Ultrastructure of the developing embryo sac of sunflower (Helianthus annuus) before and after fertilization

1991 ◽  
Vol 69 (1) ◽  
pp. 191-202 ◽  
Author(s):  
Hua Yan ◽  
Hong-Yuan Yang ◽  
William A. Jensen

The ultrastructure of the embryo sac of the sunflower (Helianthus annuus) was investigated before and after fertilization. In the young embryo sac, walls were observed that completely surrounded the egg, synergids, and the central cell. However, as maturation continued, the extent of the wall changed. By the time the embryo was mature, the chalazal portion of the walls of the egg and synergids had disappeared so these cells have a plasma membrane to plasma membrane contact. This is also true for the central cell, which has plasma membrane contact with the egg and synergids. However, the chalazal and lateral walls of the central cell become considerably thicker at this time. Before the entry of the pollen tube, the synergid that is located toward the placenta degenerates. After fertilization, a wall forms over the chalazal portion of the zygote and the persistent synergid. The endosperm appears to play an active role in this process, contributing substantial amounts of wall material. However, the wall covering the chalazal portion of the zygote is not complete by the time the zygote divides. In the proembryo, ribosome density increases and lipid bodies decrease in number. The suspensory cell has autophagic vacuoles that encircle some of the organelles. Our results support the concept that the egg, synergids, and central cell form a single functional unit, the female germ unit. Key words: sunflower, ultrastructure, embryo sac, female germ unit.


1971 ◽  
Vol 132 (4) ◽  
pp. 367-371 ◽  
Author(s):  
William Newcomb ◽  
Taylor A. Steeves


1977 ◽  
Vol 25 (6) ◽  
pp. 599 ◽  
Author(s):  
DJ Mares ◽  
BA Stone ◽  
C Jeffrey ◽  
K Norstog

In the first 4-5 days after anthesis, the central cell of the wheat embryo sac undergoes transformation from a multinucleate syncytium to the cellular endosperm. This is accomplished initially by the centripetal growth of wall projections from the central cell wall and the formation of cylindrical, highly vacuolate alveoli. Growth is mediated through the production and planar aggregation of vesicles at the distal tip of the developing projections. The innermost ends of the alveoli are closed by a thin layer of cytoplasm which is bounded on the inner side by the vacuolar membrane of the central cell. Cell wall material is not found in this thin layer of cytoplasm and the alveoli therefore are not complete cells. Following the division of the alveolar nucleus a cross-wall is laid down between the daughter nuclei by a process which is similar to normal cytokinesis and a layer of endosperm cells is formed from the peripheral portions of the alveoli. This pattern of centripetal growth of alveoli and the formation of complete cells from the proximal portions continues until cellularization is completed by the confluence of alveoli originating from opposite sides of the central cell. Further growth of the cellular endosperm is accomplished by the meristematic activity of the peripheral layer of cells. The ultrastructure of the early stages of partitioning of the central cell is discussed in relation to current views on the ontogeny of wheat endosperm.



2021 ◽  
Author(s):  
Noemi Ruiz-Lopez ◽  
Jessica Pérez-Sancho ◽  
Alicia Esteban del Valle ◽  
Richard P Haslam ◽  
Steffen Vanneste ◽  
...  

Abstract Endoplasmic reticulum-plasma membrane contact sites (ER-PM CS) play fundamental roles in all eukaryotic cells. Arabidopsis thaliana mutants lacking the ER-PM protein tether synaptotagmin1 (SYT1) exhibit decreased plasma membrane (PM) integrity under multiple abiotic stresses such as freezing, high salt, osmotic stress and mechanical damage. Here, we show that, together with SYT1, the stress-induced SYT3 is an ER-PM tether that also functions in maintaining PM integrity. The ER-PM CS localization of SYT1 and SYT3 is dependent on PM phosphatidylinositol-4-phosphate and is regulated by abiotic stress. Lipidomic analysis revealed that cold stress increased the accumulation of diacylglycerol at the PM in a syt1/3 double mutant relative to wild type while the levels of most glycerolipid species remain unchanged. Additionally, the SYT1-green fluorescent protein (GFP) fusion preferentially binds diacylglycerol in vivo with little affinity for polar glycerolipids. Our work uncovers a SYT-dependent mechanism of stress adaptation counteracting the detrimental accumulation of diacylglycerol at the PM produced during episodes of abiotic stress.



2021 ◽  
Vol 13 (3) ◽  
pp. 1128
Author(s):  
Laura Triviño-Cabrera ◽  
Elisa Isabel Chaves-Guerrero ◽  
Laura Alejo-Lozano

Studies on the adaptation from face-to-face to online teaching during lockdown show the before and after in education that faces the double challenge of promoting digital skills and public access to connectivity and electronic devices in the post-COVID-19 era. Therefore, this article contributes to these new emerging lines of educational research by presenting an educational innovation project called “Teachers Versus COVID-19”. This project aimed to verify whether the figure of teacher-prosumer, that is, consumers of media culture and creators of their own educational resources, favors the initial training of teachers during the pandemic. To this end, the following objectives were proposed: firstly, test whether the figure of the teacher-prosumer contributes to improving the adaptation of face-to-face teaching to the virtual modality of the Didactics of Social Sciences in the Degree in Primary Education during lockdown; secondly, analyze the production of content on social networks by the students in the Degree in Primary Education, according to the objectives of sustainable development. To validate our teacher-prosumer proposal, we chose the design-based research (DBR) qualitative methodology. For this, 240 students from the course in Didactics of Social Sciences of the Degree in Primary Education at the University of Malaga created 37 educational videos that teach the social sciences curriculum to children between 6 and 12 years of age from the perspective of relevant social problems and the Sustainable Development Goals. These videos were disseminated through the project’s YouTube channel. The results of this study corroborate the effectiveness of turning students into teachers-prosumers, generating the development of critical, creative, digital, and socio-emotional skills so that they feel committed to playing an active role in social changes for a sustainable world.



2009 ◽  
Vol 297 (5) ◽  
pp. R1312-R1321 ◽  
Author(s):  
Gurjit Nagra ◽  
Lena Koh ◽  
Isabelle Aubert ◽  
Minhui Kim ◽  
Miles Johnston

In some tissues, the injection of antibodies to the β1-integrins leads to a reduction in interstitial fluid pressure, indicating an active role for the extracellular matrix in tissue pressure regulation. If perturbations of the matrix occur in the periventricular area of the brain, a comparable lowering of interstitial pressures may induce transparenchymal pressure gradients favoring ventricular expansion. To examine this concept, we measured periventricular (parenchymal) and ventricular pressures with a servo-null micropipette system (2-μm tip) in adult Wistar rats before and after anti-integrin antibodies or IgG/IgM isotype controls were injected into a lateral ventricle. In a second group, the animals were kept for 2 wk after similar injections and after euthanization, the brains were removed and assessed for hydrocephalus. In experiments in which antibodies to β1-integrins ( n = 10) but not isotype control IgG/IgM ( n = 7) were injected, we observed a decline in periventricular pressures relative to the preinjection values. Under similar circumstances, ventricular pressures were elevated ( n = 10) and were significantly greater than those in the periventricular interstitium. We estimated ventricular to periventricular pressure gradients of up to 4.3 cmH2O. In the chronic preparations, we observed enlarged ventricles in many of the animals that received injections of anti-integrin antibodies (21 of 29 animals; 72%) but not in any animal receiving the isotype controls. We conclude that modulation/disruption of β1-integrin-matrix interactions in the brain generates pressure gradients favoring ventricular expansion, suggesting a novel mechanism for hydrocephalus development.



1971 ◽  
Vol 49 (12) ◽  
pp. 2067-2073 ◽  
Author(s):  
L. J. Littlefield ◽  
C. E. Bracker

The urediospores of Melampsora lini (Ehrenb.) Lev. are echinulate, with spines ca. 1 μ long over their surface. The spines are electron-transparent, conical projections, with their basal portion embedded in the electron-dense spore wall. The entire spore, including the spines, is covered by a wrinkled pellicle ca. 150–200 Å thick. The spore wall consists of three recognizable layers in addition to the pellicle. Spines form initially as small deposits at the inner surface of the spore wall adjacent to the plasma membrane. Endoplasmic reticulum occurs close to the plasma membrane in localized areas near the base of spines. During development, the spore wall thickens, and the spines increase in size. Centripetal growth of the wall encases the spines in the wall material. The spines progressively assume a more external position in the spore wall and finally reside at the outer surface of the wall. A mutant strain with finely verrucose spores was compared to the wild type. The warts on the surface of the mutant spores are rounded, electron-dense structures ca. 0.2–0.4 μ high, in contrast to spines of the wild type. Their initiation near the inner surface of the spore wall and their eventual placement on the outer surface of the spore are similar to that of spines. The wall is thinner in mutant spores than in wild-type spores.



2004 ◽  
Vol 16 (2) ◽  
pp. 226 ◽  
Author(s):  
F. Martinez-Pastor ◽  
F. Olivier ◽  
T. Spies ◽  
L. Anel ◽  
P. Bartels

Biological Resource Banks represent a potentially valuable tool for species conservation. It is, however, necessary to understand the species-specific cryopreservation process and its consequences for spermatozoa to aid in the development of assisted reproduction as a future conservation tool. The aim of this study was to assess the in vitro functionality of white rhinoceros Cerathoterium simum epididymal spermatozoa both before and after cryopreservation. Testes from a harvested white rhino bull were removed and transported at 5°C to the laboratory within 4h. The cauda epididymis was dissected out and flushed with 2mL of Tris-citrate egg yolk extender (fraction A, Biladyl, Minitüb, Germany). A 0.1mL aliquot was removed for analysis and the balance (9mL; 2mL fraction A+7mL sperm sample) mixed with an additional 27.2mL of Tris-citrate egg yolk with glycerol (fraction B, Bidadyl). The extended sample was allowed to cool to 4°C over a 6-h period before an additional 29.2mL of cooled fraction B were added (final sperm concentration=150×106mL−1). Sperm samples were loaded into 0.25-mL straws and frozen over LN2 vapor (4cm for 20min) for later assessment. Sperm straws were thawed by placing the straws in water at 37°C for 30s. Pre-freeze and post-thaw evaluations were carried out in the same manner. Media used included: HEPES for washing (20mM HEPES, 355mM sucrose, 10mM glucose, 2.5mM KOH) and HEPES saline (197mM NaCl, instead of sucrose). An aliquot was diluted with HEPES (washing) and centrifuged for 5min at 600×g; the pellet was resuspended in HEPES saline. Sperm motility (total motility %, TM;; and progressive motility %, PM) was assessed using phase contrast microscopy (×200; 37°C). Sperm plasma membrane status was assessed using the fluorescent dye, propidium iodide (50ngmL−1 in HEPES saline;; 10min, RT). Percentage of cells with plasma membranes intact (unstained;; PMI) was recorded. Mitochondrial status was assessed with the fluorescent dye, JC-1 (7.5μM in HEPES saline;; 30min, 37°C). The % of cells with an orange-stained midpiece was recorded (active mitochondria;; MIT). Resilience to hypoosmotic shock (HOS test) was assessed by diluting a sample in 100mOsm/kg HEPES saline (1:20; 15min, RT). An aliquot was stained with PI to assess plasma membrane status (HOSPMI), and the rest was fixed with formaldehyde, and % coiled tails (positive endosmosis;; HOST) was estimated using phase contrast microscopy (×400). Evaluations of PMI, MIT and HOSPMI were performed using fluorescence microscopy (×400, 450–490nm excitation filter). The results indicated that quality was good pre-freezing (TM: 60%; PMI: 86%; MIT: 100%), except for a PM value of 15%. After thawing, although there was a drop in TM (30%), there was no decrease in PM (20%). Our in vitro functional assessment indicated a loss of quality between the pre-freeze and post-thaw evaluations, but PMI and MIT maintained their pre-thaw levels (60% and 72%, respectively). The HOS test, which indicates plasma membrane integrity, decreased from the pre-freeze level (91%) to a post-thaw value of 70%. HOSTPMI was 72% pre-freeze, and decreased to 54% post-thaw. In conclusion, epididymal spermatozoa from the white rhino may retain its functionality after cryopreservation in a commerically available cryo-extender (Bidadyl). The use of assisted reproduction techniques could someday play a role in the management and conservation of the white rhinoceros and related species.



1999 ◽  
Vol 341 (2) ◽  
pp. 435-444 ◽  
Author(s):  
Jacqueline WHATMORE ◽  
Claudia WIEDEMANN ◽  
Pennti SOMERHARJU ◽  
Philip SWIGART ◽  
Shamshad COCKCROFT

Receptor-mediated phospholipase C (PLC) hydrolysis of phosphoinositides is accompanied by the resynthesis of phosphatidylinositol (PI). Hydrolysis of phosphoinositides occurs at the plasma membrane, and the resulting diacylglycerol (DG) is converted into phosphatidate (PA). Two enzymes located at the endoplasmic reticulum (ER) function sequentially to convert PA back into PI. We have established an assay whereby the resynthesis of PI could be followed in permeabilized cells. In the presence of [γ-32P]ATP, DG generated by PLC activation accumulates label when converted into PA. The 32P-labelled PA is subsequently converted into labelled PI. The formation of labelled PI reports the arrival of labelled PA from the plasma membrane to the ER. Cytosol-depleted, permeabilized human neutrophils are capable of PI resynthesis following stimulation of PLCβ (in the presence of phosphatidylinositol-transfer protein), provided that CTP and inositol are also present. We also found that wortmannin, an inhibitor of endocytosis, or cooling the cells to 15 °C did not stop PI resynthesis. We conclude that PI resynthesis is dependent neither on vesicular transport mechanisms nor on freely diffusible, soluble transport proteins. Phosphatidylcholine-derived PA generated by the ADP-ribosylation-factor-stimulated phospholipase D pathway was found to accumulate label, reflecting the rapid cycling of PA to DG, and back. This labelled PA was not converted into PI. We conclude that PA derived from the PLC pathway is selected for PI resynthesis, and its transfer to the ER could be membrane-protein-mediated at sites of close membrane contact.



1987 ◽  
Vol 88 (3) ◽  
pp. 313-325
Author(s):  
C.A. Feltkamp ◽  
H. Spiele ◽  
E. Roos

The interaction of H-2 antigens and plasma membrane-associated filaments was studied on dry-cleaved preparations of immunogold-labelled lymphoma cells. In prefixed cells, the plasma membrane-associated network was isotropic without any prevailing direction of the filaments, and the gold-labelled H-2 antigens were preferentially localized over or at a very short distance from membrane-associated filaments. Incubation of unfixed cells with anti-H-2 antibodies followed by fixation and incubation with anti-Ig, did not induce detectable redistribution of H-2 antigens or of the filament network. Notwithstanding this apparent absence of rearrangement of H-2 antigens and filaments, a detergent-resistant linkage to the cytoskeleton was induced. Before immune incubations, virtually all H-2 antigens were solubilized by extraction with Triton X-100, while after incubation with anti-H-2 antibodies about 50% of the H-2 antigens were linked to the Triton X-100-insoluble cytoskeleton. Sequential addition of anti-H-2 and anti-Ig antibodies to unfixed cells induced formation of patches and caps of H-2 antigens. Under these conditions, the majority of the H-2 antigens became linked to the detergent-resistant cytoskeleton. Redistribution into patches and caps was often accompanied by a local rearrangement of the isotropic network into bundles of parallel filaments immediately adjacent to the plasma membrane. Patches were seen to overly both isotropic networks and these parallel filaments. Large sheets of plasma membrane overlying parallel filaments were frequently devoid of gold-labelled H-2 antigens and coated pits, and thus most probably represented areas away from caps. This observation suggests that capping is accompanied by a rearrangement of filaments close to the membrane.



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