Ultrastructural evidence of endoplasmic reticulum changes during the maturation of the olive pollen grain (Olea europaea L., Oleaceae)

Characteristic features of rough endoplasmic reticulum (rER) distribution and proliferation were noted during olive pollen (Olea europaea L.) development, suggesting the physiological significance of this organelle. Initially scarce in the young microspore, ER increases as cytoplasmic vacuoles form. At the vacuolated microspore stage the cytoplasm contains numberous polysomes and elongated rER cisternae arranged preferentially in stacks, with an average intracisternal width of 0.07 µm. Stacks persist in the bicellular pollen grain but consist of fewer, shorter, dilated cisternae (mean intracisternal width 0.1 µm) containing a considerable electron-dense matrix. Cisternae in the mature grain are fragmented, leaving behind an ER of swollen pockets. Pockets of ER containing a material of greater electron density are evenly deposited along the plasmalemma, in close relation with it. A dense material is seen in the tubules of the apertural region, which was lacking in earlier stages. Our results show that ER may be involved in protein transport to the intine.     

FINE STRUCTURAL STUDIES OF ZEA MAYS POLLEN I: CELL MEMBRANES AND EXINE ONTOGENY

Electron microscopy was used to study pollen wall ontogeny in Zea mays. The initial stage of development consisted of compartmentalization of microspores within callose special walls. Microspore plasma membranes retracted and tubular elements of the endoplasmic reticulum became perpendicularly oriented to the plasma membranes. Evaginations of the endoplasmic reticulum into the microspore plasma membrane resulted in the establishment of a template or blueprint of the mature pollen wall. Sporopollenin deposition upon the template began immediately after dissolution of the callose special walls and release of the microspores into the anther locule. The columellae were the first pollen wall units to be formed; the tectum and foot layer became established shortly thereafter. The granular endexine was the last-formed unit. The relationships of membrane systems to the ontogeny of the pollen wall units and the mode of pollen wall growth are discussed.     

Sporoderm development in Liriodendron chinense (Magnoliaceae): a probable role of the endoplasmic reticulum

The developmental events in the sporoderm and the cytoplasm of Liriodendron chinense microspore from the early tetrad stage until late free microspore stage were observed. Various forms of the endoplasmic reticulum (ER) and surprising unusual aggregates of ER seen during microspore development attract special attention. Being scanty at early and middle tetrad stage, while primexine matrix (glycocalyx) acquires well-defined form, the ER becomes distinct in unusual forms at the late tetrad stage. Thin long tubules with an osmiophilic contents, which cannot be compared with the tubular smooth endoplasmic reticulum (SER), undulate through the cytoplasm. Towards the end of the tetrad period when callose begins to disintegrate, and a distinct tectate-columellate pattern of the ectexine becomes evident, two new forms of the SER occur in the cytoplasm. Instead of single tubules observed previously, 3–tubuled aggregates meander through the cytoplasm, the middle tubule contains an osmiophilic substance. The second form of the SER looks like an ordinary tubular SER, but has ampoule-like dilations with dark granular contents. Later on the tubules undergo major changes: multi-tubuled aggregates of parallel tubules overcrowd the cytoplasm, the outer tubules of each aggregate carrying ribosomes. These aggregates undulate through the cytoplasm, branch, and are associated with lipid globules. The tips of many aggregates are pressed to the plasmalemma. The ontogenetic period of time of the presence of these ER aggregates, their structure and localization in the microspore cytoplasm allow me to assume that these ER aggregates synthesize sporopollenin precursors.     

Wall projections in the vegetative cell ofParietaria officinalis L. Pollen

Summary Ultrastructural aspects of pore development inParietaria officinalis L. pollen are described. The main feature is the presence of intine wall projections formed in correspondence to the pores during the late microspore stage and persisting in the shedding pollen. This structure coexists with the tubules occurring in the pore and first appearing as evaginations of the plasma membrane.The morphological evidence and other features such as the occurrence of mitochondria next or in contact with wall projections lead us to compare the pollen vegetative cell to a transfer-cell. The possible functions of these transfer-cell type structures are discussed.     

Microspore development inBrassica napus and the effect of high temperature on division in vivo and in vitro

Summary Brassica napus cv. Topas microspores isolated and cultured near the first pollen mitosis and subjected to a heat treatment develop into haploid embryos at a frequency of about 20%. In order to obtain a greater understanding of the induction process and embryogenesis, transmission electron microscopy was used to study the development of pollen from the mid-uninucleate to the bicellular microspore stage. The effect of 24 h of high temperature (32.5 °C) on microspore development was examined by heat treating microspore cultures or entire plants. Mid-uninucleate microspores contained small vacuoles. Late-uninucleate vacuolate microspores contained a large vacuole. The large vacuole of the vacuolate stage was fragmented into numerous small vacuoles in the late-uninucleate stage. The late-uninucleate stage contained an increased number of ribosomes, a pollen coat covering the exine and a laterally positioned nucleus. Prior to the first pollen mitosis the nucleus of the lateuninucleate microspore appeared to be appressed to the plasma membrane; numerous perinuclear microtubules were observed. Microspores developing into pollen divided asymmetrically to form a large vegetative cell with amyloplasts and a small generative cell without plastids. The cells were separated by a lens-shaped cell wall which later diminished. At the late-bicellular stage the generative cell was observed within the vegetative cell. Starch and lipid reserves were present in the vegetative cell and the rough endoplasmic reticulum and Golgi were abundant. The microspore isolation procedure removed the pollen coat, but did not redistribute or alter the morphology of the organelles. Microspores cultured at 25 °C for 24 h resembled late-bicellular microspores except more starch and a thicker intine were present. A more equal division of microspores occurred during the 24 h heat treatment (32.5 °C) of the entire plant or of cultures. A planar wall separated the cells of the bicellular microspores. Both daughter cells contained plastids and the nuclei were of similar size. Cultured embryogenie microspores contained electron-dense deposits at the plasma membrane/cell wall interface, vesicle-like structures in the cell walls and organelle-free regions in the cytoplasm. The results are related to embryogenesis and a possible mechanism of induction is discussed.Abbreviations B binucleate - LU late uninucleate - LUV late uninucleate vacuolate - M mitotic - MU mid-uninucleate - RER rough endoplasmic reticulum - TEM transmission electron micrograph     

麻疯树小孢子发育的研究

用透射电镜观察了麻疯树(Jatropha curcas L.)小孢子发育的超微结构。小孢子母细胞时期内质网和质体较多;减数分裂和四分体时期,细胞处于明显的代谢活跃状态,细胞器丰富,主要有内质网、线粒体、质体、高尔基体和球状体;在小孢子发育早期和晚期,线粒体和内质网仍较丰富;小孢子经过高度的不对称分裂后,形成较大的营养细胞和较小的生殖细胞,营养细胞中细胞器数量明显减少,含大量的淀粉和脂类物质,生殖细胞中脂类物质丰富;表皮、药室内壁和中层细胞在小孢子母细胞和四分体时期淀粉粒丰富,小孢子时期明显减少,绒毡层从小孢子母细胞至小孢子发育晚期的细胞器都很丰富,主要为内质网、质体和线粒体,为二胞花粉发育奠定基础。     

Plasmodial tapetum and pollen wall development in Butomus umbellatus (Butomaceae)

This report describes the ultrastructural development of plasmodial tapetum and pollen wall of Butomus umbellatus. The tapetum contains extensive arrays of rough endoplasmic reticulum, vesicles from which are responsible for the formation of sporopollenin-like bodies. The tapetum is also involved in the formation of other forms of sporopollenin precursors. Development of pollen wall continues after microspores are released from their callosic walls; they are then enclosed by plasmodial tapetum. The activity and products of the plasmodial tapetum show substantial correlation with pollen wall development, particularly ektexine formation. In B. umbellatus, the tapetum intrudes into the anther locule at early microspore stage. This timing of plasmodial intrusion occurs at a later stage of pollen development as compared to those in the advanced monocotyledons. We report the rough endoplasmic reticulum origin of sporopollenin-like bodies and their occurrence in the plasmodial tapeta of B. umbellatus.     

The loculus content and tapetum during pollen development in Lilium

 The ratio of loculus volume to the volume of the entire anther began to increase from the microspore mother cell stage and reached 32.3% at anthesis. The content of the loculus was examined in Lilium during pollen development and two waves could be distinguished. From the premeiotic stage until the vacuolated microspore stage, the loculus consisted of neutral polysaccharides, pectins and proteins. These substances originated from tapetal activity from the premeiotic stage until the young microspore stage. Dictyosomes and rough endoplasmic reticulum seemed to be involved in tapetal secretion, although, in some mitochondria, vesicles progressively developed as early as premeiosis and increased until the young microspore stage, which could reveal their involvement in the secretion process. At this stage, numerous cytoplasmic vesticles containing material similar to the locular material fused with the plasma membrane of the tapetum so that vesicle content was in contact with the loculus. It seems that tapetal and callose wall degradation at the late tetrad stage may also have contributed to the production of material in the loculus. From pollen mitosis to anthesis, the anther loculus contained mainly the pollenkitt which was synthesized in the tapetum between the young microspore stage and the vacuolated microspore stage. At the young microspore stage, proplastids divided and developed into elaioplasts and smooth endoplasmic reticulum (SER) increased dramatically. Pollenkitt had a double origin: some droplets were extruded directly from the plastid stroma through the plastid envelopes; the others were unsaturated lipid globules, which presumably derived from the interaction between SER saccules and plastids. Received: 2 September 1997 / Revision accepted: 12 March 1998     

The tubular endoplasmic reticulum in the amoebocytes of the shell-regenerating snail,Helix pomatia L.

Summary The tubular endoplasmic reticulum has been studied in the amoebocytes which are present in the connective tissue of the hepatopancreas of the snail, Helix pomatia. The reticulum is similar to that previously described within the glandular cells of the hepatopancreas. Two distinct components are recognizable in the reticulum—central main tubules approximately 100 m in diameter and connecting tubules about 20 m in width. The profile of this tubular network in cross-sections appears as a very regular, apparently crystalline array. The tubules are intimately associated with dense granular material, dense bodies and with mitochondria. The possible function of the tubular endoplasmic reticulum is discussed.This investigation was supported by grants from the Swedish Natural Science Research Council, which are gratefully acknowledged. I am indebted to Miss G. Drugge for her technical assistance.     

Nuclear pore distribution and relation to adjacent cytoplasmic organelles in cotyledon cells of developing Vicia faba

Following a zinc iodine-osmium tetroxide fixation, nuclear pore distribution was studied in 0.3-m sections from cotyledons of developing Vicia faba L. Localised absence of nuclear pores was found to be associated with proximity of organelles to the nucleus. Golgi cisternae and mitochondria are associated with areas of pore absence while cisternal endoplasmic reticulum and tubular endoplasmic reticulum are linked with areas showing reduction in pore density. Pores were seen in the nuclear membrane adjacent to vacuoles. Pattern analysis of pore distribution indicated possible clustering within an overall regularity.Abbreviations ER endoplasmic reticulum - ZIO zinc iodine-osmium tetroxide     

Ultrastructure of tapetal cell ontogeny inBeta

Summary Tapetal cell development and degeneration in anthers ofBeta vulgaris L. were studied with the electron microscope. Tapetal cells become differentiated from sporogenous cells early in anther ontogeny. The tapetal nuclei divide mitotically; binucleate tapetal cells contain relatively little endoplasmic reticulum and otherwise resemble meristematic cells of higher plants. There follows an increase in endoplasmic reticulum and by the time the sporogenous tissue has entered meiotic prophase, the tapetal cells have differentiated the usual characteristics of secretory cells. Degenerative changes begin to appear in tapetal cells after meiosis of the sporogenous tissue. Such changes include loss of inner tangential and anticlinal walls, degeneration of tapetal nuclear envelopes, disruption of the plasmalemma, and changes in the cytoplasmic organelles. Coated tubules are associated with tapetal nucleoli during degenerative stages and the tubules persist after tapetal nuclei have degenerated. Tapetal cell cytoplasm disappears completely by the stage of microspore mitosis.     

Ultrastructure of microsporogenesis and microgametogenesis inArabidopsis thaliana (L.) Heynh. ecotype Wassilewskija (Brassicaceae)

Summary The process of microsporogenesis and microgametogenesis was studied at the ultrastructural level in wild-typeArabidopsis thaliana ecotype Wassilewskija to provide a basis for comparison with nuclear male-sterile mutants of the same ecotype. From the earliest stage studied to mature pollen just prior to anther dehiscence, microsporocyte/microspore/pollen development follows the general pattern seen in most angiosperms. The tapetum is of the secretory type with loss of the tapetal cell walls beginning at about the time of microsporocyte meiosis. Wall loss exhibits polarity with the tapetal protoplasts becoming located at a distance from the inner tangential walls first, followed by an increase in distance from the radial walls beginning at the interior edge and progressing outward. The inner tangential and radial tapetal walls are completely degenerated by the microspore tetrad stage. Unlike other members of the Brassicaceae that have been studied, the tapetal cells ofA. thaliana Wassilewskija also lose their outer tangential walls, and secretion occurs from all sides of the cells. Exine wall precursors are secreted from the tapetal cells in a process that appears to involve dilation of individual endoplasmic reticulum cisternae that fuse with the tapetal cell membrane and release their contents into the locule. Following completion of the exine, the tapetal cell plastids develop membranebound inclusions with osmiophilic and electron-transparent regions. The plastids undergo ultrastructural changes that suggest breakdown of the inclusion membranes followed by release of their contents into the locule prior to the complete degeneration of the tapetal cells.     

Structural changes during the first divisions of embryos resulting from anther and free microspore culture inBrassica napus

Summary Ultrastructural and cytochemical features of embryo development during anther and free microspore culture inBrassica napus have been followed from the late uninucleate microspore stage through the first embryonic division. On transfer to culture, the microspore cytoplasm possesses a large vacuole, often containing electron opaque aggregates, and a peripheral nucleus. Mitochondria, endoplasmic reticulum and starch-free plastids are distributed throughout the cytoplasm. The conditions of culture induce a number of major changes in the cytoplasmic organisation of the microspore. First, the central vacuole becomes fragmented allowing the nucleus to assume a central position within the cell. Secondly, starch synthesis commences in the plastids which, in turn, are seen to occupy a domain investing the nucleus. Thirdly, the cell develops a thick fibrillar wall, situated immediately adjacent to the intine of the immature pollen wall. Finally, the microspores develop large cytoplasmic aggregates of globular material. The nature of this substance remains unknown, but it remains present until the young embryos have reached the 30 cell stage. The first division of cultured microspores destined to become embryos is generally symmetrical, in contrast to the asymmetric division seen in normal development in vivo. Consideration is given to the differences observed between embryos developing from anthers and free microspores in culture.     

Ultrastructural changes in maturing pollen grains ofApium nodiflorum L. (Apiaceae), with special reference to the endoplasmic reticulum

Summary The ultrastructural events in 3-cellular pollen grains ofApium nodiflorum L. are investigated during pollen maturation. Three distinct developmental stages are distinguished from the formation of sperm cells up to anthesis, whereby the rough endoplasmic reticulum (RER) is mainly involved. The most conspicious form is the highly dilated RER in the vegetative cytoplasm of the youngest pollen grains, which changes to vesicular RER in the following stage. In mature pollen grains the RER has a narrow cisternal configuration and often forms stacks. Pollen activation is preceded by the accumulation of polysaccharide particles.     

Germination and early tube development in vitro of Lycopersicum peruvianum pollen: Ultrastructural features

Morphologic changes occurring during pollen grain activation and ultrastructural features of Lycopersicum peruvianum Mill. pollen tube during the first stages of growth in vitro have been studied. The more evident morphologic changes during activation, in comparison to those already described for mature inactive pollen, concern dictyosomes, rough endoplasmic reticulum (RER), and ribosomes. The dictyosomes are very abundant and produce large and small vesicles. Near the germinative pores both types of vesicles are present, while all along the remaining cell wall only the large type is observed. These latter react weakly to Thiéry's test and probably contain a callose precursor necessary for the deposition of a callosic layer lining at first only the inner side of the functioning pore and occasionally the other two pores, and subsequently the entire inner surface of the cell wall. The small vesicles, highly positive to Thiéry's test, are present only near the pores and could be involved in the formation of the pectocellulosic layer of the tube wall. The setting free of RER cisterns, which in the mature inactive pollen were aggregated in stacks, coinciding with polysome formation and resumption of protein synthesis, is in accord with the hypothesized role of RER cistern stacks as a reserve of synthesizing machinery. The pollen tube reaches a definitive spatial arrangement soon after the generative cell and vegetative nucleus have moved into it. At this stage four different zones that reflect a functional specialization are present. In the apical and subapical zone two types of dictysosome-originated vesicles, similar to those found in the activated pollen grain, are present. Their role in the formation of the callosic and pectocellulosic wall layers seems to be the same as in the activated pollen grain.Abbreviations ER endoplasmic reticulum - RER rough endoplasmic reticulum Research performed under CNR program Biology of Reproduction     

Fine Structure of Tapetal Cells and Ubisch Bodies in the Anther of Ophiopogon japonicus

The development of the tapetum in Ophiopogon ]aponicus is of secretory type Tapetum develops at their peak during the microspore mother cell stage. There are abundant organelles, consisting of a lot of mitochondria, dictyosomes and plastids, especially endoplasmic reticulum. Pro-Ubisch bodies e. merge as early as at the stage of microspore mother cell. At tetrad stage, a large number of pro-Ubisch bodies accumulate between inner tangential face of the plasmalemma and the cell wall. At the early microspore stage, pro-Ubisch bodies are distributed in the small embayments of the plasmalemma. As the sporopollenin begins to deposit on them, proubisch bodies develop into Ubisch bodies which consist of two types: single and aggregated. Tapetal cells degenerate completely when pollen grains reach maturity.     

Detection and quantification of rRNA by high-resolutionin situ hybridization in pollen grains

We examined changes in the localization of cytoplasmic rRNA during pollen development inNicotiana tabacum SR-1. The rRNA was visualized byin situ hybridization, and the signal intensity of rRNA in microspore, vegetative and generative cell was quantified by microphotometry. The amount of rRNA per microspore or pollen section increased about 5 times from microspore to mature pollen grain and kept increasing even in the late stage of pollen development after PMI. The increase of rRNA occur in both vegetative and generative cells. The results suggest that synthesis of rRNA occur even after PM I in both vegetative and generative cells.     

Cytochemistry of the tubular aggregates found in hepatocytes of interferon-treated suckling mice

Summary Administration of mouse interferon to neonatal mice induces the formation of tubular aggregates within hepatocytes. These aggregates are composed of networks of small tubules which are continuous with the granular endoplasmic reticulum. The tubules are bounded by trilaminar unit membranes and exhibit glucose-6-phosphatase activity. These features suggest that the tubular aggregate is an interferon-induced alteration of the endoplasmic reticulum.     

凤仙花花药发育中多糖和脂滴组织化学研究

以不同发育时期的凤仙花花药为实验材料,采用组织化学方法,对花药发育中的结构变化及多糖和脂滴物质分布进行观察。结果表明:(1)凤仙花的花药壁由6层细胞组成,包括1层表皮细胞,2层药室内壁细胞,2层中层细胞和1层绒毡层细胞。其中绒毡层细胞的形态不明显,很难与造孢细胞区分,且在小孢子母细胞时期退化。(2)在小孢子母细胞中出现了一些淀粉粒,但减数分裂后,早期小孢子中的淀粉粒消失,又出现了一些小的脂滴;随着花粉的发育,小孢子形成大液泡,晚期小孢子中的脂滴也消失;小孢子分裂形成二胞花粉后,营养细胞中的大液泡降解、消失,二胞花粉中又开始积累淀粉;接近开花时,成熟花粉中充满细胞质,其中包含了较多的淀粉粒和脂滴。(3)在凤仙花的花药发育中,绒毡层细胞很早退化,为小孢子母细胞和四分体小孢子提供了营养物质;其后的中层细胞退化则为后期花粉发育提供了营养物质。     

含笑花药发育的超微结构研究

对含笑花药发育中的超微结构变化进行观察,结果显示:(1)花粉发育中有三次液泡变化过程——第一次是小孢子母细胞在形成时内部出现了液泡,这可能与胼胝质壁的形成有关;第二次是在小孢子母细胞减数分裂之前,细胞内壁纤维素降解区域形成液泡,它的功能可能是消化原有的纤维素细胞壁;第三次是在小孢子液泡化时期,形成的大液泡将细胞核挤到边缘,产生极性。(2)含笑花粉在小孢子早期形成花粉外壁外层,花粉外壁内层在小孢子晚期形成,而花粉内壁是在二胞花粉早期形成;花粉成熟时,表面上沉积了绒毡层细胞的降解物而形成了花粉覆盖物。研究认为,含笑花粉原外壁的形成可能与母细胞胼胝质壁有关,而由绒毡层细胞提供的孢粉素物质按一定结构建成了花粉覆盖物。