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植物研究 ›› 2019, Vol. 39 ›› Issue (4): 547-556.doi: 10.7525/j.issn.1673-5102.2019.04.009

• 研究报告 • 上一篇    下一篇

84K杨4CL3/4CL5基因克隆及生物信息学分析

孙晓莎, 王遂, 赵曦阳, 曲冠证   

  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:2019-01-02 出版日期:2019-07-05 发布日期:2019-07-03
  • 通讯作者: 曲冠证 E-mail:quguanzheng@yahoo.com
  • 作者简介:孙晓莎(1991-),女,硕士研究生,主要从事林木遗传育种研究。
  • 基金资助:
    国家自然科学基金面上项目(31500549,31670675)

Cloning and Bioinformatics Analysis 4CL3/4CL5 Gene of Populus alba×P.glandulosa

SUN Xiao-Sha, WANG Sui, ZHAO Xi-Yang, QU Guan-Zheng   

  1. State Key Laboratory of Tree Genetics and Breeding, Northeast Forestry University, Harbin 150040
  • Received:2019-01-02 Online:2019-07-05 Published:2019-07-03
  • Supported by:
    National Natural Science Foundation of China(31500549,31670675)

摘要: 木质素是木材的重要组成成分,主要起机械支持、水分运输及防御病虫害等作用。4CL酶是控制木质素合成途径中的关键酶之一。利用PCR技术从84K杨幼叶cDNA中克隆得到Pag4CL3/4CL5基因,GenBank 登录号分别为MK183033(Pag4CL3)及MK183034(Pag4CL5)。利用生物信息学软件,对这两个基因的功能和特征进行分析。结果发现84K杨中4CL3/4CL5蛋白与毛果杨中4CL3/4CL5蛋白相似度高达97%;此外,均含有SSGTTGLPKGV和GEICIRG两个保守基序;亚细胞定位预测显示Pag4CL3/4CL5蛋白主要定位在内质网中,推测可能是一种膜蛋白;蛋白的亲水性预测Pag4CL3/4CL5均为亲水性蛋白。通过qRT-PCR分析Pag4CL3/4CL5在不同组织部位中的表达差异,结果发现Pag4CL3在叶及茎中表达量较高,在根和顶芽中表达量较低;Pag4CL5在叶及根中表达量较高,在茎和顶芽中表达量较低,两个同源基因表达具有组织部位的差异性。Pag4CL3和Pag4CL5可能在叶中共同行使功能,Pag4CL5主要在根中行使功能,Pag4CL3主要在茎中行使功能。

关键词: 84K杨, 4CL基因, 克隆, 生物信息学分析

Abstract: Lignin is an important component of wood, mainly for mechanical support, water transport and defense against pests and diseases. The 4CL enzyme is one of the key enzymes in controlling the pathway of lignin synthesis. The Pag4CL3/4CL5 gene was cloned from 84K poplar leaf cDNA by PCR. GenBank accession numbers are MK183033(Pag4CL3) and MK183034(Pag4CL5). The functions and characteristics of the two genes were analyzed using bioinformatics software. The similarity of the 4CL3/4CL5 proteins in 84K poplar and the 4CL3/4CL5 proteins in Populus trichocarpa were 97%,and they contained two conserved motifs of SSGTTGLPKGV and GEICIRG; subcellular localization showed the main localization of Pag4CL3/4CL5 proteins. In the endoplasmic reticulum, it may be a membrane protein and both are hydrophilic proteins. The expression of the Pag4CL3/4CL5 gene in different tissues was analyzed by qRT-PCR. The results showed that the Pag4CL3 gene was mainly expressed in leave and stem, and the expression level in root and top buds was low. The Pag4CL5 gene showed high expression level in leave and root, and low level in stem and apical buds. Pag4CL3 and Pag4CL5 may function together in the leaves, however, Pag4CL5 ismainly function in the root, and Pag4CL3 mainly function in the stems.

Key words: Populus alba×P. glandulosa, 4CL gene, cloning, bioinformatics analysis

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