Regeneration of Chinese cabbage transgenic plants expressing antibacterial peptide gene and cowpea trypsin inhibitor gene |
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| Authors: | Junliang Zhao Aihua Liang Zhen Zhu Yixiong Tang |
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| Institution: | (1) Institute of Vegetables, Shanxi Academy of Agricultural Sciences, Taiyuan, 030031, China;(2) Key Laboratory of Chemical Biology and Molecular Engineering of Ministry of Education, Shanxi University, Taiyuan, 030009, China;(3) Institute of Genetics, Chinese Academy of Science, Beijing, 100101, China;(4) Center of Biotechnology, Chinese Academy of Agricultural Science, Beijing, 100081, China |
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| Abstract: | Summary An efficient transformation system for Chinese cabbage cotyledon explants was developed using Agrobacterium tumefaciens strains LBA4404 harbouring the plasmid pMOG 411 and the plasmid pBinΩSCK respectively. Various factors affecting the transformation efficiency and subsequent regeneration were identified. The age of seedlings, growth conditions and status of Agrobacterium suspension, preculture of explants, cocultivation time, ratio between Agrobacterium and explants, acetosyringone and concentration of kanamycin had a significant influence on transformation frequency and plant regeneration. The presences of the antibacterial peptide gene and the cowpea trypsin inhibitor gene in those selected shoots in kanamycin medium were confirmed by PCR, Southern blotting and Northern blotting, The frequency of regeneration from cotyledons was analyzed in eight parental lines of Chinese cabbage and five lines were suitable for obtaining transformed plants, among which Qingmaye C and Qingmaye D were more responsive than other lines. |
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| Keywords: | Agrobacterium tumefaciens anti-bacterial peptide Chinese cabbage cowpea trypsin inhibitor transformation |
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