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     

Confocal Microscopic Observations on Microtubular Cytoskeleton Changes During Megasporogenesis and Megagametogenesis in Phaius tankervilliae (Aiton) Bl.

In nun orchid (Phaius tankervilliae (Alton) B1. ) embryo sac development follows the monosporic pattern. Changes in the pattern of organization of the microtubular cytoskeleton during megasporogenesis and megagametogenesis in this orchid were studied using the immunofluorescence technique and eonfocal microscopy. At the initial stage of development the microtubules in the arehesporium were randomly oriented into a network. Later the archesporial cell elongated to form the megasporocyte. The cytoskeleton in the elongated megasporoeyte was radially organized in which microtubules extending from the nuclear envelope to the peripheral region of the cell. The megasporoeyte then underwent meiosis 1 to form a dyad. The dyad cell at the chalazal end was larger than the cell at the micropylar end. Microtubules in the dyad cell were radially oriented. The dyad underwent meiosis to give rise to a linear array of four megaspores (i. e. tetrad formation). The chalazal-far most megaspore survived and became the functional megaspore, which contained a set of randomly oriented microtubules. The microtubules in the other 3 megaspore disappeared as the cells degenerated. The functional megaspore then underwent mitotic division giveing rise to a 2 nucleate embryo sac. The nuclei of the 2-nucleate embryo sac were separated by a set of longitudinally oriented microtubules which ran parallel to the long axis of the embryo sac. Each nucleus in the embryo sac was surrounded by a set of perinuelear microtubules. The gnucleate embryo sac again underwent mitotic division to form a 4-nucleate embryo sac. The division of the two nuclei was synchronous. But the orientation of the division plan of the two spindles was different (i. e. the spindle microtubules at the chalazal end ran parallel with the long axis of the embryo sac and those at the mieropylar end ran at right angle to the axis of the embryo sac). The 4 nuclei of the 4-nucleate embryo sac were all tightly surrounded by randomly oriented microtubules. Later the paired nuclei at the micropylr end and at the chalazal end as well underwent mitotic division in seguence. At this time when the embryo sac had reached the 8-nucleate embryo sac stage. The pattern of organization of the microtubules was very complex. Initially the nuclei were surrounded by a set of randomly oriented microtubules, but after the two polar nuclei had moved to the central region of the embryo sac, three different organizational zones of microtubules appeared, viz: a randomly oriented set of microtubules surrounding each nucleus in the chalazal zone: a set (in the form of a basket) of cortical microtubules which surrounded the vacuoles and the two polar nuclei in the central zone and a loosely knitted network of microtubules surrounding the nucleus that later became the egg cell nucleus in the micropylar zone. The two nuclei that would become the nuclei of the synergids were surrounded by a set of more densely packed mierotubules. Towards far the most micropylar end some microtubules formed thick bundles. The site of appearance of these thick bundles coincided with the site of development of the filiform apparatus. The pattern of microtubule organization after cellularization (i. e. at the beginning of embryo sac maturation) did not change much. The author's results indicated that various patterns of microtubule organization observed in the developing embryo sac of nun orchid reflected the complexity and dynamism of the embryo sac.     

EMBRYOLOGY OF CALYPSO BULBOSA. I. OVULE DEVELOPMENT

Calypso bulbosa is a terrestrial orchid that grows in north temperate regions. Like many orchids, the Calypso has ovules that are not fully developed at anthesis. After pollination, the ovule primordia divide several times to produce a nucellar filament which consists of five to six cells. The subterminal cell of the nucellar filament enlarges to become the archesporial cell. Through further enlargement and elongation, the archesporial cell becomes the megasporocyte. An unequal dyad results from the first meiotic division. A triad of one active chalazal megaspore and two inactive micropylar megaspores are the end products of meiotic division. Callose is present in the cell wall of the megaspore destined to degenerate. In the mature embryo sac the number of nuclei is reduced to six when the chalazal nuclei fail to divide after the first mitotic division. The chalazal nuclei join the polar nucleus and the male nucleus near the center of the embryo sac subsequent to fertilization.     

银杏大孢子形成的形态学研究

有关银杏（CinkgobitobaL．）生殖的研究,早在19世纪后期,Strasbuopr首先作了描述「'］,此后,在这一方面陆续有不少报道。到了本世纪50年代,对于银杏的整个生殖过程已有大致轮廓卜一'］,但是对于生殖各阶段的详细变化,无论在胚胎学或细胞学方面,都有待进一步的观?..     

赤苎无融合生殖细胞胚胎学研究

对赤苎(Boehmeria silvestrii (Pamp.)W.T.Wang)细胞胚胎学研究表明,其生殖模式属无融合生殖的二倍体孢子生殖(diplospory),但其未减数胚囊的发育途径不同于已报道的类型。大孢子母细胞的减数分裂I在到达终变期时停滞,染色体呈单价体状态并维持较长的时间。在尚未到达以核膜、核仁消失,纺锤体出现为特征的中期I前,大孢子母细胞由终变期直接“跳”入间期,从而始终保持了二倍体水平。减数分裂Ⅱ正常进行并产生二倍体二分孢子。珠孔端孢子退化,合点端孢子经3次分裂形成包括1个卵细胞、2个助细胞、2个极核和3个反足细胞的八核胚囊。胚和胚乳分别起源于卵和次生核未受精的自发分裂。胚乳属核型,其发育早于胚。     

Callose in cell walls during megasporogenesis in angiosperms

Summary Callose was detected by fluorescence microscopy in megasporogenesis in all investigated species with mono- and bisporic embryo-sac development. Callose occurs first in the meiotic prophase in the chalazal part of the megasporocyte wall and by the first meiotic metaphase the whole cell is enveloped in a callose-containing wall. Later, there is a marked decrease of callose fluorescence, usually at the chalazal end of the megasporocyte. In Oenothera, where the micropylar megaspore is active, decrease of fluorescence takes place at the micropylar pole of the megasporocyte. Callose appears centrifugally in the cell plates forming eventually the walls dividing the megaspores. It disappears from the walls of the megaspores during degeneration and differentiation.     

Rhinanthus serotinus (Schönheit) Oborny (Scrophulariaceae): immunohistochemical and ultrastructural studies of endosperm chalazal haustorium development

Chalazal endosperm haustorium in Rhinanthus serotinus consists of a single large binucleate cell. It originates from the primary endosperm cell dividing transversely into two unequal cells: a smaller micropylar cell and a larger chalazal cell. The chalazal cell undergoes a single mitotic division, then lengthens significantly during development and functions as a chalazal endosperm haustorium. In this paper, immunofluorescent techniques, rhodamine phalloidin assay, and electron microscopy were used to examine the actin and tubulin cytoskeleton during the development of the chalazal haustorium. During the differentiation stage, numerous longitudinally oriented bundles of microfilaments ran along the axis of transvacuolar strands in haustorium. Microtubules formed intensely fluorescent areas near the nuclear envelope and also formed radial perinuclear microtubule arrays. In the fully differentiated haustorium cell, the actin cytoskeleton formed dense clusters of microfilaments on the chalazal and micropylar poles of the haustorium. Numerous microfilament bundles occurred near wall ingrowths on the chalazal wall. There were numerous clusters of microfilaments and microtubules around the huge lobed polytenic haustorial nuclei. The microfilaments were oriented longitudinally to the long axis of the haustorium cell and surrounded both nuclei. The microtubules formed radial perinuclear systems which were appeared to radiate from the surface of the nuclear envelope. The early stage of degeneration of the chalazal haustorium was accompanied by the degradation of microtubules and disruption of the parallel orientation of microtubules in the chalazal area of the cell. The degree of vacuolization increased, autophagous vacuoles appeared and the number of vesicles decreased.     

开口箭大小孢子发生及雌雄配子体发育

利用石蜡切片技术,对百合科植物开口箭(Tupistra chinensis Baker)大小孢子发生及雌雄配子体发育进程进行胚胎学观察分析,以明确开口箭胚胎发育的特征,为百合科植物的研究提供生殖生物学依据。结果表明:(1)开口箭花药具有4个药室,花药壁的发育方式为基本型,由表皮、药室内壁、中层及绒毡层组成;绒毡层发育类型为分泌型,到四分体花药阶段绒毡层细胞开始解体退化,花药成熟时完全消失。(2)花粉母细胞减数分裂为连续型,依次形成二分体、四分体,四分体为左右对称形;成熟花粉为2-细胞花粉,具单萌发沟。(3)子房3室,倒生型胚珠6枚,双珠被,薄珠心;在花部的分化早期,由珠心顶端表皮下方分化出雌性孢原细胞,孢原细胞经过一次平周分裂形成周缘细胞和造孢细胞,造孢细胞发育为大孢子母细胞;大孢子母细胞第一次减数分裂后形成二分体,珠孔端的二分体孢子退化,合点端的二分体孢子继续第二次分裂,形成两个子细胞依次发育为二核胚囊、四核胚囊和八核胚囊;开口箭的胚囊发育类型为葱型。     

REPRODUCTIVE FEATURES OF DENTARIA LACINIATA AND D. DIPHYLLA (CRUCIFERAE), AND THE IMPLICATIONS IN THE TAXONOMY OF THE EASTERN NORTH AMERICAN DENTARIA COMPLEX

Reproductive features including ovule development, megasporogenesis, megagametogenesis, microsporogenesis, microgametogenesis, pollen tube growth, embryogeny, and natural seed germination were studied in a single population each of Dentaria laciniata Muhl. ex. Willd. and D. diphylla Michx. to test for possible agamospermy. The population of D. laciniata studied is sexual. The archesporial cell functions directly as the megasporocyte. It undergoes two meiotic divisions, but the micropylar cell of the dyad fails to undergo meiosis II, and a linear triplet of three cells is formed. The chalazal megaspore divides to form an eight-nucleate, seven-celled megagametophyte of the Polygonum type. Simultaneous cytokinesis follows the second meiotic division of the microsporocyte yielding a tetrahedral tetrad of microspores. A three-celled pollen grain is formed prior to anther dehiscence. Following apparent fertilization, the Capsella-variation of the Onagrad type of embryogeny results in a conduplicate embryo. Endosperm is initially nuclear, but eventually becomes cellular. Seeds readily germinate in nature. Similar events are documented in one population of D. diphylla up to the organization of the embryo-sac, which disintegrates before cellularization. These reproductive events and other data indicate that the eastern North American species of Dentaria may form a sexual polyploid complex with some sexual populations and some sterile ones.     

Embryo sac development in Arabidopsis thaliana

Summary Aspects of megasporogenesis in Arabidopsis thaliana have been investigated using a variety of histochemical techniques to visualize general cell organization, DNA and callose in whole ovules and sections by bright field, fluorescence, differential interference contrast and scanning electron microscopy. The microtubular cytoskeleton has been studied using immunofluorescence localization of tubulin in sections and whole cells. The observations deviate from reports of preceding studies in that the megasporocyte was found to undergo both meiotic divisions followed by simultaneous cytokinesis (i.e. without an intermediate dyad stage) to give a multiplanar tetrad of megaspores. This represents a variation of monosporic development not previously described. Polarized distribution of organelles prior to meiosis ensures that the functional megaspore receives the largest share. Aberrant wall formation is common between degenerating megaspores. Localized callose deposition in the tetrad separates these cells from the active megaspore. Their pattern of degeneration and displacement is extremely flexible within the embryo sac space. The microtubular cytoskeleton is extensive and largely cytoplasmic, as distinct from cortical, throughout megasporogenesis. In the megasporocyte, megaspores and one-nucleate embryo sac, randomly oriented microtubules throughout the cells may serve to maintain cytoplasmic integrity and position organelles. Numerous microtubules (MTs) associate closely with the nucleus and some radiate from it, perhaps functioning in nuclear positioning. During meiosis MTs are restricted to the spindle configurations and later to the phragmoplasts which form between daughter nuclei. The lack of interphase cortical arrays suggests that the role of internal influences on cell shape is small.     

Microtubular configurations during meiosis and megasporogenesis in Gasteria verrucosa and Chamaenerion angustifolium

Summary In Gasteria and Chamaenerion, microtubular configurations were visualized immunocytochemically during meiosis and megasporogenesis in order to study their relationship to cell development, meiotic divisions and selection of the functional megaspore. In Chamaenerion, the intensity of the fluorescence found in megaspores was weaker than that found in Gasteria. Both plants exhibited concentrations of microtubules around the meiocyte nuclei during pachytene-diplotene. Preprophase bands were not observed. In Chamaenerion, cytoplasmic microtubules radiating from meiocyte nuclei were found at late prophase, the dyad stage and in the functional megaspore; in Gasteria, they were observed only at the dyad stage and in the functional megaspore. During the second meiotic division of Gasteria, dividing cells and their nuclei exhibited differences in volumes. Also, the two microtubular spindles of the dyad cells had different widths. Fluorescence indicating the presence of the cytoskeleton diminished during maturation of the large functional megaspores in both plants, whereas in the three degenerating smaller megaspores, fluorescence intensity persisted. Our conclusion is that only an indirect relationship exists between the organization of the microtubular cytoskeleton and selection of the functional megaspore.     

Female Gametophyte Development in Maize: Microtubular Organization and Embryo Sac Polarity

The developmental stages of the maize embryo sac were correlated with the corresponding silk lengths of ear florets in the female inflorescence. The development of embryo sacs in the ovules of spikes occurs in a gradient pattern with the initiation of the embryo sac beginning at the base of the ear and progressing to the top. At the beginning of meiosis, the presence of conspicuous cortical microtubules coincides with the extensive elongation of the megasporocyte. The spindles at metaphase I and II align along the long axis of the megasporocyte leading to the linear alignment of the dyad and tetrad of megaspores. During megagametogenesis, micropylar and chalazal nuclei of the embryo sac undergo synchronized divisions and migration at the second and third mitosis. Radiate perinuclear microtubules are present during the interphase of the second and third mitosis, and inter-sister nuclear microtubules occur at the late four-nucleate embryo sac. The configuration and orientation of the spindles, phragmoplasts, and pairs of nuclei result in precise positioning of the nuclei. The fusion of the polar nuclei and the formation of a microtubule organizing center-like structure in the filiform apparatus occur right after the first division of the antipodal cells. The different patterns of organization of microtubules in the cells of the mature embryo sac reflect their structural adaptations for their future function.     

北柴胡大小孢子发生和雌雄配子体发育的研究

用石蜡切片法对北柴胡的大、小孢子发生和雌、雄配子体发育过程进行观察研究.结果显示,北柴胡的胚珠为倒生型,单珠被,薄珠心.孢原细胞不经分裂直接发育成大孢子母细胞;四分体线性,多数情况下合点端的大孢子为功能大孢子,少数情况下亚合点端的大孢子也可发育为功能大孢子,蓼型胚囊;八核胚囊时期,珠心基部和两侧的一些珠心细胞保持自己的细胞质和形状,留存较久,成为珠心座细胞,珠被内表皮细胞发育为珠被绒毡层;花药壁发育类型为基本型,绒毡层为腺质绒毡层.小孢子母细胞减数分裂的胞质分裂为同时型,产生正四面体型四分体.成熟花粉为3细胞型.     

小盐芥大孢子发生和雌配子体发育

本论文研究了小盐芥(Thellungiella halophila)大孢子发生和雌配子体发育过程及该阶段与花蕾、花、果实外部形态的相关性。结果如下: 小盐芥雌蕊由2心皮组成, 侧膜胎座, 每室胚珠多数, 弯生, 双珠被, 薄珠心。孢原细胞位于珠心表皮之下, 直接起大孢子母细胞的功能。大孢子四分体线形排列, 合点端大孢子为功能大孢子, 胚囊发育为蓼型。     

延龄草(Trillium tschonoskii Maxim.)的胚胎学研究初报Ⅰ.——大孢子的发生及雌配子体的形成

本文描述延龄草(Trillium tschonoskii Maxim.)的大孢子发生,雌配子体的形成和雄配子体的形态。胚珠为倒生型,双珠被,厚珠心型。胎座为侧膜胎座向中轴胎座的过渡类型,胶囊发育为葱型的变异型。孢原细胞直接发生于幼胚珠的珠心表皮细胞之下,孢原细胞平周分裂,形成初生周缘细胞及初生造孢细胞。初生周缘细胞分裂先于初生造孢细胞,分裂结果与珠心表皮细胞共同形成了珠心组织。初生造孢细胞进一步发育,形成大孢子母细胞。大孢子母细胞经减数第一次分裂后,即出现壁,形成二分体。一般是珠孔端二分体细胞小于合点端二分体细胞,但偶尔也见到前者大于后者的情况。在二分体形成后珠孔端二分体细胞立即退化、或经减数第二次分裂后再退化(该次分裂多为斜向的)。合点端二分体细胞发育,经二核胚囊,四核胚囊,六核胚囊阶段至成熟胚囊。一般在珠孔端的周围淀粉粒丰富,并先于合点端的核进行分裂。珠孔端由二个助细胞,一个卵细胞构成卵器,助细胞具钩突,并具丝状器,两个极核。合点端常见多核仁的大核,成熟胚囊未见八核。成熟花粉粒为二细胞的,花药壁具变形绒毡层,花粉中充满淀粉粒。沼生目型胚乳。     

Localization of Callose during the Occurrence of Megasporocyte in Gastrodia elata Blume

The structure of ovule in Gastrodia elata Blume was very simple. Functional megaspore occurred at the chalazal end. Callose was absent at megasporocyte stage. It first appeared at the chalazal wall during the first meiotic prophase and exhibited continuous fluorescence. Soon later callose fluorescence disappeared in some part of the chalazal wall and many noncallosic dark areas took place, subsequently these nonfluorescence areas became larger and the callose fluorescence appeared discontinuous granulose distribution. This fluorescence maintained until the megaspore formed. The callose of micropylar wall appeared later and usually disappeared before megaspore formation. In the cross walls between the functional and the two degenarated megaspore callose fluorescence was very strong, continued and kept for a long time. But the side walls usually lacked callose. Accoding to the morphological character of simple ovule in G. eiata and the localization of acid phosphatase and polysaecharide grains, the transfer of vegetative materials from surrounding tissues into megasporocyte mainly passing through the chalazal end of megasporocyte. Thus a continuous callose wall deposited at the ehalazal end of megasporocyte, and it in reality caused the “isolation” of meiocyte. It was possible that a reduced form of callose disposition existed in parasetic orchids.     

Embryological Studies in Glycyrrhiza uralensis Fisch.

This paper reports the studies of overall embryology of Glycyrrhiza uralensis Fisch. Development of the anther wall follows the dicotyledonous type. The cytokinesis of the microspore mother cell in meiosis is of simultaneous type. The arrangement of microspores in tetrad is tetrahedral, isobilateral and decussate. Microspores have various types of abortive to development. Mature pollen grain is of the 2-celled type. The ovule is bitegminous, crassinucellate and campylotropous. The megaspore mother cell gives rise to unequal dyad and then linear tetrad. The chalazal megaspore, the second or the third megaspore towards the micropylar end are functional megaspore. The development of the embryo sac conforms to the Polygonum type. Mature embryo sac has various types of variation. The fertilization belongs to the premitotic type of syngamy. The development of most embryoes belongs to the Onagrad type. The development of the endosperm belongs to the nuclear type and the endosperm near the chalazal end develops into haustorium.     

小盐芥大孢子发生和雌配子体发育

本论文研究了小盐芥（Thellungiella halophila）大孢子发生和雌配子体发育过程及该阶段与花蕾、花、果实外部形态的相关性。结果如下：小盐芥雌蕊由2心皮组成,侧膜胎座,每室胚珠多数,弯生,双珠被,薄珠心。孢原细胞位于珠心表皮之下,直接起大孢子母细胞的功能。大孢子四分体线形排列,合点端大孢子为功能大孢子,胚囊发育为蓼型。     

海桐大、小孢子发育及雌、雄配子体形成的研究

利用常规石蜡制片法研究了海桐大、小孢子发生及雌、雄配子体发育的过程。结果显示:(1)小孢子母细胞减数分裂过程中的胞质分裂为连续型,四分孢子为以四面体形为主,四分孢子后期部分小孢子壁皱缩;(2)花药壁由4层结构组成,由外到内为表皮、药室内壁、中层和绒毡层;(3)海桐具多个胚珠,单珠被,薄珠心,胚珠类型为倒生胚珠。大孢子母细胞减数分裂主要形成线形排列的4个大孢子,还具有少有的十字形排列,功能大孢子位于合点端;(4)胚囊发育属单孢型的蓼型,成熟的雌配子体为四细胞五核胚囊。     

小蓬草的胚胎学研究

对小蓬草(Conyza canadensis)大小孢子发生、雌雄配子体形成、受精、胚及胚乳发育过程进行了研究,主要结果如下:花药四室,药壁由表皮、药室内壁、中层和绒毡层组成.表皮退化;药室内壁宿存,细胞柱状伸长,纤维状加厚;中层细胞退化较早,在小孢子母细胞减数分裂开始时仅存残迹;绒毡层于小孢子母细胞减数第一次分裂前期开始原位变形退化,属于腺质型绒毡层;小孢子母细胞减数分裂为同时型,四分体的排列方式主要为四面体形和左右对称形;成熟花粉粒多为3细-胞花粉粒,偶见2细-胞花粉粒.子房下位,2心皮,1室,单胚珠,基生胎座;单珠被,薄珠心,倒生胚珠,具发达的珠被绒毡层.珠心表皮下分化出大孢子孢原细胞,孢原细胞直接发育为大孢子母细胞,大孢子母细胞减数分裂形成4个大孢子直线形排列,仅合点端的大孢子发育成功能大孢子母细胞,胚囊发育为蓼型.两个极核在受精前融合为次生核,珠孔受精.胚乳发育属于核型,胚胎发育为紫菀型;具胚乳吸器.