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植物研究 ›› 2020, Vol. 40 ›› Issue (4): 593-601.doi: 10.7525/j.issn.1673-5102.2020.04.014

• 研究报告 • 上一篇    

小黑杨PsnHB13基因在烟草中的遗传转化与功能分析

肖迪, 刘轶, 李开隆, 郑密, 曲冠证   

  1. 东北林业大学林木遗传育种国家重点实验室, 哈尔滨 150040
  • 收稿日期:2019-12-05 发布日期:2020-06-12
  • 通讯作者: 郑密,E-mail:zhengmi0207@163.com E-mail:zhengmi0207@163.com
  • 作者简介:肖迪(1993-),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(31700596)

Genetic Transformation and Function Analysis of PsnHB13 Gene Isolated from Populu ssimonii×P.nigra in Nicotiana tabacum

XIAO Di, LIU Yi, LI Kai-Long, ZHENG Mi, QU Guan-Zheng   

  1. State Key Laboratory of Tree Genetics and Breeding, Northeast Forestry University, Harbin 150040
  • Received:2019-12-05 Published:2020-06-12
  • Supported by:
    National Natural Science Foundation of China(31700596)

摘要: 克隆小黑杨HD-ZIP家族基因PsnHB13,对该基因进行生物信息学分析、过表达载体构建、烟草的遗传转化。结果表明小黑杨PsnHB13基因cDNA全长870 bp,编码289个氨基酸。成功构建植物过表达载体pROKⅡ-PsnHB13,并通过农杆菌介导的叶盘法将外源基因转入野生型烟草。检测结果显示PsnHB13已成功整合入烟草基因组中,并在mRNA水平表达。通过观察转基因烟草的生长,发现过表达PsnHB13基因的烟草与野生型相比出现叶面积变小、叶型变长,根系生长缓慢,花朵变小等明显表型。说明PsnHB13基因主要对烟草叶片、根系及花的生长发育起到负调控作用。本文为研究PsnHB13基因对杨树生长发育的影响提供理论基础。

关键词: PsnHB13, 烟草, 叶型, 根长

Abstract: Cloning of the gene PsnHB13 of the HD-ZIP family, bioinformatics analysis, the construction of overexpression vector and performing of tobacco genetic transformation. The result shows that the cDNA of PsnHB13 was 870 bp and encode 289 amino acids. In our study, PsnHB13 was amplified from Populu ssimonii×P.nigra cDNA by PCR. Plant overexpression vector(pROKII-PsnHB13) was constructed and transformed into wild-type tobacco by agrobacterium-mediated leaf disc transformation. The transgenic plants were confirmed by PCR, and the result showed PsnHB13 gene was integrated into tobacco genome. The result of qRT-PCR detection of T2 generation plants showed the mRNA of PsnHB13 was transcribed in transgenic tobaccos. By observation, PsnHB13 overexpressed leads the differences of leaf, flower and root in tobacco. Thus, PsnHB13 plays a negative role in the growth of tobacco leaves, roots and flowers, and provides a theoretical basis for poplar growth and development.

Key words: PsnHB13, tobacco, leaf shape, root length

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