Interfamilial cell fusion among leaf protoplasts of Populus alba, Betula platyphylla and Alnus firma: assessment of electric treatment and invitro culture conditions |
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| Authors: | Yohichi Wakita Shinso Yokota Nobuo Yoshizawa Toshio Katsuki Yoshihiko Nishiyama Toshitaka Yokoyama Mitsue Fukui and Hamako Sasamoto |
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| Institution: | (1) Greenery Research and Information Center, Hokkaido Forestry Research Institute, 079-0198 Hokkaido, Bibai, Japan;(2) Faculty of Agriculture, Department of Forest Science, Utsunomiya University, 321-8505 Tochigi, Utsunomiya, Japan;(3) Forestry and Forest Products Research Institute, 305-8687 Ibaraki, Tsukuba, Japan;(4) Faculty of Environment and Information Sciences, Yokohama National University, 240-8501 Yokohama, Japan |
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| Abstract: | The optimal conditions for fusion of leaf protoplasts of Populus alba, Betula platyphylla, and Alnus firma by electric treatment were alternate current (AC) 200 V cm−1 in 2.5 mM CaCl2 for a pearl chain formation and direct current (DC) pulse of 100 μs at 2 kV cm−1 After interfamilial cell fusion treatment, colonies were obtained using liquid media containing 2,4-D or NAA as an auxin and BA or CPPU as a cytokinin at 0.1, 1, or 10 µM in MS (Murashige and Skoog (1962) Physiol. Plant. 15: 473--497), 1/2salt MS, or NH4NO3-free MS containing 0.6 M mannitol and 3% sucrose (totaling 147 combinations). Two shoots after electric cell fusion treatment between P. alba and B. platyphylla, and 12 regenerated plants after electric cell fusion between P. alba and A. firma, were obtained from colonies induced on agar medium containing NAA, IBA, CPPU, and BA. Seven lines of the latter 12 plants which were regenerated later cultured in vitro had serrated leaves different from those of P. alba. |
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| Keywords: | Betulaceae interfamilial cell fusion protoplast Salicaceae somatic hybrid |
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