Lübberstedt T, Bolle C E, Sopory S, Flieger K, Herrmann R G, Oelmüller R
Botanisches Institut der Ludwig-Maximilians-Universität, München, Germany.
Plant Physiol. 1994 Mar;104(3):997-1006. doi: 10.1104/pp.104.3.997.
The light-regulated expression of eight nuclear-encoded genes for plastid proteins from spinach (Spinacia oleracea) (RBCS-1 and CAB-1; ATPC and ATPD, encoding the subunits gamma and delta of the ATP synthase; PC and FNR; PSAD and PSAF, encoding the subunits II and III of photosystem I reaction center) was analyzed with promoter/beta-glucuronidase (GUS) gene fusions in transgenic tobacco (Nicotiana tabacum and Nicotiana plumbaginifolia) seedlings and mature plants under standardized light and growth conditions. Unique response patterns were found for each of these promoters. GUS activities differed more than 30-fold. Strong promoters were found for the PC and PSAD genes. On the other hand, the ATPC promoter was relatively weak. Expression of the CAB/GUS gene fusion in etiolated material was at the detection limit; all other chimeric genes were expressed in the dark as well. Light stimulation of GUS activities ranged from 3- (FNR promoter) to more than 100-fold (CAB-1 promoter). The FNR promoter responded only to red light (RL) and not significantly to blue light (BL), whereas the PC promoter contained regions with different sensitivities toward RL and BL. Furthermore, different RNA accumulation kinetics were observed for the PSAF, CAB, FNR, and PC promoter/GUS gene fusions during de-etiolation, which, at least in the case of the PSAF gene, differed from the regulation of the corresponding endogenous genes in spinach and tobacco. The results suggest either that not all cis elements determining light-regulated and quantitative expression are present on the spinach promoter fragments used or that the spinach cis-regulatory elements respond differently to the host (tobacco) regulatory pathway(s). Furthermore, as in tobacco, but not in spinach, the trans-gene hardly responds to single light pulses that operate through phytochrome. Taken together, the results suggest that the genes have been independently translocated from the organelle to the nucleus during phylogeny. Furthermore, each gene seems to have acquired a unique set of regulatory elements.
利用启动子/β-葡萄糖醛酸酶（GUS）基因融合技术，在标准化光照和生长条件下，对转基因烟草（烟草属和垂花烟草）幼苗及成熟植株中菠菜（菠菜）8个质体蛋白核编码基因（RBCS - 1和CAB - 1；ATPC和ATPD，分别编码ATP合酶的γ和δ亚基；PC和FNR；PSAD和PSAF，分别编码光系统I反应中心的II和III亚基）的光调控表达进行了分析。发现这些启动子各自具有独特的响应模式。GUS活性差异超过30倍。PC和PSAD基因具有强启动子。另一方面，ATPC启动子相对较弱。CAB/GUS基因融合在黄化材料中的表达处于检测极限；所有其他嵌合基因在黑暗中也有表达。GUS活性的光刺激范围从3倍（FNR启动子）到超过100倍（CAB - 1启动子）。FNR启动子仅对红光（RL）有反应，对蓝光（BL）无明显反应，而PC启动子包含对RL和BL具有不同敏感性的区域。此外，在去黄化过程中，观察到PSAF、CAB、FNR和PC启动子/GUS基因融合的RNA积累动力学不同，至少就PSAF基因而言，这与菠菜和烟草中相应内源基因的调控不同。结果表明，要么用于研究的菠菜启动子片段上不存在所有决定光调控和定量表达的顺式元件，要么菠菜顺式调控元件对宿主（烟草）调控途径的反应不同。此外，与烟草一样，但与菠菜不同，转基因几乎不响应通过光敏色素起作用的单个光脉冲。综上所述，结果表明这些基因在系统发育过程中已从细胞器独立转移至细胞核。此外，每个基因似乎都获得了一套独特的调控元件。
