Li Ang, Chen LiangLiang, Ren HaiYun, Wang XueChen, Zhang HaiWen, Huang Rong-Feng
Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing, 100081, China.
Sci China C Life Sci. 2008 Mar;51(3):280-5. doi: 10.1007/s11427-008-0041-9.
In rice, the characterization of OsEBP-89 is inducible by various stress-or hormone-stimuli, including ethylene, abscisic acid (ABA), jasmonate acid (JA), drought and cold. Here, we report the investigation of essential DNA region within OsEBP-89 promoter for methyl jasmonic acid (MeJA) induction. PLACE analysis indicates that this promoter sequence contains multiple potential elements in response to various stimuli. First, we fused this promoter with GUS gene and analyzed its expression under MeJA treatment through Agrobacterium infiltration mediating transient expression in tobacco leaves. Our results revealed that this chimeric gene could be inducible by MeJA in tobacco leaves. To further determine the crucial sequences responsible for MeJA induction, we generated a series of deletion promoters which were fused with GUS reporter gene respectively. The results of transient expression of GUS gene driven by these mutant promoters show that the essential region for MeJA induction is positioned in the region between -1200 and -800 in OsEBP-89 promoter containing a G-box (-1127), which is distinct from the essential region containing ERE (-562) for ACC induction. In all, our finding is helpful in understanding the molecular mechanism of OsEBP-89 expression under different stimuli.
在水稻中,OsEBP - 89的特性可被多种胁迫或激素刺激所诱导,包括乙烯、脱落酸(ABA)、茉莉酸(JA)、干旱和寒冷。在此,我们报道了对OsEBP - 89启动子内茉莉酸甲酯(MeJA)诱导所必需的DNA区域的研究。PLACE分析表明,该启动子序列包含多个响应各种刺激的潜在元件。首先,我们将该启动子与GUS基因融合,并通过农杆菌介导的烟草叶片瞬时表达分析其在MeJA处理下的表达。我们的结果显示,该嵌合基因在烟草叶片中可被MeJA诱导。为了进一步确定负责MeJA诱导的关键序列,我们分别构建了一系列与GUS报告基因融合的缺失启动子。由这些突变启动子驱动的GUS基因瞬时表达结果表明,MeJA诱导的必需区域位于OsEBP - 89启动子中 - 1200至 - 800之间的区域,该区域包含一个G - 盒(- 1127),这与ACC诱导所必需的包含ERE(- 562)的区域不同。总之,我们的发现有助于理解不同刺激下OsEBP - 89表达的分子机制。