S351—1西瓜雄性不育的超微结构研究

在西瓜Ｓ３５１－１雄性不育材料遗传学及细胞分析的基础上，对其超微结构进行了观察，结果表明，次级造孢细胞在败育过程中，除细胞质壁分离、核解体消失外，细胞质中细胞器的形态与结构亦表现异常，尤其是溶酶体及液泡的数目与形态的变化。解体核周期细胞质中长丝状或封闭环状的内质网复合结构出现等，并对次级造孢细胞中液泡系的动态、内质网的结构异常与败育的关系进行了讨论。     

选择性自噬的研究进展

自噬是真核细胞中高度保守的一种亚细胞降解途径。该途径中,一些损坏、标记的大分子物质(如蛋白质等)或细胞器被自噬体包裹后送入溶酶体(动物)或液泡(酵母和植物)中降解并重新利用。长期以来,人们认为自噬在降解大量的细胞质成分时是一种非选择性的过程。然而,最近几年的研究结果表明,无论是酵母还是高等的真核生物细胞中均存在许多类型的选择性自噬。该研究主要综述了近几年选择性自噬过程与分子机制的研究进展。     

烤烟不同部位叶片超微结构与常规化学成分的关系

采用数字量化图像方法对烤烟不同部位叶片细胞超微结构进行量化,分析细胞面积、细胞周长、细胞形状因子、液泡面积、细胞壁厚度、液泡面积与细胞面积比值与烤烟化学成分的关系。结果表明:烤烟叶片表面细胞轮廓清晰,细胞壁内侧均有细胞核、叶绿体、线粒体等贴壁分布,中央大液泡填充于细胞中间;烤烟上部叶细胞面积、液泡面积、周长较小,细胞壁厚度较大;中部叶细胞面积、液泡面积、周长、细胞壁厚度较小;下部叶液泡面积、细胞壁厚度、液泡面积与细胞面积比值较大;烤烟叶片细胞壁厚度与蛋白质、氯含量呈正相关,与总糖含量呈负相关;细胞面积、细胞周长、细胞形状因子与烟碱、总氮含量呈正相关;细胞面积、液泡面积与还原糖含量呈正相关;细胞形状因子、液泡面积与细胞面积比值和蛋白质含量呈负相关。     

油菜素内酯对植物细胞钙离子分布的影响

【目的】明确油菜素内酯对Ca²⁺分布的影响,分析油菜素内酯对影响钙稳态的编码Ca²⁺通道和Ca²⁺-ATPase相关基因表达量的变化,明确油菜素内酯对钙稳态的影响。【方法】采用焦锑酸钙沉淀法对油菜素内酯（brassinosteroid,BR）处理后Ca²⁺的分布进行细胞化学定位;利用real-time PCR技术对调控细胞内Ca²⁺水平的位于细胞质膜、液泡膜和内质网上的编码Ca²⁺-ATPase的基因及位于细胞质膜、液泡膜和溶酶体上的编码Ca²⁺通道基因的表达量进行分析。【结果】在未经BR处理的拟南芥细胞中,Ca²⁺主要分布在细胞壁、细胞间隙和液泡中,细胞质和叶绿体中仅有少量Ca²⁺的分布;在1 µmol·L^-1 BR处理3 h后,Ca²⁺呈聚集状分布在液泡膜和细胞质膜附近,同时细胞质和叶绿体上的Ca²⁺分布增多;BR处理6 h后,细胞质和叶绿体中Ca²⁺分布继续增加,细胞壁中Ca²⁺分布有所减少;BR处理9 h后,细胞质和叶绿体中Ca²⁺分布减少,细胞间隙和液泡中Ca²⁺分布有所增加,但细胞壁中Ca²⁺分布明显减少,说明BR具有移除细胞壁中Ca²⁺的作用。CNGC2和CNGC12是细胞质膜上编码Ca²⁺通道的基因,在1 µmol·L^-1BR处理3 h后,CNGC2和CNGC12的表达量均明显下降;处理6 h后,CNGC2和CNGC12的表达量有所恢复;处理9 h后,CNGC2和CNGC12的表达量明显增加。TPC1和TPC2分别是液泡和溶酶体上钙离子通道相关基因,TPC1和TPC2的表达量在1 µmol·L^-1 BR处理3 h后也表现为明显下降,但TPC1的表达量在BR处理6 h后的表达量明显高于未用BR处理的对照,而TPC2的表达量直到BR处理9 h后才明显升高。可见,BR可阻滞细胞质中Ca²⁺浓度的快速上升,液泡膜上编码Ca²⁺通道基因的表达恢复早于细胞质膜和溶酶体上的Ca²⁺通道基因,说明液泡中Ca²⁺大量进入细胞质的时间早于胞外钙库和溶酶体等胞内细胞器。ACA8和ACA10是定位在细胞质膜上Ca²⁺-ATPase基因,1 µmol·L^-1 BR处理3和6 h后,ACA8和ACA10的表达量没有明显的变化;BR处理9 h后,ACA8和ACA10的表达量明显增加;ACA4和ACA11是液泡膜上编码Ca²⁺-ATPase的基因,BR处理后,ACA4和ACA11的表达量变化与质膜上的ACA8和ACA10的表达变化类似。ACA2是内质网上编码Ca²⁺-ATPase的基因,ACA2的表达量同样在BR处理9 h后出现了表达量的最高峰。可见,BR处理9 h后,Ca²⁺-ATPase表达量增加,把细胞质中高浓度的Ca²⁺泵入细胞间隙、液泡和内质网等胞外和胞内钙库中,调控细胞质中的Ca²⁺稳态。【结论】BR对第二信使Ca²⁺具有调控作用,并可通过对钙稳态调控系统的调控传递信号。     

The intracellular fate of Salmonella depends on the recruitment of kinesin

Salmonella enterica causes a variety of diseases, including gastroenteritis and typhoid fever. The success of this pathogen depends on its capacity to proliferate within host cells in a membrane-bound compartment. We found that the Salmonella-containing vacuole recruited the plus-end-directed motor kinesin. Bacterial effector proteins translocated into the host cell by a type III secretion system antagonistically regulated this event. Among these effectors, SifA targeted SKIP, a host protein that down-regulated the recruitment of kinesin on the bacterial vacuole and, in turn, controlled vacuolar membrane dynamics.     

水稻胚乳发育中ATP酶的超微细胞化学定位和功能分析

 应用磷酸铅沉淀技术 ,对水稻 (OryzasativaL .)胚乳发育中ATP酶进行了超微细胞化学定位研究。结果表明 ,在胚乳游离核期和细胞化期 ,胚囊壁、细胞核和质膜上有ATP酶活性分布。在生长分化期的早期 ,ATP酶主要定位于胚乳细胞质膜上。在灌浆高峰期 ,糊粉层细胞的质膜、胞间隙和胞间连丝上有显著的ATP酶活性 ;亚糊粉层间的质膜上ATP酶活性较高 ;淀粉胚乳细胞的质膜、衰退的细胞核上有ATP酶活性分布 ;胚乳细胞的液泡、蛋白体周围分布有ATP酶。综合观察结果 ,认为ATP酶主要参与胚乳对物质的吸收和贮藏蛋白质的合成。     

Apolipoprotein L-I promotes trypanosome lysis by forming pores in lysosomal membranes

Apolipoprotein L-I is the trypanolytic factor of human serum. Here we show that this protein contains a membrane pore-forming domain functionally similar to that of bacterial colicins, flanked by a membrane-addressing domain. In lipid bilayer membranes, apolipoprotein L-I formed anion channels. In Trypanosoma brucei, apolipoprotein L-I was targeted to the lysosomal membrane and triggered depolarization of this membrane, continuous influx of chloride, and subsequent osmotic swelling of the lysosome until the trypanosome lysed.     

Toxoplasma gondii: fusion competence of parasitophorous vacuoles in Fc receptor-transfected fibroblasts

After actively entering its host cells, the protozoan parasite Toxoplasma gondii resides in an intracellular vacuole that is completely unable to fuse with other endocytic or biosynthetic organelles. The fusion blocking requires entry of viable organisms but is irreversible: fusion competence of the vacuole is not restored if the parasite is killed after entry. The fusion block can be overcome, however, by altering the parasite's route of entry. Thus, phagocytosis of viable antibody-coated T. gondii by Chinese hamster ovary cells transfected with macrophage-lymphocyte Fc receptors results in the formation of vacuoles that are capable of both fusion and acidification. Phagocytosis and fusion appear to involve a domain of the Fc receptor cytoplasmic tail distinct from that required for localization at clathrin-coated pits. These results suggest that the mechanism of fusion inhibition is likely to reflect a modification of the vacuole membrane at the time of its formation, as opposed to the secretion of a soluble inhibitor by the parasite.     

益母草花蜜腺发育解剖学研究

益母草花蜜腺属于非淀粉型花盘蜜腺 ,环绕于子房的基部 ;由分泌表皮和泌蜜组织组成。分泌表皮分布有气孔器 ,蜜汁通过气孔向外分泌。蜜腺和雌蕊起源于共同的原始细胞 ,蜜腺是原始细胞较早形成的结构 ;花蕾膨大期细胞增大 ,细胞内有大液泡 ,后期细胞质增多 ,细胞内开始积累多糖类物质 ,泌蜜组织中形成维管束 ;花蕾露冠期除多糖类物质外 ,泌蜜组织中开始积累蛋白质 ;花蕾初放期 ,泌蜜组织内蛋白质继续增加 ;盛花期 ,蛋白质和多糖类物质的积累达到高峰 ,蛋白质的积累更明显 ,盛花期的后期蛋白质和多糖类物质相继消失 ,细胞内大液泡重新出现 ;花败期 ,泌蜜组织的细胞质和细胞核消失。     

Ultracytochemical Localization and Functional Analysis of ATPase During the Endosperm Development in Oryza sativa L.

Ultracytochemical localization of ATPase during development of rice endosperm was performed using a lead phosphate precipitation technique. The results indicated that, at the coenocyte and ceilularization stages, active ATPase was mainly distributed in an embryo sac wall, nucleus, and plasma membrane. At the early stage of development and differentiation, active ATPase was observed in the plasma membrane. At the grain filling stage, ATPase was highly active in the plasma membrane, intercellular space, and plasmodesmata in aleurone, moderately active on the plasma membrane in subaleurone. In starchy endosperm, ATPase was localized in the plasma membrane and degenerated nucleus. ATPase activity also appeared around vacuole and protein body in endosperm cell. The relationships between the ultracytochemical localization of ATPase and its function during the development of rice endosperm were discussed. Overall, ATPase was involved in the process of nutrition absorption and protein synthesis.     

作物根系镉滞留作用及其生理生化机制

一定程度的镉胁迫严重影响了作物的生长发育和农产品的产量及品质。文中全面综述了重金属镉胁迫对作物和人类的危害,以及镉在作物体内的吸收、转运和积累特征及其相关的主要调控基因和功能。简要概述了作物抗镉耐镉机制,重点讨论了其中的根系镉滞留作用的生理和生化机制。重金属镉主要通过根部吸收进入植株,在根中,Cd~(2+)首先进入由细胞间隙、细胞壁微孔以及细胞壁到质膜之间的空隙等构成的"自由空间",然后通过主动或被动吸收跨膜进入胞质,再经共质体或质外体途径运输到木质部导管中。水稻等作物主要通过下列途径来适应镉胁迫:细胞壁的滞留作用、原生质体的螯合作用、液泡的区室化作用、逆境蛋白和脯氨酸的积累、抗氧化酶系统活性的提高、根系的滞留作用。根系镉滞留作用作为一种重要的抗耐镉毒害的方式,在调控作物对镉的吸收、转运和分配积累,阻碍镉进入植株地上部和原生质体,减少镉对作物自身生长发育及农产品产量和品质的影响等方面起着非常重要的作用。主要包括根茎间低转运量导致的镉滞留、根系细胞壁滞留和液泡滞留。(1)根茎间低转运量导致的镉滞留。该种滞留作用主要受到根系木质部的镉装载能力和镉长距离运输载体——植物螯合肽(PCs)含量的影响;它们主要受到质膜上跨膜离子转运蛋白HMA2和HMA4以及细胞中的PCs合成酶及其相应基因(如HMA2、HMA4、PCs1等)的调控。这些蛋白和基因对木质部的镉滞留起到负调控作用。(2)细胞壁滞留作用。根系细胞壁滞留发生在质外体部分(包括细胞壁和胞间层),主要与质外体的组成成分和结构相关,其中起关键作用的是果胶多糖,半纤维素也起到一定作用。根据果胶和半纤维素滞留镉的作用方式的不同,细胞壁滞留作用可分为物理滞留和化学滞留。物理滞留主要与细胞壁的孔隙度和厚度有关,此二者均受到细胞壁果胶含量和果胶甲酯酶PME活性的影响。而化学滞留则是由半纤维素和低酯化果胶上的带负电荷基团,如-COO-等,与Cd2+发生静电结合作用所致。它们会受到PME14和XCD1等基因的调控。(3)液泡滞留作用。液泡滞留作用与细胞质和液泡中的PCs以及液泡膜上的转运蛋白密切相关,其对镉的滞留能力大小受到液泡分隔容量大小(VSC)的限制。在液泡的镉滞留中,不同分子量大小的PCs起到了重要作用,它们参与了胞质中镉的螯合、胞质与液泡间镉的转移及最终液泡中镉的沉积。而液泡膜上的转运蛋白则负责将胞质溶液中的低分子量PC-Cd复合物通过主动运输转移到液泡内,使镉被隔离在活跃生理区之外。作物根系中,这三种重金属滞留机制先后联合作用,降低了镉向原生质体和地上部的转移,从而减轻了镉对地上部的毒害,降低了籽实等收获器官中的镉含量。然而,由于木质部中PC-Cd占总镉比例以及细胞壁电荷总量和液泡VSC大小的有限性,从而使得根系镉滞留作用的强度和效果都存在着一定的限度。     

Effects of NaCl, Na2SO4 and mannitol on utilization of storage protein and transformation of vacuoles in the cotyledons and seedling roots of alfalfa (Medicago sativa L.)

The effects of NaCl, Na₂SO₄ and mannitol on utilization of storage protein in the cells of cotyledon mesophyll and root meristem during the seed germination of alfalfa (Medicago sativa L.) on the solutions of these substances at different concentrations and identical osmotic pressure were investigated using the transmission electron microscopy technique. Ultrastructural analysis revealed changes in the conversion of storage vacuoles to central vacuole, as well as changes in the quantity and form of residual protein bodies within vacuoles, depending on the osmotic effect of salt influence. At low concentrations corresponding to osmotic pressure of 202.6 kPa Na₂SO₄ did not affect the conversion of storage vacuoles to vegetative vacuole and the protein utilization. With increasing of concentration Na₂SO₄ and mannitol caused the retaining of considerable protein reserves, and mannitol did not disturb the transformation processes of vacuoles. NaCl did not prevent the central vacuole formation and protein utilization in cotyledon cells, but not in the root cells. When the concentration of the salt increased up to the value corresponding to the osmotic pressure of 607.8 kPa, NaCl can retard the storage protein mobilization in the root cells. These results demonstrate the sensitivity of processes of storage protein utilization during seed germination to stress factors caused by salinity.     

正常开颖授粉水稻T168和闭颖授粉水稻CL01的浆片结构比较

用OlympusAHs显微镜和日立透射电镜对闭颖授粉水稻CL01和正常开颖授粉水稻T168浆片的显微和亚微结构进行了观察。显微观察结果表明，闭颖授粉水稻材料维管束的数目和结构与正常开颖授粉水稻材料都有明显差异，闭颖授粉水稻材料浆片小且维管束数少，为8条左右，仅为正常开颖授粉水稻材料的一半。超微结构表明：开花前1d、开花当天及开花后，闭颖授粉水稻不管是液泡的形成过程还是线粒体变化过程都与开颖授粉水稻有明显的不同。闭颖授粉水稻在开花过程中没有大量长形或椭圆形线粒体积累，再加上其浆片小且维管组织不发达，从而导致了闭颖。     

Ankyrin repeat proteins comprise a diverse family of bacterial type IV effectors

Specialized secretion systems are used by many bacteria to deliver effector proteins into host cells that can either mimic or disrupt the function of eukaryotic factors. We found that the intracellular pathogens Legionella pneumophila and Coxiella burnetii use a type IV secretion system to deliver into eukaryotic cells a large number of different bacterial proteins containing ankyrin repeat homology domains called Anks. The L. pneumophila AnkX protein prevented microtubule-dependent vesicular transport to interfere with fusion of the L. pneumophila-containing vacuole with late endosomes after infection of macrophages, which demonstrates that Ank proteins have effector functions important for bacterial infection of eukaryotic host cells.     

苹果树腐烂病菌小G蛋白VmRab7基因的功能分析

苹果树腐烂病菌（Valsa mali）引起的真菌病害严重制约着苹果经济产业的发展,深入研究其致病机制,对该病害的防控十分重要。初步鉴定了小G蛋白VmRab7的功能,为解析苹果树腐烂病菌的致病机理和病害防控提供理论依据。采用同源重组的方法对VmRAB7进行基因敲除,获得ΔVmrab7敲除突变体;通过菌落直径的测量,分析VmRab7对菌丝生长速率的影响;通过离体枝条和叶片接种,分析VmRab7对致病力的影响;通过透射电镜观察,分析ΔVmrab7突变体的自噬体及液泡形态。研究发现,VmRAB7基因的缺失使菌丝生长速率降低了约40%,致病力降低了约50%,突变体丧失了产孢能力;ΔVmrab7敲除突变体具有小且多的碎片化液泡,液泡中无球状的自噬体结构。因此,VmRab7调控着苹果树腐烂病菌的营养生长、产孢、致病力、液泡融合和细胞自噬过程。     

植物硫代葡萄糖苷-黑芥子酶底物酶系统

硫代葡萄糖苷-黑芥子酶系统是十字花科植物中一种特有的底物酶系统.在植物体内,硫代葡萄糖苷存在于液泡中,而黑芥子酶分布在特定的黑芥子酶细胞中.在细胞内,黑芥子酶往往与黑芥子酶结合蛋白和黑芥子酶协助蛋白形成黑芥子酶复合体催化硫代葡萄糖苷的水解.当植物组织受到损伤时,硫代葡萄糖苷与黑芥子酶直接接触,并使硫代葡萄糖苷水解为异硫氰酸、腈等有毒物质来保护自身免遭食草动物进一步的伤害.     

Membrane recycling: vesiculation of the amoeba contractile vacuole at systole

Ultrastructural data on the protozoan Amoeba proteus support a model of membrane recycling. At systole the amoeba contractile vacuole fuses with the cell surface and expels its contents. Observations by electron microscopy indicate that, as the vacuole empties, its bounding membrane transforms into tiny (35 nanometers in diameter) vesicles, identical to the vesicles that segregate fluid and contribute to the diastolic vacuole.     

饲喂高蛋白日粮对雏鹅血清尿酸水平、肝脏和肾脏超微结构及ABCG2表达的影响

通过建立雏鹅高尿酸血症模型,研究高蛋白日粮对雏鹅血清尿酸水平、肝脏和肾脏超微结构及AB-CG2表达的影响.超微结构观察发现,对照组(CP)肝脏和肾脏细胞器结构完好,高蛋白1组(HP1)、高蛋白2组(HP2)肝脏部分线粒体开始出现肿胀、空泡变性,内质网肿胀消失,糖原减少.肾脏细胞部分线粒体开始出现肿胀、空泡变性,核膜间隙...     

蓝藻伪空胞的测定及其前处理方法研究

[目的]探讨蓝藻伪空胞的测定及其前处理方法。[方法]改进了测定伪空胞的毛细管压力法,对改进后的装置进行了检测限和精密度测试,并研究了2种浓缩方法和2种保存方法对3种蓝藻细胞数量和伪空胞含量的影响。[结果]改进后的蓝藻伪空胞测定装置其检测限达到0.001 8μl/m l藻液,同一样品测定结果相对标准偏差小于1%。采用过滤和离心2种浓缩方法时,分散单细胞的微囊藻细胞损失都大于50%,其中小群体的微囊藻采用离心的方法细胞损失较小,而丝状的颤藻采用过滤的方法损失较小;这2种方法对伪空胞含量的影响均可忽略。3种蓝藻在直接冷藏和添加鲁哥试剂保存7 d后,其细胞浓度和单位细胞的伪空胞含量均未发生明显改变,其中对于伪空胞,直接冷藏的损失更小;对于自然水体样品,宜添加鲁哥试剂进行保存。[结论]为研究蓝藻浮力调节机制及其暴发机理提供了理论依据。     

The Measurement of Membrane Potential and NO3 Activity in Root Cells Using Ion-Selective Microelectrodes

Remobilisation of nitrate in plants, especially in vacuole of plant, is mostly related to the qua-lity of agricultural products and the high nitrogen use efficiency in plants. Ion-selective microelectrodes offer anon-destructive and non-interruptive method to measure NO-3 gradients and electric potential differences acrossboth the plasma membrane and tonoplast. Thus, a double-barrelled microelectrode backfilled with a mem-brane sensor for NO-3 embedded in poly vinyl chloride (PVC) can record the NO-3 activity in cytoplasm and vac-uole of a cell. This paper presented how to make this kind of microelectrode and how to do the intracellularmeasurements on intact plants. Our result showed that nitrate activity was about 2.7 mmol L-1 in cytoplasmwhile 70 mmol L-1 in vacuole, which implicated that vacuole was a pool of nitrate in plants.