an endoplasmic reticulum-derived fraction that is associated with chloroplasts and maintains this association during isolation. This domain or fraction contains a discreet population of lipid metabolizing activities, different from that of bulk endoplasmic reticulum, as shown by that with isolated endoplasmic reticulum, acyl-CoAs strongly labelled phosphatidic acid and phosphatidylethanolamine, lipids that were never labelled in the isolated chloroplasts.     

The neurocytes of the caudal part of the tuberomamillary nucleus in Ovis aries L. (light microscopy and electron microscopy studies)

The neurons of the pars caudalis nuclei tuberomammillaris (pc-NTM) were studed light-microscopically and electron-microscopically in sheep and rams of Merino breed. In our study we observed: In the regarded neural nucleus, there is the majority of the great neurons (up to 60 microns in diameter) rich in the NISSL's bodies. When stained with the cresyl violet, the NISSL's substance is apparently stored mainly in peripheral area of the cell body and in the distant parts of numerous protoplasmic processes, what evokes an impression of the "jagged" surface of these cells. After staining with paraldehyde fuchsin, we found purple coloured lumps of irregular shape stored outside the cell bodies, in the neuropil. The less extended cells, usually with lower content of NISSL bodies, are in pc-NTM less frequent. In the electron-microscopic study we identified 3 types of neurons: Cells rich in rough endoplasmic reticulum; "light" cells, "dark" cells. The cells of the 1st type were the most frequent ones. Cisterns of rough endoplasmic reticulum in the 1st type of cells are often dilated. The protoplasmic processes of these cells are frequently stepped over by flat tubuli of endoplasmic reticulum. The 2nd type of cells is characterized by the light cytoplasmic matrix, low quantity of endoplasmic reticulum and frequent occurrence of lipofuscin bodies. The 3rd type of cells are characterized by the high density of cytoplasmic matrix, well developed GOLGI complex, and very broad cisterns of endoplasmic reticulum, forming a labyrinth, and it is bound to a broad perinuclear space.     

ELECTRON MICROSCOPIC RADIOAUTOGRAPHIC DETECTION OF SITES OF PROTEIN SYNTHESIS AND MIGRATION IN LIVER

The sites of synthesis of proteins and their subsequent migration in rat liver have been studied during a 75 min period after labeling of liver-slice proteins by exposure to leucine-H³ for 2 min. Incorporation of the label into protein began after 1 min and was maximal by 4 min. Electron microscopic radioautography showed that synthesis of proteins in hepatocytes occurs mainly on ribosomes, particularly those in rough endoplasmic reticulum and, to some extent, in nuclei and mitochondria. Most of the newly formed proteins leave the endoplasmic reticulum in the course of 40 min, and concurrently labeled proteins appear in Golgi bodies, smooth membranes, microbodies, and lysosomes. A likely pathway for the secretion of some or all plasma proteins is from typical rough endoplasmic reticulum to a zone of reticulum which is partially coated with ribosomes, to the Golgi apparatus, and thence to the cell periphery. The formation of protein by reticuloendothelial cells was measured and found to be about 5% of the total protein formed by the liver.     

Cell-free transfer and sorting of membrane lipids in spinach

Summary The donor and acceptor specificity of cell-free transfer of radiolabeled membrane constituents, chiefly lipids, was examined using purified fractions of endoplasmic reticulum, Golgi apparatus, nuclei, plasma membrane, tonoplast, mitochondria, and chloroplasts prepared from green leaves of spinach. Donor membranes were radiolabeled with [¹⁴C]acetate. Acceptor membranes were unlabeled and immobilized on nitrocellulose filters. The assay was designed to measure membrane transfer resulting from ATP-and temperature-dependent formation of transfer vesicles by the donor fraction in solution and subsequent attachment and/or fusion of the transfer vesicles with the immobilized acceptor. When applied to the analysis of spinach fractions, significant ATP-dependent transfer in the presence of cytosol was observed only with endoplasmic reticulum as donor and Golgi apparatus as acceptor. Transfer in the reverse direction, from Golgi apparatus to endoplasmic reticulum, was only 0.2 to 0.3 that from endoplasmic reticulum to Golgi apparatus. ATP-dependent transfers also were indicated between nuclei and Golgi apparatus from regression analysis of transfer kinetics. Specific transfer between Golgi apparatus and plasma membrane and, to a lesser extent, from plasma membrane to Golgi apparatus was observed at 25°C compared to 4°C but was not ATP plus cytosol-dependent. All other combinations of organelles and membranes exhibited no ATP plus cytosol-dependent transfer and only small increments of specific transfer comparing transfer at 37°C to transfer at 4°C. Thus, the only combinations of membranes capable of significant cell-free transfer in vitro were those observed by electron microscopy of cells and tissues to be involved in vesicular transport in vivo (endoplasmic reticulum, Golgi apparatus, plasma membrane, nuclear envelope). Of these, only with endoplasmic reticulum (or nuclear envelope) and Golgi apparatus, where transfer in situ is via 50 to 70 nm transition vesicles, was temperature-and ATP-dependent transfer of acetatelabeled membrane reproduced in vitro. Lipids transferred included phospholipids, mono-and diacylglycerols, and sterols but not triacylglycerols or steryl esters, raising the possibility of lipid sorting or processing to exclude transfer of triacylglycerols and steryl esters at the endoplasmic reticulum to Golgi apparatus step.