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植物研究 ›› 2014, Vol. 34 ›› Issue (4): 516-523.doi: 10.7525/j.issn.1673-5102.2014.04.015

• 论文 • 上一篇    下一篇

CYP79F1基因对西兰花的遗传转化的研究

王红波;李成;曹阳理惠;李晶*   

  1. 东北农业大学农业生物功能基因重点实验室,哈尔滨 150030
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2014-07-20 发布日期:2014-07-20
  • 通讯作者: 李晶
  • 基金资助:
     

Genetic Transformation of CYP79F1 Gene in Brassica oleracea var. italica

WANG Hong-Bo;LI Cheng;CAO Yang-Li-Hui;LI Jing*   

  1. Key Laboratory of Agricultural Biological Functional Genes,Northeast Agricultural University,Harbin 150030
  • Received:1900-01-01 Revised:1900-01-01 Online:2014-07-20 Published:2014-07-20
  • Contact: LI Jing
  • Supported by:
     

摘要: 建立有效的遗传转化体系是基因工程手段改良植物性状的重要前提。本研究对西兰花遗传转化体系进行了初步研究,成功地建立了农杆菌介导的西兰花遗传转化体系。以7日龄无菌苗的下胚轴为外植体,预培养2~3 d后进行农杆菌的侵染,共培养后将外植体转移至无筛选压力的脱菌培养基中,暗培养7 d后置于含有10 mg·L-1卡那霉素的筛选培养基中继续培养,待不定芽生长至2~3 cm时转入0.1 mg·L-1 IBA的生根培养基中诱导不定根,最后进行驯化和移栽,PCR分子鉴定及外源基因表达水平检测结果证明,我们已经成功地将芥子油苷合成关键酶基因CYP79F1基因转入西兰花商业品种“青秀”中,利用HPLC检测转基因植株中短链脂肪族芥子油苷的含量显著增加。

关键词: 西兰花, 农杆菌, 遗传转化体系, 芥子油苷, CYP79F1

Abstract: Establishment of effective transformation system is an important precondition to improve plant traits by genetic engineering. In this study, an agrobacterium-mediated transformation system of Brassica oleracea var. italica was established. Hypocotyls of 7-day-old seedlings were used as explants and preincubated for 2-3 d and infected by agrobacterium-tumefaciens. After co-incubation, the explants were transferred to degerming medium without selective agent to induce adventitious buds. Following the incubation in darkness for 7 d, the explants were transferred to selective medium containing 10 mg·L-1 kanamycin. When the adventitious buds grew 2-3 cm long, they were moved to rooting medium with 0.1 mg·L-1 IBA to induce adventitious roots. Finally, the domestication and transplantation were performed. PCR detection demonstrated that glucosinolate biosynthetic gene CYP79F1 was successfully transferred into the genome of B.oleracea var. italica commercial cultivar “Qingxiu”. HPLC determination proved that short chain aliphatic glucosinolates significantly increased in transgenic plants.

Key words: Brassica oleracea var. italica, Agrobacterium-tumefaciens, genetic transformation system, glucosinolate, CYP79F1

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