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草业学报 ›› 2022, Vol. 31 ›› Issue (9): 155-167.DOI: 10.11686/cyxb2021382

• 研究论文 • 上一篇    

多年生黑麦草LpPIL5基因特征分析及转录调控

姚佳明(), 何悦, 郝欢欢, 黄心如, 张敬(), 徐彬   

  1. 南京农业大学草业学院,江苏 南京 210095
  • 收稿日期:2021-10-28 修回日期:2021-11-29 出版日期:2022-09-20 发布日期:2022-08-12
  • 通讯作者: 张敬
  • 作者简介:Corresponding author. E-mail: nauzj@njau.edu.cn
    姚佳明(1996-),男,河北保定人,在读硕士。E-mail: 873606461@qq.com
  • 基金资助:
    江苏省农业科技自主创新资金[CX(21)3004],国家自然科学基金项目(31802117);中央高校基本科研业务费(KJQN201947)

Characterization and transcriptional regulation analysis of the LpPIL5 gene in perennial ryegrass

Jia-ming YAO(), Yue HE, Huan-huan HAO, Xin-ru HUANG, Jing ZHANG(), Bin XU   

  1. College of Agro-grassland,Nanjing Agricultural University,Nanjing 210095,China
  • Received:2021-10-28 Revised:2021-11-29 Online:2022-09-20 Published:2022-08-12
  • Contact: Jing ZHANG

摘要:

光敏色素互作因子(PIFs)属于bHLH家族的一个亚家族,在植物光信号、激素信号传导和调控植物耐逆性等方面发挥重要作用。本研究从多年生黑麦草中克隆出一个PIF基因,其与拟南芥AtPIF5的亲缘关系最近,因此命名为LpPIL5-likeLpPIL5)。LpPIL5基因编码区序列(CDS)全长为1410 bp,具有5个外显子,编码470个氨基酸,具有典型的bHLH结构域和APB功能域,且其蛋白定位于细胞核。LpPIL5的启动子区域(1500 bp)具有ABRE、MBS和G-box等多种光、激素和逆境响应顺式作用元件。表达模式分析结果显示LpPIL5在叶片中表达量较高,而在根、茎、叶鞘等部位表达量较低。LpPIL5基因的表达受昼夜节律调控,且在光下的表达量显著高于黑暗。此外,聚乙二醇(PEG)、NaCl、CdCl2和高温逆境及6-苄基腺嘌呤(6-BA)和脱落酸(ABA)等激素处理显著抑制了叶片中LpPIL5基因表达,而在根中,LpPIL5基因的相对表达量在处理的12 h后明显增加,处理24 h时,显著高于对照。

关键词: 多年生黑麦草, 碱性螺旋--螺旋, 光敏色素互作因子, 亚细胞定位, 表达分析

Abstract:

Phytochrome-interacting factors (PIFs), a subfamily of the bHLH family, play important roles in plant light and hormone signaling, and stress tolerance. In the current study, a PIF gene was cloned from perennial ryegrass (Lolium perenne), and its genetic sequence was most closely related to Arabidopsis thaliana AtPIF5, therefore, it was named LpPIL5-likeLpPIL5). The coding region sequence (CDS) of this LpPIL5 gene is 1410 bp, containing 5 exons, encoding 470 amino acids. The protein it codes for has a typical bHLH domain and APB motif and is localized in the nucleus. The promoter region (1500 bp) of LpPIL5 has various cis-elements, including ABRE, MBS, G-box, and others, related to light, hormone, and stress responses. The results of transcription analysis showed that relative expression of LpPIL5 was high in leaves, but low in roots, stems, and leaf sheaths. The transcription of LpPIL5 was also regulated by circadian rhythm, and was significantly higher in light than in dark. In addition, its relative expression levelin leaves was significantly inhibited under polyethylene glycol (PEG), NaCl, CdCl2, high temperature stresses, and by N-(phenylmethyl)-9H-purin-6-amine (6-BA) and abscisic acid (ABA) treatments. In roots, the relative expression of the LpPIL5 gene increased significantly after 12 h of these treatments, and became significantly higher than in control plants after 24 h of treatment.

Key words: perennial ryegrass, basic helix-loop-helix, phytochrome-interacting factors, subcellular localization, transcription analysis