Feinbaum R L, Storz G, Ausubel F M
Department of Genetics, Harvard Medical School, Boston, Massachusetts.
Mol Gen Genet. 1991 May;226(3):449-56. doi: 10.1007/BF00260658.
To establish a genetic system for dissection of light-mediated signal transduction in plants, we analyzed the light wavelengths and promoter sequences responsible for the light-induced expression of the Arabidopsis thaliana chalcone synthase (CHS) promoter fused to the beta-glucuronidase (GUS) marker gene. Transgenic A. thaliana lines carrying 1975, 523, 186, and 17 bp of the CHS promoter fused to the GUS gene were generated, and the expression of these chimeric genes was monitored in response to high intensity light in mature plants and to different wavelengths of light in seedlings. Fusion constructs containing 1975 and 523 bp of CHS promoter sequence behaved identically to the endogenous CHS gene under all conditions. Expression of these constructs was induced specifically in response to high intensity white light and blue light. The response to blue light was seen in the presence of the Pfr form of phytochrome. Fusion constructs containing 186 bp of promoter sequence showed reduced basal levels of expression and only weak stimulation by blue light but were induced significantly by high intensity white light. These analyses showed that the expression of the A. thaliana CHS gene is responsive to a specific blue light receptor and that sequences between -523 and -186 bp are required for optimal basal and blue light-induced expression of this gene. The experiments lay the foundation for a simple genetic screen for light response mutants.
为了建立一个用于剖析植物光介导信号转导的遗传系统,我们分析了与β-葡萄糖醛酸酶(GUS)标记基因融合的拟南芥查尔酮合酶(CHS)启动子的光诱导表达所涉及的光波长和启动子序列。构建了携带与GUS基因融合的1975、523、186和17bp CHS启动子的转基因拟南芥株系,并在成熟植株中检测这些嵌合基因对高强度光的响应,以及在幼苗中检测对不同波长光的响应。含有1975和523bp CHS启动子序列的融合构建体在所有条件下的表现与内源性CHS基因相同。这些构建体的表达在高强度白光和蓝光刺激下被特异性诱导。在存在光敏色素的Pfr形式时能观察到对蓝光的响应。含有186bp启动子序列的融合构建体表现出较低的基础表达水平,且仅受蓝光微弱刺激,但能被高强度白光显著诱导。这些分析表明,拟南芥CHS基因的表达对特定蓝光受体有响应,并且-523至-186bp之间的序列是该基因最佳基础表达和蓝光诱导表达所必需的。这些实验为光反应突变体的简单遗传筛选奠定了基础。