2个水稻PLT基因启动子的克隆及其表达分析

Cloning and expression analysis of two PLT genes promoters in rice

为研究水稻中PLT基因的表达及功能,利用同源序列分析,在水稻基因数据库中检索到2个与拟南芥PLT基因高度同源的基因序列,命名为OsPLT7和OsPLT11。根据进一步预测的PLT启动子序列设计引物,以水稻品种中花11的基因组DNA为模板,用PCR方法扩增得到PLT7、PLT11的启动子片段,克隆至含有报告基因的表达载体上并转化水稻。通过GUS染色观察T1代转基因阳性植株,发现OsPLT7和OsPLT11在水稻根部干细胞小生境和中柱部位表达,表达模式类似于其同源基因在拟南芥中的表达谱。

To study the expression and function of PLT in rice, two PLT genes in rice （OsPLT7 and OsPLT11） were firstly predicted from searching the homology of Arabidopsis PLT genes protein sequences against rice database. According to the predicted promoter sequences of OsPLT7 and OsPLT11,we designed the PCR primers and amplified the consequent promoter fragments of OsPLT7 and OsPLT11 using genomic DNA of Oryza sativa L. Zhonghua 11 as templates. Finally, we cloned these fragments into the vector containing reporter genes GUS/YFP and successfully transformed these promoter-reporter fusion vectors into rice. GUS staining observation of T1 transgenic seedlings of OsPLT7 and OsPLT11 promoter-reporter fusion vectors indicated that both OsPLT7 and OsPLT11 are expressed mainly in stem cell niche and stele area in rice root, which are very similar to the expression profile of their homologous genes in Arabidopsis. These results will be useful for the functional studies of the PLT family transcription factors.