Yokoyama R, Hirose T, Fujii N, Aspuria E T, Kato A, Uchimiya H
Center for Gene Research, Nagoya University, Japan.
Mol Gen Genet. 1994 Jul 8;244(1):15-22. doi: 10.1007/BF00280182.
The 5'-upstream region of the rolC gene of the Ri plasmid is expressed specifically in phloem cells of transgenic higher plants. In this study, we demonstrated that the rolC promoter is activated by sucrose in phloem cells of transgenic tobacco seedlings bearing rolC promoter-uidA chimeric fusion gene. Since the rolC promoter is not activated by sorbitol, sucrose metabolism rather than osmotic pressure exerted by the disaccharide may be responsible for induction. Thus, experiments using 5'-upstream deletion mutants, internal deletion mutants, and chimeric constructs with a heterologous promoter (-90 region of the cauliflower mosaic virus 35S promoter) were conducted to define the region of the rolC promoter involved in sucrose activation. The results indicated that a cis-acting sucrose responsive region of the rolC promoter is located between -135 and -94 bp with respect to the transcription initiation site. In phloem cells, high concentrations of sucrose are encountered owing to ongoing translocation of photosynthates from source to sink tissues. Therefore, sucrose as a signal molecule may regulate the phloem-specific expression of the rolC promoter.
发根农杆菌Ri质粒rolC基因的5'上游区域在转基因高等植物的韧皮部细胞中特异性表达。在本研究中,我们证明了在携带rolC启动子-uidA嵌合融合基因的转基因烟草幼苗的韧皮部细胞中,rolC启动子被蔗糖激活。由于rolC启动子不被山梨醇激活,可能是蔗糖代谢而非二糖施加的渗透压导致了诱导作用。因此,我们进行了使用5'上游缺失突变体、内部缺失突变体以及带有异源启动子(花椰菜花叶病毒35S启动子的-90区域)的嵌合构建体的实验,以确定rolC启动子中参与蔗糖激活的区域。结果表明,rolC启动子的顺式作用蔗糖响应区域相对于转录起始位点位于-135至-94 bp之间。在韧皮部细胞中,由于光合产物从源组织持续向库组织转运,会遇到高浓度的蔗糖。因此,蔗糖作为一种信号分子可能调节rolC启动子的韧皮部特异性表达。