Imoto Yusuke, Yamada Tetsuya, Kitamura Keisuke, Kanazawa Akira
Research Faculty of Agriculture, Hokkaido University.
Genes Genet Syst. 2008 Dec;83(6):469-76. doi: 10.1266/ggs.83.469.
Differentiation into specific embryo cell types correlates with the processes that lead to the accumulation of seed storage proteins in plants. The alpha subunit of beta-conglycinin, a major component of seed storage proteins in soybean, accumulates at a higher level in cotyledons than in the embryonic axis in developing embryos. To understand the mechanisms underlying this phenomenon, we characterized the upstream region of the alpha subunit gene in terms of transcriptional control using transgenic Arabidopsis thaliana plants carrying reporter gene constructs comprising the 1357-bp upstream sequence of the alpha subunit gene and the beta-glucuronidase (GUS) gene. Analysis of the time-course-dependent pattern of GUS expression revealed that the expression was first confined to the cotyledons and occurred later in the entire embryo during embryogenesis. The level of GUS expression was higher in cotyledons than in the embryonic axis throughout the period of its expression, coincident with the distribution of the alpha subunit protein in soybean embryos. By testing progressively shorter promoter fragments, the cis-acting elements responsible for transcriptional activation in the cotyledons and the embryonic axis were both localized to the region spanning -245 to -161 relative to the transcription start site. It is also concluded that the upstream region up to -245 is sufficient to control the spatial and temporal pattern of transcription, while further upstream regions influence transcription rate without affecting the transcriptional pattern. Overall, these results indicate that the unequal distribution of alpha subunit protein within the embryos is established primarily as a consequence of differential transcriptional activation controlled by a short proximal promoter region of the gene in different embryonic tissues.
分化为特定的胚胎细胞类型与导致植物种子贮藏蛋白积累的过程相关。β-伴大豆球蛋白的α亚基是大豆种子贮藏蛋白的主要成分,在发育中的胚中,其在子叶中的积累水平高于胚轴。为了了解这一现象背后的机制,我们利用携带报告基因构建体的转基因拟南芥植株,对α亚基基因的上游区域进行了转录调控方面的表征,该构建体包含α亚基基因的1357bp上游序列和β-葡萄糖醛酸酶(GUS)基因。对GUS表达的时间进程依赖性模式分析表明,在胚胎发生过程中,表达首先局限于子叶,随后出现在整个胚中。在其整个表达期间,GUS在子叶中的表达水平高于胚轴,这与α亚基蛋白在大豆胚中的分布一致。通过逐步测试越来越短的启动子片段,负责子叶和胚轴中转录激活的顺式作用元件均定位于相对于转录起始位点-245至-161的区域。还得出结论,直至-245的上游区域足以控制转录的时空模式,而更上游的区域影响转录速率但不影响转录模式。总体而言,这些结果表明,胚内α亚基蛋白的不均等分布主要是由于该基因在不同胚胎组织中的短近端启动子区域控制的差异转录激活所致。