生物技术通报 ›› 2022, Vol. 38 ›› Issue (7): 186-193.doi: 10.13560/j.cnki.biotech.bull.1985.2021-1072

• 研究报告 • 上一篇    下一篇

绞股蓝GpMIR156aGpMIR166b的克隆与功能分析

于秋琳1(), 马婧怡1, 赵盼1, 孙鹏芳1, 何玉美1, 刘世彪2, 郭惠红1()   

  1. 1.北京林业大学生物科学与技术学院,北京 100083
    2. 吉首大学生物资源与环境科学学院,吉首 416000
  • 收稿日期:2021-08-20 出版日期:2022-07-26 发布日期:2022-08-09
  • 作者简介:于秋琳,女,硕士研究生,研究方向:植物发育生物学;E-mail: YuQiulin@163.com
  • 基金资助:
    国家自然科学基金项目(31760044);国家自然科学基金项目(31870650)

Cloning and Functional Analysis of Gynostemma pentaphyllum GpMIR156a and GpMIR166b

YU Qiu-lin1(), MA Jing-yi1, ZHAO Pan1, SUN Peng-fang1, HE Yu-mei1, LIU Shi-biao2, GUO Hui-hong1()   

  1. 1. College of Biological Sciences and Biotechnology,Beijing Forestry University,Beijing 100083
    2. College of Biology and Environmental Sciences,Jishou University,Jishou 416000
  • Received:2021-08-20 Published:2022-07-26 Online:2022-08-09

摘要:

本研究旨在深入了解药用植物绞股蓝地上茎向根状茎转变过程中两个重要的miRNAs(GpmiR156a和GpmiR166b)在调控植物生长发育方面的作用。采用同源克隆的方法获得GpMIR156aGpMIR166b基因,进行了生物信息学分析。通过构建植物过表达载体并成功转化拟南芥,研究GpMIR156aGpMIR166b的过表达对植株表型的影响。GpMIR156a过表达导致拟南芥幼苗叶片数量和分枝增多,并加速了种子萌发、促进根的伸长,表明GpmiR156a能够促进营养器官发育和种子萌发,但抑制童期向成熟期发育的转变。然而,GpMIR166b过表达导致拟南芥植株矮小及早熟性状即花序茎上毛状体数量增多,并延缓种子的萌发,但促进根伸长,表明GpmiR166b在一定程度上影响营养器官的发育和延缓种子萌发,但能促进童期向成熟期发育的转变。研究结果拓宽了miR156a和miR166b在调控植物生长发育方面的认识。

关键词: 绞股蓝, GpmiR156a, GpmiR166b, 生物信息学分析, 功能分析

Abstract:

This study aims to deeply understand the role of two important miRNAs involved in the Gynostemma pentaphyllum aerial stem-to-rhizome transition,GpmiR156a and GpmiR166b,in regulating plant growth and development. GpMIR156a and GpMIR166b genes were obtained by homologous cloning and analyzed by bioinformatics. The effects of GpMIR156a and GpMIR166boverexpression on plant phenotype were investigated by constructing their overexpression vectors and then transforming Arabidopsis thaliana. The overexpression of GpMIR156a increased the number of leaves and branches of A. thaliana seedlings,and accelerated seed germination and root elongation,indicating that GpmiR156a promoted vegetative organ development and seed germination,but inhibited juvenile-to-adult transition. However,the overexpression of GpMIR166b led to the premature and dwarf plants,i.e.,increase in the number of trichomes on the inflorescence stems,as well as delayed seed germination but promoted root elongation. These findings indicated that GpmiR166b affected the normal development of vegetative organs and delayed seed germination to a certain extent,whereas promoted juvenile-to-adult transition. The results of this study broaden the understanding of miR156a and miR166b in regulating plant growth and development.

Key words: Gynostemma pentaphyllum, GpmiR156a, GpmiR166b, bioinformatics analysis, functional analysis