SummaryThe morphogenesis of lobed plant cells has been considered to be controlled by microtubule (MT) and/or actin filament (AF) organization. In this article, a comprehensive mechanism is proposed, in which distinct roles are played by... more
SummaryThe morphogenesis of lobed plant cells has been considered to be controlled by microtubule (MT) and/or actin filament (AF) organization. In this article, a comprehensive mechanism is proposed, in which distinct roles are played by these cytoskeletal components. First, cortical MT bundles and, in the case of pavement cells, radial MT arrays combined with MT bundles determine the deposition of local cell wall thickenings, the cellulose microfibrils of which copy the orientation of underlying MTs. Cell growth is thus locally prevented and, consequently, lobes and constrictions are formed. Arch‐like tangential expansion is locally imposed at the external periclinal wall of pavement cells by the radial arrangement of cellulose microfibrils at every wall thickening. Whenever further elongation of the original cell lobes occurs, AF patches assemble at the tips of growing lobes. Intercellular space formation is promoted or prevented by the opposite or alternate, respectively, arrange...
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The thallus ontogeny of Dictyopteris membranacea is the outcome of a highly co-ordinated series of cell divisions, which is completed at a long distance from the apex. Three types of “meristems” function: (a) the central apical initials,... more
The thallus ontogeny of Dictyopteris membranacea is the outcome of a highly co-ordinated series of cell divisions, which is completed at a long distance from the apex. Three types of “meristems” function: (a) the central apical initials, which give rise to the other meristems and to initial cells of the midrib; (b) the marginal apical initials, which contribute mainly to
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The thallus ontogeny of Dictyopteris membranacea is the outcome of a highly co-ordinated series of cell divisions, which is completed at a long distance from the apex. Three types of “meristems” function: (a) the central apical initials,... more
The thallus ontogeny of Dictyopteris membranacea is the outcome of a highly co-ordinated series of cell divisions, which is completed at a long distance from the apex. Three types of “meristems” function: (a) the central apical initials, which give rise to the other meristems and to initial cells of the midrib; (b) the marginal apical initials, which contribute mainly to
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Summary Centrifugation of young seedlings ofTriticum durum andTriticum aestivum for 8–10 hours at 1,500–2,000 x g causes a serious disorder of the spatial organelle relationships in the interphase as well as the preprophase and mitotic... more
Summary Centrifugation of young seedlings ofTriticum durum andTriticum aestivum for 8–10 hours at 1,500–2,000 x g causes a serious disorder of the spatial organelle relationships in the interphase as well as the preprophase and mitotic subsidiary cell mother cells (SMCs). The nucleus, most organelles and cytoplasm are displaced to the centrifugal end of the cell, while the vacuoles lie at
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Summary Centrifugation of young seedlings ofTriticum durum andTriticum aestivum for 8–10 hours at 1,500–2,000 x g causes a serious disorder of the spatial organelle relationships in the interphase as well as the preprophase and mitotic... more
Summary Centrifugation of young seedlings ofTriticum durum andTriticum aestivum for 8–10 hours at 1,500–2,000 x g causes a serious disorder of the spatial organelle relationships in the interphase as well as the preprophase and mitotic subsidiary cell mother cells (SMCs). The nucleus, most organelles and cytoplasm are displaced to the centrifugal end of the cell, while the vacuoles lie at
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The manifestation of premitotic cell polarity and the resultant structural asymmetry of the differential divisions participating in the development of stomata of Vigna sinensis vary considerably. However, two morphologically distinct... more
The manifestation of premitotic cell polarity and the resultant structural asymmetry of the differential divisions participating in the development of stomata of Vigna sinensis vary considerably. However, two morphologically distinct types of differential division were distinguished: (a) 'asymmetrical differential divisions', in which the premitotic polarization of the cell, the eccentric position of the nucleus during division and the differences in size and organization of the daughter cells are obvious; and (b) differential divisions in which the above features are inconspicuous or almost absent. The former occur in the ordinary protodermal cells, the latter in some meristemoids. The organization of a sharply demarcated preprophase microtubule band (PMB) precedes, all differential and non-differential divisions. In the first type of differential division the PMB is formed eccentrically, while in the second it may display either an approximately symmetrical or a clearly asymmetrical disposition, always indicating with surprising accuracy the sites where the succeeding cell plate will join the parent walls. The PMB foreshadowing the highly curved cell plates in meristemoids I of the mesoperigenous process, as well as in meristemoids I and II of the mesogenous one, are apposed only on one anticlinal wall and therefore do not encircle the nucleus or traverse the cell. In the symmetrical divisions of guard cell mother cells (GMC), as well as in those of protodermal cells, the PMB runs right round the internal plasmalemma surface in an equatorial position, coinciding with that of the future cell plate. In the former cells the wall abutting the cortical cytoplasm traversed by the band becomes locally thickened. The variability in the pattern of the microtubules of the band along the walls of the GMC is directly mirrored in the pattern of the thickening. It seems that in GMC the PMB mediates a directed exocytosis of dictyosome vesicles. In contrast to what is now generally accepted in dicotyledonous plants, each meristemoid I of both the mesogenous and mesoperigenous stomata in Vigna sinensis leaves does not inhibit but induces the formation of other meristemoids close to it.
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The manifestation of premitotic cell polarity and the resultant structural asymmetry of the differential divisions participating in the development of stomata of Vigna sinensis vary considerably. However, two morphologically distinct... more
The manifestation of premitotic cell polarity and the resultant structural asymmetry of the differential divisions participating in the development of stomata of Vigna sinensis vary considerably. However, two morphologically distinct types of differential division were distinguished: (a) 'asymmetrical differential divisions', in which the premitotic polarization of the cell, the eccentric position of the nucleus during division and the differences in size and organization of the daughter cells are obvious; and (b) differential divisions in which the above features are inconspicuous or almost absent. The former occur in the ordinary protodermal cells, the latter in some meristemoids. The organization of a sharply demarcated preprophase microtubule band (PMB) precedes, all differential and non-differential divisions. In the first type of differential division the PMB is formed eccentrically, while in the second it may display either an approximately symmetrical or a clearly asymmetrical disposition, always indicating with surprising accuracy the sites where the succeeding cell plate will join the parent walls. The PMB foreshadowing the highly curved cell plates in meristemoids I of the mesoperigenous process, as well as in meristemoids I and II of the mesogenous one, are apposed only on one anticlinal wall and therefore do not encircle the nucleus or traverse the cell. In the symmetrical divisions of guard cell mother cells (GMC), as well as in those of protodermal cells, the PMB runs right round the internal plasmalemma surface in an equatorial position, coinciding with that of the future cell plate. In the former cells the wall abutting the cortical cytoplasm traversed by the band becomes locally thickened. The variability in the pattern of the microtubules of the band along the walls of the GMC is directly mirrored in the pattern of the thickening. It seems that in GMC the PMB mediates a directed exocytosis of dictyosome vesicles. In contrast to what is now generally accepted in dicotyledonous plants, each meristemoid I of both the mesogenous and mesoperigenous stomata in Vigna sinensis leaves does not inhibit but induces the formation of other meristemoids close to it.
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The protodermal cells producing the 'floating' guard cell mother cells (GMCs) in three Anemia species undergo an extraordinary polarization and an unexpected shaping. During interphase an intercellular space is initiated... more
The protodermal cells producing the 'floating' guard cell mother cells (GMCs) in three Anemia species undergo an extraordinary polarization and an unexpected shaping. During interphase an intercellular space is initiated at the internal proximal end of the cell, while the polar region bulges outwards. At this stage a microtubule girdle traverses the cortical cytoplasm underneath the rims of the external periclinal wall curvature. In addition, another system of microtubules converges on a cortical site adjoining the wall delimiting the intercellular space and, or, the neighbouring region of the internal periclinal wall (internal polar cortical site, IPCS). Vacuoles are found in all regions of the cell except for that between the centrally located nucleus and the intercellular space. As the cell approaches mitosis, the growing vacuolar system retreats from the cytoplasmic region below the external periclinal wall curvature. In most cells the polarized cytoplasm forms an inclined truncated cone, the bases of which abut on the external periclinal wall curvature and the wall lining the IPCS. The organization of the cortical microtubule cytoskeleton does not change significantly during preprophase-prophase. A preprophase microtubule band (PMB) is localized in the cortex lining the rims of the external periclinal wall curvature, while some microtubules traverse the IPCS and the cytoplasm adjacent to the neighbouring wall regions. The mitotic spindle axis is diagonal, while the cell plate separating the GMCs exhibits an unusual mode of growth. It gradually encircles the proximal daughter nucleus, becoming funnel-shaped. One of its periclinal edges fuses with the external periclinal wall area lined by the PMB cortical zone and the other with the internal periclinal wall area adjoining the IPCS. The latter region seems to behave like the PMB cortical zone. The results show that the morphogenetic mechanism underlying the formation of the conical GMCs includes a series of highly integrated processes, initiated or carried out during cell polarization.
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The protodermal cells producing the 'floating' guard cell mother cells (GMCs) in three Anemia species undergo an extraordinary polarization and an unexpected shaping. During interphase an intercellular space is initiated... more
The protodermal cells producing the 'floating' guard cell mother cells (GMCs) in three Anemia species undergo an extraordinary polarization and an unexpected shaping. During interphase an intercellular space is initiated at the internal proximal end of the cell, while the polar region bulges outwards. At this stage a microtubule girdle traverses the cortical cytoplasm underneath the rims of the external periclinal wall curvature. In addition, another system of microtubules converges on a cortical site adjoining the wall delimiting the intercellular space and, or, the neighbouring region of the internal periclinal wall (internal polar cortical site, IPCS). Vacuoles are found in all regions of the cell except for that between the centrally located nucleus and the intercellular space. As the cell approaches mitosis, the growing vacuolar system retreats from the cytoplasmic region below the external periclinal wall curvature. In most cells the polarized cytoplasm forms an inclined truncated cone, the bases of which abut on the external periclinal wall curvature and the wall lining the IPCS. The organization of the cortical microtubule cytoskeleton does not change significantly during preprophase-prophase. A preprophase microtubule band (PMB) is localized in the cortex lining the rims of the external periclinal wall curvature, while some microtubules traverse the IPCS and the cytoplasm adjacent to the neighbouring wall regions. The mitotic spindle axis is diagonal, while the cell plate separating the GMCs exhibits an unusual mode of growth. It gradually encircles the proximal daughter nucleus, becoming funnel-shaped. One of its periclinal edges fuses with the external periclinal wall area lined by the PMB cortical zone and the other with the internal periclinal wall area adjoining the IPCS. The latter region seems to behave like the PMB cortical zone. The results show that the morphogenetic mechanism underlying the formation of the conical GMCs includes a series of highly integrated processes, initiated or carried out during cell polarization.
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Stomata of various sizes are produced on the primary root of Ceratonia siliqua L. Most are generated during embryogenesis, prior to seed desiccation. They can be detected on the dry embryo in a wide zone just above the root tip.... more
Stomata of various sizes are produced on the primary root of Ceratonia siliqua L. Most are generated during embryogenesis, prior to seed desiccation. They can be detected on the dry embryo in a wide zone just above the root tip. Initially, large stomata are formed. These have the ability to induce divisions of their neighbouring cells, creating particular cell patterns around them. Later, small perigenous stomata are generated. As the root grows following seed germination, the stomatal zone overlaps with that of the root hairs. Although root stomata of C. siliqua undergo a structural differentiation that seems almost identical to that of the elliptical stomata formed on leaves, they are unable to move and remain permanently open. Polarizing microscopy of fully differentiated stomata and young stomata at the stage of stomatal pore formation revealed deposition of radial cellulose microfibril systems on their periclinal walls. However, these systems were less developed than those on l...
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Stomata of various sizes are produced on the primary root of Ceratonia siliqua L. Most are generated during embryogenesis, prior to seed desiccation. They can be detected on the dry embryo in a wide zone just above the root tip.... more
Stomata of various sizes are produced on the primary root of Ceratonia siliqua L. Most are generated during embryogenesis, prior to seed desiccation. They can be detected on the dry embryo in a wide zone just above the root tip. Initially, large stomata are formed. These have the ability to induce divisions of their neighbouring cells, creating particular cell patterns around them. Later, small perigenous stomata are generated. As the root grows following seed germination, the stomatal zone overlaps with that of the root hairs. Although root stomata of C. siliqua undergo a structural differentiation that seems almost identical to that of the elliptical stomata formed on leaves, they are unable to move and remain permanently open. Polarizing microscopy of fully differentiated stomata and young stomata at the stage of stomatal pore formation revealed deposition of radial cellulose microfibril systems on their periclinal walls. However, these systems were less developed than those on l...
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Subsidiary cell generation in Poaceae is an outstanding example of local intercellular stimulation. An inductive stimulus emanates from the guard cell mother cells (GMCs) towards their laterally adjacent subsidiary cell mother cells... more
Subsidiary cell generation in Poaceae is an outstanding example of local intercellular stimulation. An inductive stimulus emanates from the guard cell mother cells (GMCs) towards their laterally adjacent subsidiary cell mother cells (SMCs) and triggers the asymmetrical division of the latter. Indole-3-acetic acid (IAA) immunolocalization in Zea mays protoderm confirmed that the GMCs function as local sources of auxin and revealed that auxin is polarly accumulated between GMCs and SMCs in a timely-dependent manner. Besides, staining techniques showed that reactive oxygen species (ROS) exhibit a closely similar, also time-dependent, pattern of appearance suggesting ROS implication in subsidiary cell formation. This phenomenon was further investigated by using the specific NADPH-oxidase inhibitor diphenylene iodonium, the ROS scavenger N-acetyl-cysteine, menadione which leads to ROS overproduction, and H2O2. Treatments with diphenylene iodonium, N-acetyl-cysteine, and menadione specifically blocked SMC polarization and asymmetrical division. In contrast, H2O2 promoted the establishment of SMC polarity and subsequently subsidiary cell formation in "younger" protodermal areas. Surprisingly, H2O2 favored the asymmetrical division of the intervening cells of the stomatal rows leading to the creation of extra apical subsidiary cells. Moreover, H2O2 altered IAA localization, whereas synthetic auxin analogue 1-napthaleneacetic acid enhanced ROS accumulation. Combined treatments with ROS modulators along with 1-napthaleneacetic acid or 2,3,5-triiodobenzoic acid, an auxin efflux inhibitor, confirmed the crosstalk between ROS and auxin functioning during subsidiary cell generation. Collectively, our results demonstrate that ROS are critical partners of auxin during development of Z. mays stomatal complexes. The interplay between auxin and ROS seems to be spatially and temporarily regulated.
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Subsidiary cell generation in Poaceae is an outstanding example of local intercellular stimulation. An inductive stimulus emanates from the guard cell mother cells (GMCs) towards their laterally adjacent subsidiary cell mother cells... more
Subsidiary cell generation in Poaceae is an outstanding example of local intercellular stimulation. An inductive stimulus emanates from the guard cell mother cells (GMCs) towards their laterally adjacent subsidiary cell mother cells (SMCs) and triggers the asymmetrical division of the latter. Indole-3-acetic acid (IAA) immunolocalization in Zea mays protoderm confirmed that the GMCs function as local sources of auxin and revealed that auxin is polarly accumulated between GMCs and SMCs in a timely-dependent manner. Besides, staining techniques showed that reactive oxygen species (ROS) exhibit a closely similar, also time-dependent, pattern of appearance suggesting ROS implication in subsidiary cell formation. This phenomenon was further investigated by using the specific NADPH-oxidase inhibitor diphenylene iodonium, the ROS scavenger N-acetyl-cysteine, menadione which leads to ROS overproduction, and H2O2. Treatments with diphenylene iodonium, N-acetyl-cysteine, and menadione specifically blocked SMC polarization and asymmetrical division. In contrast, H2O2 promoted the establishment of SMC polarity and subsequently subsidiary cell formation in "younger" protodermal areas. Surprisingly, H2O2 favored the asymmetrical division of the intervening cells of the stomatal rows leading to the creation of extra apical subsidiary cells. Moreover, H2O2 altered IAA localization, whereas synthetic auxin analogue 1-napthaleneacetic acid enhanced ROS accumulation. Combined treatments with ROS modulators along with 1-napthaleneacetic acid or 2,3,5-triiodobenzoic acid, an auxin efflux inhibitor, confirmed the crosstalk between ROS and auxin functioning during subsidiary cell generation. Collectively, our results demonstrate that ROS are critical partners of auxin during development of Z. mays stomatal complexes. The interplay between auxin and ROS seems to be spatially and temporarily regulated.
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Microtubule organization and tubulin polymerization in meristematic leaf cells of Chlorophyton comosum treated with an aqueous solution of 1 M mannitol, inducing plasmolysis, were examined with immunofluorescence and transmission electron... more
Microtubule organization and tubulin polymerization in meristematic leaf cells of Chlorophyton comosum treated with an aqueous solution of 1 M mannitol, inducing plasmolysis, were examined with immunofluorescence and transmission electron microscopy.• Hyperosmotic treatment induced disintegration of the interphase microtubule systems. Free tubulin, either liberated from the depolymerized microtubules or pre-existing as a nonassembled pool, was incorporated into a network of paracrystals. In most of the dividing cells, mitotic and cytokinetic microtubule systems were replaced by atypical spindle-like structures displaying bipolarity and atypical phragmoplasts, respectively. These atypical mitotic and cytokinetic structures consisted of large densely packed bundles of macrotubules (32 nm diameter) or macrotubules and paracrystals. Tubulin paracrystals also occurred in ectopic positions in plasmolysed mitotic and cytokinetic cells. Dividing cells displaying paracrystals only did not fo...
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Microtubule organization and tubulin polymerization in meristematic leaf cells of Chlorophyton comosum treated with an aqueous solution of 1 M mannitol, inducing plasmolysis, were examined with immunofluorescence and transmission electron... more
Microtubule organization and tubulin polymerization in meristematic leaf cells of Chlorophyton comosum treated with an aqueous solution of 1 M mannitol, inducing plasmolysis, were examined with immunofluorescence and transmission electron microscopy.• Hyperosmotic treatment induced disintegration of the interphase microtubule systems. Free tubulin, either liberated from the depolymerized microtubules or pre-existing as a nonassembled pool, was incorporated into a network of paracrystals. In most of the dividing cells, mitotic and cytokinetic microtubule systems were replaced by atypical spindle-like structures displaying bipolarity and atypical phragmoplasts, respectively. These atypical mitotic and cytokinetic structures consisted of large densely packed bundles of macrotubules (32 nm diameter) or macrotubules and paracrystals. Tubulin paracrystals also occurred in ectopic positions in plasmolysed mitotic and cytokinetic cells. Dividing cells displaying paracrystals only did not fo...
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The data presented in this work revealed that in Zea mays the exogenously added auxins indole-3-acetic acid (IAA) and 1-napthaleneacetic acid (NAA), promoted the establishment of subsidiary cell mother cell (SMC) polarity and the... more
The data presented in this work revealed that in Zea mays the exogenously added auxins indole-3-acetic acid (IAA) and 1-napthaleneacetic acid (NAA), promoted the establishment of subsidiary cell mother cell (SMC) polarity and the subsequent subsidiary cell formation, while treatment with auxin transport inhibitors 2,3,5-triiodobenzoic acid (TIBA) and 1-napthoxyacetic acid (NOA) specifically blocked SMC polarization and asymmetrical division. Furthermore, in young guard cell mother cells (GMCs) the PIN1 auxin efflux carriers were mainly localized in the transverse GMC faces, while in the advanced GMCs they appeared both in the transverse and the lateral ones adjacent to SMCs. Considering that phosphatidyl-inositol-3-kinase (PI3K) is an active component of auxin signal transduction and that phospholipid signaling contributes in the establishment of polarity, treatments with the specific inhibitor of the PI3K LY294002 were carried out. The presence of LY294002 suppressed polarization o...
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The data presented in this work revealed that in Zea mays the exogenously added auxins indole-3-acetic acid (IAA) and 1-napthaleneacetic acid (NAA), promoted the establishment of subsidiary cell mother cell (SMC) polarity and the... more
The data presented in this work revealed that in Zea mays the exogenously added auxins indole-3-acetic acid (IAA) and 1-napthaleneacetic acid (NAA), promoted the establishment of subsidiary cell mother cell (SMC) polarity and the subsequent subsidiary cell formation, while treatment with auxin transport inhibitors 2,3,5-triiodobenzoic acid (TIBA) and 1-napthoxyacetic acid (NOA) specifically blocked SMC polarization and asymmetrical division. Furthermore, in young guard cell mother cells (GMCs) the PIN1 auxin efflux carriers were mainly localized in the transverse GMC faces, while in the advanced GMCs they appeared both in the transverse and the lateral ones adjacent to SMCs. Considering that phosphatidyl-inositol-3-kinase (PI3K) is an active component of auxin signal transduction and that phospholipid signaling contributes in the establishment of polarity, treatments with the specific inhibitor of the PI3K LY294002 were carried out. The presence of LY294002 suppressed polarization o...
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Double labeling of microtubules and actin filaments revealed that in prophase subsidiary mother cells of Zea mays a monopolar prophase microtubule "half-spindle" is formed, which lines the nuclear hemisphere distal to the... more
Double labeling of microtubules and actin filaments revealed that in prophase subsidiary mother cells of Zea mays a monopolar prophase microtubule "half-spindle" is formed, which lines the nuclear hemisphere distal to the inducing guard mother cell. The nuclear hemisphere proximal to the guard mother cell is lined by an F-actin cap, consisting of a cortical F-actin patch and actin filaments originating from it. The microtubules of the "half-spindle" decline from the nuclear surface and terminate to the preprophase microtubule band. After disintegration of the latter, a bipolar metaphase spindle is organized. The polar F-actin cap persists during mitosis and early cytokinesis, extending to the chromosomes and the subsidiary cell daughter nucleus. In oryzalin treated subsidiary mother cells the prophase nuclei move away from the polar site. Cytochalasin B and latrunculin-B block the polar migration of subsidiary mother cell nuclei, but do not affect those already s...
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The leaf cells of Chlorophytum comosum seem to have the ability to regulate their protoplast volume and shape during the plasmolytic cycle. This phenomenon was morphologically expressed by the stabilization of the plasmolyzed protoplast... more
The leaf cells of Chlorophytum comosum seem to have the ability to regulate their protoplast volume and shape during the plasmolytic cycle. This phenomenon was morphologically expressed by the stabilization of the plasmolyzed protoplast volume and shape within 1-5 min after the immersion of the leaf segments in the plasmolytic fluid and temporarily at the onset of deplasmolysis. During the latter stage the plasmolyzed protoplast rounded up and assumed a perfectly convex shape and glided into the cell lumen along the cell axis. This gliding movement was active, nonsaltatory, and conducted with a constant velocity and lasted for a short time. During this movement the protoplast volume did not change appreciably. As far as we know, this movement has not been described so far. Deplasmolysis proceeded and was rapidly completed when the protoplast stopped moving. Leaf cells which have been affected by an antiactin filament drug or myosin inhibitors lost their ability to regulate the volum...
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The leaf cells of Chlorophytum comosum seem to have the ability to regulate their protoplast volume and shape during the plasmolytic cycle. This phenomenon was morphologically expressed by the stabilization of the plasmolyzed protoplast... more
The leaf cells of Chlorophytum comosum seem to have the ability to regulate their protoplast volume and shape during the plasmolytic cycle. This phenomenon was morphologically expressed by the stabilization of the plasmolyzed protoplast volume and shape within 1-5 min after the immersion of the leaf segments in the plasmolytic fluid and temporarily at the onset of deplasmolysis. During the latter stage the plasmolyzed protoplast rounded up and assumed a perfectly convex shape and glided into the cell lumen along the cell axis. This gliding movement was active, nonsaltatory, and conducted with a constant velocity and lasted for a short time. During this movement the protoplast volume did not change appreciably. As far as we know, this movement has not been described so far. Deplasmolysis proceeded and was rapidly completed when the protoplast stopped moving. Leaf cells which have been affected by an antiactin filament drug or myosin inhibitors lost their ability to regulate the volum...
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ABSTRACT The involvement of actin filaments (AFs) in polarization preceding the development of local outgrowths in protoplast-derived cells (PDCs). as well as in branch initials of Macrocystis pyrifera gametophytes was investigated after... more
ABSTRACT The involvement of actin filaments (AFs) in polarization preceding the development of local outgrowths in protoplast-derived cells (PDCs). as well as in branch initials of Macrocystis pyrifera gametophytes was investigated after AF staining with Rhodamine-Phalloidin (Rh-Ph). A well-developed network of randomly distributed AF arrays was found to traverse the cortical cytoplasm of protoplasts and thallus cells. In some thallus cells, the cortical AFs were largely reorganized and radial AF configurations were formed in the prospective sites of branch emergence. Similar AF configurations were found in PDCs at the stage of formation of rhizoid-like protrusions (RLPs), as well as in young thalli regenerated from protoplasts. The pointed-radial AF formations seemed to change into circular radial structures, from the periphery of which AFs were radiating out. These AF structures were observed at the base of the outgrowths resulting in branch or RLP formation. At later stages the above AFs disappeared and the normal AF network was developed. In parallel, AF caps were found at the tip of the developing RLPs or young branches. The sites of convergence of the radial AFs might be considered as putative AF nucleating or organizing sites, or both. Treatment with Latrunculin B and Jasplakinolide resulted in the inhibition of new RLP formation in regenerating protoplasts, showing that dynamic actin is necessary for polarity establishment and the formation of RLP. The probable mechanism(s) by which the radial AF arrays are involved in polarization of the examined cells, defining the site of cell protrusion(s), are discussed.
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ABSTRACT The involvement of actin filaments (AFs) in polarization preceding the development of local outgrowths in protoplast-derived cells (PDCs). as well as in branch initials of Macrocystis pyrifera gametophytes was investigated after... more
ABSTRACT The involvement of actin filaments (AFs) in polarization preceding the development of local outgrowths in protoplast-derived cells (PDCs). as well as in branch initials of Macrocystis pyrifera gametophytes was investigated after AF staining with Rhodamine-Phalloidin (Rh-Ph). A well-developed network of randomly distributed AF arrays was found to traverse the cortical cytoplasm of protoplasts and thallus cells. In some thallus cells, the cortical AFs were largely reorganized and radial AF configurations were formed in the prospective sites of branch emergence. Similar AF configurations were found in PDCs at the stage of formation of rhizoid-like protrusions (RLPs), as well as in young thalli regenerated from protoplasts. The pointed-radial AF formations seemed to change into circular radial structures, from the periphery of which AFs were radiating out. These AF structures were observed at the base of the outgrowths resulting in branch or RLP formation. At later stages the above AFs disappeared and the normal AF network was developed. In parallel, AF caps were found at the tip of the developing RLPs or young branches. The sites of convergence of the radial AFs might be considered as putative AF nucleating or organizing sites, or both. Treatment with Latrunculin B and Jasplakinolide resulted in the inhibition of new RLP formation in regenerating protoplasts, showing that dynamic actin is necessary for polarity establishment and the formation of RLP. The probable mechanism(s) by which the radial AF arrays are involved in polarization of the examined cells, defining the site of cell protrusion(s), are discussed.
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