The Pollen Tube Pathway in the Pistil of Lycopersicon peruvianum

The pollen tube pathway has been studied in unpollinated andpollinated pistils of Lycopersicon peruvianum using histochemicalstains for detection of proteins, lipids and arabinogalactansby bright-field microscopy, and decolourized aniline blue fordetection of pollen tubes by epifluoresence microscopy. Thepollen tube pathway is a continuous tract of mucilage from thestigma surface to the ovule micropyles, and is associated witha continuous tract of specialized, protein-rich transmittingcells comprising the stigmatic papillae, vertical files of stylartransmitting cells and the placental epithelium within the ovary.The superficial exudate of the stigma is hydrophobic and richin lipids. The mucilage of the style and ovary is hydrophilicand rich in arabinogalactans but low in proteins. Pollen tubesgrow between cells through the mucilage of the stigma and stylartransmitting tract, and across the surface of the placenta inthe ovarian mucilage. The structure of the junction of the stylartransmitting tract with the top of the ovary placenta assistseffective distribution of pollen tubes within the ovary. Lycopersicon, solanaceae, fertilization, pistil, pollen tube, pollination     

MICROPYLAR EXUDATE IN GASTERIA (ALOACEAE) AND ITS POSSIBLE FUNCTION IN POLLEN TUBE GROWTH

The micropylar exudate of Gasteria verrucosa (Mill.) H. Duval was studied using light and electron microscopic techniques. Ovules may contain micropylar exudate before stigma receptivity. During successive phases of stigma receptivity, the number of ovules with micropylar exudate and the amount of micropylar exudate per ovule increases. At the late phase of stigma receptivity, large amounts of micropylar exudate with a smooth to cauliflowerlike appearance were observed. Micropylar exudate is viscous and contains, among other components, proteins and carbohydrates. At all stages of the stigma investigated, ovules situated at the base of the ovary contain a larger quantity of micropylar exudate than those at the top. The appearance of micropylar exudate is related to the degree of development of the embryo sac and it originates primarily from the filiform apparatus. It is assumed that an uptake of water by the ovule initiates the outflow of micropylar exudate from the filiform apparatus into the micropyle. Both ovular pollen tube ingrowth and seed set mark the optimum pollination stage of the stigma, which for both events lies around the onset of stigma receptivity. When pollen tubes have reached the ovary, young micropylar exudate stimulates their growth rate. The presence of micropylar exudate seems to be a requirement for pollen tube penetration, and an interaction between the pollen tube and the micropylar exudate has been proposed. Possibly, the micropylar exudate serves as a nutritional source and, in an optimum condition, as an attractant for approaching pollen tubes.     

Tracing pollen nuclei in the ovary and ovule of Gasteria verrucosa (Mill.) H. Duval after pollination with DAPI-stained pollen

Summary The addition of DAPI particles to the stigma exudate of Gasteria results in the labelling of the pollen nuclei. By means of epifluorescence microscopy and clearing of the ovule, the labelled nuclei of the sperm cells and, subsequently, the zygotic nucleus can be observed. The method was used in a cross-pollination with low seed setting to examine different types of penetration and transport of the pollen tube nucleus and sperm cell nuclei. More than one pollen tube can penetrate, but generally only one set of sperm cell nuclei is accepted.     

Centrifugal ovule inception

Nine samples from Sattler (1973) were found to exhibit a remarkable correlation between the sequence of ovule inception and ovary structure. In every case, the sequence of ovule inception is centrifugal from its supposed final position of the floral apex. Thus, it is hypothesized that ovules are initiated centrifugally from their floral apex and that this is regulated in some mechanical way. To prove this hypothesis, ontogenetical development of the ovary was reexamined inSilene cucubalus, where the sequence of ovule inception was expected to be not simply basipetal but basipetal on the central column and acropetal on the septal protuberances. The results appear to support the hypothesis. A basipetal sequence is clearly visible on the central column. Although the ovule number is very small on the septal placenta, an acropetal sequence can be recognized, i.e., the distal primordia are obviously initiated later than the proximal ones. Two possible mechanisms for centrifugal ovule inception are suggested: (1) that it is regulated by chemical information diffusing from the floral apex or (2) that it is dependent on gradation of cell age in the placenta. The second mechanism is discussed and supporting evidence is suggested in the epidermal cell arrangement of the septal wall. Contribution from the Osaka Museum of Natural History No. 275.     

Sialic acid, electrophoretic mobility and transmembrane potentials of the Amphiuma red cell

Red cells from the giant salamander Amphiuma means are shown to contain sialic acid. The amount removed by the action of neuraminidase is equal to that released by acid hydrolysis, indicating that all of the sialic acid is present on the outer surface of the plasma membrane. These cells have a negative electrophoretic mobility and 100% enzymatic removal of sialic acid results in a 40% reduction in the mobility, suggesting that either a fraction of the sialic acid carboxyl groups are unavailable to the action of external electric fields, or other negatively charged groups contribute to the surface charge. A further reduction in mobility of normal and sialic acid-free cells is caused by an increased extracellular calcium concentration. The negative groups affected by calcium are most likely to be phosphate groups, since the isoelectric point of the cells is found to lie between the pK values for H₂PO₄⁻ groups and the carboxyl groups of sialic acid. Membrane potentials of single cells, from which 80% or more of the total sialic acid had been removed, were identical to those measured in normal cells, confirming that sialic acid plays little, if any, direct role in the maintenance of membrane potentials and ionic permeabilities.     

Cotton embryogenesis: The tissues of the stigma and style and their relation to the pollen tube

Summary The stigma of cotton (Gossypium hirsutum) is covered by unicellular hairs. The cytoplasm of these hairs degenerates before the stigma becomes receptive. The vacuole remains intact, but the hair cytoplasm becomes a mass of dark, amorphous material with only a few organelles still being visible. The rest of the stigma consists of thin-walled parenchyma cells with large vacuoles and large amounts of starch. The cells of the style are differentiated into a uniseriate epidermis, vascular tissue, a cortex of thin-walled, vacuolate parenchyma cells, and the transmitting tissue. This latter tissue occupies the center of the style and consists of thick-walled cells with few vacuoles. The cells are rich in starch, ribosomes, endoplasmic reticulum and dictyosomes. They also contain deposits of calcium salts in the form of druses. The pollen germinates on the stigmatic hairs, grows down the outside of the hair and between the cells of the stigma to the transmitting tissue of the style. There the tubes grow between the walls of the cells but do not enter the cells themselves. Some transmitting cells adjacent to the pollen tube degenerate after the tip of the pollen tube has grown past them. However, not all degenerate, and those that do show no fixed spatial relationship to one another. The cells which do degenerate follow a characteristic pattern of breakdown. No ultrastructural evidence was found for the secretion of hydrolytic enzymes by the pollen tube.     

小叶兜兰的果实生长和雌配子体发育

为了解濒危兰科植物小叶兜兰(Paphiopedilum barbigerum Tang et Wang)胚珠和雌配子体的发育过程,采用常规石蜡切片技术对其果实的生长动态进行了研究。结果表明,授粉后60~75 d的蒴果内种子数量迅速增加,到授粉后120 d时种子充满整个蒴果。授粉后40 d的胎座上分化形成多数由1层表皮细胞包被1列细胞的胚珠原基;授粉后60 d时位于胎座指状结构末端处紧靠表皮细胞下方的孢原细胞分化为大孢子母细胞。之后,大孢子母细胞经过减数分裂和有丝分裂最终形成成熟胚囊;授粉后135 d胚囊发育成熟,附着在胎座上的种子个体分化明显。小叶兜兰胚囊的发育类型为双孢子葱型,胚珠为倒生胚珠,薄珠心,单珠被,成熟胚囊为8核。这为小叶兜兰的生殖生物学及繁殖体系的建立提供理论依据。     

烟草花粉萌发和花粉管生长期间柱头和花柱中的钙分布

烟草柱头表面有两层覆盖物,其中含有少量细小的钙颗粒.花粉落到柱头上后,储存在花粉外壁中的钙被释放到覆盖层中.当花粉管穿过覆盖层长入柱头细胞之间时,花粉管顶端的细胞壁中出现了大量的细小钙颗粒.开花后22 h观察时,在花柱引导组织中形成了钙的梯度分布:花柱上部引导组织中的钙较少,而下部连接子房处的花柱引导组织中含有较多的钙颗粒.去雄花开花后1 d时,花柱上部引导组织中的钙明显增多;3 d时,连柱头细胞中也出现了较多的钙颗粒.讨论了烟草花柱引导组织中钙梯度分布和花粉管生长的关系.     

Postulated interaction between branching pollen tubes and ovules inOenothera hookeri de Vries (onagraceae)

The pollen grain germinationin vitro and progamic phase till fertilization inOenothera hookeri de Vries was observed after open and controlled pollination. The same pattern of pollen grain germination was foundin vitro and on the stigma. The pollen tubes can germinate from 1,2 or 3 poruses of the pollen grain, divide and branch during their growth in the ovary. The branches are of different length and give secondary splits. Special short branches are formed near the micropyle of the ovule. They grow into top part of integments. The pollen tubes start to branch profusely near the placental surface. In that place they are likely to react to the stimulus from mature ovules which seems to be dispersed in the exudate covering placenta.     

中国特有植物喜树的雌性不育形态解剖学研究

为阐明雌性不育与喜树（Camptotheca acuminata Decne.）开花期花序大量掉落的关系,本文采用数码体视显微镜及石蜡切片技术,对各个发育时期的喜树的花进行解剖学观察。喜树花均为两性花,雄蕊发育正常,雌蕊柱头有三种不同形态,即上位、中位和下位;柱头上位的小花,花柱伸长正常,柱头三裂,外翻,子房膨大,胚珠饱满,胚囊发育正常;柱头中位的小花,雌蕊选择性败育,柱头和花柱伸长缓慢,柱头外翻异常,局部子房细胞液泡化,胚珠退化,胚囊发育异常;柱头下位的小花,花柱退化,柱头不露,珠被细胞发育异常,大孢子母细胞未能减数分裂,胚珠萎缩,子房外形瘦长,室内中空。喜树雌性器官的发育与其柱头在子房上的表现形态有着严格的对应关系,雌性不育发生于大孢子母细胞减数分裂前后,而且雌性不育是喜树花序掉落的内部原因之一。     

Effect of calcium on the potassium flux into the exudate of excised maize roots

Summary The effect of varying calcium concentration in the medium on the potassium flux into the exudate has been studied. In media of low ionic strength (o.1 mM KCl) the potassium flux, J _K, was significantly increased by increasing the calcium concentration of the medium. But in higher ionic strength media (10 mM) KCl) there was no increase in J _K as the calcium concentration of the medium was increased. The effect of external sodium concentration on J _K was also studied. These results are discussed in relation to present theories of salt and water movement into the plant root. It is concluded that two pathways potentially exist for movement of salts to the exudate stream: firstly, via a symplasm and secondly, through the cell wall pathway. But is is further concluded that the cell wall pathway, at normal physiological ionic strengths, is not available for salt transport due to co-ion exclusion by the fixed negative charges.     

STIGMA,STYLE, AND OBTURATOR OF ORNITHOGALUM CAUDATUM (LILIACEAE) AND THEIR FUNCTION IN THE REPRODUCTIVE PROCESS

Transmitting tissue in Ornithogalum is divided into three regions corresponding to classical divisions of the gynoecium: stigma, style, and ovary. The stigma differentiates from epidermal cells of the stylar apex. These cells form the stigmal papillae and have dense cytoplasm with abundant ER and lipid bodies. Papillae have walls with small transfer-ingrowths. At floral receptivity, papillae secrete a small amount of surface exudate. Epidermal cells of the style contain numerous spherosomes and have thin filaments of cytoplasm traversing the central vacuole. The stylar cortex is composed of 3-6 layers of parenchyma cells which contain numerous spherosomes and often have secondary vacuoles. Vascular tissue in the style consists of one collateral bundle in each lobe. Cells of the epidermal layer lining the stylar canal are secretory. They are initially vacuolate but fill progressively with dense cytoplasm as their secretory activity increases. Secretory activity occurs in three phases, each characterized by a particular organelle population and secretory product. At anthesis, the canal is filled with an exudate consisting of carbohydrate, protein, and lipid. In the ovary, the obturator differentiates from cells at the base of the funiculus and the tip of the carpel margins. It forms a pad of tissue which covers most of the former placenta. The obturator is secretory and produces a surface exudate. We believe our observations on Ornithogalum support the hypothesis that all transmitting tissue is of the same morphological origin and that it provides nutritive and chemotropic factors for pollen tube growth.     

钙在被子植物受精过程中的作用

近年来,花粉管中的钙信号和生理功能的研究取得了明显的进展,同时在雌蕊系统中有关钙分布的研究也初步显示了其时、空特征与被子植物的受精作用密切相关。该文总结了花粉萌发和花粉管生长过程中外源钙和内源钙的作用机制,结合雌蕊组织中钙分布的特征,进一步探讨了钙在被子植物受精过程中的功能。     

Calcium changes in ovules and embryo sacs of Plumbago zeylanica L.

Calcium was localized in ovules of Plumbago zeylanica from 1 day before anthesis to 3 days after anthesis using potassium antimonate and transmission electron microscopy in pollinated and emasculated flowers. At 1 day before anthesis, embryo sacs (containing an egg cell, a central cell and zero to three accessory cells) appear mature and contain abundant calcium precipitates (ppts), in contrast to nucellar cells. At anthesis, the vacuoles of nucellar cells have enlarged, and micropylar cells, in particular, are heavily labeled with calcium ppts. As pollen tubes elongate through ovular tissues, ppts diminish in ovular cells and become concentrated in the pollen tube cell wall. After fertilization, the calcium ppts sharply diminish in fertilized ovules; in unfertilized ovules, calcium ppts remain abundant up to 3 days after anthesis (when unfertilized ovules are shed). The distribution of calcium in the ovule changes in apparent response to fertilization, suggesting that calcium content may be related to the attraction and receipt of the pollen tube. In contrast with conventionally-organized embryo sacs with synergids, Plumbago accumulates calcium in the egg cell. Received: 30 December 1999 / Revision accepted: 24 March 2000     

濒危植物马蹄参的大孢子发生和雌配子体发育

为探讨马蹄参(Diplopanax stachyanthus Hand.-Mazz.)濒危机制与雌性生殖发育的关系,采用石蜡切片法观察马蹄参大孢子形成和雌配子体的发育过程。结果表明,马蹄参雌蕊单心皮,子房下位,1室,1枚胚珠。胚珠横生于短片状胎座上,具单珠被,厚珠心。单孢原细胞自珠心1~2层表皮细胞处分化;大孢子四分体为直线形。成熟胚囊中,2个极核在受精前融合为次生核,3个反足细胞不发达,较早退化;二核胚囊时期出现二核分裂不均且较小核退化消失的异常发育现象。因此,马蹄参雌配子体发育过程中出现异常现象是造成其结实率低的主要原因。     

黄顶菊的大孢子发生、雌配子体与胚胎发育

用常规石蜡制片对黄顶菊(Flaveria bidentis(L.) Kuntze)大孢子发生、雌配子体和胚胎的发育过程进行了观察.黄顶菊雌蕊柱头二裂,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直列四分体...     

莴苣授粉前后柱头与花柱中钙的分布变化

莴苣柱头呈两裂片状,有接受花粉的接受面和非接受面之分。授粉前后,柱头接受面乳突细胞的细胞壁中贮存丰富的细小钙颗粒,而非接受面的表皮细胞壁中的钙颗：粒很少。在花柱组织中钙的分布具有明显特征,在同一水平上,钙主要分布在引导组织的质外体中,如细胞壁和细胞间隙中,而在引导组织外的薄壁组织中钙主要分布在的细胞内液泡和细胞壁中以及维管束导管内。在花柱不同水平上,花柱中的钙呈现出梯度分布,顶端各组织中的钙颗粒较少,基部各组织中的钙颗粒较多。授粉后1h花柱基部组织的钙颗粒增多,钙梯度分布现象增强。花柱引导组织中的钙梯度分布很可能是吸引花粉管向下生长的原因。讨论了莴苣花柱引导组织中钙梯度分布特征和花粉管在其体内生长的关系。     

Ovary,seed and fruit ofRutidea (Rubiaceae,Pavetteae)

The ovary ofRutidea is bicarpellate and incompletely bilocular (septum between locules not continuous). A solitary campylotropous ovule, ascending from a basal placenta, occurs in each locule. Based on their orientation and degree of curving, three ovule types are distinguished. As a consequence of the abortion of one ovule, the drupaceous fruits are one-seeded. The incomplete septum allows the spherical seed to fill out the entire interior of the fruit. The seeds are deeply ruminate (Spigelia type). They grow very fast, producing folds and undulations (ruminations) which invade and totally occupy the second locule, almost enveloping the aborted ovule. Comparisons with otherRubiaceae (especiallyPavetteae), show that hemianatropy and campylotropy occur more often in theRubiaceae than hitherto accepted. The study corroborates the close affinity betweenRutidea andNichallea.     

Localisation and expression of arabinogalactan-proteins in the ovaries of Nicotiana alata Link and Otto

The transmitting-tissue cells of the style of flowering plants secrete a complex extracellular matrix through which pollen tubes grow to the ovary to effect fertilisation. This matrix is particularly rich in a class of proteoglycans, the arabinogalactan-proteins (AGPs). AGPs from the ovary of Nicotiana alata were found to be developmentally regulated, as the different charge classes of AGPs altered during floral development. The AGPs from the mature ovary had charge characteristics that were distinct from those previously reported for the stigma and style. However, the concentration of AGP (0.6 g/ml fresh weight) in the ovary did not change during development, or in response to either compatible or incompatible pollination. The AGPs of the ovary are mainly associated with the epidermis of the placenta.     

Über Glykoproteideinschlüsse in den Trophoblastzellen der menschlichen Plazenta und die Frage ihres Zusammenhangs mit der Bildung von Gonadotropin

Summary In some of the basophilic trophoblastic cells of the basal plate and islands of the mature human placenta there are glycoprotein inclusions, many of which are within vacuoles. As established by the histochemical results these inclusions are composed of a neutral mucopolysaccharide and a basophilic protein with SH groups. They gradually increase in number from the 4th month of pregnancy and are not influenced by labor. From the first to the fourth month of gestation small vacuoles, initially infra-nuclear but later supranuclear, may be seen in the syncitial cells of the villi; these likewise contain glycoprotein granules. The same inclusions are to be found in some trophoblastic cells of the chorionic plate and of the chorion laeve, and in an occasional epithelial cell of the amnion.On the basis of the histochemical behavior of these inclusions, their similarity with the -granules of the hypophysis, and the obvious high activity of the trophoblastic cells which bear them, it is postulated that these inclusions contain gonadotropin. Their occurrence during pregnancy parallels the biochemical concentration and immunofluorescent histological demonstration of gonadotropin in the placenta. Thus, at the end of pregnancy the basal trophoblastic cells of the human placenta appear to form at least two different hormones: relaxin, probably represented morphologically by the acidophilic protein inclusions (described elsewhere), and gonadotropin most likely represented by the basophilic glycoprotein inclusions.