姜花花粉萌发时花粉和花粉管内微丝结构变化的共焦镜观察

用ＴＲＩＴＣ－鬼笔碱荧光显微探针入共焦激光扫描镜,观察了不同萌发阶段的姜花花粉和花粉管内肌动蛋白微丝的结构和分布格局的变化,花粉水合后在花粉内的微丝呈不规则网络状。但在靠近萌发孔周围,由於网络微丝向萌发孔汇集而延伸拉长,因此在萌发孔前缘出现荧光特别明亮区,显示微丝在此外开始密集。花粉水合后１０－３０分钟,大多数花粉管从萌发孔伸出,此时,在花粉管顶形成一个长约１０－２０μｍ的顶端区域微丝网络;在花粉     

杜仲花粉离体萌发特征及花粉管微丝骨架分布

以干燥的杜仲成熟花粉为材料,对杜仲花粉离体萌发的适宜液体培养基配方进行了筛选,并对花粉萌发特征及花粉管微丝骨架分布规律进行了研究。结果表明:(1)适宜杜仲花粉离体萌发的液体培养基组成为200g/L蔗糖+30mg/L硼酸+10mg/L Ca(NO3)2,于26℃条件下离体培养18h的花粉萌发率可达46.29%±3.75%。(2)在适宜液体培养条件下,杜仲花粉萌发率在培养6h内急剧增长,随后趋于平稳;而花粉管在培养8h内伸长较快,之后有放缓趋势,至培养48h时,花粉管长度可达363.14±30.51μm。(3)杜仲花粉属于2胞花粉,花粉萌发过程中,营养核和生殖核的移动存在一定的时序性,通常营养核先于生殖核进入到花粉管;杜仲花粉生殖核的有丝分裂发生在花粉管中,离体培养12h可逐渐观察到有丝分裂行为。(4)花粉萌发过程,微丝骨架形成束状,与花粉管伸长方向平行排布,与较为稀疏的网状微丝阵列组成连续系统,引导细胞核的运动。     

梅花花粉离体萌发和花粉管生长研究

(南京农业大学园艺学院,南京210095)摘要:研究培养基成分、pH值和培养方式对梅花花粉离体萌发和花粉管生长的影响。结果表明:不同品种梅花花粉离体萌发的最适培养基为ME3+200g.L-1PEG4000(pH5.0),品种‘淡丰后’、‘久观绿萼’、‘喧妍宫粉’和‘月光玉蝶’最高萌发率可分别达到58.6%、60.6%、85.6%和50.7%。PEG4000能显著促进梅花花粉萌发,在培养基各成分中作用最大,不可替代。低浓度(50g.L-1)蔗糖对梅花品种花粉萌发作用不显著,而高浓度(≥100g.L-1)蔗糖明显抑制花粉萌发和花粉管生长。固体和液体培养对梅花花粉离体萌发的影响差异不显著。     

百合花粉及花粉管内微丝和微管的分布

利用免疫荧光定位及荧光定位方法,结合共焦激光扫描显微镜,对百合（ＬｉｌｉｕｍｄａｖｉｄｉＤｕｃｈ．）花粉及花粉管内微丝及微管的分布进行了观察,得出了一些新的结果：１化学固定方法可以较好地保存花粉和花粉管内的微丝,从而可以在此条件下较好地进行微管和微丝的双标记,并进行两者相互关系的研究;２在距花粉管顶端１０～２０μｍ的范围内,用化学固定及ＴＲＩＴＣ鬼笔碱标记显示微丝的存在是很微弱的,基本上无法看到明显的微丝束,而同时用免疫荧光法标记却发现此部位微管很丰富,在花粉管顶端微管形成浓密的网状,而且其末端与花粉管顶端质膜相连;３在花粉管中,只有少数微丝与微管相互平行排列,而其中大多数微丝骨架与微管骨架并不存在共分布现象。为了解花粉管内微管和微丝的功能及相互关系提供了新的证据。     

花粉萌发和花粉管生长发育的信号转导

显花植物授粉是一个复杂的发育过程。从花粉落在柱头上开始,经过粘附、识别、水合、萌发,花粉管在花柱内生长,直至到达子房发生双受精作用,整个过程发生在雌、雄两性细胞和组织之间,受到严格的遗传控制和细胞控制。一方面雌雄配子的基因型决定两者是否亲和,另一方面雌雄两性细胞间发生复杂的相互作用,细胞外信号分子是这些过程的主要调控因子。当花粉或花粉管细胞感知外部信号后,必然通过信号转导级联反应,达到控制萌发、调整花粉管生长方向等目的。这一系列动力学的细胞事件,关系到受精的成败。因此研究此过程中的信号及其转换机…     

薄壳山核桃花粉离体萌发和花粉管生长特性研究

以6年生薄壳山核桃优良品种‘金华’的新鲜花粉为试材,采用离体培养法,研究了不同复水时间、不同培养基组分、不同培养温度及培养时间对薄壳山核桃花粉萌发和花粉管生长的影响。结果显示:(1)薄壳山核桃花粉萌发试验前进行4h复水处理可显著提高萌发率,萌发率可达51.78%,为对照(2.51%)的20.68倍。(2)蔗糖、H3BO3、Ca(NO3)2·4H2O在一定浓度范围内均具有促进花粉萌发和花粉管生长的作用,但浓度过高则起抑制作用。(3)正交试验结果经实验验证表明,薄壳山核桃花粉萌发和花粉管生长的最适培养基为20%蔗糖+0.02%~0.03%H3BO3+0.05%Ca(NO3)2·4H2O,最佳培养条件为25℃下恒温培养24h,此时的花粉萌发率高达74.46%,花粉管平均长度为258.84μm。     

百合远缘杂交花粉萌发及花粉管生长过程观察

利用荧光显微镜对百合远缘杂交组合Bernini×Pollyanna花粉萌发及花粉管生长过程进行观察研究,结果显示,授粉后3~30 h内花粉萌发形成花粉管,且花粉管生长速度由快到慢,48~51 h内花粉管停止生长,花粉管最后深入到花柱的1/3处,并观察到一些异常的花粉管形态;花粉管生长过程中还伴随着一系列的胼胝质反应,出现的部位依次是乳突细胞、花粉管、花柱通道细胞、胚珠中的胚囊.结果表明该杂交组合不亲和.研究认为Berni-ni柱头乳突细胞是阻碍Pollyanna花粉管生长的第一道屏障,花柱通道细胞是抑制花粉管在花柱内生长的第二道屏障.     

桔梗花粉萌发与花粉管生长研究

以2年生桔梗植株为材料,采用液体培养法研究了培养基种类、PEG、蔗糖、pH以及培养温度、培养时间对桔梗花粉离体萌发生长的影响,结果表明:(1)浓度为100~150 g.L-1的PEG可显著促进桔梗花粉萌发和花粉管的生长;200~250 g.L-1PEG显著促进花粉萌发,但对花粉管生长的作用不显著。(2)100 g.L-1的蔗糖有利于花粉萌发和花粉管生长,高浓度蔗糖(200 g.L-1)有明显抑制作用;(3)桔梗花粉离体萌发和花粉管生长的适宜培养基为ME3+BK+10%蔗糖+150 g.L-1PEG(pH5.8);(4)25~40℃条件下桔梗花粉均可较好萌发,以30℃培养1.5 h为最佳培养条件。     

花粉管中的微丝和微管

对花粉管中的微丝和微管研究的几个问题的进展进行了综述,包括微丝和微管在花粉管中的分布;微丝和微管在花粉管胞质流动、细胞器运动以及花粉管生长中的作用等几个方面,并对今后这些方面研究的重要总是进行了论述。     

Confocal Microscopic Observations of Microfilaments in germinating Pollen of Hedychium coronarium

Changes in the microfilament (actin)organization in the germinating pollen of Hedychium coronarium Koenig were followed after TRITC-phalloidin staining without fixation. Changes in the pattern of organization of the microfilaments were visualized using eonfocal microscopy. In the hydrated pollen a reticulate network of microfilament can be observed. Before the pollen tube protrudes out from the germination pore numerous microfilaments begin to converge towards the aperture. After 10–30 mins of germination,pollen tube appears. In the pollen tube a new network of microfilament forms near the tip region. Between the pollen and the pollen tube tip region there are numerous linearly arranged microfilaments. About 1 hour after germination,the pollen tube has reached a length of about 300μm Inside the pollen, tube there are numerous longitudinally oriented microfilaments. The microfilament network in the pollen tube tip region does not change much. About 2 hours after germination,the pollen tube reaches about 1000μm in length. At this stage,the pattern of distribution of microfilament in the pollen tube is very similar to that seen at the earlier stages of development ,whereas the pattern is somewhat different in the pollen. Microfilaments in the central region of the pollen grain disappear but still a parietal network in the peripheral region. About 5 hours after germination,the microfilaments in the pollen tube become abnormally variable and produce branches. Some even change into spicules, sheets and thick bundles.     

Ultrastructure of Microfilaments in Pollen and Pollen Tubes of Hosta Ventricosa (=H. voerulea)

Ultrastructure of microfilaments in pollen grains and pollen tubes of Hosta ventricosa (=H. coerulea) was investigated. Results indicate that microfilaments with conventional chemical fixation are preserved only in pollen grains, but destroyed in pollen tubes. Microfilaments treated with phalloidin before chemical fixation are found preserved in pollen tubes. In pollen grains a pronounced organization of parallel microfilaments appeared in bundles with its distribution characteristics is always restricted to their functional domains where bundles were in close contact with the vegetative nucleus. In young pollen tubes cytoplasmic bundles of microfilaments appeared also to pass close to the surface of mitochondria, plastids, endoplasmic reticulum, vesicles and small vacuoles, and always associated with lipid bodies. These findings strongly indicate that there is a relationship between microfilaments and the movement of vegetative nucleus and other organelles in the germination of pollen grains and in the growth of pollen tubes.     

钙和硼对蓝猪耳花粉萌发及花粉管生长的影响

研究了钙(Ca^2 )和硼(H3BO3)对蓝猪耳花粉萌发和花粉管生长的影响。结果表明：(1)在一定范围内Ca^2 几乎不影响花粉萌发频率,而主要影响花粉萌发速度和花粉管生长速度;低Ca^2 不利于花粉管生长,而高Ca^2 抑制花粉萌发速度和花粉管生长;在稍高于最适Ca^2 浓度的条件下,花粉管生长早期呈现波浪形。(2)硼明显影响花粉萌发频率及花粉管形态;花粉管生长必需硼,但不同浓度的硼对花粉管生长速度影响不明显;在高浓度硼条件下,较长时间内花粉管均呈现出波浪形。(3)Cooled-CCD动态跟踪观察进一步证实Ca^2 影响花粉管生长速度,而硼则不明显。     

Microfilament Orientation Constrains Vesicle Flow and Spatial Distribution in Growing Pollen Tubes

The dynamics of cellular organelles reveals important information about their functioning. The spatio-temporal movement patterns of vesicles in growing pollen tubes are controlled by the actin cytoskeleton. Vesicle flow is crucial for morphogenesis in these cells as it ensures targeted delivery of cell wall polysaccharides. Remarkably, the target region does not contain much filamentous actin. We model the vesicular trafficking in this area using as boundary conditions the expanding cell wall and the actin array forming the apical actin fringe. The shape of the fringe was obtained by imposing a steady state and constant polymerization rate of the actin filaments. Letting vesicle flux into and out of the apical region be determined by the orientation of the actin microfilaments and by exocytosis was sufficient to generate a flux that corresponds in magnitude and orientation to that observed experimentally. This model explains how the cytoplasmic streaming pattern in the apical region of the pollen tube can be generated without the presence of actin microfilaments.     

An Ultrastructural Study on Pollen Protoplasts and Pollen Tubes Germinated From Them in Gladiolus gandavensis

Large quantities of protoplasts were isolated enzymatically from the mature pollen grains in Gladiolus gandavensis. Regeneration of cell wall and germination of pollen tubes were performed during culture of purified pollen protoplasts in Ks medium supplemented with 32% sucrose, 0.1 mg/1 2,4-D, 1 mg/1 NAA and 0.2 mg/1 6-BA, with a germination rate up to 47.7%. The materials were fixed gently with gradually increasing concentration of glutaraldehyde, followed by osmium, then preembedded in a thin layer of agar and surveyed under an inverted microscope so as to select desired specimens for subsequent procedure. Small agar blocks containing specimens were dehydrated through ethanal-propylene oxide series, embedded in Araldite and ultratomed. Electron microscopic observations show that the pollen protoplasts are surrounded by a smooth plasma membrane and with ultrastructurally intact cytoplasm, a vegetative nucleus and a generative cell. After 8h of culture, wall regeneration commences resulting in a multilayered, fibrillar wall structure which is different from the intine. No exine is formed. Numerous vesicles participate actively in the wall formation. The wall is uneven in thickness around its periphery; a thickened area somewhat resembling to germ furrow is formed, from which pollen tube emerges. The tubes contain abundant plastids, mitochondria and dictyosomes. Vesicles are released out of the plasma membrane and involved in tube wall formation. After 18h of culture, the vegetative nucleus and generative cell have migrated into the tube. Technical points of preparing pollen protoplast specimens for ultastructural studies and the fearnres of wall regeneration in pollen protoplast culture are discussed.     

Visualization of Actin Filament Patterns in Pollen Tubes of Hosta caerulea Tratt. with a Non-Fixation and TRITC-Phalloidin Method

Actin filament patterns during pollen germination in Hosta caerulea Tratt. were visualized with a simple method in which there was no pre-fixation, with dimethylsulphoxide (DMSO) as a permeabilising agent and staining with TRITC-Phalloidin. The cytoplasm of the vegetative cell of the ungerminated pollen grain contained numerous crystalline fusiform bodies to constitute a storage form of actin. These bodies were transferred to the emerging pollen tube after the germination of the pollen grain. Following the growth of pollen tube, the fusiform bodies were gradually dissociated, branched, slenderized and formed a cross-linked actin network. During the further growth of the pollen tube, the preponderance of longitudinally-oriented thin actin filaments with some anastomoses to form a more complex network present always in the long pollen tube. This was the typical pattern of actin filaments in most cases. In some conditions, actin filaments were assembled to form thick actin cables near the proximate part of the pollen tube tip. The branching and connecting of the cables were probably also seen in some parts. Actin filaments were always entering to the apical region of a tube tip. The significance of the non-fixation and fluorescence-phalloidin (FI-Ph) method and the problems in the future studies are discussed     

小果短柱茶花粉离体培养系统的研究

山茶的短柱茶组是优良种质资源,有必要对小果短柱茶(Camellia confusa Chang 1941)的花粉萌发和花粉管生长的生理特性进行研究.本文研究了花粉生活力、培养温度及pH对小果短柱茶花粉萌发和花粉管生长的影响.结果表明:最适离体萌发培养基为5%蔗糖、0.003%的硼酸,0.005%的氯化钙和12%的PEG...     

微丝骨架的构成及其对花粉管极性生长的调控作用

微丝骨架是细胞骨架的重要组成部分,它由肌动蛋白和肌动蛋白结合蛋白组成,广泛存在于真核细胞中。近年来,大量研究表明植物花粉及花粉管中存在丰富的微丝骨架。目前,在微丝骨架作为信号转导途径的靶标参与对花粉管极性生长的调控、微丝骨架在花粉和花粉管中的分布及其在花粉管生长过程中与其他信号分子之间的相互作用等方面取得了一系列突破性进展。     

Chlorophyll Accumulation in Leaves of Rice Pollen Albinos

In contrast to the green plantlets, rice pollen albinos contain a little chlorophyll only, and the relative contents of protochlorophyll (ide) and carotenoids comparing with chlorophyll are much higher however. The contents of chlorophyll and related pigments in leaves of pollen albinos vary regularly with the variation of light regimes. The spectra of regenerated albinos from pollen albinos possess the same characteristics as described above, but their pigment contents are lower. It is likely that the occurrence of these characteristics of pollen albinos may be attributed to their deficiency of plastid membrane system and the latter in turn may be the result plastom mutation. In view of the variation of pigment contents among different planoets of the same variety, it is resonable to deduce that plastids of pollen albinos are heterogeneous structurally and functionally.