高等植物花色苷在液泡中的存在状态及其着色效应

综述了花色苷被摄入液泡的原因、花色苷在液泡中的存在状态及其对植物细胞的着色效应。花色苷在植物细胞质中合成后转运到液泡里是为了解除其对蛋白质和DNA等细胞功能分子的毒性。花色苷的液泡区隔化是花色苷在植物细胞中发挥正常功能的前提。在大多数植物中,花色苷在绝大多数情况下完全溶解在液泡里。但是,花色苷也能在液泡里形成颗粒,这些颗粒可以划分为花色苷体和花色苷液泡包涵体两类。花色苷体由膜包裹,其形成是液泡中小的有色囊泡逐渐合并的结果,发育完全的花色苷体为典型的球状、具比液泡更深的红色;液泡里的花色苷体具高密度,呈现为含高浓度花色苷的不溶性小球;花色苷体的存在可导致液泡的强烈色彩。花色苷液泡包涵体可能具备蛋白质基质,既无膜包裹又无内部结构,其形成是转运进液泡的花色苷与蛋白质基质结合的结果;液泡里的花色苷液泡包涵体形状不规则,象果冻;在花色苷液泡包涵体中,花色苷可能通过氢键连接于蛋白质基质的一个有限空间位点;花色苷液泡包涵体被认为是液泡中花色苷的"陷阱",优先摄取花色素3,5-二糖苷或酰化的花色苷;花色苷液泡包涵体的存在可增加液泡色彩的强度并导致"蓝化"。     

侵染细胞质膜附近小泡的来源及其作用

箭舌豌豆根瘤幼龄侵染细胞的壁和质膜比较光滑,成熟侵染细胞与此不同,不仅细胞壁厚薄均,有较多的胞间连丝,而且质膜常常内陷形成各种突起,然后离质膜形成小泡。这些位于质膜附近的小泡体积较小,多呈圆形,既可单独存在,也可多个聚在一起。在向细胞中央移动中,有的小泡靠近细胞质膜,甚至与细菌周期融合,有的小泡不民附近的小液泡融合变为较大液泡,并常用降解程度不同的细菌位于其中,在衰老侵染细胞中,细胞壁附近有小泡,     

植物R-SNARE蛋白研究进展

真核细胞中囊泡膜与靶膜的融合是囊泡运输的关键环节,由进化保守的SNARE(soluble N-ethylmaleimide-sensitive factor attachment protein receptor)蛋白家族介导完成。SNARE蛋白可被分为定位于囊泡的R-SNARE(v-SNARE)和定位于靶膜的Q-SNARE(t-SNARE)两大类。R-SNARE与QSNARE的特异性配对形成“SNARE复合物”,该复合物可介导囊泡膜与靶膜融合。与酵母和动物相比,植物R-SNARE基因在进化过程中经历了大量扩增,推测其与植物细胞特有的胞内转运途径有关。该文综述了R-SNARE参与植物发育和胁迫响应的研究进展,结合其亚细胞定位信息探讨了不同RSNARE的作用靶位和调控特点,并对该领域研究前景进行了展望。     

植物细胞核的凹入和核液泡的形成

核质互作在细胞核中形成膜囊结构首先是在动物细胞中揭示的［１］。对植物细胞超微结构的研究亦发现有类似现象的存在［２—４］。在植物细胞核中形成的膜泡认为有两种形式：一是核被膜向核基质深度凹入,形成细胞质深入细胞核的状态,称之为“假包被（ｐｓｅｕｄｏ－ｉｎｃｌｕｓｉｏｎ）”［２,５］;另一种形式是细胞核内膜或内外双层核膜向核基质深度凹入,并最终脱离核被膜,在核基质中形成囊泡结构,称之为“核液泡（ｎｕｃｌｅａｒｖａｃｕｏｌｅ）”［３,４,６,７］。对细胞核质间通过核被膜在结构上的特殊作用形式缺少像对通过…     

冬季沙冬青叶肉细胞液泡中泡状内含物的研究

用透射电镜观察了沙冬青叶肉细胞液泡中泡状内含物的形成。在早期,这种泡状内含物位于细胞质中,它由大小不等、形态各异的小泡组成,后经液泡膜内吞进入液泡。液泡中的泡状内含物主要位于两个正常叶绿体之间,附近的细胞质较多,内有丰富的内质网、高尔基体、质膜管状突起和由它们产生的小泡。也有一些液泡泡状内含物出现在解体叶绿体附近。前者主要来自内质网、高尔基体和质膜,后者则主要起源于解体的叶绿体。这种泡状内含物的形成可能与增强植物的抗冻性有关。     

甜菊愈伤组织分生区细胞中膜内含物的超微结构研究

生长在分化培养基上的甜菊（Ｓｔｅｖｉａ ｒｅｂａｕｄｉａｎａ）愈伤组织分生区细胞中存在双膜和多膜内含物。电镜观察表明,这些膜内含物是由一圈或多圈呈同民贺或卷绕状排列的内质网包围部分细胞质而形成的。双膜内含物内外层膜的靠细胞质表面有核糖体附着,而多膜内含物仅在其最外层潴泡的外膜上偶有和量核糖体附着。附着细胞液泡化程度的提高,多膜内含物通过液泡膜内陷而转移到液泡中或通过消化其中被包围的细胞质及内膜而转     

非洲狼尾草未受精无孢子生殖胚囊退化研究

为理解植物无孢子生殖胚囊未受精条件下的退化,对无孢子生殖植物非洲狼尾草未受精成熟胚囊中央细胞退化做了细胞形态学研究。没有受精的中央细胞退化时最显著的特点是细胞核产生核膜囊泡。核膜囊泡有两种类型:单层膜的囊泡和双层膜的囊泡,单层膜囊泡在细胞质中,双层膜囊泡在细胞核内。核膜囊泡有两种发生方式:1)核膜的外膜向细胞质一侧膨胀产生囊泡,囊泡进入细胞质;2)核膜向核内凹陷形成囊泡,囊泡进入细胞核。核膜囊泡类型与产生方式密切关联。核膜囊泡吞噬并消化包括线粒体在内的细胞质和核质。     

水稻胚乳细胞质膜内陷的超微结构和磷酸酶的细胞化学定位

对生长分化期水稻胚乳细胞的质膜内陷进行了超微结构和磷酸酶的细胞化学研究。结果表明 ,胚乳细胞内的小泡、内质网常与胞间连丝相连 ;质膜形态多变 ,功能活跃 ,由局部起伏的波纹状发展成明显内陷 ,深浅不一 ,多呈袋状 ,袋中包含着大小不一的泡状物 ;有些内陷脱离质膜成为胞质中的囊泡 ,表现出活跃的内吞现象。除细胞间隙中含有圆球状的内含物外 ,在质膜内陷和囊泡中常含有大量的内含物。H ATP酶定位结果显示 ,质膜及其邻近的泡状物周围有酶的分布 ;而酸性磷酸酶定位在液泡、胞间隙和其中的泡状内含物周围 ;在质膜及其内陷形成的囊泡中有G6P酶的分布。这些结果表明胞间隙和质膜内陷在物质的运输中可能起着重要作用     

拟南芥愈伤组织细胞类58K蛋白定位及多泡体的分泌途径

通常认为多泡体是原生质体和胞饮作用中的一种特殊的细胞结构, 是由细胞质膜的内陷所起源. 然而多泡体的起源、功能及其分泌途径以及与高尔基体和细胞壁的关系等问题一直存有争论. 本文应用动物细胞的58K蛋白的单克隆抗体、免疫印记、激光共聚焦显微技术在拟南芥和贯叶连翘愈伤组织中成功定位了类58K蛋白. 免疫电子显微镜的观察结果显示, 类58K蛋白存在于拟南芥愈伤组织细胞的高尔基体库槽、分泌泡、多泡体、细胞壁和液泡中. 由此我们认为多泡体与细胞壁和液泡发育的关系十分密切, 它起源于高尔基体并以分泌泡的形式将高尔基体所合成的物质运送至细胞壁和液泡. 所以多泡体的分泌途径可能是运送高尔基体合成的物质到细胞壁, 属于胞吐形式; 也有可能是运送相关物质到液泡中参与液泡的发育, 属于胞饮形式. 这主要取决于多泡体所运送物质的种类以及它将这些物质运往何处. 因此多泡体的分泌途径很可能有多种形式. 研究结果也显示应用动物细胞58K蛋白抗体可以定位植物细胞中的类58K蛋白, 说明植物细胞中存在与动物细胞相类似的蛋白.     

箭舌豌豆根瘤液泡中细菌周膜来源的研究

电镜观察结果表明,幼龄箭舌豌豆根瘤侵染细胞的细胞质较少,中央是一些体积较大的液泡。细胞质中侵入线经常可见,由侵入线释放出来的细菌均有细菌周膜。这些细菌只位于细胞质中,不出现在液泡里面。成熟根瘤中的侵染细胞与此不同,它们中有大量的成熟侵染细胞,细胞质丰富,里面充满大量细菌,中央常有一个大液泡。当中央液泡发育到一定程度时,位于其附近的细菌可通过液泡膜内吞、液泡膜与细菌周膜融合及液泡膜破裂3种途径进入液泡,后一种途径常伴有寄主细胞质。液泡中的细菌绝大部分裸露在外,只有个别细菌具有细菌周膜且多位于液泡膜的破损处附近,因此细菌周膜可能是原来就有的。     

In vivo grapevine anthocyanin transport involves vesicle-mediated trafficking and the contribution of anthoMATE transporters and GST

In cells, anthocyanin pigments are synthesized at the cytoplasmic surface of the endoplasmic reticulum, and are then transported and finally accumulated inside the vacuole. In Vitis vinifera (grapevine), two kinds of molecular actors are putatively associated with the vacuolar sequestration of anthocyanins: a glutathione-S-transferase (GST) and two MATE-type transporters, named anthoMATEs. However, the sequence of events by which anthocyanins are imported into the vacuole remains unclear. We used MYBA1 transformed hairy roots as a grapevine model tissue producing anthocyanins, and took advantage of the unique autofluorescence of anthocyanins to study their cellular trafficking. In these tissues, anthocyanins were not only visible in the largest vacuoles, but were also present at higher concentrations in several vesicles of different sizes. In the cell, small vesicles actively moved alongside the tonoplast, suggesting a vesicular trafficking to the vacuole. Subcellular localization assays revealed that anthoMATE transporters were closely related with these small vesicles, whereas GST was localized in the cytoplasm around the nucleus, suggesting an association with the endoplasmic reticulum. Furthermore, cells in hairy roots expressing anthoMATE antisense did not display small vesicles filled with anthocyanins, whereas in hairy roots expressing GST antisense, anthocyanins were accumulated in vesicles but not in the vacuole. This suggests that in grapevine, anthoMATE transporters and GST are involved in different anthocyanin transport mechanisms.     

Digestion in the peritrich ciliateOphrydium versatile

Summary Digestion in the peritrich ciliateOphrydium versatile O.F.M. involves a complex sequence of intracytotic and exocytotic membrane fusion and recycling events. Food particulates are concentrated in the lower cytopharynx which forms a fusiform-shaped food vacuole. Upon release from the cytopharynx, this food vacuole begins to condense, concentrating the food particulates. Excess membrane is removed intracytotically. These released membranes pieces form discoidal vesicles which are recycled to the base of the cytopharynx, thus providing additional membrane for subsequent food vacuole formation. In the condensed food vacuole, digestion proceeds; hydrolytic enzymes are delivered to the food vacuole via rough endoplasmic reticulum and/or by the cup-shaped coated vesicles (CSCV). As these vesicles fuse with the food vacuole, the food vacuole enlarges, digestion proceeds and an electron-dense membrane coat appears along the luminal surface of the food vacuole. Prior to defecation, the food vacuole undergoes a final condensation; irregularly-shaped, electron dense, single-membrane bound vesicles are cut-off intracytotically from the old food vacuole. These vesicles undergo condensation and invagination to form the cup-shaped coated vesicles (CSCV) which fuse with younger food vacuoles.     

Phagosome fusion vesicles of paramecium. I. Thin-section morphology

Ultrastructural studies of the digestive system of Paramecium caudatum focusing on the first 5 min of digestive-vacuole age reveal a set of vesicles, named phagosome fusion vesicles (PFVs), which fuse with the digestive vacuole just after the vacuoles are released from the cytopharynx and concomitant with vacuole acidification. Serial thin-sections of vacuoles labeled with horseradish peroxidase (HRP) and/or latex beads in pulse-chase studies were observed. PFVs, irregularly shaped, electron-translucent vesicles ranging from a small diameter to over 1 micro, are first seen in the region of the cytopharynx where they bind to the nascent vacuole membrane. Within 30 sec of vacuole release the PFVs fuse with the vacuole where they remain for a brief time connected to the vacuole by a narrow annulus. HRP-reaction product is found in vacuoles but not in PFVs before PFVs fuse with the vacuoles. After fusion with PFVs HRP is quickly inactivated. Tubular extensions of vacuole membrane then form between the fused PFVs. By 3 to 5 min both PFVs and tubules disappear from the vacuole surface and lysosomes appear in their place. We believe the tubules are pinched off as PFV membrane is being added to the vacuole. Microfilaments coat the membrane during all these dynamic events. Since the pH of the vacuole becomes acid during the first few minutes, we are now looking for a direct correlation between PFV fusion and acidification.     

Vacuole import and degradation pathway: Insights into a specialized autophagy pathway

Glucose deprivation induces the synthesis of pivotagluconeogenic enzymes such as fructose-1,6-bisphos-phatase, malate dehydrogenase, phosphoenolpyruvatecarboxykinase and isocitrate lyase in Saccharomycescerevisiae. However, following glucose replenishment,these gluconeogenic enzymes are inactivated and de-graded. Studies have characterized the mechanismsby which these enzymes are inactivated in response toglucose. The site of degradation of these proteins hasalso been ascertained to be dependent on the dura-tion of starvation. Glucose replenishment of short-termstarved cells results in these proteins being degradedin the proteasome. In contrast, addition of glucose tocells starved for a prolonged period results in theseproteins being degraded in the vacuole. In the vacuoledependent pathway, these proteins are sequestered inspecialized vesicles termed vacuole import and degra-dation (Vid). These vesicles converge with the endo-cytic pathway and deliver their cargo to the vacuolefor degradation. Recent studies have identified thatinternalization, as mediated by actin polymerization, isessential for delivery of cargo proteins to the vacuolefor degradation. In addition, components of the targetof rapamycin complex 1 interact with cargo proteins during glucose starvation. Furthermore, Tor1p dissoci-ates from cargo proteins following glucose replenish-ment. Future studies will be needed to elaborate on the importance of internalization at the plasma membrane and the subsequent import of cargo proteins into Vid vesicles in the vacuole dependent degradation pathway.     

In vitro reactions of vacuole inheritance in Saccharomyces cerevisiae

Vacuole inheritance is temporally coordinated with the cell cycle and is restricted spatially to an axis between the maternal vacuole and the bud. The new bud vacuole is founded by a stream of vacuole-derived membranous vesicles and tubules which are transported from the mother cell into the bud to form the daughter organelle. We now report in vitro formation of vacuole-derived tubules and vesicles. In semi-intact cells, formation of tubulovesicular structures requires ATP and the proteins encoded by VAC1 and VAC2, two genes which are required for vacuole inheritance in vivo. Isolation of vacuoles from cell lysates before in vitro incubation reveals that formation of tubulovesicular structures requires cytosol as well as ATP. After forming tubulovesicular structures, isolated vacuoles subsequently increase in size. Biochemical assays reveal that this increase results from vacuole to vacuole fusion, leading to mixing of organellar contents. Intervacuolar fusion is sensitive to the phosphatase inhibitors microcystin-LR and okadaic acid, suggesting that protein phosphorylation/dephosphorylation reactions play a role in this event.     

FINE STRUCTURE AND HISTOCHEMISTRY OF VESICLE CELLS OF THE RED ALGA ANTITHAMNION DEFECTUM (CERAMIACEAE)1

The ultrastructure and histochemistry of the refractile, vesiculate cells (“blasenzellen,”“cellules secretrices,”“gland cells”) of Antithamnion defectum Kylin were examined. The refringent vacuolar contents disclosed two components of differing density: an electron opaque, proteinaceous matrix material surrounding cores of irregularly shaped, less opaque material. The cores contain less protein and more unknown material than the matrix. Part or all of the vacuolar material is synthesized by abundant rough endoplasmic reticulum (ER) and deposited in smooth surfaced cisternae that swell to form vesicles. Mitochondria are usually associated with stacks of the swelling cisternae. The vesicles enlarge by continued deposition of synthesized material and coalescence with other vesicles. All vesicles eventually coalesce to form the mature vacuole. A crystalline array of fibrils develops in the cytoplasm during later stages of vacuole enlargement. The crystal contains a sulfated, acidic polysaccharidic material. The chloroplasts, if present, and nucleus degenerate at vacuole maturity. Active release of the vacuolar material does not occur, and organelles for extracellular secretion are not present. Structural evidence suggests a storage, rather than secretory, function for the cells.     

The Vid vesicle to vacuole trafficking event requires components of the SNARE membrane fusion machinery

The key gluconeogenic enzyme fructose-1,6-bisphosphatase (FBPase) is targeted to Vid vesicles when glucose-starved cells are replenished with glucose. Vid vesicles then deliver FBPase to the vacuole for degradation. A modified alkaline phosphatase assay was developed to study the trafficking of Vid vesicles to the vacuole. For this assay, FBPase was fused with a truncated form of alkaline phosphatase. Under in vivo conditions, FBPase-delta60Pho8p was targeted to the vacuole via Vid vesicles, and it exhibited Pep4p- and Vid24p-dependent alkaline phosphatase activation. Vid vesicle-vacuole targeting was reconstituted using Vid vesicles that contained FBPase-delta60Pho8p. These vesicles were incubated with vacuoles in the presence of cytosol and an ATP-regenerating system. Under in vitro conditions, alkaline phosphatase was also activated in a Pep4p- and Vid24p-dependent manner. The GTPase Ypt7p was identified as an essential component in Vid vesicle-vacuole trafficking. Likewise, a number of v-SNAREs (Ykt6p, Nyv1p, Vti1p) and homotypic fusion vacuole protein sorting complex family members (Vps39p and Vps41p) were required for the proper function of Vid vesicles. In contrast, the t-SNARE Vam3p was a necessary vacuolar component. Vid vesicle-vacuole trafficking exhibits characteristics similar to heterotypic membrane fusion events.     

甜菊愈伤组织细胞中的液泡膜内突和液泡内囊泡

对生长在分化培养基上的甜菊愈伤组织分生区细胞的液泡膜内突和液泡内囊泡,进行了超微结构和酸性磷酸酶细胞化学研究。在不同液泡化时期的细胞中,都存在不同大小和形态的液泡膜内突,它们有的缺乏明显的内含物;有的含有许多小泡或复杂膜系;有的含有一个较大的具许多小泡或复杂膜系的膜束缚囊泡。在液泡内还存在一些游离的液泡内囊泡,它们通常具有两层紧贴的界膜或为多层同心膜,推测它们来自液泡膜内突。ＡｃＰａｓｅ定位结果显     

Involvement of smooth and coated vesicles in the function of the contractile vacuole complex of some cryptophycean flagellates

The contractile vacuole complex of cryptophycean flagellates comprises the contractile vacuole, a pore and a vesicular spongiome. A minority of spongiome vesicles bear a 15-nm coat on the cytoplasmic surface of the membrane. The coat superficially resembles a clathrin coat. The majority of vesicles are smooth surfaced. Both types of vesicles are found at the same time. Smooth vesicles can be seen in profile suggesting vesicle-vesicle and vesicle-vacuole fusion. It is suggested that smooth vesicles are involved in the segregation of fluid from the cytoplasm and in filling the vacuole. Coated elements exist only as independent vesicles and as coated pits in the contractile vacuole membrane. There is no evidence of fusion of coated vesicles. It is suggested that coated vesicles function to retrieve specific membrane components from the contractile vacuole.