Wang Zhong, Yang Jinchu, Gao Qian, He Shun, Xu Yongming, Luo Zhaopeng, Liu Pingping, Wu Mingzhu, Xu Xin, Ma Lanxin, Zhang Zhan, Yang Yongfeng, Yang Jun
China Tobacco Gene Research Center, Zhengzhou Tobacco Research Institute of CNTC, Zhengzhou 450001, China.
Technology Center, China Tobacco Henan Industrial Co., Ltd., Zhengzhou 450000, China.
Plant Sci. 2023 Sep;334:111772. doi: 10.1016/j.plantsci.2023.111772. Epub 2023 Jun 16.
The AP2/ERF (APETALA2/ETHYLENE RESPONSE FACTOR) transcription factors play multiple roles in modulating the biosynthesis of diverse specialized metabolites in response to various environmental stresses. ERF13 has been shown to participate in plant resistance to biotic stress as well as in repressing the synthesis of fatty acid. However, its full roles in regulating plant metabolism and stress resistance still remains to be further studied. In this study, we identified two NtERF genes from N. tabacum genome that belong to Ⅸa subgroup of ERF family. Over-expression and knock-out of NtERF13a showed that NtERF13a could enhance plant resistance to salt and drought stresses, as well as promoted the biosynthesis of chlorogenic acid (CGA), flavonoids, and lignin in tobacco. Transcriptome analysis between WT and NtERF13a-OE plants revealed 6 differentially expressed genes (DEGs) that encode enzymes catalyzing the key steps of phenylpropanoid pathway. Chromatin immunoprecipitation, Y1H, and Dual-Luc assays further clarified that NtERF13a could directly bind to the fragments containing GCC box or DRE element in the promoters of NtHCT, NtF3'H, and NtANS genes to induce the transcription of these genes. Knock-out of NtHCT, NtF3'H, or NtANS in the NtERF13a-OE background significantly repressed the increase of phenylpropanoid compound contents caused by over-expression of NtERF13a, indicating that the promotion of NtERF13a on the phenylpropanoid compound contents depends on the activity of NtHCT, NtF3'H, and NtANS. Our study demonstrated new roles of NtERF13a in promoting plant resistance to abiotic stresses, and provided a promising target for modulating the biosynthesis of phenylpropanoid compounds in tobacco.
AP2/ERF(APETALA2/乙烯响应因子)转录因子在响应各种环境胁迫时,对多种特殊代谢产物的生物合成具有多种调节作用。ERF13已被证明参与植物对生物胁迫的抗性以及抑制脂肪酸的合成。然而,其在调节植物代谢和抗逆性方面的完整作用仍有待进一步研究。在本研究中,我们从烟草基因组中鉴定出两个属于ERF家族Ⅸa亚组的NtERF基因。NtERF13a的过表达和敲除表明,NtERF13a可以增强植物对盐胁迫和干旱胁迫的抗性,还能促进烟草中绿原酸(CGA)、类黄酮和木质素的生物合成。野生型和NtERF13a过表达植株之间的转录组分析揭示了6个差异表达基因(DEG),这些基因编码催化苯丙烷途径关键步骤的酶。染色质免疫沉淀、酵母单杂交和双荧光素酶实验进一步表明,NtERF13a可以直接结合到NtHCT、NtF3'H和NtANS基因启动子中含有GCC盒或DRE元件的片段上,从而诱导这些基因的转录。在NtERF13a过表达背景下敲除NtHCT、NtF3'H或NtANS,显著抑制了NtERF13a过表达导致的苯丙烷类化合物含量的增加,这表明NtERF13a对苯丙烷类化合物含量的促进作用依赖于NtHCT、NtF3'H和NtANS的活性。我们的研究证明了NtERF13a在促进植物对非生物胁迫抗性方面的新作用,并为调控烟草中苯丙烷类化合物的生物合成提供了一个有前景的靶点。