欢迎访问《植物研究》杂志官方网站,今天是 分享到:

植物研究 ›› 2016, Vol. 36 ›› Issue (3): 388-394.doi: 10.7525/j.issn.1673-5102.2016.03.011

• 研究报告 • 上一篇    下一篇

拟南芥AtPI基因植物表达载体的构建及其在烟草中的遗传转化

刘彩霞, 郑唐春, 代丽娟, 刘轶, 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:2015-11-25 出版日期:2016-05-15 发布日期:2016-05-20
  • 通讯作者: 曲冠证,E-mail:quguanzheng@yahoo.com E-mail:quguanzheng@yahoo.com
  • 作者简介:刘彩霞(1990-),女,硕士研究生,主要从事林木遗传育种研究。
  • 基金资助:
    教育部新世纪优秀人才支持计划项目(NCET-12-0808);国家自然科学基金项目(31370661)资助

Construction of Plant Expression Vector and Genetic Transformation Analysis of Arabidopsis thaliana AtPI Gene in Nicotiana tabacum

LIU Cai-Xia, ZHENG Tang-Chun, DAI Li-Juan, LIU Yi, QU Guan-Zheng   

  1. State Key Laboratory of Tree Genetics and Breeding, Northeast Forestry University, Harbin 150040
  • Received:2015-11-25 Online:2016-05-15 Published:2016-05-20

摘要: 花是被子植物主要的繁殖器官,在繁育后代的过程中,发挥着极其重要的作用,PISTILLATA(PI)基因作为控制花器官发育的B类功能基因中的一员,在花器官发育中起到重要的作用。为探究PI基因在花瓣和雄蕊发育中的功能,本文以拟南芥(Arabidopsis thaliana)中的PI基因作为研究对象,利用PCR技术从拟南芥花序cDNA扩增出AtPI基因,构建植物表达载体(pROKⅡ-AtPI)并进行烟草(Nicotiana tobacum)转化。转基因植株的PCR检测结果表明,AtPI基因已经整合到了烟草基因组中。在T2代植株中,通过实时定量荧光PCR检测显示,AtPI在mRNA水平也均有表达。过量表达PI的转基因烟草在花器官中存在明显表型,与野生型相比主要表现为转基因植株花冠变小,雄蕊缩短,果实畸形且子房基部比野生型长5~10mm,上述结果表明AtPI基因是特异性参与雄蕊和花瓣的发育并起着至关重要的作用。

关键词: 拟南芥, PISTILLATA(PI), 烟草, 雄蕊, 花瓣, 子房

Abstract: The flower is the main reproductive organs of the plants, plays an extremely important role in the process of breeding and the PISTILLATA(PI) gene, and as a member of the B functional gene, plays an important role in floral organ development. In order to explore the function of PI gene in the development of petals and stamens, the PI gene in Arabidopsis thaliana was used as the research object, and the AtPI gene was amplified from cDNA by PCR, and the plant expression vector(pROKⅡ-AtPI) was constructed and transformed into tobacco. By PCR, the AtPI gene was integrated into the tobacco genome. The T2 generation plants were detected by qRT-PCR, and AtPI was also expressed at the level of mRNA. Transgenic tobacco plants exist obvious phenotype compared to wild-type tobacco, such as smaller corolla, shorter stamens, and abnormal ovary, which is 5-10 mm longer than wild-type tobacco. The specific participation in the development of stamens, petals and ovary meanwhile plays a crucial role is gene AtPI.

Key words: Arabidopsis thalian, PISTILLATA(PI), Nicotiana tabacum, stamen, petal, ovary

中图分类号: