植物学报 ›› 2012, Vol. 47 ›› Issue (5): 462-473.DOI: 10.3724/SP.J.1259.2012.00462

• 研究报告 • 上一篇    下一篇

落叶松体胚发育中5个miRNA前体与成熟体的表达

张俊红1,2, 张守攻1, 吴涛1, 韩素英3, 杨文华1, 齐力旺1*   

  1. 1中国林业科学研究院林业研究所细胞生物学实验室, 北京 100091;
    2浙江农林大学亚热带森林培育国家重点实验室培育基地, 临安 311300;
    3中国林业科学研究院森林生态环境与保护研究所, 北京 100091
  • 收稿日期:2012-04-06 修回日期:2012-05-24 出版日期:2012-09-01 发布日期:2012-07-24
  • 通讯作者: 齐力旺
  • 基金资助:

    国家自然科学基金项目;国家“973”计划项目

Expression Analysis of Five MiRNAs and Their Precursors During Somatic Embryogenesis in Larch

Junhong Zhang1,2, Shougong Zhang1, Tao Wu1, Suying Han3, Wenhua Yang1, Liwang Qi1*   

  1. 1Laboratory of Cell Biology, Research Institute of Forestry, Chinese Academy of Forestry, Beijing 100091, China;

    2The Nurturing Station for the State Key Laboratory of Subtropical Silviculture, Zhejiang Agricultural and Forestry University, Lin’an 311300, China

    3Research Institute of Forest Ecology, Environment and Protection, Chinese Academy of Forestry, Beijing 100091, China
  • Received:2012-04-06 Revised:2012-05-24 Online:2012-09-01 Published:2012-07-24
  • Contact: Liwang Qi

摘要: 利用同源比对或RACE克隆了5个落叶松(Larix leptolepis) miRNA前体。结果显示, 在各物种miRNA前体间, 成熟序列高度相似, 但其它序列相似度差异大, 序列相似度与亲缘关系有关。采用qRT-PCR分析了5个miRNA、前体和靶基因在落叶松体胚8个发育阶段的表达变化。结果显示, miRNA表达最高峰出现在后期子叶胚, 暗示与促进胚胎休眠有关; 表达次高峰出现时期不同, 表现为miR397和miR408在PEMIII, miR398在早期单胚, miR156和miR166在早期子叶胚, 表明其与保持薄细胞壁、质子传递、顶端分生组织形成等调控有关。miRNA成熟体表达与前体含量不呈线性相关, 可能受多重调控。研究结果对于阐明MIR基因进化、表达调控及在体胚发育中的调控功能具有重要理论意义。

Abstract: We used the homolog search strategy or RACE to obtain 5 precursor microRNAs (miRNAs) of larch, Larix leptolepis. Mature miRNAs shared high similarity among plant species, and non-mature sequences showed distinct similarity related to genetic relationships. Expression profiles of the 5 precursor and mature miRNAs were examined during 8 developmental stages with quantitative RT-PCR. The expression of precursor and mature miRNAs changed dynamically during developmental stages, with major peaks in expression for mature miRNAs in late cotyledonous embryos, which might be associated with promoting embryo dormancy. However, miR397 and miR408 showed minor peaks in expression at PEMIII, miR398 at the early single embryo stage, and miR156 and miR166 at the early cotyledonous embryo stage, which might be related to regulation to maintain a thin cell wall, proton transfer between photo systems, and formation of shoot meristem. However, the expression of mature miRNAs was not linearly associated with that of precursor miRNAs, which might be regulated at several levels. Our study sheds light on miRNA evolution, expression regulation of mature miRNAs, and regulation of somatic embryogenesis in larch.