利用RNAi技术沉默大麦B-Hordein基因的研究

为限制大麦籽粒中贮藏蛋白的积累,降低其蛋白质含量,以大麦幼胚为受体材料,采用农杆菌介导的转化方法,将大麦B组醇溶蛋白基因gp4/antisense gp4片段与Ubi启动子相连导入大麦,经PCR检测证实,含有gp4反向重复基因片段的RNAi抑制表达框架已成功转入大麦。RT-PCR结果表明,转基因株系中B-Hordein mRNA积累受到明显抑制。收获T1代籽粒,经近红外谷物分析仪测定表明,2个转基因大麦株系籽粒蛋白质含量(分别为11.4%、11.7%)较对照(12.3%)降低。因此,利用RNA干涉技术降低大麦蛋白质含量是可行的。

Silence of B Hordein Gene in Barley Induced by RNAi Strategy

In order to restrict the accumulation of storage protein in the grain of malting barley (Hordeum vulgare L.), the inverted repeat sequence of B Hordein gene with Ubi promoter was introduced into barley via Agrobecterlum infection using the immature embryos as receptor tissue. Transgenic barley plants containing the inverted repeat structure were confirmed by PCR. The results of RT PCR demonstrated that the mRNA of B Hordein in transgenic barley was significantly inhibited. The transgenic grains harvested were analyzed for protein content (%) using a near infrared spectroscopy analyzer and the results indicated that two transgenic lines had lower protein content than the control. Results suggested it was possible to inhibit protein content of barley through RNA interference technology.