百合花粉母细胞间细胞融合期间腺苷三磷酸酶活性的细胞化学定位及其与染色质胞间转移的关系

用标准的磷酸铅沉淀的细胞化学方法,对百合花粉母细胞间染色质穿壁运动期间及其前后三个时期中的腺苷三磷酸酶(ATP 酶)活性进行了超微结构的定位。结果表明:(1)在穿壁前,ATP 酶活性主要定位于质膜、胞间连丝及细胞间隙;在内质网、高尔基体、质体和某些局部的基质(groundplasm)中,也表现有 ATP 酶活性反应的产物;但在染色质和核仁中,一般都没有这种反应。(2)在穿壁时,染色质从一个细胞穿壁转移到另一个相邻细胞,同时看到染色质和核仁内出现密集的 ATP 酶活性反应产物;在内质网和高尔基体的腔内以及质体的片层上也产生明显的 ATP 酶活性反应;而在质膜、胞间连丝及细胞间隙内 ATP 酶活性明显降低,甚至看不到明显的活性反应。(3)在穿壁后,质膜及细胞间隙中又产生明显的 ATP 酶活性反应产物,但核内染色质上的 ATP 酶活性则显著降低,而核仁内则仍有较高的活性。同前二个时期一样,内质网、高尔基体和质体上的 ATP 酶仍表现明显的活性反应。最后讨论了三个不同发育时期 ATP 酶活性及其分布部位的改变与染色质胞间转移的关系。     

烟草花粉母细胞细胞融合过程中细胞骨架的超微结构观察

应用普通电镜和DGD去包埋技术 ,研究了烟草花粉母细胞中的细胞融合现象及细胞融合过程中细胞骨架的变化。观察发现 ,处于凝线期的花粉母细胞 ,其内含物 ,包括细胞器和染色质 ,主要通过胞质通道向相邻细胞发生转移。DGD去包埋观察发现 ,花粉母细胞中核骨架与细胞质内及胞间连丝和胞质通道内胞质骨架连接成一个整体。在整个细胞融合过程中 ,均有核骨架纤维与染色质相连。本文讨论了细胞骨架在细胞融合过程中的作用     

西瓜柱头乳突细胞分泌活动期间ATP酶活性超微结构定位

研究了西瓜柱头乳突细胞ＡＴＰ酶活性的超微结构定位。分泌活动旺盛的细胞中,质膜、内质网、质体的内部片层、胞间连丝以及多数大液泡的膜上面都有大量ＡＴＰ酶活性反应产物,线粒体和小泡上只有少量酶活性反应产物。分泌活动停止后处于解体状态的细胞内,反应产物主要定位于液泡膜上。分泌旺盛的乳突细胞质膜具有高的ＡＴＰ酶活性表明分泌物运出需要大量能量,内质网ＡＴＰ酶活性强可能意味着该细胞参与分泌物合成。     

在三维结构上对百合花粉母细胞actin的免疫定位

传统的切片仅仅能够显示样品的平面结构,不能用于细胞中三维网络结构的研究。笔者在DGD(diethylene glycol distearate)包埋去包埋的基础上,结合电镜免疫胶体金技术对大卫百合花粉母细胞胞间及胞内细胞的骨架系统进行了研究,观察到高反差细胞微梁结构的三维网络,actin这一细胞骨架的主要成员被定位在该微梁结构纤维上。三维结构上的研究表明,actin不但是植物细胞核及细胞质骨架的成员,而且也存在于胞间连接结构(胞质桥和胞间连丝)中,推测它可能与细胞融合有关。实验结果同时表明,三维结构免疫胶体金技术对于细胞骨架和核基质的结构蛋白研究是行之有效的。     

Changes of Microtubular Skeleton in the Generative Cell in Pollen Tube During Mitosis of Lilium davidii

Previous observations indicated that division of the generative cell (GC) in some plant genura such as Lilium and Tradescantia is characterized by several unusual features, including persistence of surrounding microtubule (MT) bundles during mitosis, lacking a matephase plate, the cytokinesis is completed with furrow. The authors have further studied the changes of MT organizations and the chromosome (CHs) behavior in the GC during mitosis using electron microscopy and method of tubulin localizations. No MTs in the GC before GC division and during prophase was seen under electron microscopy. However, there was tubulin in the GC with antitubulin staining. During promatephase to matephase, the CHs appeared and arranged in a complexed tangled pattern lengthwise along the cell. Correspond- ingly, transverse pairs of kinetochores were located along the length and depth of the cell. They stacked successively like the rungs of a ladder. In this phase, a large mount of MTs appeared in the GC, which distributed in the cortex of the cell and among the CHs and along the whole length of the CHs. In the beginning, one or two kinetochore pairs changed from transversely to longitudinally situated in each cell. MTs ended on the kinetochore to form kinetochore MTs (KMTs). With the electron microscopy, authors did not find the image of lateral connection between the MTs and the kinetochores as previous reported with immunofluorescent method. As karyokinesis proceeded, more transverse kinetochore pairs gradually became longitudinal, and KMTs gradually increased in number. However, a distinct spindle was not evidenced. During anaphase, CHs seperation started at various positions along the length of the cell. The distribution of MTs in the GC was similar to that of promatephase to matephase. In late anaphase, the CHs segregated as two groups. Most MTs disappeared but only some remained in the polar regions and the interzone. Authors also measured and compared the lengths of the CHs and indirectly identified the existing anaphase B. During late tolephase, the MTs increased in number gradually in the region between the two newly formed sperm nuclei. The region might be the MT interdigitating zone visualized with antitubulin localization. The MTs disappeared after the cell plate (CP) appeared.     

辐照大麦M~1花粉母细胞染色体畸变及其与M~2 性状变异的关系

大麦种子经5种不同剂量γ射线辐照后, M1植株的花粉母细胞出现多种染色体畸变类型,例如后Ⅰ、后Ⅱ中出现桥、落后,后Ⅰ中出现不对称分裂,后Ⅱ中出现不同步分裂,中期Ⅰ出现环状四价体,末期Ⅰ中有三极分裂和微核,四分体中出现微核且有的微核独立于四分体,从而形成多分体。花粉母细胞的染色体畸变率随剂量的增高而升高,二者呈极显著直线相关关系。 M1花粉母细胞的染色体畸变率与M2的叶绿素突变率及农艺性状变异率呈显著或极显著指数函数关系。 M2花粉母细胞的染色体畸变率低于M1。 Abstract:Barley seeds were irradiated with five different doses of 60 Co-γ ray.There were many kinds of chromosome aberration in pollen mother cells,such as bridge,laggards in anaphase I and anaphase II,asymmetrical division in anaphase I and asynchronous division in anaphase II,ring quadrivalent in metaphase I,three pole division and micro-nuclei in telophase I,micro-nuclei in tetrad and some micro-nucleus separates from the tetrad to form polyad.The frequency of chromosome aberration in pollen mother cells of M1 increased with the increase of dose.There was a significant linear relationship between them.The frequencies of agronomic character mutation and chlorophyll mutation in M2 significantly correlated with the frequencies of chromosome aberration in pollen mother cells of M1 and showed an exponential function.The chromosome aberration frequency in pollen mother cells of M2 was lower than that of M1.     

The Relationship Between the Intercellular Chromatin Migration of Pollen Mother Cells and the Changes of Chromosome Numbers During the Genesis of Male Gametes in Allium Cepa

During the genesis of microspores and male gametes, the percentages of cells with an increas- ed and decreased chromosome number at metaphase Ⅱ, mitotic metaphase of microspores and generative cells approximated to the one at previous stage (metaphase Ⅰ, metaphase Il, and microspores) respectively, and the one at metaphase 1 again approximated to the percentages of chromatin intercellular migration of type 2.3→1 and 1→2.3 respectively, which had no significant diversity statistically. According to this fact, it was considered that the changes of chromosome numbers were related to the chromatin intercellular migration at synizesis stage and that pollen mother cells with an abnormal chromosome number at metaphase Ⅰ basically were able to continue developing untill the formation of male gametes.     

黑芥与花椰菜体细胞杂种花粉母细胞的减数分裂不同步及染色体行为异常

本研究对黑芥与花椰菜体细胞杂种及其双亲的花粉母细胞减数分裂行为进行了观察和比较,结果表明黑芥与花椰菜的花粉母细胞减数分裂同步性很高,而其体细胞杂种表现出明显的减数分裂不同步,各杂种不同株系间减数分裂不同步的程度与跨度各异.减数分裂不同步发生是由于为部分花粉母细胞在减数分裂某一时期发生停滞或推迟,这一时期(或第一个停滞时期)主要发生在第一次减数分裂的前期.花粉母细胞发育虽不同步,但最后均完成减数分裂形成花粉粒,所形成的花粉粒的活性相差甚远.此外,研究结果还显示,体细胞杂种有着较高的减数分裂染色体行为异常频率,尤其是在中期Ⅰ后.主要表现为减数分裂中期Ⅰ的染色体的早迁、后期Ⅰ和后期Ⅱ的染色体滞后及染色体不均等分离、四分体时期的异常等现象.花粉粒的活性与减数分裂时期染色体行为异常程度有关,染色体行为异常及微核现象出现的频率越高,相对的花粉粒活力也越低.     

Ultrastructural Observation on the Intra- and Intercellular Microtrabecular Network of the Pollen Mother Cells in Onion (Allium cepa)

Using DGD embedment-free electron microscopy, ultrastructural observation on the intra- and intercellular microtrabecular network (MN) of the pollen mother cells (PMC) of the whole meiotic prophase Ⅰ in onion ( Allium cepa L.) was performed. Complex nuclear MN was observed in the nucleus of PMCs, spreading throughout the nuclear region. The nucleolus and chromosomes were connected with the MN filament network. The uniformity of nuclear MN changed with the development of the PMCs. A lamina-like structure surrounded the nucleus and joined the MN in nucleus and in cytoplasm, but it disassembled at the end of prophase Ⅰ. There was also a complex cytoplasmic MN in PMCs, without obvious variation during the prophase Ⅰ. Furthermore, MN in cytoplasmic connections (plasmodesmata and cytoplasmic channels) was noticed to link the frameworks in two neighboring PMCs into one entity. Cytomixis was observed at synizesis of prophase Ⅰ in this experiment, and MN in cytoplasm and in nucleus was noticed to distribute in these granules which migrated from one PMC into its neighboring cell. At this time the nucleus moved aside from center of the PMC, but the rest of the cell was still fulfilled with MN filaments. The relationships of nuclear MN with nucleolus and chromosomes, lamina with nucleus, as well as intra- and intercellular MN with cytomixis are discussed in this paper.     

洋葱花粉母细胞细胞内、细胞间微梁骨架的超微结构观察(英文)

以洋葱(AlliumcepaL.)花粉母细胞为材料,采用DGD包埋去包埋原位技术,对花粉母细胞不同发育时期的细胞内、细胞间微梁骨架的超微结构进行了电镜观察。结果发现,花粉母细胞核内存在粗细不等的微梁骨架,与核仁和染色体紧密相连,随着发育的推移,其均一性发生改变。在核周有核纤层样的结构存在,与细胞核和胞质中的微梁骨架紧密相连,到前期结束时解体。洋葱花粉母细胞内具有发达的胞质微梁骨架,这种结构在减数分裂前期Ⅰ变化不明显。在胞间连接(胞间连丝和胞质通道)内,也有精细的微梁骨架分布,并且与两端细胞中的骨架相连。在凝线期的花粉母细胞中观察到细胞融合现象,有胞质或核内微梁骨架与穿壁转移的胞质小球和核小球内骨架相连。此时细胞核偏向一边,但细胞的其余部位仍充满了胞质微梁骨架。初步探讨了核微梁骨架与核仁和染色体之间的关系,核纤层与细胞核之间的关系,以及细胞内、细胞间微梁骨架与细胞融合之间的关系     

Ultrastructural Localization of Adenosine Triphosphatase Activity in Stigma Cells of Populus lasioearpa and Its Changes During Development

The ultrastructural localization of adenosine triphosphatase (ATPase) activities in stigmatic cells in various developmental stages of PopUlus lasiocarpa was investigated using the cytochemical method Of read phosphate precipitation. The results show as follows: 1. Lead deposits which marked the ATPase activities were observed on the pellicle of stigmas. The ATPase activities greatly increased in receptive stage, but they were few or even absent in young and old stages. The changes Of pellicle ATPase strongly exhibited that ATPase was correlated with the pollen-stigma interaction. 2. In the stigma ceils inreceptive stage, ATPase was mainly located at mitochondria cristae, chloroplast lamellae and endoplasmic reticulum. Lead deposits were also visible on the plasmalemma, plasmodesma, nuclear membrane and in nucleoli. No lead deposits were found on dictyosome and vacuole membrane. 3. During the degeneration of stigmatic ceils; the location of ATPase changed. The distribution of ATPase was in vacuole membrane, digestive vesicle, mitochondrium envelop, chloroplast envelop, and digested fragment. The mitochondrium cristae and chloroplast lamellae where ATPase was the most active in previous stage now lost their ATPase activities.     

Effects of High Temperature on the Development of Pollen Mother Cells and Microspores in Barley Hordeum vulgare L.

Received 15 May 2000/ Accepted in revised form 26 August 2000     

Combining Magnetic Sorting of Mother Cells and Fluctuation Tests to Analyze Genome Instability During Mitotic Cell Aging in Saccharomyces cerevisiae

Saccharomyces cerevisiae has been an excellent model system for examining mechanisms and consequences of genome instability. Information gained from this yeast model is relevant to many organisms, including humans, since DNA repair and DNA damage response factors are well conserved across diverse species. However, S. cerevisiae has not yet been used to fully address whether the rate of accumulating mutations changes with increasing replicative (mitotic) age due to technical constraints. For instance, measurements of yeast replicative lifespan through micromanipulation involve very small populations of cells, which prohibit detection of rare mutations. Genetic methods to enrich for mother cells in populations by inducing death of daughter cells have been developed, but population sizes are still limited by the frequency with which random mutations that compromise the selection systems occur. The current protocol takes advantage of magnetic sorting of surface-labeled yeast mother cells to obtain large enough populations of aging mother cells to quantify rare mutations through phenotypic selections. Mutation rates, measured through fluctuation tests, and mutation frequencies are first established for young cells and used to predict the frequency of mutations in mother cells of various replicative ages. Mutation frequencies are then determined for sorted mother cells, and the age of the mother cells is determined using flow cytometry by staining with a fluorescent reagent that detects bud scars formed on their cell surfaces during cell division. Comparison of predicted mutation frequencies based on the number of cell divisions to the frequencies experimentally observed for mother cells of a given replicative age can then identify whether there are age-related changes in the rate of accumulating mutations. Variations of this basic protocol provide the means to investigate the influence of alterations in specific gene functions or specific environmental conditions on mutation accumulation to address mechanisms underlying genome instability during replicative aging.     

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.