Different effects of four salts and pHs on protoplast cultures of a mangrove, Bruguiera sexangula suspension cells, Populus alba leaves and tobacco BY-2 cells

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  • FUKUMOTO Takeshi
    Faculty of Environment and Information Sciences, Yokohama National University
  • NAKAMURA Tatsuo
    Faculty of Environment and Information Sciences, Yokohama National University
  • SUZUKI Masashi
    Faculty of Environment and Information Sciences, Yokohama National University
  • OGITA Shinjiro
    Biotechnology Research Center, Faculty of Engineering, Toyama Prefectural University
  • MIMURA Tetsuro
    Department of Biology, Faculty of Science, Kobe University
  • SASAMOTO Hamako
    Faculty of Environment and Information Sciences, Yokohama National University

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Effects of a wide range of concentrations of four salts (NaCl, KCl, MgCl2, CaCl2) and different pHs (pH 4.5, 5.0, 5.7, 6.2) on colony formation, were investigated in protoplast culture of mangrove, Bruguiera sexangula, suspension cells. The results were compared to those obtained with protoplasts of tobacco BY-2 cells and leaf protoplasts of Populus alba. A 96-multiwell culture plate method was used and only 250 to 2500 protoplasts in 55 μl medium were sufficient to test the effect of each treatment. The effects of four salts were always found to inhibit colony formation of Populus and tobacco protoplasts at pH 5.7. In contrast, MgCl2 and CaCl2 stimulated further development of B. sexangula protoplasts, while KCl had an inhibitory effect. The highest number of colony formation was obtained at pH 5.7 without salt additions. However, at lower pHs, some stimulatory effects were found at low concentrations of all four salts tested. Depending on the pHs, different patterns of stimulation and inhibition of four salts were obtained in B. sexangula protoplasts.

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