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  • 1
    ISSN: 1365-3040
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: We show here that, within 1–2 min of application, systemin triggers a transient increase of cytoplasmic free calcium concentration ([Ca2+]c) in cells from Lycopersicon esculentum mesophyll. The systemin-induced Ca2+ increase was slightly but not significantly reduced by L-type Ca2+ channel blockers (nifedipine, verapamil and diltiazem) and the Ca2+ chelator [ethylene glycol tetraacetic acid (EGTA)], whereas inorganic Ca2+ channel blockers (LaCl3, CdCl2 and GdCl3) and compounds affecting the release of intracellular Ca2+ from the vacuole (ruthenium red, LiCl, neomycin) strongly reduced the systemin-induced [Ca2+]c increase. By contrast, no inhibitory effect was seen with the potassium and chloride channel blockers tested. Unlike systemin, other inducers of proteinase inhibitor (PI) and of wound-induced protein synthesis, such as jasmonic acid (JA) and bestatin, did not trigger an increase of cytoplasmic Ca2+. The systemin-induced elevation of cytoplasmic Ca2+ which might be an early step in the systemin signalling pathway, appears to involve an influx of extracellular Ca2+ simultaneously through several types of Ca2+ permeable channels, and a release of Ca2+ from intracellular stores sensitive to blockers of inositol 1,4,5-triphosphate (IP3)- and cyclic adenasine 5’-diphosphoribose (cADPR)-mediated Ca2+ release.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Calcium signalling is little understood in filamentous fungi largely because easy and routine methods for calcium measurement in living hyphae have previously been unavailable. We have developed the recombinant aequorin method for this purpose. High levels of aequorin expression were obtained in Neurospora crassa, Aspergillus niger and Aspergillus awamori by codon optimization of the aequorin gene. Three external stimuli (mechanical perturbation, hypo-osmotic shock and high external calcium) were found transiently to increase [Ca2+]c. Each of the calcium signatures associated with these physico-chemical treatments was unique, suggesting the involvement of three distinct calcium-mediated signal transduction pathways. The fungal calcium channel blocker KP4 inhibited the [Ca2+]c responses to hypo-osmotic shock and high external calcium, but not to mechanical perturbation. The divalent cation chelator BAPTA inhibited [Ca2+]c responses to mechanical perturbation and hypo-osmotic shock. The calcium agonists A23187 and cyclopiazonic acid increased [Ca2+]c levels.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1573-9368
    Keywords: calcium ; moss ; aequorin ; mutant ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The gene for apoaequorin has been used previously to indicate cytosolic calcium changes in higher plants. Here we report the transformation of the mossPhyscomitrella patens with the cDNA for apoaequorin. Stable transformants were obtained in the wild type which reconstitute the calcium-sensitive luminescent protein aequorinin vivo after incubation in coelenterazine, and continue to grow normally. The wild type responds to cold-shock (0–10°C) with increases in cytosolic calcium. Mechanical perturbation, in the form of touch, also induces transient increases in cytosolic calcium. A smaller response to pH, distinct from the touch response and exhibiting different kinetics, can also be detected.
    Type of Medium: Electronic Resource
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