| 代玉华;王锡锋;李莉;周广和.一种适合水稻农杆菌转化的RNAi载体的构建和潮霉素对水稻转化的影响[J].植物保护,2007,33(2):37-40. |
| 一种适合水稻农杆菌转化的RNAi载体的构建和潮霉素对水稻转化的影响 |
| Construction of a RNAi vector and effects of hygromycin B (Hyg) on rice transformation |
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| DOI： |
| 中文关键词: 水稻条纹病毒 载体构建 潮霉素 |
| 英文关键词:Rice stripe virus(RSV) vector construction hygromycin B(Hyg) |
| 基金项目: |
| 代玉华;王锡锋;李莉;周广和 |
| 中国农业科学院植物保护研究所植物病虫害生物学国家重点实验室 北京100094 |
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| 中文摘要: |
| 以感染水稻条纹病毒(RSV)水稻叶片的总RNA为模板,利用RT-PCR扩增出了RSV的CP基因。分别将正反向的CP基因连接到载体pMCG161,再将此载体和pCAMBIA1301载体同时双酶切,产物回收后连接,成功构建了适合农杆菌介导的水稻转化RNAi载体。在农杆菌水稻转化过程中,比较了分化阶段是否加潮霉素对愈伤组织分化的影响,结果表明,分化阶段去除潮霉素的筛选可以提高愈伤组织的分化程度但同时降低获得水稻植株的阳性率。 |
| 英文摘要: |
| The CP gene of Rice stripe virus(RSV)was amplified by reverse transcription polymerase chain reaction(RT-PCR)from the total RNA purified from infected rice leaves.Sense and antisense of RSV CP gene were cloned into the pMCG161 plasmid.This vector and pCAMBIA1301 were both digested with BamHI and HindⅢ,and then the RNAi vector was obtained that could be transformed into rice by Agrobacterium tumefaciens.The effect of Hyg on callus differentiation during the process of rice transformation was analyzed.The results showed that the extent to which the callus differentiated increased,but the positive rates of transgenic plants decreased if Hyg was not added in the medium. |
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