甜菊组织培养物中叶绿体的超微结构与脱分化

含有叶绿体的甜菊愈伤组织细胞转移至亲鲜培养基后,导致光合片层的逐渐减少或消失,最后叶绿体脱分化形成原体样的结构。超微结构观察表明,光合片层的减少或消失与降解及叶绿体分裂特别是不均等缢缩分裂而致基质组分和类囊体膜稀释有关,这一过程并不完全同步,一些质体含有少量正常的片支厕国一些质体有退化的片层对片导全消失。细胞的一个明显特点是细胞器大多聚细胞核附近,细胞质增加并向细胞中央伸出细胞质丝,同时可观察到质     

烟草叶片外植体脱分化与再分化过程中FtsZ蛋白的表达（简报）

在植物叶肉细胞的脱分化、再分化过程中伴随着叶绿体与质体相互转化的过程。已高度分化的叶肉细胞脱分化为分生状态细胞时．其中的原质体主要由叶绿体出芽生殖产生．偶尔可以看到某些叶绿体分裂或分裂与出芽同时出现的情况。此外,叶绿体在出芽产生原质体的同时自身逐渐被巨大的淀粉粒所充满．从而转变为淀粉体。     

甜菊叶愈伤组织诱导过程中叶绿体的超微结构变化

观察了甜菊(Stevia rebaudiana Bertoni)叶外植体愈伤组织诱导过程中叶绿体的超微结构变化。结果表明,当叶外植体转移到培养基上培养后,叶绿体的片层结构逐渐退化。在叶绿体发生退化的过程中伴有叶绿体出芽和原质体的形成。推测新产生的原质体来自叶绿体产生的芽状体。而叶绿体本身最后完全解体消失。叶绿体超微结构的这种变化与高度液泡化的叶肉细胞脱分化至分生状态是平行的。随着培养的进行,分生状态的细胞发生液泡化变为薄壁细胞时,在愈伤组织表层的细胞中,质体重新形成片层结构,而内部细胞的质体则充满淀粉粒。     

质体

质体是植物细胞特有的一类细胞器,藻类的质体类型比较单一,一般只有叶绿体一种,到了被子植物质体的类型才增多,而且被子植物细胞的发育,很大程度表现在质体的分化上。自从本世纪60年代确证质体含有自己的DNA以来,有人将质体看作是细胞内的细胞,这对理解质体有一定意义。在分生组织中质体呈未分化状态,叫做原质体。随着细胞的分化,原质体的结构和功能都发生深刻的变化,并形成各种质体。按照惠特利(Whatley 1978)的意见,质体大体可分     

甜菊质体发育过程的超微结构研究

观察了甜菊(Steviarebaudiana Bertani)质,本发育过程的超微结构变化、黄化幼芽质体的原片层体,为小管组成的网状“晶格体”,“质体中心”逐渐弥散,形成放射状排列的片层结构。质体的被膜出现突起的芽体,可能是质体增殖的一种方式。至幼叶转绿后,“质体中心”弥散消失,类囊体基本形成,原片层体转化为片层结构。随着叶片的成长变绿,基粒片层和基质片层的明显分化,基粒垛叠层增多,光合膜系十分发达。在幼叶和正在伸展的叶片细胞内,可以观察到叶绿体的分裂成为哑铃形,这是叶绿体增殖的主要途径。     

油菜叶片及其脱分化和再分化中质体的电镜观察

我们用电镜观察了油菜叶片植株再生中质体的超微结构变化。在油菜叶肉细胞中,叶绿体的基粒,基质片层发育良好,偶尔有淀粉粒。在来自叶片的愈伤组织细胞中,质体体积变小,类囊体已经消失或部分消失,有的质体含有淀粉粒,但很少有质体小球。经培养分化后的愈伤组织,特别是在表层细胞中,质体数量急剧增多,形态变化很大,贮藏淀粉明显减少。基质内已有泡状或管状结构。有的质体已出现长的基质片层,但未见到基粒;质体中常有质体球。由此可见,质体是一个十分敏感的细胞器,它的变化与细胞分化有关,变化最大的部分是片层系统,贮藏淀粉,质体小球。片层系统中尤以基粒片层变化最为显著。     

甜菊愈伤组织生长、分化与甜菊糖苷积累的关系

耐菊（Steviarebaudiana）愈伤组织中甜菊糖苷的积累与愈伤组织的生长呈负相关、与愈伤组织细胞的组织化及转绿呈正相关。愈伤组织芽的分化并不是积累较高水平甜菊糖苷的必要前提。绿色、质地致密、生长缓慢的愈伤组织,不论有芽分化或无芽分化时,其甜菊糖苷含量均较高。在电镜下观察到,这两种愈伤组织细胞具有类似的超微结构特征：细胞高度液泡化;叶绿体发育成熟,光合膜系统结构发达,基质浓厚且含有质体小球;微体具有典型的晶格结构,常与叶绿体紧密相靠。黄色、质地致密、生长缓慢的愈伤组织中甜菊糖苷含量较低,其细胞内质体富含淀粉粒,只有少量分散的片层结构,有的质体甚至完全被淀粉粒所充塞。黄色、质地疏松、生长快速的愈伤组织中甜菊糖苷含量最低,其细胞内质体结构简单,片层稀少。质体的发育和液泡的分化与甜菊糖苷的积累密切相关。愈伤组织具有较高的甜菊糖苷含量在于愈伤组织细胞的组织化以及细胞的高度液泡化并具有发育成熟的叶绿体。     

珊瑚豆果实成熟过程中叶绿体转化为杂色体的研究

珊瑚豆 (Solanum pseudo- capsicum var.diflorum (Vell.) Bitter)果实成熟过程中 ,果实颜色的变化和叶绿素含量降低及类胡萝卜素含量增长相符合。对果实中叶绿体转化为杂色体进行了电镜观察。早期绿色果实的特点是叶绿体具典型的基粒 -基粒间类囊体结构。在黄绿色果实时期叶绿体类囊体系统解体 ,代之以少数非叶绿素的单个类囊体和积累大的嗜锇的质体小球。质体转变为所谓的原质体。这表明叶绿体在果实成熟中的脱分化过程。当果实达到黄色阶段 ,这些质体所含的质体小球开始从中央形成质体小管的结构。最初质体小球中央变为半透明 ,认为是质体累积胡萝卜素的开始。随着质体小球的延长 ,小管从小球中伸出。这些小管围以电子致密的膜 ,中央是半透明的轴心。与此同时 ,在质体基质中出现一系列发育不同阶段的小泡 ,似乎是形成新的质体小球的过程。在成熟的橙色和橙红色果实中的杂色体中只包含无数小管和小的质体小球。质体小管在数量和长度上增长 ,充满成熟的杂色体。无数质体小球分布在小管之间的空间中。成熟杂色体从脱分化的原质体的重建是真正的再分化过程。可以作出结论 ,珊瑚豆果实叶绿体转化为杂色体实质上是一个脱分化和再分化过程     

马占相思子叶离体培养中细胞早期变化的研究

用光学和电子显微镜观察马占相思子叶在离体培养中的细胞早期变化,结果表明,未培养的子叶薄壁细胞内充满大量贮藏物质,在脱分化过程中,贮藏物质逐渐减少,细胞核体积增大,液泡蛋白体出现,质体转化为原质体,线粒体,内质网等细胞器数量增加,马脱分化启动后的细胞分裂方式进行了讨论。     

盐胁迫对玉米叶片叶肉细胞生物膜超微结构的影响

研究了NaCl胁迫对玉米叶肉细胞生物膜超微结构的影响. 结果表明：NaCl胁迫破坏了玉米叶片叶肉细胞生物膜的正常结构,50 mmol·L^-1 NaCl处理胁迫下,玉米叶肉细胞核膜,线粒体膜,细胞膜,叶绿体膜,液泡膜都受到不同程度的破坏,叶绿体基粒类囊体膨胀,间质片层空间增大,片层紊乱。100 mmol·L^-1 NaCl处理胁迫下,质膜,液泡膜,线粒体,叶绿体都受到严重的破坏。细胞质膜破坏,破损的叶绿体充斥在细胞间隙中;叶绿体外膜破坏,甚至解体消失,叶肉细胞中充满膜结构,基粒排列方向改变,垛叠层数减少,基粒和基质片层界限模糊不清,有的基粒解体消失,甚至叶绿体完全解体;核膜破坏、解体,核中的染色质高度凝缩;线粒体的数量增多,线粒体膜破坏,脊的数量减少,甚至整个线粒体破损解体;液泡膜破坏;由于各种生物膜的破坏,使细胞内充满许多囊状小泡、多泡体或斑层小体;叶肉细胞发生严重的质壁分离,严重时发生细胞壁断裂;甚至整个细胞溶解。     

AN ULTRASTRUCTURAL STUDY OF CHLOROPLAST STRUCTURE AND DEDIFFERENTIATION IN TISSUE CULTURES OF STREPTANTHUS TORTUOSUS (CRUCIFERAE)

Cells of Streptanthus tortuosus callus tissue contain chloroplasts when cultured in a liquid medium in the light. Similar cells grown in the dark contain proplastids that fail to develop prolamellar bodies but do contain a complex of loosely-associated membranes. When green, light-grown cultures are cut into small pieces and subcultured to a fresh culture medium, they become bleached even though maintained under the same illumination. The fine structure of the chloroplasts and the chlorophyll content of the cells indicate a dedifferentiation of the chloroplasts to a proplastid state during the early culture period. The changes in the ultrastructure of the plastids are paralleled by a dedifferentiation of the vacuolate cells to a less differentiated, meristematic state. Subsequent growth in the light results in a re-formation of chloroplasts and an increase in the chlorophyll content of the cells. The period of chloroplast redevelopment is associated with the re-formation of large central vacuoles in the cultured cells. Invaginations of the inner membrane of the plastid envelope occur at all stages of plastid development and are not lost during the period of grana degeneration. The proplastids formed from the dedifferentiation of the chloroplasts contain a large number of these invaginations and the redevelopment of grana is associated with a change in the electron density of the invaginating membranes. The degradation of the chlorophyll-containing membranes of the grana occurs during a period of rapid cytoplasmic synthesis induced by the fresh supply of nutrients in the culture medium. These results suggest that the high levels of nutrients may act directly on the chloroplasts and cause their dedifferentiation or that the rapid cell growth induced by the nutrients may cause a degradation of the membrane proteins in the grana of the chloroplasts and an incorporation of the released amino acids into non-plastid components of the cytoplasm.     

THE ULTRASTRUCTURAL MORPHOLOGY AND ONTOGENY OF OAT COLEOPTILE PLASTIDS

Structurally similar proplastids occur in the shoot, scutellum, and root of the oat embryo at the start of germination. These proplastids follow several pathways of differentiation, depending on their location within an organ and on previous exposure to light. During the first 24 hr of germination morphologically similar amyloplasts are formed from the preexisting proplastids in most of the cells of the seedling. After about 24 hr in the light, unique chloroplasts begin to develop in a subepidermal ring of small cortical parenchyma cells in the coleoptile and give the organ a pale green color. At 48 and 72 hr the coleoptile chloroplasts and etioplasts are conspicuously different from the corresponding leaf plastids in morphology and ontogeny but contain typical photosynthetic grana and prolamellar bodies. Study of the ontogeny of plastids in the epidermal and nongreening parenchymal regions of dark grown coleoptiles shows that these plastids undergo significant losses in starch content, and some increase of membranes within the plastid, related to the age of the cell. Light has little effect on the structure of these plastids. It is suggested that the ontogeny of all the plastid types of the oat seedling begins with a common precursor—a relatively simple proplastid that is present at the time of germination. Starch grains showing two distinct types of erosion, apparently enzymatic, were observed in oat coleoptile plastids. In one type (grooved appearance) the starch grains are consistently associated with plastid membranes, while in the other type (irregular, spiny appearance) the starch grains are associated with the plastid stroma only. We suggest that there are two enzyme systems for metabolizing starch in oat plastids—one membrane-bound and the other free in the stroma.     

ULTRASTRUCTURE OF CARPOSPOROGENESIS IN THE PARASITIC RED ALGA FAUCHEOCOLAX ATTENUATA SETCH. (RHODYMENIALES,RHODYMENIACEAE)

Carpospore differentiation in Faucheocolax attenuata Setch. can be separated into three developmental stages. Immediately after cleaving from the multinucleate gonimoblast cell, young carpospores are embedded within confluent mucilage produced by gonimoblast cells. These carpospores contain a large nucleus, few starch grains, concentric lamellae, as well as proplastids with a peripheral thylakoid and occasionally some internal (photosynthetic) thylakoids. Proplastids also contain concentric lamellar bodies. Mucilage with a reticulate fibrous substructure is formed within cytoplasmic concentric membranes, thus giving rise to mucilage sacs. Subsequently, these mucilage sacs release their contents, forming an initial reticulate deposition of carpospore wall material. Dictyosome vesicles with large, single dark-staining granules also contribute to wall formation and may create a separating layer between the mucilage and carpospore wall. During the latter stages of young carpospores, starch is polymerized in the perinuclear cytoplasmic area and is in close contact with endoplasmic reticulum. Intermediate-aged carpospores continue their starch polymerization. Dictyosomes deposit more wall material, in addition to forming fibrous vacuoles. Proplastids form thylakoids from concentric lamellar bodies. Mature carpospores are surrounded by a two-layered carpospore wall. Cytoplasmic constituents include large floridean starch granules, peripheral fibrous vacuoles, mature chloroplasts and curved dictyosomes that produce cored vesicles which in turn are transformed into adhesive vesicles. Pit connections remain intact between carpospores but begin to degenerate. This degeneration appears to be mediated by microtubules.     

Heterotrophic tobacco cell cultures during greening

Structural changes in cells and plastids are described that occur during the greening of an initially dark-grown cell suspension of tobacco ( Nicotiana tabacum L. cv. Xanthi). The pattern of cell growth during greening, expressed in dry weight or cell number, showed a classical sigmoid curve with a lag phase (T₀–T₂), an exponential phase (T₃–T₉) and a stationary phase (T₁₀–T₂₁). Achlorophyllous vacuolated cells (T₀), obtained after 3 culture cycles in the dark, contained amyloplasts devoid of lamellae. Exposure to light brought about an enrichment in cytoplasm and an amyloplast to proplastid transformation (starch loss) accompanied by chlorophyll synthesis. By T₃, many cells appeared meristematic and contained dividing proplastids with rudimentary single lamellae typical of those in intact meristematic leaf cells. As cell division occurred (T₃ to T₉), plastids replicated and their internal membrane system developed progressively into defined grana-intergrana thylakoids. By the stationary phase of cell growth (T₁₄), the lamellar system had reached a highly structured grana-intergrana network typical of higher plant chloroplasts. We have emphasized the analogies between the sequence of events (proplastid to chloroplast transition) during the greening of tobacco cells and that in developing intact leaves; in this respect the cell cultures provide a useful material for studies dealing with the biogenesis of structural or physiological events.     

Plastid Structure and Development in Green Callus Tissues of Oxalis dispar

Cultured callus tissues derived from endosperm of Oxalis disparare shown to contain virescent amyloplasts. In darkness, proplastidsdevelop into typical amyloplasts, starch being deposited assingle or multiple grains. In light, amyloplasts are transformedinto chloroplasts. Thylakoid formation begins in spaces aroundand between existing starch grains. As thylakoids are assembledinto grana, starch slowly disappears; the plastids increasein size and the photosynthetic apparatus enlarges to fill thewhole of the plastid. Slight carotenoid synthesis takes placeas amyloplasts are laid down, but there is no chlorophyll synthesis.All pigments accumulate rapidly during the early stages of granaldevelopment, but slowly, and at a declining rate, during thelater stages. Treatment of the tissues with auxins suppressesthe development of thylakoid membranes, but has no effect uponthe development of amyloplast membranes. The possible significanceof this observation is discussed. Greening is accompanied by a marked decline in the rates ofboth cell division and cell expansion. This is attributed inpart to the diversion of nitrogen from the normal growth channelsinto the synthesis of thylakoid proteins.     

Plastids with densely staining thylakoid contents inNymphoides indica

Summary Most chloroplasts of the lower epidermal and sub-epidermal cells in mature floating leaves ofNymphoides indica have densely staining thylakoid loculi which contrast markedly with electron translucent loculi of chloroplasts elsewhere in the leaves. Examination of leaf primordia (< 1 mm long) showed that granal membranes arise within the plastid stroma and already at that stage the stainable substance can be seen associated with the lamellae. In slightly enlarged primordia (5 mm long), nearly all plastids of the young leaf contain thylakoids with stained loculi, but by the time the leaf reaches 25 mm in length, the staining is restricted to the lower epidermal layers.     

DEVELOPMENT OF CHLOROPLAST FINE STRUCTURE IN ASPEN TISSUE CULTURE

Chloroplast ontogeny has been examined in 42-day etiolated triploid aspen callus (Populus tremuloides Michx.) subjected to two different light conditions. White and low-intensity red illumination showed little differences in their stimulatory effects on plastid development, the red light-irradiated plastids developing only slightly more slowly. Asynchronous plastid development was noted in both lighting systems. Etioplasts contained an interconnected tubular net, phytoferritin aggregates, electron-transparent vesicles which seem to invaginate from the inner plastid membrane, membrane-bound homogeneous spheroids and starch grains. Irradiation caused various morphological changes within the proplastids; the tubular complex became transformed into the more ordered prolamellar body-like structure from which radiated membrane-bound sacs filled with electron-dense material. These sacs, characterized as thylakoid precursors, were transformed into a thylakoidal system typical of mature chloroplasts. This ontogenetic scheme represents an additional pathway for the development of photosynthetic lamellae. Other light-induced changes in the developing plastid include disappearance of phytoferritin particles and homogeneous spheroids, decrease in starch content, and appearance of osmiophilic droplets.     

Ultrastructure of the mesophyll in scots pine and Norway spruce: Seasonal variation and molarity of the fixative buffer

Summary Resolution of the ultrastructure of the needles of both Scots pine (Pinus silvestris L.) and Norway spruce [Picea abies (L.) Karst.] is strongly influenced by the molarity of the buffer used in fixation. When 0.2 M or 0.1 M buffer is used in fixation during the summer, the constituents of the cytoplasm are precipitated, resulting in poor resolution of the membranes and lamellae and often in negative staining. The tannin in the central vacuole appears as a thick ribbon. By using correct molarities of buffer during each season (0.1 M for autumn and winter and ca. 0.05 M for the growing season), the best possible resolution will be achieved. With good resolution the tannin in the central vacuole appears in granular form throughout the year, and the cytoplasm and its organelles are clearly distinguishable during every season. During the growing season, the chloroplasts in the needles of Scots pine are spread to the cell walls and have large starch grains; the stroma and grana lamellae are well developed; the stroma and cytoplasm are rich in polysomes. Mitochondria and microbodies can be clearly resolved. During hardening and afterwords throughout the winter, the chloroplasts, which at this time contain no starch, and other cytoplasmic organelles aggregate in the corners of the cells. The chloroplast envelopes and the stroma and grana lamellae stay intact. The cytoplasm is netlike and rich in ribosomes, mitochondria and microbodies, all of which are intact and clearly distinguishable. During spring activation the structure returns to that described for the growing season.     

Leucoplasts: a distinct kind of organelles lacking typical 70S ribosomes and free thylakoids

Non-pigmented plastids were observed in fully differentiated cells from leaves and stem tissues of various species. Although showing important differences in size and shape, these plastids exhibit permanent structural features which allow to get them together as a distinct kind of organelles: the leucoplasts. Leucoplasts are distinct from the proplastids and every intermediate stage of plastid differentiation, from white chromoplasts and tuber amyloplasts. Mature leucoplasts do not contain an autonomous central system of thylakoids structurally independent from the envelope and, therefore, are never green. However, the envelope inner membrane invaginates within the plastid a cisternal or tubular stroma reticulum connected with the intermembrane space of the envelope. In addition, the leucoplast stroma is often less dense than chloroplasts stroma and contain several nucleoids with DNA fibrils. However, 70S ribosomes either scattered in the stroma or attached to the stroma reticulum or the envelope are not visible in ultrathin sections of leucoplasts stained with uranyl and lead. The existence of more discrete particles as dense as ribosomes is suggested. The relationship between the absence of ribosomes and thylakoids is discussed. Except for their specific role in C10 monoterpene synthesis in glandular cells, the functions of leucoplasts in plant cells remains largely up to now a matter of conjecture.     

Chloroplast ultrastructure,photosynthetic apparatus activities and production of steviol glycosides in <Emphasis Type="Italic">Stevia rebaudiana in vivo</Emphasis> and <Emphasis Type="Italic">in vitro</Emphasis>

The accumulation of steviol glycosides (SGs) in cells of Stevia rebaudiana Bertoni both in vivo and in vitro was related to the extent of the development of the membrane system of chloroplasts and the content of photosynthetic pigments. Chloroplasts of the in vitro plants, unlike those of the intact plants, had poorly developed membrane system. The callus cells grown in the light contained proplastids of almost round shape and their thylakoid system was represented by short thylakoids sometimes forming a little number of grana consisting of 2–3 thylakoids. In cells of the etiolated in vitro regenerants and the callus culture grown in the dark, only proplastids practically lacking the membrane system were observed. All the chloroplasts having developed thylakoids and forming at least a little number of grana were equipped with photochemically active reaction centers of photosystems 1 and 2. Leaves of in vivo plants accumulated greater amount of the pigments than leaves of the in vitro plants. In both the callus culture grown in the light and the etiolated in vitro regenerants, the content of the pigments was one order of magnitude lower than that in leaves of the intact plants. The callus tissue grown in the dark contained merely trace amounts of the pigments. Leaves of the intact and the in vitro plants did not exhibit any significant differences in photosynthetic O₂ evolution rate. However, photosynthetic O₂ evolution rate in the callus cells was much lower than that in the differentiated plant cells. The in vitro cell cultures containing merely proplastids did not practically produce SGs. However, after transferring these cultures in the light, both the formation of chloroplasts and the production of SGs in them were detected.