生物技术通报 ›› 2021, Vol. 37 ›› Issue (12): 13-21.doi: 10.13560/j.cnki.biotech.bull.1985.2021-0497

• 研究报告 • 上一篇    下一篇

紫花苜蓿转录因子基因MsAP2的克隆及转化

石欣玥(), 尚骁尧, 周玲芳, 张铁军, 晁跃辉()   

  1. 北京林业大学草业与草原学院,北京100083
  • 收稿日期:2021-04-15 出版日期:2021-12-26 发布日期:2022-01-19
  • 作者简介:石欣玥,女,硕士研究生,研究方向:草地植物遗传育种与生物技术;E-mail: shi_xinyue@163.com
  • 基金资助:
    国家自然科学基金面上项目(31772656)

Cloning and Transformation of MsAP2 Gene in Medicago sativa

SHI Xin-yue(), SHANG Xiao-yao, ZHOU Ling-fang, ZHANG Tie-jun, CHAO Yue-hui()   

  1. College of Grassland Science of Beijing Forestry University,Beijing 100083
  • Received:2021-04-15 Published:2021-12-26 Online:2022-01-19

摘要:

紫花苜蓿(Medicago sativa)为重要的豆科牧草植物,研究紫花苜蓿转录因子基因MsAP2的功能,为探究MsAP2的信号转导网络提供理论指导和材料基础,也为紫花苜蓿生物技术育种提供一定的参考和借鉴。运用RT-PCR技术克隆MsAP2,利用DNA重组技术构建3302-3flag-AP2植物表达载体,并利用农杆菌介导法对紫花苜蓿进行遗传转化,对再生植株进行转基因鉴定、基因表达量分析、内源激素的测定(脱落酸、细胞分裂素、生长素和赤霉素)和表型鉴定。结果表明,成功获得转MsAP2紫花苜蓿植株,与野生型相比,转基因植株的MsAP2表达量和激素水平均发生明显改变,转基因植株呈现早衰性状,叶片形态和大小发生改变,根系生长受到抑制,分枝数发生改变。

关键词: 紫花苜蓿, AP2/ERF转录因子, MsAP2, 转基因

Abstract:

Alfalfa(Medicago sativa)is an important leguminous forage plant. Studying the function of MsAP2 provides theoretical guidance and material basis for exploring the signal transduction network of MsAP2,and also provides some reference for alfalfa biotechnology breeding technology. The MsAP2 was cloned by RT-PCR,and the 3302-3flag-AP2 plant expression vector was constructed by DNA recombination technology. The M. sativa was transformed by Agrobacterium-mediated method,and the regenerated plants were identified by transgene detection,gene expression analysis,determination of endogenous hormones(abscisic acid,cytokinin,auxin and gibberellin)and phenotype identification. The results showed that the MsAP2 transgenic M. sativa plants were successfully obtained. Compared with the wild type,the MsAP2 gene expression and hormone levels of transgenic plants were significantly changed. The transgenic plants demonstrated premature senescence,the leaf shape and size changed,the root growth was inhibited and the number of branches changed.

Key words: Medicago sativa L., AP2/ERF transcription factor, MsAP2 gene, transgene