Samach A, Broday L, Hareven D, Lifschitz E
Department of Biology, Technion-Israel Institute of Technology, Haifa.
Plant J. 1995 Sep;8(3):391-406. doi: 10.1046/j.1365-313x.1995.08030391.x.
The gene coding for threonine deaminase (TD), the enzyme which catalyzes the first committed step in the biosynthesis of isoleucine, was isolated from tomato as a consequence of its unusual 500-fold upregulation in floral organs. It was subsequently shown that TD is induced in potato leaves in response to wounding, abscisic acid and methyl jasmonate (MeJa). Detailed analysis presented here, reveals an intricate developmental regulation pattern of gene expression in flowers that is operating solely in parenchyma territories. Yet, despite its high pre-existing expression level, TD in flowers can be further induced by MeJa. Induction of TD in flowers as well as in leaves is effective only in the parenchyma domains, irrespective of the prior expression levels. TD is neither expressed nor induced in epidermal, vascular or sporogenous tissues. Promoter analysis in transgenic tomato plants indicates that induction of TD follows identical kinetics in flowers and leaves. Furthermore, the 'conditioning' of developmental upregulation in flowers, the response to MeJa in flowers and leaves, and the parenchyma-specific expression are all mediated by the cis-elements within the proximal 192 bp of the promoter. Promoter elements regulating the correct organ-specific expression are located, however, further upstream. The promoter constructs used in this study can serve as useful tools for expressing extremely high levels of transgenes in specific cells. A scheme explaining tissue-specific response to MeJa, in conjunction with developmental control, is discussed.
编码苏氨酸脱氨酶(TD)的基因,该酶催化异亮氨酸生物合成中的第一个关键步骤,因其在花器官中异常上调500倍而从番茄中分离出来。随后发现,TD在马铃薯叶片中受到创伤、脱落酸和茉莉酸甲酯(MeJa)的诱导。此处进行的详细分析揭示了花中基因表达的复杂发育调控模式,该模式仅在薄壁组织区域起作用。然而,尽管花中TD的预先存在的表达水平很高,但仍可被MeJa进一步诱导。花和叶中TD的诱导仅在薄壁组织区域有效,与先前的表达水平无关。TD在表皮、维管或孢子发生组织中既不表达也不被诱导。转基因番茄植株中的启动子分析表明,花和叶中TD的诱导遵循相同的动力学。此外,花中发育上调的“调节”、花和叶中对MeJa的反应以及薄壁组织特异性表达均由启动子近端192 bp内的顺式元件介导。然而,调节正确器官特异性表达的启动子元件位于更上游。本研究中使用的启动子构建体可作为在特定细胞中表达极高水平转基因的有用工具。讨论了一个结合发育控制来解释对MeJa的组织特异性反应的方案。