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作 者:易自力 Wang Li Cao Shouyun Yuan Longping Guan Chunyun Zhau Puhan Tian Wenzhang Chu Chengcai
机构地区:[1]Hunan Agricultural University, Changsha 410128,P.R.China Institute of Genetics, Chinese Academy of Sciences, Beijing 100101, P.R.China [2]Institute of Genetics, Chinese Academy of Sciences, Beijing 100101, P.R.China [3]Hunan Agricultural University, Changsha 410128,P.R.China
出 处:《High Technology Letters》2003年第1期6-10,共5页高技术通讯(英文版)
基 金:the Chinese 863 High-Technology Program(NO.863-101-01-01-01)and Chinese Post-Doctoral Science Foundation(2001-14)
摘 要:In order to improve the frequency of indica rice transformation by biolistic bombardment, suitable culture conditions for embryonic calli,an optimal selection scheme for resistant calli and seedlings, and optimum bombardment parameters a investigated by using 14 commercially important indica rice cultivars. The main results show that the CC medium with 36g/L mannitol is a scheme subculture medium in which the browning of indica rice calli can be mitigated significantly; The concentration of 30~40mg/L Hyg or 150~200mg/L G418 or 10~20 mg/L Basta is suitable for selection of resistant calli; The transformation parameters of 100μg gold powder absorbing 0.2μg DNA per shot and 900 psi helium pressure and 6 cm bombardment distance and bombarded twice for each plate give the best result; Keeping the target calli on osmotic medium containing 60g/L mannitol from 12 ~24h before bombardment to 24~48h after it can increase the efficiencies of transformation . Furthermore, some transgenic indica rice plants are obtained using this optimized transformation system.In order to improve the frequency of indica rice transformation by biolistic bombardment, suitable culture conditions for embryonic calli, an optimal selection scheme for resistant calli and seedlings, and optimum bombardment parameters a investigated by using 14 commercially impor-tant indica rice cultivars. The main results show that the CC medium with 36g/L mannitol is a scheme subculture medium in which the browning of indica rice calli can be mitigated significant-ly; The concentration of 30-40mg/L Hyg or 150-200mg/L G418 or 10-20 mg/L Basta is suitable for selection of resistant calli; The transformation parameters of 100μg gold powder ab-sorbing 0. 2μg DNA per shot and 900 psi helium pressure and 6 cm bombardment distance and bombarded twice for each plate give the best result; Keeping the target calli on osmotic medium containing 60g/L mannitol from 12 - 24h before bombardment to 24 - 48h after it can increase the efficiencies of transformation . Furthermore, some transgenic indica rice plants are obtained using this optimized transformation system.
关 键 词:indica rice biolistic bombardment TRANSFORMATION CALLUS
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