鹤顶兰胚囊发育的超微结构观察

运用电子显微镜技术对鹤顶兰(Phaius tankervilliae(Aiton)BI.)胚囊发育过程中功能大孢子、二核胚囊、四核胚囊、成熟胚囊的超微结构进行观察,捕捉到了功能大孢子的三个阶段、成熟胚囊的两个阶段,进一步积累了鹤顶兰生殖生物学研究的基础资料.在功能大孢子、四核胚囊时期的合点端壁上可观察到胞间连丝,与体细胞间有物质及信息的交换,胚囊发育并非处于完全“隔离”状态.功能大孢子早期可见明显大液泡,随后进入第一次有丝分裂时大液泡消失,移向两极的染色体之间可见大量体积较小的液泡,成熟胚囊前期助细胞及卵细胞内也可见明显液泡,但当助细胞解体时,卵细胞内的大液泡也消失,液泡形态的变化可能是细胞生理状态发生改变的结果.     

鹤顶兰胚囊发育过程中微管变化的共焦显微镜观察

光镜的观察确定了鹤顶兰（Ｐｈａｉｕｓ ｔａｎｋｅｒｖｉｌｌｉａｅ （Ａｉｔｏｎ） Ｂｌ．）胚囊发育属单孢子蓼型。应用免疫荧光标记技术及共焦镜观察了胚囊发育过程中微管分布的变化。当孢原细胞初形成时,细胞内的微管呈网状分布。之后,孢原细胞体积增大发育为大孢子母细胞。大孢子母细胞延长,进入减数分裂Ⅰ。微管由分裂前的网状分布变为辐射状排列。二分体的两个细胞内的微管分布一样,呈辐射状。四分体的近珠孔端的３ 个大孢子解体,细胞内的微管消失。靠合点端的功能大孢子内有许多微管呈网状分布。当功能大孢子进入第一次有丝分裂时,细胞内的微管由网状变为辐射状,从核膜伸展至周质。再经两次有丝分裂形成八核胚囊。在核分裂之前微管一般是呈网状分布并紧包围着核。在分裂期间二核和四核胚囊都呈极性现象,微管系统也呈极性分布。微管在八核胚囊内的分布变化情形特别复杂。首先,八核分别作不同程度的移动,其中两个核移向胚囊中央,珠孔端和合点端的３ 个核分别互相靠拢,形成３ 个区,即中央区、反足区和卵器区。胚囊未形成区时,８ 个核都被网状分布的微管包围着。当胚囊明显分成区时,反足区内的微管仍作网状分布。中央区的微管分布则趋疏松,形成篮形结构,包围着液泡和两个极核。在     

甘蓝型油菜授粉前后珠孔和胚囊中钙分布的超微细胞化学定位

用焦锑酸盐沉淀法对甘蓝型油菜（ＢｒａｓｉｃａｎａｐｕｓＬ．）授粉前后珠孔和胚囊中的钙进行了超微细胞化学定位。Ｘ射线能谱微区定性分析（ＥＤＸＡ）表明前法所得沉淀确系焦锑酸钙。同时利用图像处理系统对胚囊成员细胞中钙沉淀颗粒的体密度和大小进行了定量分析。结果显示,授粉前后内、外珠孔处钙沉淀明显较珠被其它部位为多,钙主要分布在细胞壁和胞间基质等质外体系统中。授粉前２个助细胞中均含有较其它细胞为多的钙沉淀,约为卵细胞的２５倍,中央细胞的１９倍;但沉淀颗粒的直径仅为它们的２／３左右。助细胞中含钙最多的部位是丝状器和细胞核。授粉后１对姊妹助细胞均明显退化,钙含量显著增高,约为授粉前的２４倍;钙颗粒明显减小,不足授粉前的１／３。讨论了钙的分布与助细胞的退化及其功能的关系。     

陆地棉雌蕊的花粉管生长途径中钙分布的超微细胞化学定位

用焦锑酸盐沉淀法对陆地棉(Gossypium hirsutum L.)授粉前后的柱头、花柱、珠孔与珠心组织中的钙进行了超微细胞化学定位。X射线波谱与能谱分析证明所定位的沉淀确系焦锑酸钙。观察结果表明:在整个花粉管生长途径中的雌蕊组织,钙分布均较其它相邻组织密集;钙主要分布在细胞壁与胞间基质等质外体系统中。在雌蕊中生长的花粉管,其尖端细胞器区也有丰富的钙。     

小麦受精过程中酸性磷酸酶的超微细胞化学定位

小麦（Ｔｒｉｔｉｃｕｍ ａｅｓｔｉｖｕｍ ）受精前成熟胚囊,除胚囊中央细胞的合点端细胞质中有酸性磷酸酶外,其余部位均未发现酸性磷酸酶。受精时期,以下部位存在酸性磷酸酶活性：卵细胞的细胞核内一部分染色质和细胞质中大部分线粒体;精、卵核融合时两核的核周腔内;退化助细胞合点端细胞质和一些液泡内;进入雌性细胞中的两个精核;胚囊各成员细胞的细胞壁及胚囊周围珠心细胞的细胞壁。二细胞原胚中未见有酸性磷酸酶。早期胚乳游离核染色质上有酸性磷酸酶。小麦受精过程酸性磷酸酶的分布特点可能与卵细胞生理状态的变化和细胞质中线粒体的改组、助细胞的退化、精核的生理状态以及精核与卵核的核膜融合等有关。     

向日葵柱头,花柱和珠孔中钙分布的超微细胞化学定位

用焦锑酸盐沉淀法对向日葵（ＨｅｌｉａｎｔｈｕｓａｎｎｕｕｓＬ．）授粉前后柱头、花柱和珠孔中的钙进行了超微细胞化学定位。同时还运用Ｘ射线能谱（ＥＤＸ）和波谱（ＷＤＸ）两种方法进行了Ｘ射线定性分析,证明了前法所得沉淀确系焦锑酸钙。观察表明,花粉萌发和花粉管生长所经的柱头接受面,花柱引导组织和珠孔引导组织中含钙较柱头、花柱和珠孔的其它部位明显地多。柱头乳突细胞的表面和花柱引导组织的胞间基质中、尤其胞间基质与细胞壁外层相接之处钙很密集。在珠孔外端引导组织中,以角质层为界,钙主要分布于其近珠柄侧。花粉管壁果胶质层中有相当多的钙。结合向日葵中已有的研究和其他文献,讨论了钙的分布与花粉管生长的关系     

小麦珠心细胞衰退过程酸性磷酸酶的超微细胞化学定位(英文)

采用磷酸铅盐沉淀技术对小麦（ Ｔｒｉｔｉｃｕｍ ａｅｓｔｉｖｕｍ Ｌ．） 珠心细胞衰退过程进行了酸性磷酸酶的超微细胞化学定位研究。结果显示,在未有明显衰退迹象的一些珠心细胞中,酸性磷酸酶只出现在细胞核轻微凝聚的染色质上。随珠心细胞衰退程度的逐渐增大,其衰退特征越来越明显,酸性磷酸酶依次在细胞质中较小液泡、细胞壁、线粒体、质体以及内质网等结构上出现活性反应。紧连胚囊的珠心细胞衰退程度最大,细胞严重变形,酸性磷酸酶定位于细胞绝大部分结构中,但此时变形的细胞核则无酸性磷酸酶活性反应。研究结果表明,小麦珠心细胞的衰退过程中,酸性磷酸酶存在一个有规律的变化,支持珠心细胞的衰退是属于细胞程序性死亡类型的观点     

小麦珠心细胞衰退过程中ATP酶的超微细胞化学定位

采用磷酸铅沉淀技术对小麦（Ｔｒｉｔｉｃｕｍ ａｅｓｔｉｖｕｍ ）珠心细胞衰退过程进行了ＡＴＰ酶的超微细胞化学定位。初始衰退的珠心细胞,ＡＴＰ酶只定位于细胞膜上,其它部位未见有ＡＴＰ酶活性。衰退中期的珠心细胞,细胞膜上ＡＴＰ酶活性减弱并逐渐消失;细胞核染色质和细胞质中一些细胞器上存在ＡＴＰ酶活性。在严重衰退的珠心细胞中,只在细胞核染色质上存在ＡＴＰ酶活性。珠心细胞的细胞核以两种方式衰退。衰退的细胞核染色质碎片仍存在ＡＴＰ酶活性,并向胚囊方向转移。推测小麦珠心细胞衰退过程中细胞膜上ＡＴＰ酶变化反映了珠心细胞生理状态转变;细胞核染色质上ＡＴＰ酶与其形态变化和运动等有关     

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。     

西瓜胚乳吸器的发育及ATP酶的超微细胞化学定位

报道了西瓜（Citrullus lanatus)胚乳吸器发育过程,并对胚乳吸器细胞中的ATP酶进行了超微细胞化学定位,球形胚早期,胚囊合点端的壁伸长发育成一管状胚乳吸器,进而吸器靠近乳本体端膨大为囊状,球形胚晚期吸器自珠孔端向合点端逐渐细胞化,胚分化出子叶时,胚乳吸器自合点端向珠孔端退化,在刚形成的胚乳吸器细胞中,ATP酶活性反应主要分布在细胞的核膜,内质网上,胞间连丝和吸器细胞壁内的小球状物上也有较强的ATP酶活性反应;在开始退化的吸器细胞中,核膜上的ATP酶性的反应减弱较早,内质网稍晚,进一步退化的胚乳吸器细胞中,ATP酶主要集中分布在细胞壁,细胞间隙内,核上几乎没有ATP酶性反应,内质网上仅有微弱的ATP酶反应。     

鹤顶兰花粉管在子房中的生长途径

运用扫描电镜对鹤顶兰(Phaiustankervilliae(Aiton)Bl.)花粉管在子房内的生长途径进行了观察。结果表明:花粉管在子房中的生长途径可以分为3个阶段:(1)沿子房壁轴向生长阶段,从授粉开始至大孢子母细胞四分体时期,花粉管经过合蕊柱到达子房,经由胎座基部沿子房壁轴向生长;(2)沿子房径向生长阶段,二核胚囊之后,花粉管在胚珠之间穿梭,以径向生长为主;(3)朝珠孔定向生长阶段,胚囊成熟时,花粉管朝珠孔定向生长进入胚囊。实验结果说明花粉管的定向生长受胚珠的分子信号调控。     

Ultracytochemical Localization of Calcium in Micropyle and Embryo Sac of Brassica napus Before and After Pollination

Potassiam antimonate was used to localize Ca2+ in the micropyle and embryo sac of Brassica napus L. before and after pollination. To identify the nature of the pyroantimonate deposits, energy-dispersive X-ray microanalysis (EDXA) was employed and the deposits were proved to contain calcium pyroantimonate. Image processing system was employed to measure the volume density and the diameter of the deposits. Before and after pollination, calcium was more abundant in the exostome and endostome as compared with the other regions of the integuments, and was concentrated at the apoplast system, i.e. the intercellular matrix of the micropyle canal and the cell wall. Before pollination, each of the two sister synergids accumulated more calcium than the other embryo sac cells. Although the mean diameter of the deposits in the synergid was only two-thirds as that in the egg cell and central cell, the volume density of the deposits in the synergid was about 2.5 times and 1.9 times as that in the egg cell and the central cell respectively. The filiform apparatus and the nucleus had the most abundant calcium within a synergid. After pollination both sister synergids degenerated conspicuously and were characterized by much more deposited calcium (about 2.4 times more than before); and the diameter of the deposits decreased dramatically, which was less than one-third as before. The relationship between calcium distribution and synergid degeneration as well as its functions was discussed.     

The microtubular cytoskeleton during megasporogenesis in the Nun orchid, Phaius tankervilliae

This study examines the microtubular cytoskeleton during megasporogenesis in the Nun orchid, Phaius tankervilliae . The subepidermal cell located at the terminal end of the nucellar filament differentiates first into an archesporial cell and then enlarges to become the megasporocyte. The megasporocyte undergoes the first meiotic division, giving rise to two dyad cells of unequal size. Immunostaining reveals that microtubules become more abundant as the megasporocyte increases in size. Microtubules congregate around the nucleus forming a distinct perinuclear array and many microtubules radiate directly from the nuclear envelope. In the megasporocyte, prominent microtubules are readily detected at the chalazal end of the cell cytoplasm. After meiosis I, the chalazal dyad cell expands in size at the expense of the micropylar dyad cell. At this stage, new microtubule organizing centres can be found at the corners of the cells. The appearance of these structures is stage-specific and they are not found at any other stages of megasporogenesis. The functional dyad cell undergoes the second meiotic division, resulting in the formation of two megaspores of unequal size. The chalazal megaspore enlarges and eventually gives rise to the embryo sac. As the functional megaspore expands, the microtubules again form a distinct perinuclear array with many microtubules radiating from the nuclear envelope. A defined cortical array of microtubules has not been found in P. tankervilliae during the course of megasporogenesis.     

Study on Degeneration of Unfertilized Aposporous Embryo Sac in Pennisetum squamulatium (Gramineae)

To understand the degeneration of unfertilized aposporous embryo sac in an aposporous species Pennisetum squamulatum, the central cell in the unfertilized embryo sac was characterized ultrastructurally . The most prominent sign of degeneration is that the central cell nucleus produced nuclear vacuoles . These nuclear vacuoles can be classified into singleanddouble- layered types . Single- layered nuclear vacuoles are found in the cytoplasm, while the double-layered are inside the nucleus . There are two ways to produce nuclear vacuoles . One is that the outer membrane of the nuclear envelope distends towards the cytoplasm to form nuclear vacuoles ; and the other is the double-layered nuclear envelope derives vacuoles by depressing inwards . Nuclear envelope types are in relation to the ways they are produced . All nuclear vacuoles absorb cytoplasm or nuclear matrix , and trap organelles such as mitochondria .     

多花地宝兰胚囊和胚发育的细胞学研究

为探讨多花地宝兰(Geodorum recurvum)胚胎发育的系统分类学意义,采用石蜡制片法对多花地宝兰胚囊和胚的发育进行解剖学观察。结果表明,在开花前,多花地宝兰胚珠原基发育缓慢,开花授粉后胚珠原基快速发育成"树状二杈分枝结构",随后在"分枝结构"末端形成孢原细胞,开始胚囊发育。多花地宝兰的胚囊发育属于单孢蓼型胚囊,胚珠具有双层珠被。孢原细胞形成后,经过细胞膨大延长发育形成胚囊母细胞,胚囊母细胞经过减数分裂形成线性四分体,在珠孔端形成1个功能大孢子,功能大孢子经过3次有丝分裂形成8核胚囊。多花地宝兰的胚发育具有藜型和紫苑型两种方式。双受精完成后,多花地宝兰合子进行一次橫裂后形成基细胞和顶细胞;基细胞经过多次分裂形成细胞团,细胞团中的细胞向不同方向膨大延长形成多个胚柄细胞;顶细胞有两种分裂方式,一种是横裂形成藜型胚,一种是纵裂形成紫苑型胚。因此,推测多花地宝兰在兰科植物系统分类学上属于较为原始种。     

中国鹤顶兰属(兰科)一新记录种

报道了中国鹤顶兰属Phaius一新记录种——中越鹤顶兰P.tonkinensis(Aver.)Aver.。该种与紫花鹤顶兰P.mishmensis(Lindl.et Paxton)Rchb.f.相似,不同在于该种的花萼和花瓣均为象牙白色,唇盘的脊上无毛,脊两侧具稀疏白色长毛。该文提供了该新记录种的形态描述和墨线图。     

非洲狼尾草未受精无孢子生殖胚囊退化研究

为理解植物无孢子生殖胚囊未受精条件下的退化,对无孢子生殖植物非洲狼尾草未受精成熟胚囊中央细胞退化做了细胞形态学研究。没有受精的中央细胞退化时最显著的特点是细胞核产生核膜囊泡。核膜囊泡有两种类型:单层膜的囊泡和双层膜的囊泡,单层膜囊泡在细胞质中,双层膜囊泡在细胞核内。核膜囊泡有两种发生方式:1)核膜的外膜向细胞质一侧膨胀产生囊泡,囊泡进入细胞质;2)核膜向核内凹陷形成囊泡,囊泡进入细胞核。核膜囊泡类型与产生方式密切关联。核膜囊泡吞噬并消化包括线粒体在内的细胞质和核质。     

The Use of a Whole Stain-Clearing Technique for Observations on Embryo Sac,Embryo, Endosperm and Embryoid

A new stain-clearing procedure has been developed for embryological observations on whole mounted specimens. Ovules of Helianthus annus and Nicotiana tabacum as well as ovaries of Oryza sativa were stained with diluted Ehrlich's hematoxylin for a proper short time, followed by steps of washing and dehydration, and finally cleared and mounted in methyl salicylate. When observed by ordinary bright-field microscopy, the embryo sacs before fertilization and the embryos and endosperms after fertilization were clearly visible. The gynogenic embryoids induced in unpollinated rice ovaries in vitro were also finely detectable. The Ehrlich's hematoxylin-methyl salicylate technique has the merits of rapidity in specimen preparation, high contrast and three dimensional view, needlessness of phase- or interference-contrast equipment, and the feasibility for a wide range of materials. The special significance of this technique for in vitro embryological studies is emphasized.     

An Ultrastructural Study on the Development of Egg- apparatus in Rice Embryo Sac

The development of the egg-apparatus (consisted of an egg cell and two synergids) of rice ( Oryza sativa L. ) was studied at the uhrastructural level. The walls of the egg cell and synergids, immediately after their formation, possessed numerous plasmodesmata. Plasmodesmata were also present on walls between the egg cell and synergids. During the enlargement phase of the egg cell and synergids, the walls at the tip region began to loosen and vesiculate. By the time the embryo sac became mature, the part of the wall of the egg cell and synergids, facing the chalaza, disappeared. Consequently, the tip regions of the egg cell and synergids were only protected by a plasma membrane. When the embryo sac reached full maturity, the upper and middle region of the wall of the synergids broke up into pieces. At that time one synergid began to degenerate. Plasmodesmata persisted at the hook region of the wall of both the egg cell and synergids. Most plastids in the egg cell contained starch grains that persisted throughout the period of the embryo sac development. Starch grains in the plastids of the synergids appeared only before the time when the two polar-nuclei moved into the region above the egg-apparatus. They then disappeared and did not appear again until the embryo sac had reached full maturity. The size and location of the vacuoles in the egg cell were different from those in the synergids. The time of formation was also different. Vacuoles in the egg cell formed late in comparison with the synergids. Vacuoles in the chalazal region of the egg cell (especially at the early stage of the embryo sac development) were much larger than those in the micropylar region. Vacuoles in the synergids tended to concentrate mainly in the chalazal region. There was a peak period of lipid formation in the two synergids. The peak appeared when the embryo sac neared maturity. At the early stage of development, the nuclei of the synergids were elliptical in shape and were situated at the central region near the micropyle. The shape of the nuclei at the late stage of development became less regular and tended to move more towards the micropylar region. Changes in the uhrastructure of the egg cell and synergids of rice appeared to be closely related to the metabolic processes controlling the embryo sac formation and development.