小麦1B/1R易位系由于具有高产和适应性广等优点,在我国的小麦生产中得到了广泛的应用,但这类品种有一个共同的缺点,就是加工品质比较差,ω-黑麦碱被认为是影响其加工品质的一个重要因素.通过RNA干扰途径沉默ω-黑麦碱基因的表达,是改良小麦1B/1R易位系加工品质的一个重要策略.使用由ω-黑麦碱基因自身启动子驱动的RNA干扰表达载体,可使转入的基因在需要的部位和需要的时间表达,提高分子育种的效果.为获得有活性的ω黑麦碱基因的启动子,本研究设计了覆盖启动子区和部分编码区的一对特异引物,以小麦(Triticum aestivum)1B/1R易位系兰考906为试材,通过PCR克隆得到了与3个有转录活性的ω-黑麦碱基因有对应关系的5个启动子A9-1、H2-1、H7-1、C11和F11-1,选择与其中2个有转录活性的ω-黑麦碱基因分别有对应关系的启动子A9-1和F11-1构建以GUS为标记基因的表达载体,并用基因枪对小麦幼嫩种子进行了轰击,通过对GUS基因的瞬时表达分析,证实其中一个启动子F11-1具有活性.研究结果为构建由ω黑麦碱基因自身启动子驱动的RNA干扰表达载体提供了基础资料.%Wheat 1B/IR translocation lines have been widely used in China because of their high yield and wide adaptability.However they have a common defect that is bad grain processing quality, ω-secalin is believed to be an important factor for the bad quality.It is a good strategy to silence these ω-secalin genes through RNAi.Using ω-secalin gene's promoter to construct the RNAi expression vector can make the introduced gene to express in needed tissue and needed time and thus raise the effect of molecular breeding.In order to get a active promoter from ω-seclin genes, one pair of specific primers was designed.Promoter cloning was carried out with Lankao 906 as the material, a wheat (Triticura aestivum) 1B/1R translocation line.Five promoters A9-1, H2-1, H7-1, C11 and F11-1 corresponding to three active ω-seclin genes were obtained.Two promoters A9-1 and F11-1 corresponding to two active ω-seclin genes were chosen to make expression vectors with GUS as the marker gene.Transient expression analysis of the GUS gene was made by bombarding young wheat seeds and one promoter F11-ls activity was confirmed.The results provide the basic data for the construction of RNA interfering expression vector driven by the ω secalin gene itself promotor.
展开▼
