摘要： 水稻黑条矮缩病是由水稻黑条矮缩病毒引起的水稻病害，该病可导致病株生长迟缓、矮化、抽穗结实困难，严重影响了水稻的生产。在水稻黑条矮缩病毒基因组不同双链RNA序列的基础上，分别设计了5对特异性引物，通过RT—PCR获得了相应片段，结合基因沉默的方法，分别构建RNA干扰载体，并进一步将发夹结构构建入以玉米泛素Ubi为启动子、以bar基因为选择标记的改造后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，成功获得了5个含有水稻黑条矮缩病干扰片段的植物双元表达载体，经农杆菌介导获得了部分水稻转化植株。旨在为下一步对水稻黑条矮缩病的研究及抗病水稻新品种的培育提供材料。

关键词: 水稻黑条矮缩病, RNA干扰载体, Ubi启动子, bar基因, 抗病水稻新品种培育

Abstract: Rice black streaked dwarf disease caused by rice black streaked dwarf virus, lead to dwarf, strong green color of rice, which could seriously influence the yield of rice and food safety. In order to solve the problem, the objective of this study is to use RNA inference （RNAi） strategy to improve the rice resistance ability to virus. Here 5 pairs of black streaked dwarf virus primers were designed according to the published virus sequences in da- tabase and related literature, and 5 interference fragment were obtained by RT-PCR, these interference fragments were then built into the modified pUC19 RNA interference vector,and the whole RNA interference fragments were further integrated into the modified binary vector pCAMBIA1300,which is using ubiquitin as promoter and herbicide resistance Bar gene as a selectable marker. Enzyme digestion and PCR analysis showed that the recombined plasmid were successfully constructed and the transgenic rice seedlings were obtained by agrobacterium-mediated transforma- tion. Our research will shed some light for further investigation of quality and resistance ability improvement of rice to rice black streaked dwarf disease resistance breeding.

Key words: Rice black-streaked dwarf virus, RNA interference vector, Ubiquitin promoter bar gene, Cultiva- tion of new germplasm of rice disease resistance

中图分类号: 

• Q78