National Agriculture and Food Research Organization (NARO), Institute of Floricultural Science, Tsukuba, Ibaraki 305-8519, Japan.
Research Fellow of Japanese Society for the Promotion of Science (JSPS), Tokyo 102-0083, Japan.
Sci Rep. 2016 Mar 29;6:23609. doi: 10.1038/srep23609.
Chlorophyll (Chl) degradation occurs during leaf senescence, embryo degreening, bud breaking, and fruit ripening. The Chl catabolic pathway has been intensively studied and nearly all the enzymes involved are identified and characterized; however, the molecular regulatory mechanisms of this pathway are largely unknown. In this study, we performed yeast one-hybrid screening using a transcription factor cDNA library to search for factors controlling the expression of Chl catabolic genes. We identified ANAC046 as a common regulator that directly binds to the promoter regions of NON-YELLOW COLORING1, STAY-GREEN1 (SGR1), SGR2, and PHEOPHORBIDE a OXYGENASE. Transgenic plants overexpressing ANAC046 exhibited an early-senescence phenotype and a lower Chl content in comparison with the wild-type plants, whereas loss-of-function mutants exhibited a delayed-senescence phenotype and a higher Chl content. Microarray analysis of ANAC046 transgenic plants showed that not only Chl catabolic genes but also senescence-associated genes were positively regulated by ANAC046. We conclude that ANAC046 is a positive regulator of Arabidopsis leaf senescence and exerts its effect by controlling the expression of Chl catabolic genes and senescence-associated genes.
叶绿素(Chl)在叶片衰老、胚胎褪绿、芽休眠和果实成熟过程中降解。Chl 分解代谢途径已得到深入研究,几乎所有涉及的酶都已被鉴定和表征;然而,该途径的分子调控机制在很大程度上尚不清楚。在这项研究中,我们使用转录因子 cDNA 文库进行酵母单杂交筛选,以寻找控制 Chl 分解代谢基因表达的因子。我们鉴定出 ANAC046 是一个共同的调节剂,它可以直接结合 NON-YELLOW COLORING1、STAY-GREEN1(SGR1)、SGR2 和 PHEOPHORBIDE a OXYGENASE 的启动子区域。与野生型植物相比,过表达 ANAC046 的转基因植物表现出衰老提前的表型和较低的 Chl 含量,而功能缺失突变体表现出衰老延迟的表型和较高的 Chl 含量。ANAC046 转基因植物的微阵列分析表明,不仅 Chl 分解代谢基因,而且衰老相关基因也被 ANAC046 正向调控。我们得出结论,ANAC046 是拟南芥叶片衰老的正调节剂,通过控制 Chl 分解代谢基因和衰老相关基因的表达来发挥其作用。