水稻受精前后胚囊内钙调素分布的变化：免疫金电镜观察

用胶体金免疫电镜技术观察了水稻 (Oryzasativasubsp .japonica)受精前后胚囊内钙调素的分布变化。授粉后 ,卵细胞、助细胞和中央细胞内的钙调素较授粉前均有所增加。中央细胞内钙调素的增加要比卵细胞中约早 2h ,退化助细胞与宿存助细胞之间的钙调素含量无明显差异。授粉到受精期间 ,钙调素的主要分布形式由分散的单颗粒转变为聚集颗粒 ,受精完成后再变为分散的单颗粒形式。胚囊壁及珠心细胞的细胞壁和胞间隙中也观察到钙调素的分布和数量变化。初步讨论了胞内和胞外钙调素在水稻受精与合子形成中的作用。     

莴苣助细胞发育过程中钙的分布研究

用焦锑酸盐沉淀法对莴苣助细胞中的钙分布进行了观察。结果表明,开花前3天刚形成的助细胞中的钙颗粒很少:开花前2天助细胞壁中的钙颗粒增加;开花前1天助细胞珠孔端细胞壁加厚,其中积累了许多钙颗粒:开花当天助细胞珠孔端的丝状器中聚集了大量的钙颗粒。授粉后1h时两个助细胞的结构和钙分布发生差异,一个呈退化状,其中的钙颗粒明显增多,另一宿存助细胞中的钙分布与授粉前相似。去雄不授粉1天后两个助细胞均保持完好,且两助细胞中的钙分布没有明显差异,表明由花粉管引起一个助细胞中钙含量增加进而导致了助细胞退化。退化助细胞在卵细胞与中央细胞之间形成一薄层。助细胞退化后不同部位的钙颗粒呈现出与受精作用密切有关的变化:授粉后1h时,钙主要聚集在近合点端部位;授粉后2.5h卵细胞即将受精,这时许多细小的钙颗粒主要聚集在卵细胞与中央细胞之间的薄层中;授粉后4h精、卵细胞已融合,这时退化助细胞合点端的钙颗粒明显减少,而在其珠孔端又聚集了较多的钙。上述助细胞中的钙含量变化与吸引花粉管进入胚囊和促使精卵细胞融合密切有关。     

水稻雌蕊与胚囊中钙的超微细胞化学定位

用焦锑酸盐沉淀法对水稻（OryzasativaL．）授粉前后雌蕊和胚囊中的钙进行了超微细胞化学定位。结果表明,柱头乳突细胞表面和花柱薄壁细胞中均含钙沉淀;开花前1d,整个胚囊中含钙较少,两个助细胞中钙分布无差异;临近开花时,1个助细胞已退化,其钙含量明显增加;开花后6h,胚囊已受精,退化助细胞中钙含量进一步增加;受精前卵细胞中钙主要分布在液泡中,核和胞质中较少;受精后,其钙含量明显增加,主要分布于核中。重点讨论了钙与助细胞退化和卵细胞激活的关系。     

莴苣卵细胞、合子与原胚细胞中钙的分布

用焦锑酸盐沉淀法对莴苣开花前后的卵细胞、合子与原胚细胞中的钙颗粒分布变化进行了观察。结果表明,开花前三天,刚形成的卵细胞内钙颗粒很少,开花前二天的卵细胞内钙颗粒开始增多,开花前一天的卵细胞形成了大液泡,建立了极性,细胞内的钙颗粒又减少。开花后、受精前的卵细胞的钙颗粒主要聚集在细胞核中。受精后合子中的钙颗粒又明显增多,在核质中分布一些较大的钙颗粒,在珠孔端大液泡中聚集了较多的絮状钙。二胞原胚中的钙颗粒又开始减少,多胞原胚细胞中的钙进一步减少,但原胚表面分布一层丰富的钙颗粒。探讨了钙在卵细胞分化成熟、受精以及原胚发育初期中的作用。     

受精前后烟草卵细胞内玉米素和三类酸性植物激素分布的免疫金电？…

用胶体免疫金电镜技术观察了受精前后烟草（Ｎｉｃｏｔｉａｎａ ｔａｂａｃｕｍ ｖｅｒ．ｍａｒｏｐｈｙｌｌａ）卵细胞内玉米素（ｔ－Ｚ）、ＧＡ７与ＧＡ４、（＋）ＡＢＡ与ＩＡＡ分布的变化。受精前卵细胞内有大量ｔ－Ｚ,主要位于细胞核、内质网与线粒体上;与卵细胞相邻的助细胞合点端及中央细胞珠孔端亦有较多ｔ－Ｚ。受精后,例子与宿存助细胞ｔ－Ｚ显著减少,正加厚的合子细胞壁中ｔ－Ｚ分布。未受精卵细胞内ＧＡ７与ＧＡ４     

西瓜不同发育时期的助细胞超微结构的变化

被子植物胚囊的“雌性生殖单位”,已在多种植物上进行了超微结构的观察,但大多都以卵细胞受精前后的结构变化为主要研究内容。对于“雌性生殖单位”中的另一重要成员——助细胞,在不同发育状态下其结构变化的详细资料不多,尤其是助细胞退化后的物质去向,少见报道。本研究主要观察了西瓜不同发育时期(受精前后)、不同发育状态(柱头授粉和未授粉)的助细胞超微结构,以期为研究助细胞在双受精中所起作用提供新的资     

受精前后烟草卵细胞内玉米素和三类酸性植物激素分布的免疫金电镜观察

用胶体免疫金电镜技术观察了受精前后烟草（Nicotianatabacumvarmacrophylla）卵细胞内玉米素（t－Z）、GA7与GA4、（＋）ABA与IAA分布的变化。受精前卵细胞内有大量t－Z,主要位于细胞核、内质网与线粒体上;与卵细胞相邻的助细胞合点端及中央细胞珠孔端亦有较多t-Z。受精后,合子与宿存助细胞t－z显著减少,正加厚的合子细胞壁中有t－Z分布。未受精卵细胞内GA7与GA4及（＋）ABA减少,IAA更少。合子内GA7与GA4及（＋）ABA略有增加,IAA仍然很少。初步讨论了上述植物激素与受精作用的关系。     

多胚水稻ApⅢ的胚胎发生:受精前后卵器的结构和组织化学研究

对高频率多胚水稻(Oryza sativa L.) ApⅢ受精前后卵器的细胞结构和组织化学变化进行了观察,并同已报道的正常水稻和多胚水稻大至相同发育时期的卵器进行了比较,结果表明: ApⅢ的2 932个幼嫩子房中,每个子房只有一个胚囊.没有看到含有一对胚囊和每个胚囊里有一套卵器的现象.除解体的和含胚的胚囊外, 1 655个胚囊中,含1个卵细胞和2个助细胞组成的正常卵器为1 643个 (99.27%), 含2个卵细胞和2个助细胞的4细胞卵器为12个(0.73%).没有观察到大量4细胞卵器、5细胞卵器(即由3个卵细胞和2个助细胞组成)和卵状细胞,以及其他4卵、5卵卵器的变异类型.卵细胞位于对着子房壁维管束一侧.细胞质含丰富的蛋白质和多糖颗粒;细胞核位于细胞中下部,少有偏远轴端的,直到合子分裂前由蛋白质物质和多糖颗粒聚成的环所包裹. 成熟胚囊中常见2个助细胞.助细胞位于珠孔端靠子房壁维管束一侧,多数为长颈烧瓶状,少有长形和星月形的.其珠孔端壁内侧丝状器发达,细胞质的结构,蛋白质物质和多糖颗粒的积累、分布及消长,细胞核的大小、组织化学反应和周围物质的动态与卵细胞的相同.此外,ApⅢ的2个助细胞存留时间较长.当花粉管进入助细胞的早期,助细胞的丝状器和帽颈端被花粉管损伤,中下部细胞质和核所在区仍保持完好.由以上结果得出结论: 多胚水稻ApⅢ高频率的额外胚(1或2个)主要来自3细胞正常卵器,极少来源于4细胞卵器; ApⅢ的助细胞除在受精和胚胎发生早期具特殊功能外,与卵细胞相似的细胞质结构、物质代谢过程以及崩溃较晚可能与胚胎发生有关; 在ApⅢ的少数胚囊中,接受助细胞可能有发生胚的潜能.     

水稻双受精过程的细胞形态学及时间进程的观察

应用常规石蜡切片和荧光显微镜观察水稻(Oryz a sativa)受精过程中雌雄性细胞融合时的形态特征及时间进程, 确定合子期, 为花粉管通道转基因技术的实施提供理论依据。结果表明: 授粉后, 花粉随即萌发, 花粉管进入羽毛状柱头分支结构的细胞间隙, 继续生长于花柱至子房顶部的引导组织的细胞间隙中, 而后进入子房, 在子房壁与外珠被之间的缝隙中向珠孔方向生长, 花粉与花粉管均具有明显的绿色荧光。花粉管经珠孔及珠心表皮细胞间隙进入一个助细胞, 释放精子。精子释放前, 两极核移向卵细胞的合点端; 两精子释放于卵细胞与中央细胞的间隙后, 先后脱去细胞质, 然后分别移向卵核和极核, 移向卵核的精核快于移向极核的精核; 精核与两极核在向反足细胞团方向移动的过程中完成雌雄核融合。大量图片显示了雌雄性核融合的详细过程以及多精受精现象。水稻受精过程经历的时间表如下: 授粉后, 花粉在柱头萌发; 花粉萌发至花粉管进入珠孔大约需要0.5小时; 授粉后0.5小时左右, 花粉管进入一个助细胞, 释放精子; 授粉后0.5-2.5小时, 精卵融合形成合子; 授粉后约10.0小时, 合子第1次分裂, 合子期为授粉后2.5-10.0小时; 授粉后1.0-3.0小时, 精核与两极核融合; 授粉后约5.0小时, 初生胚乳核分裂。     

普通小麦与鸭茅状摩擦禾的远缘杂交 Ⅲ受精和早期胚胎的发育

用石蜡制片法和整体解剖法,对小麦品种Ｆｕｋｕｈｏ经鸭茅状摩擦禾花粉授粉后不同时间固定的子房样品进行了细胞胚胎学观察。结果表明,观察的１６８个授粉后１２０小时（５天）以内的小麦子房中有３２１％、０６％和２６２％分别发生了卵细胞单受精、极核单受精和卵细胞、极核双受精现象。发生卵细胞受精、极核受精和总受精频率分别为５８３％、２６８％和５８９％。对１１６个授粉后９天的小麦子房进行整体解剖,检测到有５００％的子房有幼胚。报道了小麦与鸭茅状摩擦禾杂交的受精过程和早期胚胎发育的情况。为小麦与鸭茅状摩擦禾杂交提供了高频率受精和胚形成的细胞胚胎学证据。     

EMBRYO SAC LACKING ANTIPODAL CELLS IN ARABIDOPSIS THALIANA (BRASSICACEAE)

Ultrastructure of the embryo sac lacking antipodals in prefertilization stages in Arabidopsis thaliana has been examined 2 hr before and 5 hr after manual cross pollination. The cytoplasm of both synergids before fertilization is rich in ribosomes, mitochondria, and rough endoplasmic reticulum, and also contains several microbodies and spherosomes. The filiform apparatus includes electron-dense material and a fibrous part. Many cortical microtubules appear in the filiform apparatus area. One of the two synergids degenerates before fertilization. The synergids, the egg cell, and central cell have a rich cytoskeleton of microtubules; only the synergids appear to contain microfilaments. At the chalazal end, the antipodals are initially present but degenerate by the time of pollination in most embryo sacs in the starchless line studied. The embryo sac is completely surrounded by a wall containing an electron-dense layer, separating it from the nucellus, including the chalazal end. When the antipodals have degenerated, the electron-dense layer disappears at the chalazal end only, and the wall between the central cell and the nucellus is homogeneous. Between the central cell and nucellar cells no plasmodesmata are found. The membranes of both antipodal cells at the chalazal end of the embryo sac appear sinuous, like those of transfer cells. The central cell has plastids preferentially distributed around the nucleus, but the other organelles are randomly distributed. The central cell in the embryo sac and the adjacent chalazal nucellar cells show a transfer-cell function in the embryo sac after the antipodals degenerate.     

Ultrastructure of the Developing and Fertilized Embryo Sac of Amaranthus hypochondriacus L.

Amaranthus hypochondriacus embryo sac development was investigatedbefore and after fertilization. During the early stages of development,the young embryo sac displays three antipodal cells at the chalazalpole that degenerate very early in the maturation process, beforethe synergids and egg cell are completely differentiated. Themature embryo sac is composed only of the female germ unit.The synergid cells organize a filiform apparatus accompaniedby the presence of mitochondria and dictyosomes with numerousvesicles. The involvement of the synergids in transport andsecretory functions related to pollen tube attraction and guidance,are discussed. The egg cell is located at the micropylar polenear the synergids and displays exposed plasma membranes atthe chalazal pole. The fertilized egg cell does not exhibitmarked changes after fertilization except for the closure ofthe cell wall. The central cell is the largest cell of thisvery long embryo sac. The fused nucleus is close to the eggapparatus before fertilization and displays a remarkable chalazalmigration after gamete delivery. The ultrastructure of the centralcell cytoplasm and the numerous wall ingrowths around this cellsuggest an important role in nutrient transportation. Aftergamete delivery, the embryo sac displays electron dense bodiesthat aggregate within the intercellular space between the synergids,egg cell and central cell. These bodies, that appear in theembryo sac of several plants, are probably involved in gametedelivery for double fertilization. The possibility of biparentalinheritance of mitochondria in this plant is also discussed.Copyright 1999 Annals of Botany Company Amaranthus hypochondriacus, grain amaranth, embryo sac, fertilization.     

东北地区野豌豆属VICIAL．物种生物学研究Ⅷ．幼苗形体结构动态和种间界限

在野外居群调查的启示下,本文以组件观点对柳叶野豌豆复合种和歪头菜幼苗亚单位的时序变化与开花关系进行了分析。结果发现在柳叶野豌豆复合种栽培居群中存在打破物种间形体结构特征的个体,即在复叶由一对小叶组成的植株就已开花而进入生殖时期。另外,在歪头菜的野生居群中发现由三或四枚小叶组成复叶的个体,因此,我们推测这种形体结构的变化可能暗示着柳叶野豌豆复合种和歪头菜有着共同的祖先。     

Structure of Embryo Sac Before and After Fertilization and Distribution of Transfer Cells in Ovules of Green Gram

The structure of embryo sac before and after fertilization, embryo and endosperm development and transfer cell distribution in Phaseolus radiatus were investigated using light and transmission electron microscopy. The synergids with distinct filiform apparatus have a chalazal vacuole, numerous mitochondria and ribosomes. A cell wall exists only around the micropylar half of the synergids. The egg cell has a chalazally located nucleus, a large micropylar vacuole and several small vacuoles. Mitochondria and plasrids with starch grains are abundant. No cell wall is present at its chalazal end. There are no plasma membranes between the egg and central cell in several places. The zygote has a complete cell wall, abundant mitochondria and plastids containing starch grains. Both degenerated and persistent synergids migh.t serve as a nutrient supplement to proembryo. The wall ingrowths occur in the central cell, basal cell, inner integumentary cells, suspensor cells and endosperm cells. These transfer cells may contribute to embryo nutrition at different developmental stages of embryo.     

向日葵胚囊的超微结构和雌性生殖单位问题

本文对向日葵胚囊中卵细胞、助细胞与中央细胞开花前和传粉后的超微结构变化进行了研究。着重报道了不同发育时期这三种细胞之间特定区域的界壁的消长动态。在此基础上结合现有文献资料探讨了由三者共同组成“雌性生殖单位”以适应受精作用的问题。     

天竺葵雌性生殖单位的超微结构

应用透射电镜研究了临近受精时天竺葵(Pelargonium hortorum Bailey)胚囊中的卵细胞、助细胞和中央细胞的结构。证明了卵细胞与助细胞以及助细胞与助细胞之间从合点端至珠孔端有很大的面积以质膜分界,仅珠孔端少部分以壁分隔。卵细胞与中央细胞之间同样缺乏细胞壁。在卵细胞的合点端,两质膜不同程度地分离形成宽窄相间的间隙。在间隙的絮状基质中存在小泡,这些小泡的产生似与卵和中央细胞中周质内质网的活动有关。推测小泡为多糖性质,可能为合子新壁的建造提供物质。卵细胞质中含巨大线粒体,质体和内质网也较丰富。基于超微结构的特征,可认为卵细胞具高度的生理合成活动的潜能。中央细胞极核位于珠孔端与卵器细胞毗邻,有利于在双受精作用中同时发生精细胞与卵细胞和精细胞与中央细胞核的融合。中央细胞的侧壁在珠孔端形成内突,具传递细胞的特点,表明这是雌配子体向孢子体摄取营养的重要部位。助细胞的细胞质含丰富的细胞器,这与多数植物中的相似,但具几个明显的特征,即核中存在微核仁,内质网形成圆球体或脂体,线粒体富集在丝状器的附近。传粉后花粉管进入胚囊之前,两个助细胞中一个退化。     

Early Events in the Penetration of the Embryo Sac in Torenia fournieri(Lind.)

At maturity, Torenia fournieri(Lind.) has an embryo sac whichprotrudes through the micropyle placing the synergids, egg celland part of the central cell within the ovary locule adjacentto the placenta. The present study utilized this unique attributein combination with confocal and light microscopy to characterizethe timing and associated structural changes during pollinationevents leading to double fertilization. The observation of spermnuclei in living gametophyte tissue is an important advancein the identification, in real time, of stages leading to fertilizationin angiosperms. A continuum of fertilization occurred between12 and 16 h after pollination (hap), with peak frequency ofegg and sperm fusion at 14 hap (43%). Movement of the spermcells through the degenerated synergid took several hours andfusion between sperm and their respective female nuclei occurredsimultaneously. Changes in embryo sac structure were also documented.Cell walls in the region between the synergids and egg cellwere poorly developed prior to pollen tube penetration. Thickenedcell walls were observed around the periphery of the synergidsand egg cell following pollination, and in the central cellwhere it lay within the body of the ovule. Starch was observedin the cells of the embryo sac, although the number and distributionof granules varied before and after pollination. These temporaland spatial observations of the embryo sac inTorenia fournieriprovide a basis for further research to determine control mechanismsoperating during specific double fertilization events in angiosperms.Copyright 2000 Annals of Botany Company Double fertilization, embryo sac, sperm nuclei, Hoechst, Torenia fournieri     

Ultrastructures of the Egg Apparatus and the Central Cell of Calystegia hederacea (Convolvulaceae) Before and After Fertilizati on

Ultrastructures of the mature embryo sac of Calystegia hederacea Wall. and its changes after fertilization are described. The positional organization of the egg cell, the two synergids and the central cell, as well as their interrelationships were studied. Some regions of the cell boundaries between the egg cell and the central cell, as well as between the egg cell and the synergids were devoid of typical cell wall before fertilization, displayed a feature quite similar to the characteristic absence of the cell wall in the fertilization target zone occurred in most angiosperms. Besides the genera ultrastructural characteristics of the egg apparatus and the central cell, there were several unusual aspects in C. hederacea, such as the egg nucleus located above the large vacuole near the chalazal end of the cell, many polyribosomes in the cytoplasm of the egg cell and wall ingrowths on both sides of the hooks of the central cell. All these unusual characteristics seemed to be closely associated with the short duration of the fertilization and the absence of antipodal cells in the mature embryo sac. It is concluded that the female germ unit of C. hecleracea is considered to be a topographical and physiological unit to realize their functions for successful double fertilization.     

The Ultracytochemical Localzation of Acid Phosphatase Activity in Wheat During Fertilization

No acid phosphatase activity was observed in the mature embryo sac of wheat (Triticum aestivum) except the chalazal cytoplasm Of the central cell before fertilization. During fertilization, acid phosphataseactivity was observed in the following loci: part of chromatin of the egg nucleus and most of the mitochondria in the egg cytoplasm; the perinuclear spaces of the egg and sperm nuclei at the fusion of the egg and sperm nuclei; the chalazal cytoplasm and some vacuoles of the degenerated synergid; two sperm nuclei within the cytoplasm of female cells; the cell wall of each cell of the embryo sac and that of the nucellar cells surrounding the embryo sac. No acid phosphatase was observed in the two-celled proembryo. Dense enzyme reaction product was localized in the chromatin of the free nuclei at early stage of the endosperm. The characteristic of acid phosphatase distribution during fertilization may be associated with the physiological change of the egg Cell, the reorganization of mitochondria in the egg cell cytoplasm, the degeneration of one of the two synergids, the physiological state of the sperm nuclei and the nuclear membrane fusion of the egg and sperm nuclei.