In situ Localization and Isolation of Actin Filaments from Pollen Tubes of Amaryllis vittata Ait

ＩｎｓｉｔｕＬｏｃａｌｉｚａｔｉｏｎａｎｄＩｓｏｌａｔｉｏｎｏｆＡｃｔｉｎＦｉｌａｍｅｎｔｓｆｒｏｍＰｏｌｌｅｎＴｕｂｅｓｏｆＡｍａｒｙｌｌｉｓｖｉｔｔａｔａＡｉｔＣＡｌＸｕｅ（蔡雪）;ＤＯＮＧＹｕｎ－ｚｈｏｕ（董云洲）（ＣｏｌｌｅｇｅｏｆＬｉｆｅＳ...     

朱顶红精细胞超微结构观察

精细胞是双受精作用的直接参与者,是生殖生物学中的重点研究对象之一。以往的研究表明,应用连续超薄切片和计算机辅助三维重组技术,结合免疫荧光定位,发现两个精细胞在体积和细胞器含量上存在着差异,即精子的二型性,而且与营养细胞核三者构成紧密功能单位即雄性生殖单位（ＭＧＵ）,微管对精细胞的性状的确定、运动和维持ＭＧＵ的动态结构稳定具有重要的作用。本文应用透射电镜,详细观察了朱顶红花粉管中精细胞的超微结构,并着重微管结构及其分布的观察。朱顶红成熟花粉为两细胞型。成熟花粉于２６℃、黑暗条件下,在液体培养基（含１０％蔗糖和１００ｐｐｍ硼酸）中培养１３－１８小时,然后收集花粉管,固定,供电镜观察并照相。朱顶红成熟花粉培养１３小时后,生殖细胞在花粉管中完成核分裂和胞质分裂等两个过程。形成两个精细胞。初形成的两个精细胞前后排列,营养核前导并靠近花粉管顶端。领头的精细胞的细胞质以很大的表面与营养核相互贴合（图版Ⅰ－１,２）,有时营养核与两个精细胞彼此穿插、缠绕（图版Ⅰ－３）。两精细胞之间的共同壁上具有很多胞质通道和含均质电子密度中等的基质（图版Ⅱ－４）。精细胞质在核与共同壁之间的区域染色较深,经高倍放大,观察到此处含丰富的微管,基     

Positional Shift Between the Vegetative Nucleus and the Generative Cell in Amaryllis Pollen Tube

A modified technique, FITC-tubulin immunofluorescence and DAPI localization to demonstrate simultaneously both the generative cell (GC) and the vegetative nucleus (VN) in the pollen tube under ultra-violet excitation, was developed sucessfully. During the germination of the pollen tube of Amaryllis vittata Ait. the GC and the VN, either being the first one, entered the tube within the first 1—2 h from the pollen grain. However, before the time of GC division, the VN was always positioned distally near the tip of the tube. In case when the GC entered the tube first, then the VN must have a positional shift in order to pass over the GC. The detail processes of positional shift between the GC and the VN were observed. Three basic processes were demonstrated: 1. The anterior end of the VN first reached the vicinity of the posterior attenuated extension of the GC about 2 h following germination forming a temporal physical association. Sometimes their both ends could be inserted into one another for certain extent. 2. The whole VN moved forward and contacted in parallel with the GC until they became twisted together and 3. The VN passed over the GC and greatly elongted lengthwise. Its posterior part became inserted into the anterior end of the GC. The behavior of positional shift between the VN and the GC in the pollen tube seems to be an adjustment of their diameters to fit the narrow tube. A conclusion may be drawn that the rate of movement between the VN and the GC was apparently different during the passage through the tube. Such difference may presumably be accompanied by the independent motive mechanism and structural difference between the VN and GC themselves, which provide their motive force for movement in the tube.     

Structure and Development of Stigmatic Branches and Style and Their Relation to Pollen Tube Growth in Wheat

The style of wheat divides into 2 branches, separated from its base and covered with a large number of slender stigmatic branches. The stigma is of dry type. The style is solid. There is no transmitting tissue differentiated in the style. Young stylar cells appear polygonal in transverse sections and elongated in longitudinal sections with an increase in length of the cells from periphery towards center. In transverse sections, mature stylar cells look extremely irregular. They are contorted and mosaicked with one another. During their development, stylar cells elongated vigorously with intrusive growth. The wall of stylar cells is thin, except at the corners where cells connect, that slight thickening of the cell wall occurs. Stylar cells start vacuolation at the earlier stages and gradually become highly vacuolated, but still remain rich in organelles, such as mitochondria, endoplasmic reticulum, dictyosomes and chloroplasts, the amount of which varied with the development stages of the style. Stigmatic branches are differentiated from the stylar epidermal cells, composed of 4 files of cells which link end to end with one another. Not long before anthesis, wall material in the intercellular corners becomes loose and porous. After pollination, pollen tubes grow along the intercellular spaces among the 4 files of cells in the stigmatic branches and then enter the style. Pollen tubes may pass through any intercellular corner throughout the 2 branches of the style, except for the lateral-outer portion which is composed of larger stylar cells. Eventually, pollen tubes enter the ovary.     

钙调素对花粉萌发和花粉管生长的效应

牛脑和玉米胚ＣａＭ能显著促进花粉萌发和花粉管生长（图１）,而ＣａＭ抑制剂ＴＦＰ、ＣＰＺ及另外两个专一性更强的抑制剂Ｃｏｍｐｏｕｎｄ４８／８０和Ｗ７均严重抑制甚至阻止花粉的萌发（图２,３）。用对ＣａＭ亲和性较低的Ｗ７同系物Ｗ５,在与Ｗ７同样浓度下,对花粉萌发和花粉管生长无明显影响。此外,Ｗ７对花粉萌发和花粉管生长的抑制效应可被外源ＣａＭ所消除（图４）。在花粉萌发过程中,其内源ＣａＭ含量显著上升,在花粉萌发率接近最大值时,花粉ＣａＭ含量达最高水平（图５）。上述结果表明ＣａＭ对花粉萌发和花粉管生长的调控起重要作用。     

桃花粉离体萌发和花粉管生长特性研究

采用花粉离体萌发法研究不同培养基组分和培养条件对桃花粉萌发和花粉管生长的影响,同时对不同贮藏温度下的桃花粉寿命进行研究.结果表明:固体培养基与液体培养基对桃的花粉萌发率和花粉管长度影响差异不显著;10%蔗糖是大多数桃品种花粉的最适萌发条件;硼能提高桃花粉的萌发率,但对花粉管的生长没有促进作用;桃花粉在20℃～25℃的培养温度下萌发率最高,花粉管最长;桃花粉萌发率和花粉管长度在培养前3 h内上升最快,3～5 h上升趋势减弱,5 h后基本停止;随着贮藏温度的升高和贮藏时间的延长,花粉生活力呈降低的趋势.     

Axial Distribution of Glycosidases in Relation to Cellular Growth and Ageing in Pisum sativum Root

Tanimoto, E. 1985. Axial distribution of glycosidases in relationto cellular growth and ageing in Pisum sativum root.—J.exp. Bot. 36: 1267–1274. Eight glycosidase activities were measured in relation to cellulargrowth and ageing along the axis of pea roots. The highest activities of ß-D-fucosidase and ß-D-glucosidasewere in the apical 1.0 mm segment containing the root cap andmeristem. Activities of -L-arabinosidase, -D-mannosidase, ß-D-galactosidase, -D-glucosidase and ß-D-xylosidase werehighest in the segment between 1 and 2 mm from the tip and containingyoung elongating cells with high growth potential. The activityof -D-galactosidase was high between 1 and 2 mm from the roottip, decreasing to a minimum 4-5 mm from the tip and increasingagain up to the base of root. This distribution of enzyme activitiesrelative to the root tip remained unchanged during 28 h whilethe root length doubled. No -L--fucosidase, ß-L-fucosidaseand -D-xylosidase activities were detected. Key words: Pisum sativum, root growth, glycosidase, galactosidase, mannosidase     

不同培养条件对黄连木花粉萌发和花粉管生长的影响

以黄连木花粉为试材,采用离体培养法研究了培养基组分和植物生长调节物质对黄连木花粉萌发和花粉管生长的影响.结果表明:花粉萌发和花粉管生长的适宜蔗糖浓度为15%,适宜培养温度为25℃;该培养条件下,花粉萌发率和花粉管长度分别达最大值63.3%和412.1 μm.硼酸、赤霉素(GA3)和吲哚乙酸(IAA)在一定浓度范围内,可以促进黄连木花粉萌发和花粉管生长,浓度过高时起抑制作用;最适宜黄连木花粉萌发和花粉管生长的硼酸浓度、赤霉素(GA3)和吲哚乙酸(IAA)浓度分别为100、50和15 mg/L.     

朱顶红鳞茎芽诱导及植株再生体系的建立

以朱顶红杂交品种红孔雀(Amaryllis vittatacv. Red Peacock)鳞茎盘切块为外植体,对影响鳞茎芽直接萌发增殖及生根的主要因素进行了研究,以建立其高效再生体系.结果表明:(1)1/2MS基本培养基有利于鳞茎芽的萌发和生长;切分方式不同对朱顶红鳞茎增殖具显著效应,偏离切分鳞茎可显著提高增殖系数,最高增殖系数可达6,平均增殖系数为3.6;而对半切分或未切分的增殖系数较低.(2)1/2MS 0.1 mg/L NAA 1.5 mg/L 6-BA 2.0 g/L活性炭的培养基利于鳞茎增殖;活性碳可显著提高鳞茎生根率,在1/2MS 0.1 mg/L NAA 2.0 g/L活性炭的培养基上生根率可达100%,而未加活性炭的生根率较低;再生植株经25 d的自然光照驯化后进行移栽,成活率达90%以上.     

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…     

Pistil Receptivity and Pollen Tube Growth in Relation to the Breeding System of Eucalyptus woodwardii (Symphyomyrtus: Myrtaceae)

Pistil structure, stigma receptivity and pollen tube growthwere investigated in relation to seed set of Eucalyptus woodwardii.Self-pollination resulted in reduced capsule retention and seeddevelopment as compared with cross-pollination. The pistil consistedof an ovary with five locules, a long style with a canal extendingfor two-thirds of its length, and a papillate stigma. Therewas no change in style length with time after anthesis, butboth stigma secretion and ability to support pollen germinationand tube growth increased to reach a peak at 7 d. Pollen germinatedon the stigma surface and in the stylar canal, but most tubegrowth occurred intercellularly in the transmitting tissue surroundingthe canal. At the base of the style the pollen tubes split intofive groups following the transmitting tissue strands to theovary. Each group grew through a septum dividing two loculesand entered the placenta. The tubes then emerged from the placentato penetrate the ovules at between 10 and 20 d after pollination.Fewer ovules were penetrated following self- than cross-pollination. Eucalyptus woodwardii Maiden, Lemon-flowered gum, Pistil receptivity, Pollen tube growth, Breeding system, Self-incompatibility     

果梅花粉离体萌发及花粉管生长特性研究

研究了果梅(Prunus mumeSieb.1et Zucc.)花粉在不同培养基组分、花粉不同培养密度和不同温度及培养时间的离体萌发和花粉管生长特性.结果表明:细叶青花粉萌发及花粉管生长最适宜的液体培养基为30mmol/L MES(pH 6.5)缓冲液中含20%蔗糖,0.01%硼酸,20%PEG-4000,0.03?(NO3)2?4H2O,0.02%Mg-SO4?7H2O;萌发率达45.03%,花粉管长度达597.2μm.适宜于果梅花粉萌发和花粉管生长的花粉粒密度为20~80粒/μL.培养温度过高或过低都不利于果梅花粉的萌发和生长,25℃时花粉萌发和花粉管生长最好.细叶青"、月世界"、莺宿"3个品种的平均萌发率为48.6%,平均花粉管长度为762.3μm.果梅花粉在不同培养温度下,萌发及生长不同,在25℃条件下花粉管生长速度最快,集中在0~12 h内,3个品种花粉管平均生长速度为58.5μm/h.     

肌动蛋白在丝瓜花粉管顶端生长中的作用

用非固定荧光标记的鬼笔环肽作为肌动蛋白探针观察并证明了丝瓜未萌发的花粉粒和不同生长时期花粉管中肌动蛋白纤丝的分布及其形态变化。又用细胞松弛素B（CB）、氯两嗪（CPZ）及N-乙酰马来酰胺（NEM）证明了丝瓜花粉管伸长与肌动蛋白既有密切的关系,也受Ca2 的调节。     

花柱和花粉胞外钙调素对花粉萌发和花粉管伸长的影响

以烟草为材料,通过半体内实验,就花柱和花粉胞外钙调素对花粉萌发和花粉管伸长的影响进行了观察。发现用EGTA及钙调素抗血清处理柱头或花粉均可抑制花粉在柱头上的萌发;向花柱引导组织中显微注射纯化钙调素可促进花粉管束伸长,而注射钙调素抗血清可抑制花粉管束伸长;同时证实玉米花柱和花粉细胞壁中均存在钙调素及钙调素结合蛋白,而且花粉和花柱细胞壁中钙调素结合蛋白的种类有差异。结果表明存在于花粉和花柱细胞外的钙调素对花粉萌发和花粉管伸长均有促进作用。     

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.     

Inhibition of Camellia japonica Pollen Tube Growth by Maltose

Various oligosaccharides were studied with regard to their effecton the in vitro growth of Camellia japonica pollen tube. Sucrose,raffinose, melezitose, cellobiose, turanose and isomaltose,especially the first four, promoted pollen tube growth, whilemaltotriose, trehalose, gentiobiose, palatinose, melibiose,lactose and lactulose had little effect. Maltose strongly inhibitednot only the tube growth on sugar-free medium but also sugar-stimulatedgrowth, except in the case of sucrose stimulation. Glycosidaseactivities toward the growth-stimulating oligosaccharides weredetected in the extract of sucrose-grown pollen, but the activitiesof -glucosidase and -galactosidase were much lower than thoseof ß-fructosidase and ß-glucosidase. Maltosesuppressed the increase in UDP-glucose level of the glucose-grownpollen but not that of the sucrose-grown one. These resultssuggest that maltose acts, directly or indirectly, somewherein the pathway from glucose to UDP-glucose via glucose-1-phosphate,but does not interfere with the direct conversion of sucroseto UDP-glucose. (Received December 1, 1984; Accepted May 24, 1985)     

Distribution of Glycosidases and Acid Invertase Activities in Relation to Elongation Growth in Pearl Millet Internode

The mean cell length along a differentiating internode and alliedchanges in the activities of ß-glucosidase, - andß-galactosidase. -mannosidase and acid invertase,together with the contents of reducing and non-reducing sugars,were examined in pearl millet (Pennisetum americanum L. Leekecv. BJ-104). The specific activities of cytoplasmic -mannosidase,wall ß-glucosidase, and cytoplasmic and wall acidinvertase showed close relationships with the rate of cell elongation.The linear regressions of the rate of cell elongation, and thespecific activities of wall ß-glucosidase and cytoplasmicand wall invertase showed significant positive correlations(P<0·05), whereas cytoplasmic -mannosidase was negativelycorrelated (P<0·01). The results are discussed in the light of cell wall looseningand the provision of carbon substrates for cell elongation. Key words: Glycosidases, acid invertase, sugars, cell elongation, Pennisetum americanum L., Leeke