Changes in growth patterns and intracellular calcium concentrations in Aspergillus awamori treated with amphotericin B
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Growth patterns and intracellular Ca2+ concentrations in the mutant strain Aspergillus awamori 66A containing a recombinant aequorin gene were studied in the presence of a permeabilizing fungicidal agent amphotericin B. The cell response, i.e., changes in the growth and development of the fungus (initiation of spore germination, mycelial growth, and intensity of sporulation) was dose-dependent. Low concentrations of amphotericin B (2.5 μM) stimulated spore germination: the number of germinating spores was 2–3 times higher than in the control (without the fungicide). At higher amphotericin concentrations (20 μM) spore germination was inhibited. Amphotericin B had a dose-dependent effect on mycelial growth and sporulation intensity on solid Vogel medium. Intracellular Ca2+ concentrations in the presence of amphotericin B were investigated using the luminescence of the photoprotein aequorin. High concentrations of amphotericin B (10 and 20 μM) were shown to cause an instantaneous increase in Ca2+ concentrations compared to the control and lower amphotericin concentration (2.5 μM). Ca2+ concentrations remained elevated throughout the experiment and correlated with the inhibition of mycelial growth and development.
KeywordsAspergillus awamori recombinant aequorin permeabilizing fungicide amphotericin B cell response sporulation Ca2+ signaling Ca2+ dynamics
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