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Bulletin of Botanical Research ›› 2014, Vol. 34 ›› Issue (4): 516-523.doi: 10.7525/j.issn.1673-5102.2014.04.015

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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
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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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