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植物研究 ›› 2019, Vol. 39 ›› Issue (4): 521-528.doi: 10.7525/j.issn.1673-5102.2019.04.006

• 研究报告 • 上一篇    下一篇

84K杨PagC3H3基因表达模式分析

樊二勤, 刘彩霞, 付鹏跃, 杨传平, 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:2019-01-03 出版日期:2019-07-05 发布日期:2019-07-03
  • 通讯作者: 杨传平 E-mail:yangcp@nefu.edu.cn
  • 作者简介:樊二勤(1992-),男,硕士研究生,主要从事林木遗传育种研究。
  • 基金资助:
    国家自然科学基金项目(31670675);教育部新世纪优秀人才支持计划项目(NCET-12-0808)

Expression Pattern of PagC3H3 Gene in Populus alba×P.glandulosa

FAN Er-Qin, LIU Cai-Xia, FU Peng-Yue, YANG Chuan-Ping, QU Guan-Zheng   

  1. State Key Laboratory of Tree Genetics and Breeding, Northeast Forestry University, Harbin 150040
  • Received:2019-01-03 Online:2019-07-05 Published:2019-07-03
  • Supported by:
    National Natural Science Foundation of China(31670675);Ministry of Education New Century Excellent Talent Support Program(NCET-12-0808)

摘要: 木质素作为木材的主要组成成分,通常是由3种单体聚合而成,在其生物合成过程中,共有10个酶家族参与负责将苯丙胺酸转化为单体木质素,其中C3H是在对-香豆酰辅酶A(p-coumaroyl CoA)到咖啡酰辅酶A(caffeoyl CoA)的羟基化过程和G/S单体形成中的关键控制酶类,探究PagC3H3基因表达模式,对于进一步了解该基因功能具有重要意义。该研究通过定量PCR对PagC3H3基因的组织特异性表达进行分析;克隆得到了长度为2 035 bp的PagC3H3的启动子序列,预测含有多个顺式作用元件;同时,将获得的PagC3H3的启动子序列构建植物表达载体pBI121-PagC3H3pro::GUS,进行拟南芥瞬时转化,结果显示PagC3H3基因在84K杨的根、中部茎节和基部茎节中的表达量较高;瞬时转化拟南芥,GUS染色表明:在下胚轴和根中GUS活性较强,由此推测PagC3H3基因在木质素合成过程中发挥作用。

关键词: 84K杨, PagC3H3, 启动子, 拟南芥, 瞬时转化, GUS检测

Abstract: As the main component of wood, lignin is usually polymerized from three monomers. During its biosynthesis, a total of ten enzyme families are involved in the conversion of phenylalanine to monomeric lignin, of which C3H is the key control enzymes in the hydroxylation process of p-coumaroyl CoA to caffeoyl CoA and in the formation of G/S monomer. Exploring the expression pattern of PagC3H3 gene is of great significance for further understanding of the function of this gene. In this study, the tissue-specific expression of PagC3H3 gene was analyzed by quantitative PCR; the promoter sequence of PagC3H3 with a length of 2 035 bp was cloned, and it was predicted to contain multiple cis-acting elements. At the same time, the promoter sequence of PagC3H3 was constructed to construct a plant expression vector pBI121-PagC3H3pro::GUS for transient transformation of Arabidopsis thaliana. The expression of PagC3H3 gene was higher in the roots, middle stem segments and basal stem segments of Populus alba×P. grandulos; transient transformation of A. thaliana showed that GUS activity was stronger in hypocotyls and roots, suggesting that PagC3H3 gene plays a role in lignin synthesis.

Key words: Populus alba×P. grandulos, PagC3H3 gene, promoter, Arabidopsis thaliana, transient transformation, GUS detection

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