Ong Wen-Dee, Okubo-Kurihara Emiko, Kurihara Yukio, Shimada Setsuko, Makita Yuko, Kawashima Mika, Honda Kaori, Kondoh Yasumitsu, Watanabe Nobumoto, Osada Hiroyuki, Cutler Sean R, Sudesh Kumar, Matsui Minami
Synthetic Genomics Research Group, Biomass Engineering Research Division, RIKEN Center for Sustainable Resource Science, Yokohama, Kanagawa, Japan.
Ecobiomaterial Research Laboratory, School of Biological Sciences, Universiti Sains Malaysia, Minden, Penang, Malaysia.
Plant Cell Physiol. 2017 Jan 1;58(1):95-105. doi: 10.1093/pcp/pcw181.
Plants have a remarkable ability to perceive and respond to various wavelengths of light and initiate regulation of different cascades of light signaling and molecular components. While the perception of red light and the mechanisms of its signaling involving phytochromes are largely known, knowledge of the mechanisms of blue light signaling is still limited. Chemical genetics involves the use of diverse small active or synthetic molecules to evaluate biological processes. By combining chemicals and analyzing the effects they have on plant morphology, we identified a chemical, 3-bromo-7-nitroindazole (3B7N), that promotes hypocotyl elongation of wild-type Arabidopsis only under continuous blue light. Further evaluation with loss-of-function mutants confirmed that 3B7N inhibits photomorphogenesis through cryptochrome-mediated light signaling. Microarray analysis demonstrated that the effect of 3B7N treatment on gene expression in cry1cry2 is considerably smaller than that in the wild type, indicating that 3B7N specifically interrupts cryptochrome function in the control of seedling development in a light-dependent manner. We demonstrated that 3B7N directly binds to CRY1 protein using an in vitro binding assay. These results suggest that 3B7N is a novel chemical that directly inhibits plant cryptochrome function by physical binding. The application of 3B7N can be used on other plants to study further the blue light mechanism and the genetic control of cryptochromes in the growth and development of plant species.
植物具有非凡的能力,能够感知并响应各种波长的光,并启动对不同光信号级联和分子成分的调控。虽然红光的感知及其涉及光敏色素的信号传导机制在很大程度上已为人所知,但蓝光信号传导机制的相关知识仍然有限。化学遗传学涉及使用各种活性小分子或合成分子来评估生物过程。通过结合化学物质并分析它们对植物形态的影响,我们鉴定出一种化学物质3-溴-7-硝基吲唑(3B7N),它仅在持续蓝光下促进野生型拟南芥下胚轴伸长。对功能缺失突变体的进一步评估证实,3B7N通过隐花色素介导的光信号传导抑制光形态建成。微阵列分析表明,3B7N处理对cry1cry2基因表达的影响远小于野生型,这表明3B7N以光依赖的方式特异性地干扰隐花色素在幼苗发育控制中的功能。我们使用体外结合试验证明3B7N直接与CRY1蛋白结合。这些结果表明,3B7N是一种通过物理结合直接抑制植物隐花色素功能的新型化学物质。3B7N的应用可用于其他植物,以进一步研究蓝光机制以及植物物种生长发育中隐花色素的遗传控制。