玉米ZmPR4启动子的克隆及其在小麦幼胚愈伤组织中的活性分析

Cloning and Activity Analysis of Zea mays ZmPR4 Promoter in Wheat Immature Embryonic Calli

以玉米B73基因组DNA为模板,通过特异PCR扩增,克隆出玉米启动子ZmPR4序列。序列分析表明,该启动子与AJ969166序列同源性为100%。构建了ZmPR4或玉米泛素基因(Ubiquitin)启动子控制的报告基因GUS的表达载体。通过基因枪介导法转化小麦幼胚愈伤组织。瞬间表达实验表明,在小麦幼胚愈伤组织中,玉米ZmPR4启动子的本底表达活性明显比Ubi启动子的低,但经纹枯病菌诱导后,ZmPR4启动子控制的GUS基因的表达明显增强。PCR检测结果证实ZmPR4启动子在小麦愈伤组织中具有表达活性,能够驱动GUS基因的表达。因此,玉米ZmPR4启动子在小麦抗病基因工程育种中具有潜在的应用价值。

The ZmPR4 promoter was cloned from genomic DNA of maize inbred line B37 through specific PCR amplification. The promoter cloned had the same sequence as the fragment registered in GenBank under the accession number of AJ969166. Expression vectors driven by ZmPR4 promoter or Ubiquitin promoter were constructed, which harbor GUS gene. The vectors were bombarded into wheat immature embryogenic calli. Histochemical assays of GUS activity, the ZmPR4 promoter was obviously expressed in wheat immature embryonic calli, but the expression activity was lower than that of Ubi Promoter. The ZmPR4 promoter could be induced by infection of Rhizoctonia cerealis. PCR assay confirmed the expression of GUS gene driven by ZmPR4 promoter. These results suggest that ZmPR4 promoter has a use potential in molecular breeding of resistance in wheat.