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     

Peculiarities of cytomixis in pollen mother cells of transgenic tobacco plants (Nicotiana tabacum L.) with mutant phenotype

The frequency characteristics and cytological picture of cytomixis in the course of male meiosis are described in transgenic tobacco plants (Nicotiana tabacum L.) with altered flower morphology and male sterility. Effects of cytomixis on qualitative composition of meiotic products are studied (formation of cytoplasts and polyads). Doubling of the chromosome number was established to increase frequency of cytomixis in the studied plants.     

Role of microtubular cytoskeleton and callose walls in the manifestation of cytomixis in pollen mother cells of tobacco Nicotiana tabacum L.

The structure and dynamics of microtubular cytoskeleton and of callose walls in normal pollen mother cells (PMC) of tobacco N. tabacum L. and in cells with intercellular translocation of nuclear material (cytomictic) was studied in the course of the cell cycle. The microtubular cytoskeleton was established as playing no obvious role in the process of cytomixis. The elevated level of cytomictic seems to be due to disturbances of synthesis of callose walls as a result of their attenuation and perforation. Possible causes of cytomictic in tobacco PMC at the cellular level are discussed.     

洋葱卵细胞的分离

将洋葱的胚珠置于酶液中酶解50-110 min后剥去其珠被,可清楚地看到珠心中的胚囊轮廓。用解剖针将珠心从中部横切,然后挤压其珠孔部位,卵器细胞从胚珠的切口处逸出。再用显微操作仪的玻璃针将卵细胞和两个助细胞分开,达到分离洋葱卵细胞的目的。酶对分离卵细胞具有重要作用,在最佳的酶液浓度[0.02%果胶酶Y23、0.08%果胶酶（Serva）、0.05%纤维素酶和0.05%半纤维素酶]下酶解胚珠110 min后,解剖1 h可从24个胚珠中分离出10个卵细胞（41.67%）。随着胚囊的发育,两个助细胞的体积出现明显的二形性。洋葱生活卵细胞的分离为开展洋葱离体受精建立了基础,也为研究洋葱卵器细胞的发育创造了条件。     

Somatic embryogenesis and plant regeneration from immature embryo cultures of onion (Allium cepa L.)

Somatic embryos were obtained and plants regenerated from immature embryos of onion following culture on embryogenic induction media. Highest rates of somatic embrogenesis resulted from 0.5- to 1.5-mm immature embryos cultured on media containing 5 mg/l of picloram. Somatic embryos formed either directly on the surface of embryos or developed from compact cultures. The production of somatic embryos was significantly affected by the addition of auxin, embryo size and cultivar. The potential of somatic embryogenic cultures for plantlet regeneration has been maintained for over 1 year in some lines. Three types of immature-embryo-derived cultures were characterized by histology. Some cultures were morphologically similar to immature-embryo-derived embryogenic cultures of other monocotyledonous species. Cultures such as these have proven to be useful target tissues in transformation studies. Received: 16 December 1997 / Revision received: 23 February 1998 / Accepted: 13 March 1998     

Ultrastructural Aspects and Possible Origin of Cytoplasmic Channels Providing Intercellular Connection in Vegetative Tissues of Anthers1

Cytoplasmic channel represents a huge intercellular connection other than plasmodesma. They are proposed to play an essential role in controlling cell-to-cell trafficking of macromolecules. The present study ultrastructurally examined the occurrence, structure, and formation of this intercellular path in somatic tissues of wheat, tobacco, and onion anthers as well as their differences from those present in anther generative tissue. It was shown that cytoplasmic channels existed not only in the pollen mother cells, but also in both epidermis and vascular parenchyma of the anthers. In somatic tissues, they appeared as plasmallema-lined tubes 400–750 nm wide filled with nuclear or cytoplasmic material, the latter including cytoplasmic matrix, ribosomes, and filamentous structures. Their biogenesis seems to result from the thinning of the local portions of cell wall containing multiple plasmodesmata and the fusion of plasmodesmata in such regions induced by the wall-digesting enzymes released by nearby located vesicles. In contrast to the channels existing in the pollen mother cells of tobacco, the cytoplasmic channels in the epidermal or vascular parenchyma cells of wheat, onion, and tobacco anthers usually do not appear in groups, but are single. Perhaps this is the way for nuclear material to migrate from cell to cell via a single channel and then form a single chromatin spherical body in the adjoining cell. An individual cell could not accept the nuclear material from another cell while extruding its own to the third cell. Cell-to-cell migration of organelles through the cytoplasmic channels was not shown in the somatic tissues.     

洋葱小孢子母细胞减数分裂过程中微管分布变化

应用间接免疫荧光标记技术和激光共聚焦扫描显微镜成像技术观察洋葱小孢子母细胞减数分裂过程中微管分布变化。减数分裂之前,小孢子母细胞中的微管较短,呈辐射状,由细胞核表面向四周扩散。减数分裂开始后,细胞质中的一部分微管蛋白聚集成纺锤体微管,控制染色体的分布。进入减数分裂I后期,纺锤体微管变为牵引染色体移向两极的着丝粒微管和连接纺锤体两极的极丝微管。之后,所有微管集中在两个核之间,构成成膜体。然后,微管解聚成微管蛋白弥散在细胞质中。减数分裂I完成后,二分体2个子细胞中的微管蛋白又聚集成2个纺锤体微管,开始减数分裂II过程。经过减数分裂II中期,2个二分体细胞中的微管再次集中在2个细胞核之间形成成膜体,隔离2个细胞核。此后,微管蛋白解聚,弥散分布在小孢子细胞质中。     

洋葱花药发育中钙离子分布特征

采用焦锑酸钾沉淀钙离子技术,对洋葱（Alliumcepa）花药发育中Ca^2＋分布进行了研究。在小孢子母细胞时期,小孢子母细胞中的钙沉淀颗粒很少,但绒毡层细胞的内切向壁已出现明显的钙沉淀颗粒。在四分体时期,四分体小孢子的胼胝质壁中出现较多的钙沉淀颗粒;绒毡层细胞内切向壁的钙沉淀颗粒消失,而在外切向壁和径向壁部位的钙沉淀颗粒增加。在小孢子早期,小孢子中也出现了钙沉淀颗粒,而绒毡层细胞内切向壁表面出现了很多絮状物,其上附有细小钙沉淀颗粒。到小孢子晚期,小孢子中出现一些小液泡,细胞质中的钙沉淀颗粒有所下降。此时绒毡层细胞已明显退化,但在绒毡层膜上仍有一些乌氏体和钙沉淀颗粒。在二胞花粉早期,营养细胞中的液泡收缩、消失,细胞质中又出现了较多的钙沉淀颗粒,在质体和其内部的淀粉粒表面上附有较多的钙沉淀颗粒。到二胞花粉晚期,花粉中的钙沉淀颗粒已明显下降,仅在花粉外壁中还有一地钙沉淀颗粒．     

两种山羊豆属植物花粉形态与花粉母细胞减数分裂行为观察

采用扫描电镜与1%醋酸洋红压片的方法,观察山羊豆属两种植物的花粉形态及花粉母细胞减数分裂行为,从而为山羊豆属的遗传学及分类学提供依据。两种山羊豆属植物立体形状为近球形或扁球形,均具三孔沟,穴—网状纹饰,极面观为近三角形,赤道面观为圆形。东方山羊豆的大小为:赤道轴×极轴=14.3（13.5～15.0）×12.6（12.1～13.7）μm,山羊豆的大小为：赤道轴×极轴=13.7（12.3～14.8）×12.4（10.6～13.0）μm.花粉母细胞减数分裂为同时型胞质分裂,终变期染色体构型为8Ⅱ,没有出现落后染色体及微核现象。小孢子四分体为四面体型。减数分裂过程有一定规律,与花蕾大小,颜色有密切关系。减数分裂进程存在一定差异,同一个花蕾中的不同花药花粉母细胞分裂速度基本相同,所处的时期及观察的分裂相一致,在同一个花药中的不同花粉母细胞中,可见各种不同时期的细胞。     

Ascorbate free radical enhances vacuolization in onion root meristems

Abstract. Ascorbate free radical (AFR) induced cell elongation in merislems of Allium cepa roots by promoting a high vacuolization as shown by the increased vacuole volume, vacuole volume density, tonoplast surface and tonoplast surface density. Accordingly, both plasma membrane- and tonoplast-associated ATPases and vacuole soluble acid phosphatase of meristematic cells were also increased. Neither the other subcellular organelles nor cell proliferation appeared to be significantly affected. It is suggested that AFR may be involved in some plasma membrane events related to the initiation of plant cell elongation.     

洋葱花粉发育过程中ATP酶的分布特征(简报)

在真核细胞中,除了线粒体和叶绿体ATPase的功能是合成ATP外,其余部位ATPase是水解ATP以获取生物能量的代谢酶,在生物体细胞内广泛存在。探索ATPase在细胞中的分布状态是研究细胞生理状态的一种重要手段。ATPase在细胞中的多少可反映出细胞当时的生活状态,这一特征已被初步用于探索小麦和水稻雄性不育的细胞生物学研究中,希望通过比较可育花药和不育花药中ATPase的分布差异寻找雄性不育的机理,发现     

In Vitro Regeneration of Onion through Repetitive Somatic Embryogenesis

A reliable protocol for the regeneration of onion through repetitive somatic embryogenesis was established. Embryogenic callus was derived from mature seeds on Murashige and Skoog (MS) medium supplemented with 2 mg dm-3 2,4-dichlorophenoxyacetic acid (2,4-D). Somatic embryos aroused on the surface of calli cultures and formed plantlets after the removal of 2,4-D or its substitution with 1 mg dm-3 kinetin (Kin). Reculturing the somatic embryos on 2,4-D containing medium led to secondary embryos formation. The embryogenic cultures which were preserved for five months on maintenance medium containing 2 mg dm-3 2,4-D + 0.5 mg dm-3 Kin have retained their ability for regeneration, while those kept on 2,4-D only, failed to form plantlets. Electrophoretic analysis of total soluble proteins revealed that the competence for successful conversion of somatic embryos into plantlets is associated with the expression of new set of proteins (112, 58 and 30 kD). The regenerated plants were successfully transferred to the soil.     

激光微束切割小冰麦异附加系染色体的研究

利用激光微束,并选用适当的功率密度可将小冰麦异附加系花粉细胞染色体切割成2～3个片段,这个技术的建立为激光微束应用于染色体片段DNA微克隆及基因定位提供可能。     

紫斑牡丹栽培品种小孢子发育过程的细胞遗传学研究

本文对紫斑牡丹栽培品种的花平细胞减数分裂过程进行了系统研究,结果发现紫斑牡丹品种约６８．９６％的花平细胞在减数分裂过程表现正常,约有３１．０４％的花粉母细胞在减数分裂的比线期、中期Ⅰ、后期Ⅰ、中期Ⅱ、后期Ⅱ及四分体时期观察到染色体行为异常。本实验表明,在小孢子形成过程中,多数小孢子发育政党,但有约３１．０４％的花粉母细胞减数分裂异常,导致了花粉的败育。     

罗宾根螨的初步研究

罗宾根螨于1988年发现于哈密死亡洋葱植株上,其后发生不断扩大,危害不断加重,严重处洋葱植株死亡腐烂率达39.52％。 罗宾根螨可用尼索朗、灭扫利和久效磷等杀螨剂,加水浇灌防治。     

Immunolcocalization of actin in intact and DNA—and histone—depleted nuclei and chromosomes of allium cepa

The presence of actin in eukaryotic nuclei and chromosomes,and especially in higher plant nuclei and chromosomes,has not been well established.We detected actin in meristematic cells of Allium cepa with indirect immunofluorescence technique and observed bright fluorescence in the intact nuclei and chromosomes,indicating that actin is present in the nuclei and chromosomes of the higher plant.We labeld sections of the meristematic cells of A.cepa with immunogold technique,gold parti cles were concentrated in condensed chromatin and nucleoli,confirming the results of the immunofluoresence observations.We traeated the nuclei and chromosomes of A.cepa with DNase I and 2M NaCl and obtained DNA-and histone-depleted nuclei and chromosomes.Indirect immunofluorescence tests showed that the DNA-and histonedepleted nuclei and chromosomes reacted positively with the anti-actin antibodies.These results demonstrate that the anti-actin antibodies.These results demonstrate that actin exists not only in intact nuclei and chromosomes,but also in DNA-and histone-depleted nuclei and chrmosomes of the plant.In addition,our immuno-fluorescence tests indicate that tropomyosin is present in the nuclei and chromosomes of A.cepa.     

木立芦荟小孢子母细胞减数分裂与花粉育性关系的初步研究

文章从减数分裂过程、小孢子发育两方面,探讨了木立芦荟（Aloe arboresens Mill.）花粉败育的原因。木立芦荟花粉母细胞染色体数目为2n=14,由四对长染色体和三对短染色体组成,属二型性核型。其减数分裂异常,发现存在单价体和多价体、染色体桥、落后染色体、不均等分离、微核等。同时观察到木立芦荟染色体具有极度的粘质性,使减数分裂各阶段的染色体不易散开。统计各种异常现象出现的频率并分析了这些异常现象形成的可能机制及对正常小孢子形成的影响,推测染色体间的丝状粘连可能是木立芦荟小孢子母细胞减数分裂异常并导致败育花粉产生的主要因素。成熟花粉粒中90%以上为败育花粉,属碘败型。