Embryology of Brassica campestris: the entrance and discharge of the pollen tube in the synergid and the formation of the zygote

1992 ◽  
Vol 70 (8) ◽  
pp. 1577-1590 ◽  
Author(s):  
M. J. Sumner
Keyword(s):  
Endoplasmic Reticulum ◽  
Cell Wall ◽  
Plasma Membrane ◽  
Pollen Tube ◽  
Brassica Campestris ◽  
Periodic Acid ◽  
Outer Integument ◽  
Central Cell ◽  
Egg Cell ◽  
Intense Staining

The postanthesis synergids and zygote of Brassica campestris cv. Candle were examined using techniques of light, fluorescence, and electron microscopy. The pollen tube enters the degenerate synergid by way of the filiform apparatus. A degeneration of one of the two synergids occurs after anthesis and is independent of pollination. The first sign of synergid degeneration is a more intense staining of one of the synergids, followed by a loss of organelle membrane integrity. There is a disappearance of the plasma membrane and dictyosome cisternae; however, profiles of degenerate synergid mitochondria, plastids, and dilated endoplasmic reticulum remain along with dictyosome vesicles that contain periodic acid – thiocarbo-hydrazide – silver proteinate positive substances. The zygote, shortly after fertilization, is reduced in size and lacks the large micropylar vacuole characteristic of the mature unfertilized egg cell. Plastids and mitochondria are concentrated around the centrally located nucleus of the zygote, and dictyosomes, active in vesicle production, are located in the lateral and chalazal regions of the cell, adjacent to the cell wall. The lateral cell walls are periodic acid – Schiff's and Calcofluor positive, while the ampulliform chalazal tip of the cell is weakly periodic acid – Schiff s positive and Calcofluor negative. Microtubules, with the long axis perpendicular to the long axis of the zygote, are abundant in the ampulliform chalazal tip of the cell. Following fertilization the central cell becomes highly vacuolate. There is continuity between the zygote – central cell plasma membrane, the central cell vacuole tonoplast, and membranes of the central cell endoplasmic reticulum. Central cell wall projections, of the transfer cell type, are located in the lateral regions of the megagametophyte adjacent to the developing zygote cell and are positioned adjacent to the region of inner and outer integument starch. Key words: Brassica, ultrastructure, synergid, megagametophyte, pollen tube, zygote.

The development of the central cell of Brassica campestris prior to fertilization

1990 ◽  
Vol 68 (12) ◽  
pp. 2553-2563 ◽  
Author(s):  
M. J. Sumner ◽  
L. van Caeseele
Keyword(s):  
Endoplasmic Reticulum ◽  
Cell Wall ◽  
Brassica Campestris ◽  
Periodic Acid ◽  
Light And Electron Microscopy ◽  
Lateral Wall ◽  
Polar Nucleus ◽  
Central Cell ◽  
Transfer Cell ◽  
Egg Apparatus

The development of the central cell of Brassica campestris cv. Candle (canola-rapeseed) was examined using techniques of light and electron microscopy and cytochemistry. The mature central cell is devoid of the large central vacuole characteristic of the early cellular stage of megagametophyte development. Prior to anthesis, cell wall projections, of the transfer cell type, develop on the lateral wall of the central cell. These central cell wall projections extend from the midregion of the megagametophyte to the egg apparatus and are immediately adjacent to the starch-containing region of the inner and outer integuments. The cell wall projections are periodic acid – thiocarbohydrazide – silver proteinate positive as are the contents of dictyosome vesicles that appear to contribute to their formation. Mitochondria are associated with the wall projections as is a network of central cell endoplasmic reticulum that extends from the wall projections to the egg apparatus. Microtubules are associated with the migrating chalazal polar nucleus. The two polar nuclei partially fuse prior to double fertilization, united by nuclear bridges and endoplasmic reticulum interconnections. Proplastids are a characteristic feature of the immature cellular megagametophyte. By anthesis, the proplastids of the mature central cell develop into chloroplasts with stacked thylakoids and starch deposits. Microbodies are frequently found associated with lipid bodies, and polysomes with the endoplasmic reticulum of the mature central cell. Key words: Brassica, central cell, megagametophyte, ovule, transfer cell.

A study of the mature megagametophyte of Stipa elmeri

1975 ◽  
Vol 53 (24) ◽  
pp. 2958-2977 ◽  
Author(s):  
Jack Maze ◽  
Shu-Chang Lin
Keyword(s):  
Electron Microscopy ◽  
Endoplasmic Reticulum ◽  
Plasma Membrane ◽  
Pollen Tube ◽  
Endosperm Development ◽  
Pollen Grain ◽  
Central Cell ◽  
Filiform Apparatus ◽  
Large Area ◽  
Dense Cytoplasm

In Stipa elmeri Piper & Brodie ex Scribn., the pollen tube enters at the filiform apparatus of the degenerated synergid. The degenerated synergid has electron-dense cytoplasm in which organelles are not discernible. All other cells of the mature megagametophyte have nuclei, endoplasmic reticulum, plastids, mitochondria, dictyosomes, and vacuoles. Starch is found in the persistent synergid (in minute quantities), egg, and central cell. Lipids occur in the persistent synergid, central cell, and antipodals. The filiform apparatuses of the two synergids are hypothesized to perform different functions. In the degenerated synergid, the filiform apparatus serves to increase the surface area of the plasma membrane and thereby to offer a large area for pollen-tube-growth-directing compounds to diffuse out of the synergid. In the persistent synergid, the filiform apparatus is part of a suite of features which indicate that the persistent synergid is involved in the transference of materials into the megagametophyte. Another possible function of the persistent synergid is to aid in establishing the polarity of the egg. The pollen grain and tube have distinctive polysaccharide spheres that serve to delimit the pollen tube cytoplasm after discharge into the degenerated synergid. Associated with the degenerated synergid are bodies of dense materials as seen under electron microscopy, and bodies of RNA and protein as determined histochemically. These are probably the same thing and come from the degenerating synergid. The antipodals are the most cytologically active cells of the megagametophyte. They have some features which are characteristic of transfer cells and possibly function in the transference of materials into the megagametophyte. Other studies (Brink and Cooper 1944) have indicated that grass antipodals are involved in the control of endosperm development. The active cytoplasm of the antipodals may reflect the synthesis or transference of growth-controlling substances.

scholarly journals The haustorial synergids of Cortaderia (Gramineae) at maturity

2014 ◽  
Vol 50 (1-2) ◽  
pp. 151-160 ◽  
Author(s):  
Melva N. Philipson
Keyword(s):  
Cell Wall ◽  
Plasma Membrane ◽  
Pollen Tube ◽  
Pollen Tube Growth ◽  
Central Cell ◽  
Transfer Cell ◽  
Tube Growth ◽  
Transport Activity ◽  
Cell Cytoplasm ◽  
Cortaderia Selloana

The ultrastructure of synergids which extend through the micropyle as haustoria and lie against 'the ovary wall are described in <em>Cortaderia selloana</em> and its F<sub>l</sub> hybrid with <em>C. araucana</em>. These haustoria bear typical transfer cell wall invaginations closely associated with the plasma membrane and with mitochondria. Their function seems to be one involved in the absorption and conduction of nutrients to the synergids which are atypical in their highly vacuolate structure, degenerate nuclei and few organelles. The synergids appear to act as repositories of nutrients which are, readily accessible to the central cell by virtue of deep intrusions made into them by the central cell cytoplasm. Enzymatic secretion could also be a function of the distal end of the haustorial synergids, both in facilitating tissue peneration during its outward growth and in directing pollen tube growth. At anfhesis, the haustorium - synergid complex appears to be past its peak of absorption and transport activity, and to be involved in a seeretory or degenerative phase.

Fertilization in Plumbago zeylanica: entry and discharge of the pollen tube in the embryo sac

1982 ◽  
Vol 60 (11) ◽  
pp. 2219-2230 ◽  
Author(s):  
Scott D. Russell
Keyword(s):  
Cell Wall ◽  
Pollen Tube ◽  
Female Gametophyte ◽  
Embryo Sac ◽  
Central Cell ◽  
Vegetative Nucleus ◽  
Egg Cell ◽  
Ultrastructural Organization ◽  
Plumbago Zeylanica ◽  
Male Gametes

The ultrastructural organization of the megagametophyte of Plumbago zeylanica, which lacks synergids, was examined in chemically and physically fixed ovules after entry of the pollen tube. Similar to angiosperms with conventionally organized megagametophytes, the pollen tube enters the ovule through a micropyle, formed by the inner integument, and approaches the female gametophyte by growing between nucellar cells. Unlike other described female gametophytes, however, continued pollen tube growth results in direct penetration of the base of the egg through cell wall projections forming a filiform apparatus and is completed between the egg and central cell without disrupting either of these cells' plasma membranes. A terminal pollen tube aperture forms when the pollen tube reaches an area of strong curvature near the summit of the egg; this results in the release of two sperm cells, the vegetative nucleus, and a limited amount of pollen cytoplasm. The formerly continuous chalazal egg cell wall is locally disrupted near the tip of the pollen tube and apparently is thus modified for reception of male gametes. Discharged pollen cytoplasm rapidly degenerates between the egg and central cell, but unlike pollen tube discharge in conventionally organized megagametophytes, it is unassociated with the degeneraton of any receptor cell within the female gametophyte. Sperm nuclei are transmitted, one to the egg and the other to the central cell, to effect double fertilization by nuclear fusion with their respective female reproductive nuclei. The vegetative nucleus and discharged pollen cytoplasm degenerate between the developing embryo and endosperm during early embryogenesis. The emerging concept that the egg of Plumbago possesses combined egg and synergid functions is supported by the present study and suggests that the megagametophyte of this plant displays a highly specialized egg apparatus composed exclusively of a single, modified egg cell.

THE ORGANIZATION OF CYTOPLASMIC COMPONENTS DURING THE PHASE OF CELL WALL THICKENING IN DIFFERENTIATING CAMBIAL DERIVATIVES OF ACER RUBRUM

1965 ◽  
Vol 43 (11) ◽  
pp. 1401-1407 ◽  
Author(s):  
James Cronshaw
Keyword(s):  
Endoplasmic Reticulum ◽  
Cell Wall ◽  
Plasma Membrane ◽  
Osmium Tetroxide ◽  
Acer Rubrum ◽  
Middle Layer ◽  
Wall Thickening ◽  
Cytoplasmic Microtubules ◽  
Derivatives Of ◽  
Peripheral Cytoplasm

Cambial derivatives of Acer rubrum have been examined at stages of their differentiation following fixation in 3% or 6% glutaraldehyde with a post fixation in osmium tetroxide. At early stages of development numerous free ribosomes are present in the cytoplasm, and elements of the endoplasmic reticulum tend to align themselves parallel to the cell surfaces. The plasma membrane is closely applied to the cell walls. During differentiation a complex system of cytoplasmic microtubules develops in the peripheral cytoplasm. These microtubules are oriented, mirroring the orientation of the most recently deposited microfibrils of the cell wall. The microtubules form a steep helix in the peripheral cytoplasm at the time of deposition of the middle layer of the secondary wall. During differentiation the free ribosomes disappear from the cytoplasm and numerous elements of rough endoplasmic reticulum with associated polyribosomes become more evident. In many cases the endoplasmic reticulum is associated with the cell surface. During the later stages of differentiation there are numerous inclusions between the cell wall and the plasma membrane.

scholarly journals Four distinct secretory pathways serve protein secretion, cell surface growth, and peroxisome biogenesis in the yeast Yarrowia lipolytica.

1997 ◽  
Vol 17 (9) ◽  
pp. 5210-5226 ◽  
Author(s):  
V I Titorenko ◽  
D M Ogrydziak ◽  
R A Rachubinski
Keyword(s):  
Endoplasmic Reticulum ◽  
Cell Wall ◽  
Plasma Membrane ◽  
Protein Secretion ◽  
Peroxisome Biogenesis ◽  
Peroxisomal Membrane ◽  
Mycelial Form ◽  
Secretory Pathways ◽  
Associated Proteins ◽  
Yeast Yarrowia Lipolytica

We have identified and characterized mutants of the yeast Yarrowia lipolytica that are deficient in protein secretion, in the ability to undergo dimorphic transition from the yeast to the mycelial form, and in peroxisome biogenesis. Mutations in the SEC238, SRP54, PEX1, PEX2, PEX6, and PEX9 genes affect protein secretion, prevent the exit of the precursor form of alkaline extracellular protease from the endoplasmic reticulum, and compromise peroxisome biogenesis. The mutants sec238A, srp54KO, pex2KO, pex6KO, and pex9KO are also deficient in the dimorphic transition from the yeast to the mycelial form and are affected in the export of only plasma membrane and cell wall-associated proteins specific for the mycelial form. Mutations in the SEC238, SRP54, PEX1, and PEX6 genes prevent or significantly delay the exit of two peroxisomal membrane proteins, Pex2p and Pex16p, from the endoplasmic reticulum en route to the peroxisomal membrane. Mutations in the PEX5, PEX16, and PEX17 genes, which have previously been shown to be essential for peroxisome biogenesis, affect the export of plasma membrane and cell wall-associated proteins specific for the mycelial form but do not impair exit from the endoplasmic reticulum of either Pex2p and Pex16p or of proteins destined for secretion. Biochemical analyses of these mutants provide evidence for the existence of four distinct secretory pathways that serve to deliver proteins for secretion, plasma membrane and cell wall synthesis during yeast and mycelial modes of growth, and peroxisome biogenesis. At least two of these secretory pathways, which are involved in the export of proteins to the external medium and in the delivery of proteins for assembly of the peroxisomal membrane, diverge at the level of the endoplasmic reticulum.

scholarly journals Intercellular trafficking via plasmodesmata: molecular layers of complexity

2020 ◽  
Author(s):  
Ziqiang Patrick Li ◽  
Andrea Paterlini ◽  
Marie Glavier ◽  
Emmanuelle M. Bayer
Keyword(s):  
Endoplasmic Reticulum ◽  
Cell Wall ◽  
Plasma Membrane ◽  
Plant Cells ◽  
Functional Domains ◽  
Dynamic Interactions ◽  
Intercellular Trafficking ◽  
Multiple Routes ◽  
Molecular Components ◽  
Molecular Layers

Abstract Plasmodesmata are intercellular pores connecting together most plant cells. These structures consist of a central constricted form of the endoplasmic reticulum, encircled by some cytoplasmic space, in turn delimited by the plasma membrane, itself ultimately surrounded by the cell wall. The presence and structure of plasmodesmata create multiple routes for intercellular trafficking of a large spectrum of molecules (encompassing RNAs, proteins, hormones and metabolites) and also enable local signalling events. Movement across plasmodesmata is finely controlled in order to balance processes requiring communication with those necessitating symplastic isolation. Here, we describe the identities and roles of the molecular components (specific sets of lipids, proteins and wall polysaccharides) that shape and define plasmodesmata structural and functional domains. We highlight the extensive and dynamic interactions that exist between the plasma/endoplasmic reticulum membranes, cytoplasm and cell wall domains, binding them together to effectively define plasmodesmata shapes and purposes.

scholarly journals Subcellular localization of cellulases in auxin-treated pea.

1976 ◽  
Vol 69 (1) ◽  
pp. 97-105 ◽  
Author(s):  
A K Bal ◽  
D P Verma ◽  
H Byrne ◽  
G A Maclachlan
Keyword(s):  
Endoplasmic Reticulum ◽  
Cell Wall ◽  
Plasma Membrane ◽  
Major Part ◽  
Osmium Tetroxide ◽  
Close Association ◽  
Cellulase Activity ◽  
Cytochemical Localization ◽  
Tissue Sections ◽  
Inner Surface

Two forms of cellulase, buffer soluble (BS) and buffer insoluble (BI), are induced as a result of auxin treatment of dark-grown pea epicotyls. These two cellulases have been purified to homogeneity. Antibodies raised against the purified cellulases were conjugated with ferritin and were used to localize the two cellulases. Tissue sections were fixed in cold paraformaldehyde-glutaraldehyde and incubated for 1 h in the ferritin conjugates. The sections were washed with continuous shaking for 18 h and subsequently postfixed in osmium tetroxide. Tissue incubated in unconjugated ferritin was used as a control. A major part of BI cellulase is localized at the inner surface of the cell wall in close association with microfibrils. BS cellulase is localized mainly within the distended endoplasmic reticulum. Gogli complex and plasma membrane appear to be completely devoid of any cellulase activity. These observations are consistent with cytochemical localization and biochemical data on the distribution of these two cellulases among various cell and membrane fractions.

Cytoskeletal organisation and modification during pollen tube arrival, gamete delivery and fertilisation in Plumbago zeylanica

Zygote ◽  
1993 ◽  
Vol 1 (2) ◽  
pp. 143-154 ◽  
Author(s):  
Bing-Quan Huang ◽  
Elisabeth S. Pierson ◽  
Scott D. Russell ◽  
Antonio Tiezzi ◽  
Mauro Cresti
Keyword(s):  
Pollen Tube ◽  
Embryo Sac ◽  
Central Cell ◽  
Egg Cell ◽  
Target Cells ◽  
Sperm Cells ◽  
Plumbago Zeylanica ◽  
Rhodamine Phalloidin ◽  
Male Gametes ◽  
Pollen Tube Penetration

The cytoskeletal organisation of the isolated embryo sac and egg cells of Plumbago zeylanica was examined before, during and after pollen tube penetration into the embryo sac to determine the potential involvement of microtubules and actin filaments in fertilisation. Material was singly and triply stained using Hoechst 33258 to localise DNA, fluorescein isothiocyanate (FITC)-labelled anti- α-tubulin to detect microtubules and rhodamine-phalloidin to visualise F-actin. Microtubules in the unfertilised egg cell are longitudinally aligned in the micropylar and mid-lateral areas, aggregating into bundles near the filiform apparatus. In the perinuclear cytoplasm of the egg cell, microtubules become more or less randomly aligned. F-actin bundles form a longitudinally aligned mesh in the chalazal cytoplasm of the egg cell. In the central cell, microtubules and F-actin are distributed along transvacuolar strands and are also evident in the perinuclear region and at the periphery of the cell. During pollen tube penetration, sparse microtubule bundles near the pathway of the pollen tube may form an apparent microtubular ‘conduit’ surrounding the male gametes at the delivery site. Actin aggregates become organised near the pathway of the pollen tube and at the delivery site of the sperm cells. Subsequently, actin aggregates form a ‘corona’ structure in the intercellular region between the egg and central cell where gametic fusion occurs. The corona may have a role in maintaining the close proximity of the egg and central cell and helping the two sperm cells move and bind to their target cells. The cytoskeleton may also be involved in causing the two nuclei of the egg and central cell to approach one another at the site of gametic fusion and transporting the two sperm nuclei into alignment with their respective female nucleus. The cytoskeleton is reorganised during early embryogenesis.

The ultrastructure and cytochemistry of the egg apparatus of Brassica campestris

1989 ◽  
Vol 67 (1) ◽  
pp. 177-190 ◽  
Author(s):  
M. J. Sumner ◽  
L. Van Caeseele
Keyword(s):  
Plasma Membrane ◽  
Cell Walls ◽  
Brassica Campestris ◽  
Positive Response ◽  
Uv Light ◽  
Egg Cell ◽  
Filiform Apparatus ◽  
Egg Apparatus ◽  
Large Numbers ◽  
Acidic Polysaccharides

The egg apparatus of Brassica campestris L. cv. Candle (canola-rapeseed) is composed of an egg and two synergids juxtaposed at the extreme micropylar end of the megagametophyte with the egg cell displaced in a chalazal direction. The cell walls of the synergids and egg are uniformly PAS and PA–TCH–SP-positive, but contained β-linked glucans only in the micropylar region. The number and development of the cytoplasmic organelles suggested that the egg cell is relatively inactive metabolically while the synergid cells are active. The synergids contain large numbers of dictyosomes with PA–TCH–SP-positive vesicles at the maturing face. These vesicles appear to fuse with the plasma membrane in the region of the filiform apparatus. The filiform apparatuses of the synergids are micropylar finger-like projections that extend into the cytoplasm of the synergid. These are PAS and PA–TCH–SP-positive, fluoresce in uv light when stained with Calcofluor, and show a positive response for acidic polysaccharides when stained with alcian blue. After treatment with cellulase, fluorescence was not observed. The incipient degenerate synergid was intensely stained by cationic dyes 24–36 h after anthesis.

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