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植物研究 ›› 2020, Vol. 40 ›› Issue (4): 575-582.doi: 10.7525/j.issn.1673-5102.2020.04.012

• 研究报告 • 上一篇    下一篇

毛果杨固有无序蛋白质基因克隆及胁迫响应分析

董实伟, 杨宇宁, 王乃锐, 张含国, 李淑娟   

  1. 林木遗传育种国家重点实验室, 东北林业大学, 哈尔滨 150040
  • 收稿日期:2019-12-29 出版日期:2020-07-05 发布日期:2020-06-12
  • 通讯作者: 李淑娟,E-mail:lishujuan@126.com E-mail:lishujuan@126.com
  • 作者简介:董实伟(1993-),女,硕士研究生,主要从事林木遗传育种研究。
  • 基金资助:
    黑龙江省科学基金项目(C2017008)

Gene Cloning and Stress Response Analysis of Natural Disorder Protein in Populus trichocarpa

DONG Shi-Wei, YANG Yu-Ning, WANG Nai-Rui, ZHANG Han-Guo, LI Shu-Juan   

  1. State Key Laboratory of Tree Genetics and Breeding, Northeast Forestry University, Harbin 150040
  • Received:2019-12-29 Online:2020-07-05 Published:2020-06-12
  • Supported by:
    Heilongjiang Science Fund Project(C2017008)

摘要: 为了解毛果杨中固有无序蛋白PtrIDP1(Potri.010G161200.1)基因的相关信息,探究该基因在毛果杨的不同组织、不同逆境胁迫下的表达特性,本研究根据Phytozome数据库中得到的基因全长序列设计引物,克隆得到该基因的目的片段。该基因完整的CDs区序列长度为423 bp,共编码140个氨基酸。构建亚细胞定位表达载体,瞬时转化洋葱表皮细胞,在激光共聚焦显微镜下观察显示,PtrIDP1定位在细胞核内。利用实时定量RT-PCR技术分析了PtrIDP1基因在毛果杨不同组织中的表达特异性和应对非生物胁迫的表达特性。结果表明:PtrIDP1基因在毛果杨的根、茎、叶中均有表达,其中在根部表达量最低,在茎和叶中相对表达量较高。PtrIDP1基因在高盐和干旱胁迫诱导时其表达量的变化模式不同,初步分析认为PtrIDP1基因参与了毛果杨非生物逆境胁迫的响应过程。

关键词: 毛果杨, 基因克隆, 逆境胁迫响应, 固有无序蛋白质

Abstract: In order to know the related information of PtrIDP1(Potri.010G161200.1) gene in Populus trichocarpa and to explore the expression characteristics of the gene in different tissues and under different stress, primers were designed according to the full-length sequence of the gene obtained from Phytozome database, and the target fragment of the gene was cloned. The complete CDs region of the gene is 423 bp long and encodes 140 amino acids. The expression vector of subcellular localization was constructed, and the epidermal cells of the onion were transiently transformed. The result showed that the PtrIDP1 was located in the cell nucleus under the laser confocal microscope. The expression specificity of PtrIDP1 gene in different tissues of P.trichocarpa and its response to abiotic stress were analyzed by real-time quantitative RT-PCR. The results showed that PtrIDP1 gene was expressed in the roots, stems and leaves of P.trichocarpa, but the lowest expression was in the roots, whereas the relative expression was higher in the stems and leaves. The expression patterns of PtrIDP1 gene were different under the induction of high salt and drought stress, and the preliminary analysis suggested that PtrIDP1 gene was involved in the process of responding to abiotic stress in P.trichocarpa.

Key words: Populus trichocarpa, gene cloning, stress response to adversity, intrinsically disordered proteins

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