National Key Laboratory for Rice Biology, Institute of Biotechnology, Zhejiang University, Hangzhou, 310058, China.
BMC Plant Biol. 2019 Jun 25;19(1):278. doi: 10.1186/s12870-019-1883-y.
NAC (NAM, ATAF and CUC) transcriptional factors constitute a large family with more than 150 members in rice and several members of this family have been demonstrated to play crucial roles in rice abiotic stress response. In the present study, we report the function of a novel stress-responsive NAC gene, ONAC066, in rice drought and oxidative stress tolerance.
ONAC066 was localized in nuclei of cells when transiently expressed in Nicotiana benthamiana and is a transcription activator with the binding ability to NAC recognition sequence (NACRS) and AtJUB1 binding site (JBS). Expression of ONAC066 was significantly induced by PEG, NaCl, HO and abscisic acid (ABA). Overexpression of ONAC066 in transgenic rice improved drought and oxidative stress tolerance and increased ABA sensitivity, accompanied with decreased rate of water loss, increased contents of proline and soluble sugars, decreased accumulation of reactive oxygen species (ROS) and upregulated expression of stress-related genes under drought stress condition. By contrast, RNAi-mediated suppression of ONAC066 attenuated drought and oxidative stress tolerance and decreased ABA sensitivity, accompanied with increased rate of water loss, decreased contents of proline and soluble sugars, elevated accumulation of ROS and downregulated expression of stress-related genes under drought stress condition. Furthermore, yeast one hybrid and chromatin immunoprecipitation-PCR analyses revealed that ONAC066 bound directly to a JBS-like cis-elements in OsDREB2A promoter and activated the transcription of OsDREB2A.
ONAC066 is a nucleus-localized transcription activator that can respond to multiple abiotic stress factors. Functional analyses using overexpression and RNAi-mediated suppression transgenic lines demonstrate that ONAC066 is a positive regulator of drought and oxidative stress tolerance in rice.
NAC(NAM、ATAF 和 CUC)转录因子构成了一个拥有超过 150 个成员的大家族,在水稻中,该家族的几个成员已被证明在非生物胁迫响应中起着关键作用。本研究报告了一个新的应激响应 NAC 基因 ONAC066 在水稻耐旱和抗氧化胁迫中的功能。
ONAC066 在瞬时表达于黄花烟时定位于细胞核中,是一种转录激活子,具有与 NAC 识别序列(NACRS)和 AtJUB1 结合位点(JBS)结合的能力。ONAC066 的表达在 PEG、NaCl、HO 和脱落酸(ABA)的处理下显著上调。ONAC066 在转基因水稻中的过表达提高了耐旱和抗氧化胁迫耐受性,并增加了 ABA 敏感性,同时降低了失水率,提高了脯氨酸和可溶性糖的含量,减少了活性氧(ROS)的积累,并上调了干旱胁迫下的应激相关基因的表达。相比之下,RNAi 介导的 ONAC066 抑制削弱了耐旱和抗氧化胁迫耐受性,并降低了 ABA 敏感性,同时增加了失水率,降低了脯氨酸和可溶性糖的含量,增加了 ROS 的积累,并下调了干旱胁迫下的应激相关基因的表达。此外,酵母单杂交和染色质免疫沉淀-PCR 分析表明,ONAC066 直接结合到 OsDREB2A 启动子中的一个 JBS 样顺式元件上,并激活了 OsDREB2A 的转录。
ONAC066 是一种定位于细胞核的转录激活子,可以响应多种非生物胁迫因子。利用过表达和 RNAi 介导的抑制转基因系进行的功能分析表明,ONAC066 是水稻耐旱和抗氧化胁迫的正调控因子。