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  • 1
    Electronic Resource
    Electronic Resource
    Overexpression of the Ca2+-binding protein calreticulin in the endoplasmic reticulum improves growth of tobacco cell suspensions (Nicotiana tabacum) in high-Ca2+ medium (2005)
    Oxford, UK; Malden, USA : Munksgaard International Publishers
    Physiologia plantarum 123 (2005), S. 0 
    Details
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Calreticulin (CRT) is a eukaryotic, highly conserved, Ca2+-binding protein predominantly located in the endoplasmic reticulum (ER) lumen. In addition to being involved in the regulation of cellular Ca2+, calreticulin is a key quality control element during protein folding in the ER lumen. Tobacco (Nicotiana tabacum L.) suspension cells overexpressing a maize CRT (CRT1a) were used here to examine the properties of CRT in growing plant cells with respect to stress exposure. The endogenous CRT gene was induced rapidly after subculturing of the cells to new medium. In accordance, the CRT protein levels increased, peaking at days 3–4. At day 5, when the CRT transcript levels had levelled off, a further increase in endogenous CRT expression was obtained when the cells were treated with excess Ca2+ or the N-linked glycosylation inhibitor tunicamycin. Whereas the response to Ca2+ occurred within 30 min, the induction by tunicamycin took several hours to be established. Transforming tobacco cells with maize CRT1a, under a constitutive mannopine synthase promoter, resulted in a stable level of expressed CRT1a during the growth cycle compared with endogenous CRT. The CRTs showed differences in attached glycans, but both contained the high mannose-rich-type glycans characteristic of ER proteins. In agreement with an ER location, both tobacco CRT and the transgene product CRT1a codistributed with the ER marker NADH cytochrome c reductase after density gradient centrifugation of microsomal fractions from tobacco cells. Increased production of CRT, as was obtained in the transgenic tobacco cell lines, made cells more tolerant than wild-type cells to high Ca2+ during growth. These data suggest that overexpression of CRT1a in plant cells results in a more efficient calcium buffering capacity in the ER.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1399-3054.2004.00434.x
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  • 2
    Electronic Resource
    Electronic Resource
    The effect of fluorescent labeling on calcium-induced fusion of fusogenic carrot protoplasts (1986)
    Springer
    Plant cell reports 5 (1986), S. 252-255 
    Details
    ISSN: 1432-203X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Various fluorescent compounds — carboxyfluorescein, scopoletin, fluorescein isothiocyanate (FITC), rhodamine B isothiocyanate (RITC), rhodamine 123, and rhodamine B ethyl ester — were used to study their effects on calcium-induced fusion of fusogenic carrot (Daucus carota L.) protoplasts. These protoplasts normally fused at a high percentage (50–60%) in response to 10 mM calcium, pH 6.0; however, if cells had been labeled with scopoletin, FITC, or RITC, fusion was greatly reduced. In contrast, labeling with carboxyfluorescein, rhodamine 123, or rhodamine B ethyl ester had no detectable effect on calcium-induced fusion. The two rhodamine dyes are shown to be localized in mitochondria.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00269814
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  • 3
    Electronic Resource
    Electronic Resource
    The polyphosphoinositides, phosphatidylinositol monophosphate and phosphatidylinositol bisphosphate, are present in nuclei isolated from carrot protoplast (1989)
    Springer
    Protoplasma 151 (1989), S. 62-72 
    Details
    ISSN: 1615-6102
    Keywords: Phosphatidylinositol ; Lysophosphatidylinositol ; Phosphoinositides ; Nuclei ; Membrane
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Nuclei were isolated from carrot protoplasts and the distribution of [3H]inositol-labeled phospholipids was analyzed by thinlayer chromatography. Phosphatidylinositol (PI), lysophos-phatidylinositol (LPI), phosphatidylinositol monophosphate (PIP), lysophosphatidylinositol monophosphate (LPIP), and phosphatidylinositol bisphosphate (PIP2) were 55.7%, 12.3%, 5.0%, 11.5%, and 3.6% of the respective [3H]inositol-labeled lipids recovered from the nuclear fraction. While both the plasma membrane and nuclear fraction contained polyphosphoinositides, the distribution of the phosphoinositides and the amount of inositol-labeled lipid were distinct. For example, the nuclear fraction had a higher percentage of LPI and PIP2 and less PI and LPIP than the plasma membrane fraction. The amount of [3H]inositol-labeled lipid recovered from the nuclear fraction per mg protein was an order of magnitude lower than that recovered from either the plasma membrane of lower phase fraction isolated by aqueous two-phase partitioning, or from whole cells and protoplasts. In addition, when the ratio of the [3H]inositol-labeled lipid was compared to total [14C]myristate-labeled lipid recovered there was three to ten fold less [3H] relative to [14C] in the nuclear fraction. These data indicate that while the polyphosphoinositides are a relatively high percentage of the inositol lipid in the nuclear fraction, the inositol lipid was only a small portion of the total lipid in the nuclei. Despite this low concentration of inositol lipid, when [γ 32P]-ATP was added to the isolated nuclei,32P-labeled PIP and PIP2 were synthesized. Thus, the carrot nuclei contained PI and PIP kinase as well as the polyphosphoinositides.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01403302
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  • 4
    Electronic Resource
    Electronic Resource
    Handbook of plant cell culture. Volume 1: Techniques for propagation and breeding. Edited by D. A. Evans, W. R. Sharp, P. V. Ammirato, and Y. Yamada. New York: Macmillan, 1983. 970 pp. $49.50 (1984)
    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 5 (1984), S. 59-61 
    Details
    ISSN: 0192-253X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/dvg.1020050105
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