Callus proliferation from leaf protoplasts using phenylurea type cytokinin, 4-PU,inBetula platyphylla VAR.Japonica |
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Authors: | Yohichi Wakita Hamako Sasamoto Shinso Yokota Nobuo Yoshizawa |
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Institution: | (1) Forestry and Forest Products, Department of Forest Science, Faculty of Agriculture, Utsunomiya University, Utsunomiya 321, Tochigi, Japan;(2) Forestry and Forest Products Research Institute, Ministry of Agriculture, Forestry and Fisheries, Tsukuba 305, P.O. Box 16, Ibaraki, Japan |
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Abstract: | Protoplasts were isolated from leaves ofBetula platyphylla var.japonica using a 0.6M mannitol solution containing 1% Cellulase Onozuka R-10 and 1% Driselase. The cell division and colony formation were largely
enhanced using Murashige and Skoog (1962) liquid medium at half strength (1/2 MS), containing 0.6M mannitol, 0.09M sucrose, and factorial combinations of 0.1–30 μM N-(2-chloro-4-pyridyl)-N′-phenylurea (4-pu) and 0.1–10 μM 1-naphthaleneacetic acid (NAA) or 0.1–30 μM 2,4-dichlorophenoxyacetic acid (2,4-D). The optimal protoplast density was 5–7 × 104/ml. Continuous callus proliferation from protoplasts was achieved by transferring colonies to fresh 1/2 MS agar medium containing
1 μM NAA and 1 μM 4-pu with no mannitol. It appeared that supplementation of the medium with phenylurea type cytokinin, 4-pu gave the successful
callus proliferation from the protoplasts ofB. platyphylla. |
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Keywords: | Betula platyphylla var japonica callus 4-pu protoplasts |
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