水稻黑条矮缩病毒RNA干扰载体的构建及遗传转化  

Construction and Genetic Transformation of RNA Interference Vectors for Rice Black-Streaked Dwarf Virus

作　　者：张贵林[1,2] 李臻[2] 王庆国[2] 柳絮[2] 姚方印[2] 王莹莹[2] 刘炜[1,2] 

机构地区：[1]山东师范大学生命科学学院,山东济南250014 [2]山东省农业科学院高新技术研究中心/山东省作物遗传改良与生态生理重点实验室,山东济南250100

出　　处：《山东农业科学》2013年第1期19-24,共6页Shandong Agricultural Sciences

基　　金：国家转基因植物新品种培育重大专项(编号:2009ZX08001-010B)

摘　　要：针对水稻黑条矮缩病毒S6、S10两条RNA双链,参考相关文献,设计了2对引物,通过RT-PCR获得了2条干扰片段,将这些干扰片段分别构建到RNA干扰载体中,进一步将整个RNA干扰片段构建入以玉米泛素Ubi为启动子、以生物安全性的bar基因为选择标记的植物双元表达载体pCA1300-Ubi中,经农杆菌介导对黄淮稻区主栽水稻品种圣稻13进行遗传转化,为下一步对水稻黑条矮缩病的研究及抗性品种的培育提供候选材料。Two pairs of specific primers were designed according to the rice black - streaked dwarf virus sequences of $6 and SIO in this research. Two interference fragments were obtained by RT - PCR and were constructed into RNA interference vectors using modified pUC19. Then the complete interference fragments were further digested and integrated into the modified pCAMBIA1300 vector which used ubiquitin as promoter and bar gene as selectable marker. The transgenic rice seedlings were obtained by Agrobacterium - mediated transformation to rice variety Shengdao 13. This research could provide candidate materials for the cultivation of new germplasm with rice black - streaked dwarf virus resistance.

关 键 词：水稻黑条矮缩病 RNA干扰(RNAI) Ubi启动子 BAR基因 

分 类 号：Q781[生物学—分子生物学]