小麦胚胎分化时营养吸收一种可能方式的细胞学论据

以光学与电子显微镜术结合荧光标记物示踪,对小麦（ＴｒｉｔｉｃｕｍａｅｓｔｉｖｕｍＬ．）胚胎分化时胚根鞘的形态结构特征的变化与营养吸收相联系进行了实验观察。发现在分化前期胚根鞘顶端的球状突起处和颈区朝向颖果背面一侧的特定位点上出现了结构的变更;外沿局部细胞的衰退瓦解导致这些区段内侧存活细胞分界壁上出现大量残存的胞间连丝。不透膜的荧光物质、分子量达４３ｋＤ的ＦＩＴＣｄｅｘｔｒａｎ可进入球状突起和颈区多处细胞,表明残存胞间连丝仍具有共质传输功能,是分化幼胚摄取外围介质中营养物质的一种方式。分化进程中颈区的短时期局部缺损与随后的修复以及衰退区周围多层细胞胞间通透性的显著增大,既有利于外源物质的适时与大量掺入和在细胞间的快速运转,并仍保持了胚根鞘的结构完整,与分化前期胚胎对养分的大量需求正相适应,显示出胚根鞘的颈区比之球状突起处在营养吸收上占有更重要的地位。     

The Structural Changes of the Basal Region of Wheat Proembryo in Relation to Nurture Absorption

There are some cellular fail and degeneration in the parietal area of the basal region of developing wheat proembryo. Electron microscopic studies reveal that the envelopment of peripheral wall to the proembryo is partly ruptured in this area and the disassembled protoplasm of the degenerated cells mixes with the disintegrated constituents of adjacent endosperm cells. Hence, in the limited area a direct communication between the inner surviving proembryo cells and the surrounding medium is established. A number of ectodesma-like plasmodesmata and open channels appear at the boundary wall, various nutrients may enter the proembryo via symplastic pathway or by endocytosis. The surrounding macromolecules (disassembled nuclei, mitochondria, cytoplasmic granules and vesicles packed with fibrils) appear to traverse across the wall continually, and it seems that this is'an important mode of nurture translocation. Also, within the proembryo some of the densely distributed plasmodesmata undergo modification and become fully opened for macromolect, les traversing, which is in favor of re-distribution of cell contents amongst proembryo cells. Presumably, the structural changes occurred in the basal region is a special kind of differentiation which results in function of this local area as apparatus of nurture absorption. Evidently, it would enhance the incorporation of external materials into the proembryo, and then the normal proliferation, development and differentiation of proembryo cells would be ensured.     

Incorporation of Exogenous Macromolecules and Impermeable Fluorescent Dyes into the Developing Proembryo in Triticum aestivum

Our recent work has uncovered that a direct communication is established between the developing wheat proembryo and the surrounding medium at the parietal area of the basal region, in which a few cells were undergoing structural deformation and disintegration. With fluorescence microscopy and electron microscopy a series of experiments were conducted by using cationized ferritin and lucifer yellow CH as tracers to investigate the above mentioned “direct communication” in relation to the absorption of external materials. Evidences obtained show that either the macromolecule, ferritin, or the fluorescent dye, lucifer yellow actually can enter into the proembryo cells through non-transmenbrane pathway. Ultrastructural studies confirm that the incorporation of these external chemicals is realized by traversing a special route (ectodesma-like plasmodesmata and open channels) at the boundary wall of the local area of the basal region.     

Callose deposition at plasmodesmata

Summary The transport of ions and metabolites through plasmodesmata has been thought to be controlled at the neck region where the cytoplasmic annulus is constricted and where callose has also been localised. In order to determine the possible structural and functional effects of callose, its deposition was inhibited through incubation of the plant tissue with 2-deoxy-D-glucose (DDG) for 1 h prior to fixation in 2.5% glutaraldehyde. The inhibition of callose formation was monitored through aniline blue-induced fluorescence of callose. The neck region of the plasmodesmata fromAllium cepa L. roots treated with DDG exhibited a funnel-shaped configuration. This is in contrast to the plasmodesmata from tissue not incubated with DDG, which exhibited constricted necks similar to those previously reported. Both initial dissection and glutaraldehyde fixation induced neck constriction in plasmodesmata, however, dissection of tissue increased the frequency of constrictions. The inhibition of callose formation by chemical means showed that the neck constrictions and raised collars in this area are artefacts due to physical wounding and glutaraldehyde fixation. The external electron-dense material observed when tannic acid is included in the primary fixative appears to be unrelated to the deposition of callose at the neck region.Abbreviations DDG 2-deoxy-D-glucose     

Are there symplastic connections between the endosperm and embryo in some angiosperms?—a lesson from the Crassulaceae family

It is believed that there is symplastic isolation between the embryo (new sporophyte) and the endosperm (maternal-parental origin tissue, which nourishes the embryo) in angiosperms. However, in embryological literature there are rare examples in which plasmodesmata between the embryo suspensor and endosperm cells have been recorded (three species from Fabaceae). This study was undertaken in order to test the hypothesis that plasmodesmata between the embryo suspensor and the endosperm are not so rare but also occur in other angiosperm families; in order to check this, we used the Crassulaceae family because embryogenesis in Crassulaceae has been studied extensively at an ultrastructure level recently and also we tread members of this family as model for suspensor physiology and function studies. These plasmodesmata even occurred between the basal cell of the two-celled proembryo and endosperm cells. The plasmodesmata were simple at this stage of development. During the development of the embryo proper and the suspensor, the structure of plasmodesmata changes. They were branched and connected with electron-dense material. Our results suggest that in Crassulaceae with plasmodesmata between the endosperm and suspensor, symplastic connectivity at this cell-cell boundary is still reduced or blocked at a very early stage of embryo development (before the globular stage). The occurrence of plasmodesmata between the embryo suspensor and endosperm cells suggests possible symplastic transport between these different organs, at least at a very early stage of embryo development. However, whether this transport actually occurs needs to be proven experimentally. A broader analysis of plants from various families would show whether the occurrence of plasmodesmata between the embryo suspensor and the endosperm are typical embryological characteristics and if this is useful in discussions about angiosperm systematic and evolution.     

FINE STRUCTURE OF THE ZYGOTE AND EARLY EMBRYO IN QUERCUS GAMBELII

This investigation begins with the late zygote and traces ultrastructural development to the late globular stage of the embryo. Two nucleoli and satellite nucleoli sometimes occur in the zygote nucleus. Mitochondria, dictyosomes, cytoplasmic ribosomes, rough ER, and lipid bodies are numerous in the zygote. Microbodies are occasionally seen. The cell wall becomes well developed before the first division. No plasmodesmata occur in the zygote wall. The basal cell of the proembryo and the suspensor cells of the later embryo have very dense cytoplasm with a high concentration of cytoplasmic ribosomes. The nuclei are very electron opaque. The terminal cell and the cells of the embryo proper have a fine structure similar to that of the zygote. Plastids increase in number, size, starch content, and amount of thylakoid lamellae as the embryo develops. Mitochondria are numerous and appear active at all stages. Dictyosome activity, ribosomal aggregation, and the amount of ER are highest during the late globular stage. Lipid bodies are present up to the early globular stage, then disappear. The inner cell walls of the embryo are thin and have many plasmodesmata. These walls begin to thicken at the late globular stage, and at this time the size of the embryo begins to show an increase over that of the zygote. The results show a corresponding increase in the amount and activity of the metabolic machinery as the development of the embryo progresses. Lipids are probably more important as a nutrient source in the zygote and early embryo; starch becomes more important in the late stages. Absorption of nutrient material into the embryo sac and developing embryo appears to be from the chalazal end.     

The formation of symplasmic domains by plugging of plasmodesmata: a general event in plant morphogenesis?

Summary The plasmodesmal network was examined in multicellular protoplast-derived calluses of the dicotyledonSolanum nigrum which had not yet formed any visible adventitious organs and in globular proembryogenic structures developed from scutellar calluses of the monocotyledonMolinia caerulea. Electron microscopical analyses revealed that both calluses and proembryos consisted of small, undifferentiated cells. The interconnecting plasmodesmata at many cell interfaces were structurally inconspicuous in both systems; in particular cell walls, however, all plasmodesmata were occluded with an osmiophilic, dense material. As the blocking material was obviously located in the microchannels of the plasmodesmal cytoplasmic sleeves, the plugged plasmodesmata can be assumed to be nonfunctional. Thus, selective occlusion of all the plasmodesmata in specific cell walls resulted in the symplasmic disconnection of particular adjacent cells. Complex patterns of symplasmic continuity and discontinuity were established within the developing tissues. Some cells or groups of cells were entirely symplasmically disconnected from the surrounding cells by plugged plasmodesmata and might function as independent domains. However, blockage of plasmodesmata was achieved by the surrounding cells rather than by those cells belonging to the isolated domains. The demarcation of symplasmic domains might be a general prerequisite for differential morphogenesis, since they were found to be established very early in the course of morphogenetic processes.     

Filament Disruption in Funaria Protonemata: Occlusion of Plasmodesmata

Plasmodesmata are occluded when Funaria chloronemata are fragmented by the development of tmema cells (TCs). The TC deposits a new wall layer along the cross wall toward the neighbouring non-sister cell (NC). This wall layer cuts off the plasmodesmata and its connection with the cross wall is soon lost. The plasmodesmata become isolated when the NC forms a new wall layer along the former cross wall. At the end of TC development, before its disintegration, the sister cell (SC) also deposits a new wall layer along the cross wall toward the TC, cutting off the plasmodesmata. For some time the plasmalemma of the plasmodesmata remains connected to the NC or the TC, whereas the desmotubule soon disappears. Relicts of the plasmalemma remain even after the isolation of the plasmodesmata and the disintegration of the TC. During the decay of the plasmodesmata, a cylinder of electron-dense material is frequently formed along the border of the plasmodesmatal channel. This may extend over the surface of the cell wall. Eventually, the plasmodesmatal channel is filled with wall material. Callose is only observed around functional plasmodesmata and does not seem to play a role in their occlusion.     

Localization of a myosin‐like protein to plasmodesmata

Myosin has been localized to plasmodesmata in root tissues of Allium cepa, Zea mays and Hordeum vulgare using a polyclonal antibody to animal myosin in both fluorescence and electron microscopy. Labelling was also observed throughout the cytoplasm, mainly associated with the endoplasmic reticulum and plasma membrane. On Western blots, bands of 180 and 110 kDa were consistently labelled in all three species. These bands were also labelled when the blot was incubated in actin prior to staining with antibodies to actin, raising the possibility that either of these proteins (180 kDa or 110 kDa) may be present in plasmodesmata. Pre-treatment of the tissue with 2,3-butanedione monoxime (BDM), an inhibitor of actin–myosin motility, resulted in a strong constriction of the neck region of plasmodesmata. These results indicate that a myosin-like protein may be present in plasmodesmata and may also play a role in the regulation of transport at the neck region.     

Analysis of chlorophyll fluorescence reveals stage specific patterns of chloroplast-containing cells during Arabidopsis embryogenesis

The basic body plan of a plant is established early in embryogenesis when cells differentiate, giving rise to the apical and basal regions of the embryo. Using chlorophyll fluorescence as a marker for chloroplasts, we have detected specific patterns of chloroplast-containing cells at specific stages of embryogenesis. Non-randomly distributed chloroplast-containing cells are seen as early as the globular stage of embryogenesis in Arabidopsis. In the heart stage of embryogenesis, chloroplast containing cells are detected in epidermal cells as well as a central region of the heart stage embryo, forming a triangular septum of chloroplast-containing cells that divides the embryo into three equal sectors. Torpedo stage embryos have chloroplast-containing epidermal cells and a central band of chloroplast-containing cells in the cortex layer, just below the shoot apical meristem. In the walking-stick stage of embryogenesis, chloroplasts are present in the epidermal, cortex and endodermal cells. The chloroplasts appear reduced or absent from the provascular and columella cells of walking-stick stage embryos. These results suggest that there is a tight regulation of plastid differentiation during embryogenesis that generates specific patterns of chloroplast-containing cells in specific cell layers at specific stages of embryogenesis.     

Capsella embryogenesis: The suspensor and the basal cell

Summary The suspensor and basal cell ofCapsella were examined with the electron microscope and analyzed by histochemical procedures. The suspensor cells are more vacuolate and contain more ER and dictyosomes, but fewer ribosomes and stain less intensely for protein and nucleic acids than the cells of the embryo. The end walls of the suspensor cells contain numerous plasmodesmata but there are no plasmodesmata in the walls separating the suspensor from the embryo sac. The lower suspensor cells fuse with the embryo sac wall and the lateral walls of the lower and middle suspensor cells produce finger-like projections into the endosperm. At the heart stage the suspensor cells begin to degenerate and gradually lose their ability to stain for protein and nucleic acids.The basal cell is highly vacuolate and enlarges to a size of 150 X 70. An extensive network of wall projections develops on the micropylar end wall and adjacent lateral wall. The nucleus becomes deeply lobed and suspended in a strand of cytoplasm traversing the large vacuole. The cytoplasmic matrix darkens at the late globular stage and histochemical staining for protein becomes very intense. The basal cell remains active after the suspensor cytoplasm has degenerated. It is proposed that the suspensor and basal cell function as an embryonic root in the absorption and translocation of nutriments from the integuments to the developing embryo.Research supported by NSF grant GB 3460 and NIH grant 5-RO 1-CA-03656-09.     

Embryogenic cells in Dactylis glomerata L. (Poaceae) explants identified by cell tracking and by SERK expression

 Single mesophyll cells in leaf explants of Dactylis glomerata L. (Dactylis) that were competent to form somatic embryos directly or through callus were identified by semi-automatic cell tracking. These competent cells were a subpopulation of small, isodiametric, cytoplasm-rich cells located close to the vascular bundles. Using whole mount in situ hybridization, we showed that a similar subpopulation of cells expressed the Somatic Embryogenesis Receptor-like Kinase (SERK) gene during the induction of embryogenic cell formation. In both leaf explants and suspension cultures, a transient pattern of SERK gene expression was found during early embryo development, up to the globular stage. In later embryo stages, SERK mRNA was present in the shoot apical meristem, scutellum, coleoptile and coleorhiza. Received: 14 May 1999 / Revision received: 27 August 1999 / Accepted: 8 September 1999     

The neck constriction in plasmodesmata

Simple plasmodesmata between mesophyll and bundle sheath cells in actively expanding leaves of Salsola kali L. and roots of Epilobium hirsutum L. are shown to possess specialized structures, called sphincters, around their neck regions. The sphineters are made visible by the combined effects of tannic acid and heavy metal staining; they are localized just outside that area of the plasmalemma, which forms the collar around the entrance to each plasmodesmos. This localization corresponds to a very active area of the plasmodesmos/olasmalemma complex (i.e. enzyme activity and/or presence of strongly reducing substances).Evidence is presented that these ring structures are structural equivalents to hypothetical sphincters performing some valve function; i.e. participating in the control of rates and directions of symplastic transport of solutes through plasmodesmata. The middle layer of the plasmalemma in the neck region is composed of closely-packed, globular subunits appearing in negative contrast. Apparently, these subunits correspond to particle clusters observed at the plasmodesmatal entrance in freeze-fracture preparations. They appear similar to particle clusters in animal tight junctions, and their possible function in providing electrical coupling via low resistance junctions between plant cells is discussed.     

Histology of somatic embryogenesis in cultured immature embryos of maize (Zea mays L.)

Summary The developmental histology of somatic embryo (=embryoid) formation in cultured immature embryos of hybrid maize cultivars (Zea mays L.) is described. Embryos cultured on media containing 2% sucrose formed distinct globular embryoids. These embryoids arose either directly by divisions confined to the epidermal and the subepidermal cells at the coleorhizal end of the scutellum or from a soft and friable embryogenic callus produced by them. On media containing 6% sucrose divisions were initiated in the cells adjacent to the procambium of the cultured embryos. Subsequently, zones of meristematic cells also were observed in the region of the node and in the basal portion of the scutellum. Mature, well organized somatic embryos as well as a compact nodular type of embryogenic callus were produced as a result of localized meristematic activity along the tip of the scutellum toward the coleorhiza. Some embryos formed only the compact type of callus, and shoot primordia were organized later in the surface layers of this callus.Abbreviations CH casein hydrolysate - MS Murashige and Skoog's nutrient medium - 2,4-D 2,4-dichlorophenoxyacetic acid     

Cell number,cell growth,antheridiogenesis, and callose amount is reduced and atrophy induced by deoxyglucose in <Emphasis Type="Italic">Anemia phyllitidis</Emphasis> gametophytes

Fluorescence staining and morphometrical measurements revealed that callose was a component of newly formed cell plates of symmetrically dividing cells and asymmetrically dividing antheridial mother cells during gibberellic acid-induced antheridiogenesis as well as in walls of young growing cells of Anemia phyllitidis gametophytes. Callose in cell walls forms granulations characteristic of pit fields with plasmodesmata. 2-deoxy-d-glucose (DDG), eliminated callose granulations and reduced its amount estimated by measurements of fluorescence intensity. This effect was accompanied by reduction of antheridia and cell numbers as well as size and atrophy of particular cells and whole gametophytes. It is suggested that inhibition of glucose metabolism and/or signalling, might decrease callose synthesis in A. phyllitidis gametophytes leading to its elimination from cell plates of dividing cells and from walls of differentiating ones as well as from plasmodesmata resulting in inhibition of cytokinesis, cell growth and disruption of the intercellular communication system, thus disturbing developmental programs and leading to cell death.     

Ultrastructural characterization of apospory in Panicum maximum

The nucellar ultrastructure of apomictic Panicum maximum was analyzed during the meiocytic stage and during aposporous embryo sac formation. At pachytene the megameiocyte shows a random cell organelle distribution and sometimes only an incomplete micropylar callose wall. The chalazal nucellar cells are meristematic until the tetrad stage. They can turn into initial cells of aposporous embryo sacs. The aposporous initials can be recognized by their increased cell size, large nucleus, and the presence of many vesicles. The cell wall is thin with few plasmodesmata. If only a sexual embryo sac is formed, the nucellar cells retain their meristematic character. The aposporous initial cell is somewhat comparable to a vacuolated functional megaspore. It shows large vacuoles around the central nucleus and is surrounded by a thick cell wall without plasmodesmata. In the mature aposporous embryo sac the structure of the cells of the egg apparatus is similar to each other. In the chalazal part of the egg apparatus the cell walls are thin and do not hamper the transfer of sperm cells. Structural and functional aspects of nucellar cell differentiation and aposporous and sexual embryo sac development are discussed.     

Ultrastructural characteristics of « Diplotaxis erucoides (L.) DC. » suspensor

Abstract

The development and general morphology of Diplotaxis erucoides (L.) DC. suspensor is of the « Onagrad Type », « Alyssum Variation ». Maximum growth of the suspensor occurs from the globular to the early heart stage of embryo development. The suspensor starts then to degenerate disintegrating shortly after the torpedo stage of the embryo.

The wall ingrowths of the long, tapering, basal cell are especially abundant at the cell's micropilar pole which is closely surrounded by well developed wall ingrowths formed by the endosperm. Wall ingrowths and plasmodesmata are present on the suspensor cells cross walls with the exception of the cell closest to the embryo. No such structures in fact are present on the walls separating this last cell both from the embryo and from the rest of the suspensor. Wall ingrowths are generally associated with numerous, large, mitochondria.

The morphological data seem to indicate that absorption and transport of nutrients from the surrounding tissues is a main function of the suspensor. The possibility of an elaborative and secretory function of this structure is discussed.     

The Effects of X-rays on the Embryonal Growth and Development of Hordeum vulgare L.

The effects of softer X-rays as an external agent on the developingembryos were manifested by a number of abnormalities such asenlargement and vacuolation of cells, disturbances in cell walls,disintegration of cells leading to the formation of cavities,disturbances in distribution of meristematic tissues, a numberof proliferations in different parts of the embryo, and retardedgrowth of the embryo during the early post-radiation period.In addition to these, deeply stained globular cytoplasmic bodieswere observed in radiated embryonic cells. Very similar kindsof abnormalities were produced by softer X-rays starting from400 r. and including 800 r. The frequency of occurrence anddegree of proliferations in embryos irradiated with 800 r.,however, were relatively higher than those produced by 400 r.and 500 r. Treatments with 300 r. units of irradiation inducedonly the appearance of deeply stained globular bodies withincytoplasm and no effects were visible with dosages below 300r. Comparative measurements of cell- lengths of the treatedand control embryos at undifferentiated as well as at differentiatedstages showed an increase in cell-lengths within radiated embryos.Radiated embryos showed defective pattern of germination. HarderX-rays were found to be less effective in inducing embryonicabnormalities than softer rays when equal calculated dosagesof the two were applied to the flowering spikes of Hordeum vulgareL.     

Occurrence of plasmodesmata between differentiating vessels and other xylem cells inSorbus torminalis L. Crantz and their fate during xylem maturation

Summary The development of pit-pairs between differentiating xylem cells has been examined by transmission electron microscopy in young shoots ofSorbus torminalis. In some vessel-to-tracheid pits, as well as in previously studied intertracheid pits, a thickening of the pit membrane containing branched plasmodesmata was observed. A secondary wall-like cap was deposited over the thickening prior to cytoplasmic autolysis; some plasmodesmata, parallel to the plane of section, appeared to perforate the cap. At the end of the cell maturation stage, the central part of the primary wall thickening was hydrolysed, while the cap, including plasmodesmata remnants, appeared unaltered. In half-bordered pit-pairs between a parenchyma cell and a vessel or a tracheid, similar structures could be observed beside the conducting elements. When the vessel or tracheid matured, sealing of the pit membrane plasmodesmata resulted from the formation of a protective layer on the parenchyma-side rather than from the deposition of a cap on the conducting cell-side. These observations provide the first information on the presence of symplasmic connections in pits between differentiating vessels and neighbouring xylem cells. InS. torminalis, xylem differentiation is probably highly coordinated within a symplasmic domain; the persistence of such connections may account for the lack of specialization ofSorbus wood.     

Integument cell differentiation in dandelions (<Emphasis Type="Italic">Taraxacum</Emphasis>, Asteraceae,Lactuceae) with special attention paid to plasmodesmata

The aim of the paper is to determine what happens with plasmodesmata when mucilage is secreted into the periplasmic space in plant cells. Ultrastructural analysis of the periendothelial zone mucilage cells was performed on examples of the ovule tissues of several sexual and apomictic Taraxacum species. The cytoplasm of the periendothelial zone cells was dense, filled by numerous organelles and profiles of rough endoplasmic reticulum and active Golgi dictyosomes with vesicles that contained fibrillar material. At the beginning of the differentiation process of the periendothelial zone, the cells were connected by primary plasmodesmata. However, during the differentiation and the thickening of the cell walls (mucilage deposition), the plasmodesmata become elongated and associated with cytoplasmic bridges. The cytoplasmic bridges may connect the protoplast to the plasmodesmata through the mucilage layers in order to maintain cell-to-cell communication during the differentiation of the periendothelial zone cells.