RIKEN Plant Science Center, Yokohama, Kanagawa 230-0045, Japan.
Plant Physiol. 2010 Jul;153(3):906-14. doi: 10.1104/pp.110.154013. Epub 2010 May 20.
We previously showed that the VASCULAR-RELATED NAC-DOMAIN6 (VND6) and VND7 genes, which encode NAM/ATAF/CUC domain protein transcription factors, act as key regulators of xylem vessel differentiation. Here, we report a glucocorticoid-mediated posttranslational induction system of VND6 and VND7. In this system, VND6 or VND7 is expressed as a fused protein with the activation domain of the herpes virus VP16 protein and hormone-binding domain of the animal glucocorticoid receptor, and the protein's activity is induced by treatment with dexamethasone (DEX), a glucocorticoid derivative. Upon DEX treatment, transgenic Arabidopsis (Arabidopsis thaliana) plants carrying the chimeric gene exhibited transdifferentiation of various types of cells into xylem vessel elements, and the plants died. Many genes involved in xylem vessel differentiation, such as secondary wall biosynthesis and programmed cell death, were up-regulated in these plants after DEX treatment. Chemical analysis showed that xylan, a major hemicellulose component of the dicot secondary cell wall, was increased in the transgenic plants after DEX treatment. This induction system worked in poplar (Populus tremula x tremuloides) trees and in suspension cultures of cells from Arabidopsis and tobacco (Nicotiana tabacum); more than 90% of the tobacco BY-2 cells expressing VND7-VP16-GR transdifferentiated into xylem vessel elements after DEX treatment. These data demonstrate that the induction systems controlling VND6 and VND7 activities can be used as powerful tools for understanding xylem cell differentiation.
我们之前表明,编码 NAM/ATAF/CUC 结构域蛋白转录因子的 VASCULAR-RELATED NAC-DOMAIN6(VND6)和 VND7 基因是木质部导管分化的关键调节因子。在这里,我们报告了 VND6 和 VND7 的糖皮质激素介导的翻译后诱导系统。在该系统中,VND6 或 VND7 作为融合蛋白表达,与疱疹病毒 VP16 蛋白的激活结构域和动物糖皮质激素受体的激素结合结构域融合,并且该蛋白的活性通过用地塞米松(DEX)诱导,一种糖皮质激素衍生物。在用 DEX 处理后,携带嵌合基因的转基因拟南芥(Arabidopsis thaliana)植物表现出各种类型的细胞向木质部导管元素的转分化,并且植物死亡。DEX 处理后,这些植物中许多参与木质部导管分化的基因,如次生壁生物合成和程序性细胞死亡,被上调。化学分析表明,在 DEX 处理后,转基因植物中半纤维素的主要成分二糖次生细胞壁的木聚糖增加。该诱导系统在杨树(Populus tremula x tremuloides)树木和拟南芥和烟草(Nicotiana tabacum)细胞的悬浮培养物中起作用; 在 DEX 处理后,表达 VND7-VP16-GR 的超过 90%的烟草 BY-2 细胞转分化为木质部导管元素。这些数据表明,控制 VND6 和 VND7 活性的诱导系统可作为理解木质部细胞分化的有力工具。