Plant A L, van Rooijen G J, Anderson C P, Moloney M M
Department of Biological Sciences, University of Calgary, Alberta, Canada.
Plant Mol Biol. 1994 May;25(2):193-205. doi: 10.1007/BF00023237.
Progressive deletions of the 5'-flanking sequences of an Arabidopsis oleosin gene were fused to beta-glucuronidase (GUS) and introduced into Brassica napus plants using Agrobacterium-mediated transformation. The effect of these deletions on the quantitative level of gene expression, organ specificity and developmental regulation was assessed. In addition, the influence of abscisic acid (ABA), jasmonic acid (JA), sorbitol and a combined ABA/sorbitol treatment on gene expression was investigated. Sequences that positively regulate quantitative levels of gene expression are present between -1100 to -600 and -400 to -200 of the promoter. In addition, sequences present between -600 and -400 down-regulate quantitative levels of expression. In transgenic B. napus plants, the oleosin promoter directs seed-specific expression of GUS which is present at early stages of seed development and increases throughout seed maturation. Sequences present between -2500 and -1100 of the promoter are involved in modulating the levels of expression at early stages of embryo development. Histochemical staining of embryos demonstrated that expression is uniform throughout the tissues of the embryo. Sequences involved in the response to ABA and sorbitol are present between -400 and -200. The induction of GUS activity by a combined ABA/sorbitol treatment is additive suggesting that ABA is not the sole mediator of osmotically induced oleosin gene expression. A response to JA was only observed when the oleosin promoter was truncated to -600 suggesting that the reported effect of JA on oleosin gene expression may be at a post-transcriptional level.
将拟南芥油质蛋白基因5'-侧翼序列的逐步缺失片段与β-葡萄糖醛酸酶(GUS)融合,并通过农杆菌介导的转化方法导入甘蓝型油菜植株中。评估了这些缺失对基因表达定量水平、器官特异性和发育调控的影响。此外,还研究了脱落酸(ABA)、茉莉酸(JA)、山梨醇以及ABA/山梨醇联合处理对基因表达的影响。在启动子的-1100至-600以及-400至-200之间存在正向调控基因表达定量水平的序列。此外,在-600至-400之间的序列会下调表达的定量水平。在转基因甘蓝型油菜植株中,油质蛋白启动子指导GUS在种子发育早期存在的种子特异性表达,并且在种子成熟过程中表达量增加。启动子-2500至-1100之间的序列参与调节胚胎发育早期的表达水平。胚胎的组织化学染色表明,表达在胚胎的所有组织中是均匀的。参与对ABA和山梨醇响应的序列存在于-400至-200之间。ABA/山梨醇联合处理对GUS活性的诱导是累加的,这表明ABA不是渗透诱导油质蛋白基因表达的唯一介质。仅当油质蛋白启动子截短至-600时才观察到对JA的响应,这表明所报道的JA对油质蛋白基因表达的影响可能发生在转录后水平。