Nagatani A, Kay S A, Deak M, Chua N H, Furuya M
Laboratory of Plant Biological Regulation, Frontier Research Program, RIKEN Institute, Wako City, Saitama, Japan.
Proc Natl Acad Sci U S A. 1991 Jun 15;88(12):5207-11. doi: 10.1073/pnas.88.12.5207.
We have examined the biological activity of rice type I phytochrome (PI) in transgenic tobacco seedlings. The progeny of four independent transformants that expressed the rice PI gene segregated 3:1 for shorter hypocotyl length under dim white light (0.04 W/m2). By contrast, this phenotype was not observed either in the dark or under white light at higher intensity (6.0 W/m2). This suggests that the phenotype is dependent not only on light but also on light intensity. The increased light sensitivity cosegregated with the kanamycin-resistance marker as well as with the rice PI polypeptides, indicating that this phenotype is directly related to the expression of the transgene. The transgenic plants showing short hypocotyls exhibited a reduced growth rate throughout the elongation period, and the resulting shorter hypocotyl length was attributable to shorter epidermal cell length but not to reduced cell number. Furthermore, successive pulse irradiations with red light elicited short hypocotyls similar to those obtained under dim white light, and the effect was reversed by immediate far-red light treatment, providing a direct indication that the phenotype is caused by biologically active rice PI. Therefore, the far-red-absorbing form of the introduced rice PI appears to regulate the hypocotyl length of the transgenic tobacco plants through endogenous signal-transduction pathways. This assay system will be a powerful tool for testing the biological activity of introduced phytochrome molecules.
我们检测了水稻I型光敏色素(PI)在转基因烟草幼苗中的生物活性。在昏暗白光(0.04 W/m²)下,四个表达水稻PI基因的独立转化体的后代中,短胚轴长度的分离比例为3:1。相比之下,在黑暗中或更高强度白光(6.0 W/m²)下未观察到这种表型。这表明该表型不仅依赖于光,还依赖于光强度。光敏感性增加与卡那霉素抗性标记以及水稻PI多肽共分离,表明该表型与转基因的表达直接相关。表现出短胚轴的转基因植物在整个伸长阶段生长速率降低,胚轴长度缩短是由于表皮细胞长度缩短而非细胞数量减少。此外,连续用红光脉冲照射会引发类似于在昏暗白光下获得的短胚轴,并且通过立即进行远红光处理可逆转这种效应,这直接表明该表型是由具有生物活性的水稻PI引起的。因此,导入的水稻PI的远红光吸收形式似乎通过内源性信号转导途径调节转基因烟草植物的胚轴长度。该检测系统将成为测试导入光敏色素分子生物活性的有力工具。