| 番茄果实ABA合成酶NCED基因的克隆及其RNAi遗传转化体系的建立 |
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| 引用本文： | 王玲,孙亮,冷平,王永军,徐丽哲.番茄果实ABA合成酶NCED基因的克隆及其RNAi遗传转化体系的建立[J].中国农业大学学报,2009,14(5). |
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| 作者姓名： | 王玲 孙亮 冷平 王永军 徐丽哲 |
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| 作者单位： | 1. 中国农业大学,农学与生物技术学院,北京,100193
2. 北京市平谷区,科学技术委员会,北京,100035 |
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| 基金项目： | 北京市科委重大资助项目(D0706002000091) |
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| 摘 要： | 以番茄为材料,根据ABA合成关键酶编码基因NCED的保守序列设计引物,通过RT-PCR方法从番茄果实中克隆了2个NCED基因片段,Le NCED1和Le NCED2。通过农杆菌介导法进一步建立了‘嘉宝’番茄Le NCED1基因的RNAi再生和转化体系,并研究影响番茄遗传转化的4个因素外植体类型、预培养时间、激素组合、农杆菌侵染时间。结果表明:番茄子叶预培养2 d,用农杆菌LBA4404侵染5 min后,避光共培养2 d,在加入1mg/L 6-BA+0.05 mg/LIAA+5 mg/L潮霉素的MS培养基上可以诱导出芽,转入添加0.1 mg/LIAA+7 mg/L潮霉素的1/2 MS培养基上可以使芽生根。本实验共获得26株抗性植株。GUS染色证明,有9株抗性植株成功转入含有NCED基因RNAi载体的T-DNA,为深入了解ABA促进番茄果实成熟的机理提供了方法和材料。
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| 关 键 词： | 番茄 NCED基因 ABA 再生 转化 RNAi |
Cloning of ABA biosynthesis key enzyme NCED gene from tomato fruit and its RNA interference genetic transformation |
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| WANG Ling,SUN Liang,LENG Ping,WANG Yong-jun,XU Li-zhe.Cloning of ABA biosynthesis key enzyme NCED gene from tomato fruit and its RNA interference genetic transformation[J].Journal of China Agricultural University,2009,14(5). |
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| Authors: | WANG Ling SUN Liang LENG Ping WANG Yong-jun XU Li-zhe |
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| Institution: | WANG Ling1,SUN Liang1,LENG Ping1,WANG Yong-jun2,XU Li-zhe2(1.College of Agronomy , Biotechnology,China Agricultural University,Beijing 100193,China,2.Municipal Science , Technology Commission in Pinggu,Beijing 100035,China) |
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| Abstract: | We designed primers based on the conserved sequence of ABA biosynthesis key enzyme gene,NCED and cloned two NCED gene fragments(LeNCED1 and LeNCED2) from tomato fruit by RT-PCR.The gene,LeNCED1 RNAi construct was introduced into 'jiabao' tomato plant via Agrobacterium tumefaciens-mediated method: four factors affecting tomato genetic transformation,explant type,preculturing time,hormone combination and A.tumefaciens dipping time,were examined.The result suggested that cotyledons precultured on medium(MS sal... |
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| Keywords: | tomato NCED gene ABA regeneration transformation RNAi |
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