| 根癌农杆菌介导寒富苹果转化体系的建立 |
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| 引用本文： | 杨越,张志宏,刘月学,何平,周宇,代红艳,赵桂玲. 根癌农杆菌介导寒富苹果转化体系的建立[J]. 果树学报, 2010, 0(2) |
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| 作者姓名： | 杨越 张志宏 刘月学 何平 周宇 代红艳 赵桂玲 |
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| 作者单位： | 沈阳农业大学园艺学院; |
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| 基金项目： | 辽宁省高等学校创新团队项目(2009T089) |
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| 摘 要： | 以寒富苹果组培苗叶片为外植体,利用植物表达载体上含潮霉素抗性基因的根癌农杆菌EHA105(pCAMBI-A1301)和含卡那霉素抗性基因的根癌农杆菌EHA105(pCAMBIA2301)对影响寒富遗传转化效率的因素进行系统研究。结果表明,寒富叶片对潮霉素反应敏感,在附加潮霉素的培养基上寒富叶片褐化较为严重。潮霉素和卡那霉素适宜的筛选质量浓度分别为4 mg.L-1和25 mg.L-1。农杆菌介导寒富苹果转化体系的建立以MS+TDZ 2.0 mg.L-1+NAA 0.5 mg.L-1培养基为叶片离体再生芽培养基。适宜的转化条件为:菌液浓度D 600nm=0.5、叶片外植体在菌液浸泡8 min、共培养时间为3～4 d、推迟4 d进行筛选培养。抗性基因对转化效率具有明显的影响,EHA105(pCAMBIA2301)的平均抗性芽再生率(0.96%)比EHA105(pCAMBIA1301)的平均抗性芽再生率(0.58%)高出几乎50%,EHA105(pCAMBIA2301)的抗性芽再生率最高达1.87%。GUS组织化学染色和PCR鉴定结果表明本研究获得了寒富苹果转基因植株。
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| 关 键 词： | 苹果 寒富 农杆菌 抗性基因 遗传转化 |
Establishment of Agrobacterium tumefaciens-mediated transformation system for Hanfu apple |
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| YANG Yue,ZHANG Zhi-hong,LIU Yue-xue,HE Ping,ZHOU Yu,DAI Hong-yan,ZHAO Gui-ling. Establishment of Agrobacterium tumefaciens-mediated transformation system for Hanfu apple[J]. Journal of Fruit Science, 2010, 0(2) |
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| Authors: | YANG Yue ZHANG Zhi-hong LIU Yue-xue HE Ping ZHOU Yu DAI Hong-yan ZHAO Gui-ling |
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| Affiliation: | College of Horticulture;Shenyang Agricultural University;Shenyang;Liaoning 110866 China |
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| Abstract: | Several factors that affect the efficiency of the Agrobacterium tumefaciens-mediated transformation of Malus do-mestica cv.Hanfu were studied using leaves of in vitro plants and Agrobacterium strain EHA105 containing a binary vector pCAMBIA1301 inserted the hygromycin(Hyg) resistance gene or a binary vector pCAMBIA2301 inserted kanamycin(Kan) resistance gene.The result showed that the leaves of Hanfu apple cultivar browned seriously in medium with Hyg which indi-cated that they are sensitive to Hyg.The suit... |
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| Keywords: | Apple Hanfu Agrobacterium Resistant gene Genetic transformation |
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