Laboratory of Plant Molecular Physiology, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Bunkyo-ku, Tokyo, Japan.
Mol Genet Genomics. 2010 Sep;284(3):173-83. doi: 10.1007/s00438-010-0557-0. Epub 2010 Jul 15.
The transcription factor OsNAC5 in rice is a member of the plant-specific NAC family that regulates stress responses. Expression of OsNAC5 is induced by abiotic stresses such as drought, cold, high salinity, abscisic acid and methyl jasmonic acid. Transactivation assays using rice protoplasts demonstrated that OsNAC5 is a transcriptional activator, and subcellular localization studies using OsNAC5-GFP fusion proteins showed that it is localized to the nucleus. Pull-down assays revealed that OsNAC5 interacts with OsNAC5, OsNAC6 and SNAC1. To analyze the function of OsNAC5 in rice plants, we generated transgenic plants that overexpressed OsNAC5. The growth of these plants was similar to that of control plants, whereas the growth of OsNAC6-overexpressing transgenic plants was retarded. OsNAC5-overexpressing transgenic plants also had improved tolerance to high salinity compared to control plants. By microarray analysis, many stress-inducible genes, including the "late embryogenesis abundant" gene OsLEA3, were upregulated in rice plants that overexpressed OsNAC5. By gel mobility shift assay, OsNAC5 and OsNAC6 were shown to bind to the OsLEA3 promoter. Collectively, our results indicate that the stress-responsive proteins OsNAC5 and OsNAC6 are transcriptional activators that enhance stress tolerance by upregulating the expression of stress-inducible rice genes such as OsLEA3, although the effects of these proteins on growth are different. Furthermore, because OsNAC5 overexpression did not retard growth, OsNAC5 may be a useful gene that can improve the stress tolerance of rice without affecting its growth.
水稻转录因子 OsNAC5 是植物特异性 NAC 家族的一员,调节应激反应。OsNAC5 的表达受非生物胁迫如干旱、寒冷、高盐、脱落酸和茉莉酸甲酯诱导。使用水稻原生质体的转录激活测定表明,OsNAC5 是一种转录激活剂,使用 OsNAC5-GFP 融合蛋白的亚细胞定位研究表明它定位于细胞核。下拉测定显示 OsNAC5 与 OsNAC5、OsNAC6 和 SNAC1 相互作用。为了分析 OsNAC5 在水稻植株中的功能,我们生成了过表达 OsNAC5 的转基因植物。这些植物的生长与对照植物相似,而 OsNAC6 过表达转基因植物的生长则滞后。与对照植物相比,过表达 OsNAC5 的转基因植物对高盐也有更好的耐受性。通过微阵列分析,许多应激诱导基因,包括“晚期胚胎丰富”基因 OsLEA3,在过表达 OsNAC5 的水稻植物中上调。通过凝胶迁移率变动测定,表明 OsNAC5 和 OsNAC6 与 OsLEA3 启动子结合。总的来说,我们的结果表明,应激响应蛋白 OsNAC5 和 OsNAC6 是转录激活剂,通过上调应激诱导的水稻基因如 OsLEA3 的表达来增强应激耐受性,尽管这些蛋白对生长的影响不同。此外,由于 OsNAC5 的过表达不会减缓生长,因此 OsNAC5 可能是一个有用的基因,可以在不影响其生长的情况下提高水稻的应激耐受性。