小麦Vp-1基因RNA干扰表达载体的构建及遗传转化  

Construction of the RNAi Vector of Vp-1 Gene from Wheat and Its Genetic Transformation

作　　者：高东尧[1,2] 夏兰琴[2] 马有志[2] 徐兆师[2] 徐惠君[2] 杜丽璞[2] 聂丽娜[2] 李彦舫[3] 原亚萍[3] 李连城[2] 陈明[2] 孙金海[1,4] 

机构地区：[1]吉林大学农学部畜牧兽医学院,长春130062 [2]中国农业科学院作物科学研究所/农作物基因资源与基因改良国家重大科学工程/农业部作物遗传育种重点开放实验室,北京100081 [3]吉林大学农学部植物科学学院,长春130062 [4]青岛农业大学动物科技学院,青岛266109

出　　处：《植物遗传资源学报》2009年第1期9-15,共7页Journal of Plant Genetic Resources

基　　金：国家高技术研究发展计划("863"计划)项目(2006AA10Z115;2007AA10ZA130)

摘　　要：小麦成熟期穗发芽是世界性的自然灾害,严重影响小麦的产量和品质。Viviparous-1(Vp-1)是促进胚成熟和休眠的主要转录调节因子,与小麦穗发芽抗性有着密切的关系。本实验根据小麦Vp-1基因序列,以植物表达载体pAHC25为基础,成功构建了含有反向重复序列的RNA干扰表达载体pAHC-WVpRi。采用基因枪法轰击小麦品种新春9号幼胚材料1825个,共获得34株T0再生植株。利用Bar基因引物和干扰片段特异引物对再生植株进行PCR检测,获得Bar基因和干扰片段均为阳性的植株3株,转化率为0.16%。本研究为深入分析Vp-1基因功能,进而通过分子育种进行小麦穗发芽抗性的遗传改良提供了科学依据。Pre-harvest sprouting （PHS）, as a worldwide natural disaster, seriously affects the quality and yield of wheat. Vp-1 （ Viviparous- 1 ） gene is an important regulator which promotes embryo maturation and dormancy, and play an important role in wheat PHS resistance. In this study, according to the Vp-1 sequence from wheat, the RNAi expression vector of Vp-1 gene named pAHC-WVpRi was successfully constructed and transformed into wheat variety Xinchun 9. In total, 1825 embryos were bombarded. A total of 34 regenerated plants with resistance to Bialaphos were obtained. Among them, three transgenic lines both with Bar gene and interference fragment were identified, with the transformation frequency of 0.16%. This laid a basis for the function analysis of Vp-1 gene and wheat improvement for PHS resistance through molecular breeding.

关 键 词：成熟期穗发芽 Vp-1基因 RNAI 遗传转化 

分 类 号：S512.1[农业科学—作物学]