Callus regeneration from cotyledon protoplasts ofChamaecyparis obtusa (Hinoki cypress) |
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Authors: | Hamako Sasamoto Akira Kondo Yoshihisa Hosoi Hisae Maki Keiji Odani |
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Affiliation: | (1) Forestry and Forest Products Research Institute, Ministry of Agriculture, Forestry and Fisheries, 305 Tsukuba, Ibaraki, Japan;(2) Forestry and Forest Products Research Institute, 434 Shizuoka Prefecture, Hamakita, Shizuoka, Japan |
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Abstract: | Summary Protoplasts were isolated from cotyledons of 1- to 1.5-mo.-old seedlings ofChamaecyparis obtusa using 1% driselase or 0.25% pectolyase Y-23 in combination with 1% cellulase RS in 0.6M mannitol solution. Cell division and colony formation were induced efficiently in liquid Murashige and Skoog’s (MS) medium containing 0.6M mannitol and 10 to 30 μM 2,4-dichlorophenoxyacetic acid or 1 μM of naphthaleneacegic acid at the cell density of 1 to 2×103 ml. Continued callus proliferations was observed by transferring tissue to fresh medium of the same composition as the induction medium without mannitol. Campbell and Durzan’s medium and ammonium nitrate-free MS medium were less effective than MS medium. High concentration of benzyladenine (1 or 10 μM) was inhibitory to cell division. |
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Keywords: | protoplast culture callus regeneration conifers |
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