Kyozuka J, McElroy D, Hayakawa T, Xie Y, Wu R, Shimamoto K
Plantech Research Institute, Yokohama, Japan.
Plant Physiol. 1993 Jul;102(3):991-1000. doi: 10.1104/pp.102.3.991.
A previously isolated rice (Oryza sativa) rbcS gene was further characterized. This analysis revealed specific sequences in the 5' regulatory region of the rice rbcS gene that are conserved in rbcS genes of other monocotyledonous species. In transgenic rice plants, we examined the expression of the beta-glucuronidase (gusA) reporter gene directed by the 2.8-kb promoter region of the rice rbcS gene. To examine differences in the regulation of monocotyledonous and dicotyledonous rbcS promoters, the activity of a tomato rbcS promoter was also investigated in transgenic rice plants. Our results indicated that both rice and tomato rbcS promoters confer mesophyll-specific expression of the gusA reporter gene in transgenic rice plants and that this expression is induced by light. However, the expression level of the rice rbcS-gusA gene was higher than that of the tomato rbcS-gusA gene, suggesting the presence of quantitative differences in the activity of these particular monocotyledonous and dicotyledonous rbcS promoters in transgenic rice. Histochemical analysis of rbcS-gusA gene expression showed that the observed light induction was only found in mesophyll cells. Furthermore, it was demonstrated that the light regulation of rice rbcS-gusA gene expression was primarily at the level of mRNA accumulation. We show that the rice rbcS gene promoter should be useful for expression of agronomically important genes for genetic engineering of monocotyledonous species.
对先前分离得到的水稻(Oryza sativa)rbcS基因进行了进一步表征。该分析揭示了水稻rbcS基因5'调控区中的特定序列,这些序列在其他单子叶植物物种的rbcS基因中是保守的。在转基因水稻植株中,我们检测了由水稻rbcS基因2.8 kb启动子区域指导的β-葡萄糖醛酸酶(gusA)报告基因的表达。为了研究单子叶和双子叶rbcS启动子调控的差异,还在转基因水稻植株中研究了番茄rbcS启动子的活性。我们的结果表明,水稻和番茄rbcS启动子在转基因水稻植株中均赋予gusA报告基因叶肉特异性表达,且这种表达受光诱导。然而,水稻rbcS - gusA基因的表达水平高于番茄rbcS - gusA基因,这表明在转基因水稻中,这些特定的单子叶和双子叶rbcS启动子的活性存在定量差异。对rbcS - gusA基因表达的组织化学分析表明,观察到的光诱导仅在叶肉细胞中发现。此外,已证明水稻rbcS - gusA基因表达的光调控主要在mRNA积累水平。我们表明,水稻rbcS基因启动子对于单子叶植物物种基因工程中农艺重要基因的表达应该是有用的。