Zhang J Z, Santes C M, Engel M L, Gasser C S, Harada J J
Division of Biological Sciences, University of California, Davis 95616, USA.
Plant Physiol. 1996 Apr;110(4):1069-79. doi: 10.1104/pp.110.4.1069.
We analyzed DNA sequences that regulate the expression of an isocitrate lyase gene from Brassica napus L. during late embryogenesis and during postgerminative growth to determine whether glyoxysomal function is induced by a common mechanism at different developmental stages. beta-Glucuronidase constructs were used both in transient expression assays in B. napus and in transgenic Arabidopsis thaliana to identify the segments of the isocitrate lyase 5' flanking region that influence promoter activity. DNA sequences that play the principal role in activating the promoter during post-germinative growth are located more than 1,200 bp upstream of the gene. Distinct DNA sequences that were sufficient for high-level expression during late embryogenesis but only low-level expression during postgerminative growth were also identified. Other parts of the 5' flanking region increased promoter activity both in developing seed and in seedlings. We conclude that a combination of elements is involved in regulating the isocitrate lyase gene and that distinct DNA sequences play primary roles in activating the gene in embryos and in seedlings. These findings suggest that different signals contribute to the induction of glyoxysomal function during these two developmental stages. We also showed that some of the constructs were expressed differently in transient expression assays and in transgenic plants.
我们分析了在胚胎发育后期和发芽后生长过程中调控甘蓝型油菜异柠檬酸裂解酶基因表达的DNA序列,以确定乙醛酸循环体功能是否在不同发育阶段由共同机制诱导。β-葡萄糖醛酸酶构建体用于甘蓝型油菜的瞬时表达分析和转基因拟南芥中,以鉴定影响启动子活性的异柠檬酸裂解酶5'侧翼区域片段。在发芽后生长过程中激活启动子起主要作用的DNA序列位于该基因上游1200多个碱基对处。还鉴定出了在胚胎发育后期足以实现高水平表达但在发芽后生长过程中仅为低水平表达的不同DNA序列。5'侧翼区域的其他部分在发育中的种子和幼苗中均增强了启动子活性。我们得出结论,多种元件共同参与调控异柠檬酸裂解酶基因,不同的DNA序列在激活胚胎和幼苗中的该基因时起主要作用。这些发现表明,在这两个发育阶段,不同的信号促成了乙醛酸循环体功能的诱导。我们还表明,一些构建体在瞬时表达分析和转基因植物中的表达不同。