The Ultracytochemical Localization of ATPase Activity in the Ovules of Antirrhinum majus L.

The ultracytochemical localization of ATPase activity was carried out by the method of lead precipitation in the ovules of Antirrhinum majus L. No ATPase activity is observed in the egg apparatus, but some in the polar nuclei, cytoplasm and plasma membrane of the central cell. Between the embryo sac wall and the cuticle surrounding it, there is a gap where some filamentand vesicle-like structures were demonstrated by conventional staining method, and much of ATPase activity is found there. At the chalaza of the ovule, a lot of ATPase particles are found irt the nuclei, plasma membranes and the thick and loose wall of the hypostase cells. The particles of ATPase in the hypostase and those in the gap surrounding embryo sac are continuously distributed through the intervals of the cuticle at the chalazal end of the embryo sac. Some of ATPase particles are found on the plasma membranes and plasmadesmata of integument ceils, noticeably much more in the nucleoplasm of the integumentary tapetum. According to the ATPase distribution pattern in the ovules, we suggest that the function of the integumentary tapetum and hypostase is secretion, and that the gap surrounding the embryo sac may be an apoplastic ehannal for nutrient flow into the embryo sac.     

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。     

金鱼草珠被绒毡层壁囊的分离与鉴定

由金鱼(Antirrhinum majus L.)草开花后的胚珠中分离出抗乙酰解、抗酶解、对强氧化剂有一定抗性但不抗二乙醇胺的细胞壁囊状结构。切片原位观察表明,它是珠被绒毡层的内壁,能被苏丹 IV 染色与金胺 O 荧光染色。它全面地包围胚囊内正在发育的胚和胚乳,仅在合点与珠孔两端开口,而在此二处分别有胚乳吸器与胚柄与外相通。这一结构的功能可能是保证营养物质定向导入胚囊以及防止珠被绒毡层分泌的水解酶侵蚀胚囊等。     

宁夏枸杞大孢子发生和雌配子体发育的细胞学研究

采用半薄切片技术和组织化学染色法对宁夏枸杞大孢子发生和雌配子体发育过程中的细胞结构变化及营养物质积累特征进行了观察。结果表明,(1)宁夏枸杞为中轴胎座,多室子房,倒生胚珠,单珠被,薄珠心类型。(2)位于珠心表皮下的孢原细胞可直接发育为大孢子母细胞,减数分裂后形成直线型大孢子四分体,合点端第一个大孢子发育为功能大孢子,胚囊发育类型为蓼型,具有珠被绒毡层。(3)初形成的胚囊外周组织中没有营养物质积累,成熟胚囊时期出现了大量的淀粉粒且呈珠孔端明显多于合点端的极性分布特征。(4)助细胞的珠孔端具有明显的丝状器结构,呈PAS正反应表现出多糖性质,成熟胚囊具有承珠盘结构。     

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.     

山茶科核果茶属和石笔木属的胚胎学研究

观察了短叶核果茶,石笔木,粗毛石笔木和屏石笔木的大小孢子和雌雄配子体的发生和发育过程,４个种的胚胎学特征高度相似,均为基本型药壁发育,腺质线毡层,同时型小孢子母细胞胞质分裂,四面体形小孢子四分体,二细胞成熟花粉,倒生胚珠,双珠被,薄珠心,单孢原,蓼胚囊,卵细胞与助细胞区分明显,均具有珠被绒毡层和承珠盘,以蓼型胚囊区别于邻近的山茶属。     

Ultrastructure of endothelium in ovules of Penstemon gentianoides Poir. (Scrophulariaceae) at mature embryo sac phase

In this study ultrastructural differences between endothelial cells of different location in Penstemon gentianoides have been examined with electron microscope at mature embryo sac phase. Embryo sac is of the Polygonum type and surrounded by endothelium except the micropylar region. The cuticle is located primarily around the chalazal three-fourths of the embryo sac. Endothelium cells around the chalaza and toward the micropylar region are rich in cytoplasmic organelles. The cytoplasm of endothelial cells near the central cell has large vacuoles and few organelles. There are also plasmodesmas on the anticlinal walls of endothelial cells. The endothelium and the micropylar integumentary cells play a role in transport of metabolites into the embryo sac.     

Embryo sac development in Arabidopsis thaliana II. The cytoskeleton during megagametogenesis

The microtubular and actin cytoskeletons have been investigated during megagametogenesis in Arabidopsis thaliana using immunofluorescence labelling of isolated coenocytic and mature embryo sacs. We found both actin and microtubules (MTs) to occur in abundance throughout megagametogenesis and in all constituent cells of the mature embryo sac. During many stages, the patterns of distribution of these cytoskeletal elements are congruent and may prove to be co-aligned. Many changes in the arrays of MTs and microfilaments take place and indicate varying roles of the cytoskeleton in the different stages and cell types of megagametogenesis. Two major populations of MTs recur throughout embryo sac formation: (1) Elaborate nuclear-based networks are found during the two-nucleate and four-nucleate developmental stages as well as in the egg cell. These arrays may function in positioning the nuclei. (2) Cytoplasmic MTs in longitudinal orientation in the two-nucleate embryo sac, synergids and part of the egg cell, or in a reticulate pattern in the four-nucleate embryo sac, egg and central cell probably participate in organization of the cytoplasm. Synergid MTs converge at the filiform apparatus. Preprophase bands of MTs are absent throughout megagametogenesis but phragmoplast arrays occur during cellularization of the embryo sac. Well developed arrays of cortical MTs are restricted to the antipodal cells. A large concentration of MTs in the part of the egg cell adjacent to the synergids is well placed for being involved with sperm cell movement within the degenerative synergid. On the basis of the morphology of the cytoskeleton, we concur with views that the shape of megagametophyte is largely determined by the surrounding tissues, including the integumentary tapetum.     

Changes of calcium distribution in ovules ofTorenia fournieri during pollination and fertilization

Summary Calcium distribution in ovules ofTorenia fournieri was studied by electron energy loss spectroscopy and transmission electron microscopic visualization of calcium antimonate precipitates. High calcium levels were found in the ovules ofT. fournieri. Calcium is situated mainly in extracellular regions before fertilization, including the surface of embryo sac, in the mucilage, and among the cells of the egg apparatus. Intracellular calcium was found only in the nucellar cells around the embryo sac and in the epidermis of the central axis and funiculus. After pollination, a labyrinthine structure (coralloid-like cell wall formation) develops on the micropylar surfaces of the egg apparatus that contain high levels of calcium. Calcium levels increase in the degenerating synergid after the penetration of the pollen tube. Calcium-antimonate precipitates are abundant in vacuoles of the disrupted synergid and pollen tube cytoplasm.Abbreviations EELS electron energy loss spectroscopy - EDX energy-dispersive X-ray microanalysis - LS labyrinthine structure     

Self-sterility in Ipomopsis aggregata (Polemoniaceae) is due to prezygotic ovule degeneration

Based on previous studies, extreme (>99%) self-sterility in scarlet gilia (Ipomopsis aggregata) appears to be involved in late-acting ovarian self-incompatibility (OSI). Here, we confirm this suggestion by comparing structural events that follow from cross- vs. self-pollinations of I. aggregata. Growth of cross- and self-pollen tubes in the style at 11 h and growth in the ovary at 24 h was equivalent. Nonetheless, by 24 h, cross-pollen effected a significantly higher percentage of both ovule penetration and fertilization. Ovules in self-pollinated flowers showed pronounced changes, including an absence of embryo sac expansion and reduced starch in the integument, by 11 h post-pollination, well before pollen tube entry into the ovary. In addition, the integumentary tapetum and adjacent 1-3 cell layers exhibited abnormal cell division, pronounced deposition of thick, pectin-rich cell walls, and cellular collapse. Ovules and embryo sacs from cross-pollinated flowers rarely showed such features. Developmental changes in ovules from self-pollinated flowers eventually resulted in integument and embryo sac collapse, a process not observed in ovules of unpollinated flowers. We suggest that OSI involves long-distance signaling between self-pollen or self-pollen tubes and carpel tissue that reduces availability of receptive ovules for fertilization before pollen tubes arrive in the ovary.     

狗枣猕猴桃果实发育的解剖学观察

对狗枣猕猴桃果实发育进行了显微和超微结构观察。其主要过程可分为：1. 早期胚胎发育：多心皮合生子房,通常具16个心室,中轴胎座。中轴上着生大量倒生胚珠,单珠被,具珠被绒毡层。胚囊蓼型。授粉后约2～4h花粉萌发。授粉后约120h花粉管到达胚珠。受精后,初生胚乳核分裂先于合子。胚乳发育为细胞型。2. 果壁：果壁可分为果皮、果肉和果心三部分。果皮较薄,由2～3层薄壁细胞组成。外表面光滑无毛,但覆有角质层,气孔下陷,分布于其中。果肉的大部分由薄壁细胞组成,分大、小两种细胞 ,小细胞含较多淀粉粒,淀粉粒的水解是果实软化的原因之一。果肉薄壁组织细胞还含有叶绿体及两种异细胞,一种异细胞内含物为结晶,普遍存在;另一种含被番红染成红色的絮团状物。果心由维管束和薄壁细胞组成,维管束不发达。3. 种子：种皮外表面呈蜂窝状,种皮较硬,由两层细胞构成。线性直立胚,胚乳发达,主要贮藏物是蛋白质。     

Embryological Studies on Narcissus tazetta var. chinensis

Chinese narcissus (Narcissus tazetta var.chinensis Roem) blooms but has no seeds.Embryological studies on the species were conducted to discover the causes of its sterility.Its anther wall is composed of four layers of cells,and its tapetum is of the secretory type.The cytokinesis of microspore mother cells is of the successive type,and the tetrad is tetrahedral.During meiosis of microspore mother cells,some chromosomes lagged,and several micronuclei were found in tetrads.Only 27.7% of the pollen grains contained full cytoplasm,and 1.3% of them germinated in culture medium.No pollen grain,however,could germinate on the stigma.The ovary is trilocular with axile placenta,and the ovules are bitegmic,tenuinucellate,and anatropous.Its embryo sac is of the polygonum type.Most embryo sacs degenerated,and only about 4.5% of the ovules contained a normal embryo sac with an egg cell,two synergids,three antipodal,and a central cell containing two polar nuclei.One reason for the sterility of Chinese narcissus is the abnormality of microsporogenesis and megasporogenesis,in which only a few functional pollen grains and embryo sacs are produced.The other reason is that the pollen grains cannot germinate on the stigma.     

The immunolocalization of plasma membrane H+-ATPase in the transfer cell region of Brassica napus (Brassicaceae) ovules

Unfertilized mature ovules of Brassica L. contain an abundance of starch in the integument cells from the micropyle to a plane approximately at the level of the central cell polar nuclei. Inside the embryo sac central cell, in the coinciding region, there are transfer cell-like wall projections with plasma membranes appressed to their inner surfaces. H⁺-ATPase is present along the inner surfaces of the wall projections as indicated by reactivity with antibody raised against plasma membrane H⁺-ATPase. A number of mitochondria are in close association with wall projections in the region of the egg apparatus. Antibody raised against corn plasma membrane H⁺-ATPase cross reacts with a protein of the same size in extracts of Brassica napus indicating that the two species contain a similar plasma membrane H⁺-ATPase.     

Megasporogenesis,Microsporogenesis and Development of Gametophytes in Eleutherococcus senticosus (Araliaceae)

This paper describes megasporogenesis, microsporogenesis, and development of female and male gametophytes in Eleutherococcus senticosus. The main results are as follows: Flowers of E. senticosus are epigynous, pentamerous. Anthers are 4 -microsporangiate. An ovary has 5 loculi. Each ovary loculus has 2 ovules: the upper ovule and the lower ovule. The upper one is orthotropous and degenerates after the formation of archesporial cell, while the lower one is anatropous, unitegmic and crassinucellar, and able to continue developing. In male plants, microsporogenesis and development of male gametophytes took place in regular way, but a series of abnormal phenomena were found in megasporogenesis and development of female gametophytes. The microspore mother cells gave rise to tetrahedral tetrads by meiosis. Cytokinesis was of the simultaneous type. The mature pollen was 3-celled and shed singly. The anther wall formation belonged to the dicotyledonous type. At the stage of microspore mother cell, the anther wall consisted of four layers, i.e. epidermis, endothecium, middle layer, and tapetum. The tapetum was of glandular type and its most cells were binucleate. When microspores were at the uninucleate stage, the tapetum began to degenerate in situ. When microspores developed into 3-celled pollen grains, the tapetum had fully degenerates. In the lower ovule of male flower, the megaspore mother cell gave rise to a linear or “T” -shaped tetrad. In some cases, a new archesporial cell over the tetrad or two tetrads parallel or in a series were observed. Furthermore, the position of functional megaspore was variable; any one or two megaspores might be functional, or one megaspore gave rise to a uninucleate embryo sac, but two other megaspores also had a potentiality of developing into the embryo sac. In generally, on the day when flowers opened, female gametophytes contained only 4 cells: a central cell, two irregular synergids and one unusual egg cell. In female plants, microspore mother cells and secondary sporogenous cells were observed. But at the stage of secondary sporogenous cell, the newly differentiated tapetum took the appearance of degeneration. Later, during the whole stage of meiosis, the trace of degenerative tapetum could be seen. At last, the microsporangium degenerated and no tetrad formed. On the blossom day, all anthers shriveled without pollen grains. In female flowers, megasporogenesis and development of female gametophytes were normal: the tetrad of megaspores was linear or “T”-shaped; the chalazal megaspore was usually functional; the development of embryo sac was of the Polygonum type. On the blossom day, most embryo sacs consisted of 7 cells with 8 nuclei or 7 cells with 7 nuclei; but the egg apparatus was not fully developed. In hermaphroditic plants, microsporogenesis was normal but the development of male gametophytes was partially abnormal. When the hermaphroditic flowers blossomed, there were more or less empty pollen grains in the microsporangium and these pollen grains were quite different in size. The development of most gynoecia was normal but numerous abnormal embryo sacs could be seen. On the blossom day, female gametophytes were mainly 7-celled with 8-nuclei or with 7-nuclei or 4-celled with antipodal cells degenerated; the egg apparatus wasnot fully developed either.     

Anatomy of ovary and ovule in dandelions (Taraxacum, Asteraceae)

The genus Taraxacum Wigg. (Asteraceae) forms a polyploid complex within which there are strong links between the ploidy level and the mode of reproduction. Diploids are obligate sexual, whereas polyploids are usually apomictic. The paper reports on a comparative study of the ovary and especially the ovule anatomy in the diploid dandelion T. linearisquameum and the triploid T. gentile. Observations with light and electron microscopy revealed no essential differences in the anatomy of both the ovary and ovule in the examined species. Dandelion ovules are anatropous, unitegmic and tenuinucellate. In both sexual and apomictic species, a zonal differentiation of the integument is characteristic of the ovule. In the integumentary layers situated next to the endothelium, the cell walls are extremely thick and PAS positive. Data obtained from TEM indicate that these special walls have an open spongy structure and their cytoplasm shows evidence of gradual degeneration. Increased deposition of wall material in the integumentary cells surrounding the endothelium takes place especially around the chalazal pole of the embryo sac as well as around the central cell. In contrast, the integumentary cells surrounding the micropylar region have thin walls and exhibit a high metabolic activity. The role of the thick-walled integumentary layers in the dandelion ovule is discussed. We also consider whether this may be a feature of taxonomic importance.     

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。     

芒苞草形态学和胚胎学研究：Ⅱ.花药和胚珠发育的研究

芒苞草成熟胚珠为倒生型,薄珠心,双珠被。胎座为侧膜胎座向中轴胎座的过渡类型。胚囊发育为单孢蓼型。 成熟胚囊由印器,具二极核的中央细胞及三个反足细胞组成。助细胞呈倒梨形,极性不明显,珠孔端壁有角状的丝状器。中央细胞的二极核在受精前融合为次生核。 花药具二个小孢子囊,花药壁层为单子叶型,具分泌型绒毡层,小孢子母细胞减数分裂时,胞质分裂为连续型,四分体是左右对称式排列,成熟花粉粒为二细胞的。 在花药与胚珠发育过程中,多糖物质的消长是有规律的变化。     

BIOCHEMICAL DETERMINATION OF Mg2+, K+-DEPENDENT,DIETHYLSTILBESTROL-INHIBITED PHOSPHATASE ACTIVITY IN THE OVULES OF SAINTPAULIA IONANTHA BEFORE FERTILIZATION

The occurrence of phosphatase activity that was dependent on Mg²⁺ and K⁺ and inhibited by diethylstilbestrol has been detected in unfertilized African violet ovules with the use of biochemical techniques. Since cation-dependent phosphatase activity is believed to be involved in ion transport across cell membranes, these results are interpreted as lending indirect support to cytochemical evidence that suggests the innermost integumentary cells of African violet ovules are involved in active transport of solutes to the embryo sac.     

Studies on the Development of Male and Female Gametophytes in Sinojakia xylocarpa

Observed in this paper was the development of the microspore and megaspore, male and female gametophytes in Sinojakia xylocarpa, which is endemic to China. The anther comprises four microsporangia. Microspore wall forms simultaneously after meiotic division in PMCs. The arrangment of microspore in a tetrad is tetrahedral. Bicel lular pollen grains appear at the shedding stage. ‘They are 3-colporate, with irregular min ute-faveolate exine sculpture. The anther wall development is of the dicotyledonous type, and its endothecitum develops slight fibrous thickenings, which also form on some epidermal cells. The tapetum is glandular. The pistil with hollow style is composed of three carpels, and its ovary contains several anatropous ovules. The ovule is unitegmic, tenuinucellar, but no obturator was observed. The archesporial cell functions directly as the megaspore mother cell which forms a linear tetrad, but T-shaped tetrad was found in a few ovules. A Polygonum type embryo sac forms from the functional chalazal megaspore. In the mature embryo sac, the synergids are elongate with a large vacuole at the chalazal end, but the distrihution of vacuoles in the egg cell appears random. Two polar nuclei remain in contact with each other for a spell before the fertilization and the 3 antipodal cells may persist into early postfertilzation stages. Numerous starch gra ins occur in the embryo sac. According to the present embryological studies on Sinojakia xylocarpa and the works on embryogenesis by some early embryologist, authors consider that Styracaceae, Symplocaceae, Sapotaceae and Ebenaceae are rather closely related, and we alsoconsider it reasonable to put the 4 families mentioned above in Ebenales.     

Cytopathology of Mature Ovaries from Lettuce Plants Infected by Lettuce Mosaic Potyvirus

The location of lettuce mosaic potyvirus (LMV) in mature ovaries of lettuce plants ( Lactuca sativa ) was studied by conventional thin-section electron microscopy and immunogold cytochemistry. Flexuous filamentous particles and inclusion bodies characteristic of LMV infection were observed in the integumentary tapetum, integument and ovary wall of the mature ovary but not in the embryo sac.