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  • 1
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Plant, cell & environment 17 (1994), S. 0 
    ISSN: 1365-3040
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: During plasmolysis of onion epidermal cells, the contracting protoplast remains connected to the cell wall by an intricate, branched system of plasma membrane (PM) ‘Hechtian strands’ which stain strongly with the fluorescent probe DiOC6. In addition, extensive regions of the cortical endoplasmic reticulum (ER) network remain anchored to the cell wall during plasmolysis and do not become incorporated into the contracting protoplast with the other cell organelles. These ER profiles become tightly encased by the PM as the latter contracts towards the centre of the cell. Thus, although the cortical ER is left outside the main protoplast body, it is nonetheless still bound by the PM of the cell. As well as being anchored to the wall, the cortical ER remains intimately linked with plasmodesmata and retains continuity between cells via the central desmotubules which become distended during plasmolysis. The PM also remains in close contact with the plasmodesmatal pore following plasmolysis. It is suggested that plasmodesmata, although sealed, may not be broken during plasmolysis, their substructure being preserved by continuity of both ER and PM through the plasmodesmatal pore. A structural model is presented which links the behaviour of PM, ER and plasmodesmata during plasmolysis.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Plant, cell & environment 10 (1987), S. 0 
    ISSN: 1365-3040
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Abstract An experimental system has been developed for studying efflux of 14C assimilates in growing potato tubers. Small wells are cut into the phloem-rich perimedulla and filled with trap solutions of varying composition which inhibit or promote assimilate efflux. One well on each tuber acts as the treatment while a second well acts as the control. Movement of 14C into wells occurred at comparable rates to that found in intact tissue, harvested from importing tubers in the form of microcores. Sucrose was the predominant translocated sugar in the stolon and was not hydrolysed in either the wells or the microcores following unloading. Efflux into wells containing agar traps was stimulated 40-fold relative to buffer controls by the addition of 20 mol m−3 EGTA to the agar. This was interpreted as passive efflux to the apoplast due to increased membrane permeability in the pathway between the sieve elements and the collecting wells. The EGTA stimulation was reversed by addition of Ca2+. 14C efflux into buffered solutions was inhibited significantly by both DNP and PCMBS, suggesting the involvement of active and carrier-mediated transport components. However, it was not possible to determine whether these compounds acted at the site of unloading only, or on the short-distance transfer step between phloem and collecting wells. The rate of tracer efflux was not significantly different when 1 mol m−3 and 300 mol m−3 sucrose were applied to the wells, indicating insensitivity of solute movement to low apoplastic solute concentrations. However, raising the solute concentration to 800 mol m−3 caused a severe inhibition of tracer efflux. These results were duplicated with mannitol as the osmoticum. It is suggested that plasmolysis prevented further efflux by disruption of a predominantly symplastic transport pathway between the phloem and collecting wells.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Planta 180 (1990), S. 555-561 
    ISSN: 1432-2048
    Keywords: Allium ; Apoplast ; Endocytosis (fluidphase) ; Epidermis ; Plasmolysis/deplasmolysis ; Vesicle (endocytic)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A transient plasmolysis/deplasmolysis (plasmolytic cycle) of onion epidermal cells has been shown to induce the formation of fluid-phase endocytic vesicles. Plasmolysis in the presence of the membrane-impermeant fluorescent probes Lucifer Yellow CH (LYCH) and Cascade Blue hydrazide resulted in the uptake of these probes by fluid-phase endocytosis. Following deplasmolysis, many of the dye-containing vesicles left their parietal positions within the cell and underwent vigorous streaming in the cytoplasm. Vesicles were observed to move within transvacuolar strands and their movements were recorded over several hours by video-microscopy. Within 2 h of deplasmolysis several of the larger endocytic vesicles had clustered around the nuclear membrane, apparently lodged in the narrow zone of cytoplams surrounding the nucleus. In further experiments LYCH was endocytically loaded into the cells during the first plasmolytic cycle and Cascade Blue subsequently loaded during a second plasmolytic cycle. This resulted in the introduction of two populations of endocytic vesicles into the cells, each containing a different probe. Both sets of vesicles underwent cytoplasmic streaming. The data are discussed in the light of previous observations of fluid-phase endocytosis in plant cells.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1432-2048
    Keywords: Apoplast ; Endocytosis ; Solanum (tuber, transport) ; Symplast ; Tuber-vacuole
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The fluorescent dye Lucifer Yellow CH (LYCH) was introduced directly into the symplast of potato (Solanum tuberosum L.) tuber storage parenchyma by microinjection and also into the apoplast through cuts made in the stolon cortex. Microinjected LYCH moved away rapidly from a single storage cell and spread radially via the symplast. When the microinjected tissue was subsequently fixed in glutaraldehyde and sectioned the dye was seen clearly to be localised in the cytoplasm but not in the vacuole. In comparison, when LYCH was introduced into cuts made in the stolon cortex the dye entered the tuber by the xylem and subsequently spread apoplastically. No movement of dye was observed in the phloem. In glutaraldehyde-fixed tissues, in which LYCH was introduced to the apoplast, the dye was found within xylem vessels, in the cell walls and in intercellular spaces. Wall regions, possibly associated with plasmodesmata, became stained by the dye as it moved through the apoplast. Three hours after introduction of the dye to the stolon, intense deposits of LYCH were found in the vacuoles of all cells in the tuber, many aligned along the tonoplast. Differentiating vascular parenchyma elements contained large amounts of dye within enlarging vacuoles. However, with the exception of plasmolysed and-or damaged cells, LYCH was absent from the cytoplasm following its introduction to the plasmalemma it is suggested that the most likely pathway from the cell wall to the vacuole was by endocytosis, the dye being transported across the cytoplasm in membrane-bound vesicles. Clathrin-coated vesicles were abundant in the storage cells, providing a possible endocytotic pathway for dye movement. The significance of these observations is discussed in relation to the movement of LYCH in plant tissues and to the movement of solutes within and between storage cells of the tuber.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1432-2048
    Keywords: Enzyme activity (potato tuber) ; Sink isolation (sugar uptake) ; Solatium (tuber) ; Starch synthesis ; Sugar transport
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Import into potato (Solarium tuberosum L. cv. Record) tubers was terminated by removing the sink at its connection with the stolon. The ability of discs of storage tissue from the excised tubers to take up exogenous sugars and convert them to starch was compared with that of discs from untreated tubers from the same plant population. In rapidly-growing control tubers, glucose and fructose were taken up to a greater extent than sucrose, 77% of the glucose being converted to starch within 3 h (compared with 64% and 27% for fructose and sucrose, respectively). These values fell as the tubers aged but the ranking (glucose 〉 fructose 〉 sucrose) was maintained, emphasising a severe rate-limiting step following the import of sucrose into the growing tuber. Sink isolation had little effect on the ability of the storage cells to take up exogenous sucrose across the plasmalemma for up to 7 d after sink isolation. However, the ability of the same cells to convert the sucrose to starch was severely inhibited within 24 h, as was the sensitivity of starch synthesis to turgor. In the case of glucose, sink isolation inhibited both the uptake and the conversion to starch, the latter being inhibited to a greater degree. A detailed metabolic study of tubers 7 d after excision showed that, with sucrose as substrate, 94% of the radioactivity in the soluble sugar pool was recovered in sucrose following sink isolation (92% in control tubers). However, with glucose as substrate, 80% of the radioactivity was recovered as sucrose following tuber excision (28% in control tubers), providing evidence that sucrose synthesis acts as a major alternative carbon sink when starch synthesis is inhibited. In the same tubers, sucrose-synthase activity decreased by 70% following sink isolation, compared with a 45% reduction in ADP-glucose pyrophosphorylase. Activities of UDP-glucose pyrophosphorylase, starch phosphorylase, starch synthase nd both PPi- and ATP-dependent phosphofructokinases remained unchanged. Acid-invertase activity increased fivefold.
    Type of Medium: Electronic Resource
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  • 6
    ISSN: 1432-2048
    Keywords: Endocytosis ; Hordeum (endocytosis) ; Lucifer Yellow (uptake, root) ; Root (endocytosis) ; Vacuole (dye uptake)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Intact barley (Hordeum vulgare L.) roots have been shown to take up the highly fluorescent dye Lucifer Yellow CH (LYCH) into their cell vacuoles. In the apical 1 cm of root tip, differentiating and dividing cells showed a prolific uptake of LYCH into their provacuoles. The LYCH was retained during fixation, apparently becoming bound to electron-dense material in the vacuoles. The dye freely entered the apoplast of roots in which the Casparian band was not developed, being taken up into the vacuoles of cells in both the cortex and stele. However, when LYCH was applied to a 1-cm zone approx. 6 cm behind the root tip the Casparian band on the radial walls of the endodermis completely prevented the dye from entering the cells of the stele, only the cell walls and vacuoles of the cortical cells taking up the dye. The inability of LYCH to cross the plasmalemma of the endodermal cells and enter the stele via the symplast substantiates previous claims that the dye is unable to cross the plasmalemma of plant cells. The results are discussed in the light of recent demonstrations that LYCH is a particularly effective marker for fluid-phase endocytosis in animal and yeast cells. A calculation of the energetic requirements for LYCH uptake into barley roots supports the contention that LYCH is taken up into the vacuoles of plant cells by fluid-phase endocytosis.
    Type of Medium: Electronic Resource
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  • 7
    ISSN: 1615-6102
    Keywords: Green fluorescent protein ; Virus vectors ; Coat protein ; Fusion proteins ; Potato virus X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The green fluorescent protein (GFP) was introduced into plant cells using potato virus X as a vector. The GFP was produced at high levels within virus-infected cells by utilising a duplication of the viral coat protein subgenomic RNA promoter sequence to direct transcription of mRNA encoding the GFP. We also exploited the ability of GFP to retain its fluorescence when fused to other proteins by fusing it to the PVX coat protein. The resultant fluorescent virus became systemic and its movement from cell to cell was traced using confocal laser scanning microscopy. Using PVX as the vector, additional fusions of the GFP were made to the movement protein of tobacco mosaic virus (TMV). The fluorescent fusion protein produced was targeted to specific wall sites thought to be plasmodesmatal pit fields. The utility of virus-based vectors for the delivery and targeting of GFP in living plant cells is discussed.
    Type of Medium: Electronic Resource
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