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Flow cytometry of the differentiation of Physarum polycephalum myxamoebae to cysts
Authors:J Fry  H R Matthews
Institution:1. CNR-IMEM (National Council of the Researches — Institute of Materials for Electronics and Magnetism), Parco Area delle Scienze 37A, 43124 Parma, Italy;2. Department of Physics and Earth Science, University of Parma, Viale Usberti 7A, 43124 Parma, Italy
Abstract:Myxamoebae of Physarum polycephalum, strain Cld, were grown on agar lawns on live bacteria. Myxamoebae were harvested, fixed and stained with propidium iodide. Flow cytometry showed that, as in the case of Physarum plasmodia, there is no G1 phase during rapid exponential growth. However, an apparent G1 phase was observed at the end of exponential growth when the culture arrested with the G1 DNA content for about a day between growth and differentiation. Most myxamoebae differentiated into cysts, but some formed microplasmodia and others appeared to lose DNA. The cysts possessed the G2 phase DNA content and there was an S phase connecting the G1-arrested state with the encysted state. Encystment was blocked by hydroxyurea (HU) suggesting that DNA synthesis is essential for encystment. The natural temporary synchronization in G1 phase may provide the basis of a method for selecting mutants with a conditional block in G2 or M phases.
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