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Direct adventitious shoot regeneration system of Euonymus fortunei var. radicans and its genetic transformation mediated by Agrobacterium tumefaciens

Published online by Cambridge University Press:  01 October 2008

Shang Ai-Qin
Affiliation:
Department of Ornamental Horticulture and Landscape Architecture, China Agriculture University, Beijing 100094, China College of Horticulture, Hebei Agricultural University, Baoding 071001, China
Chen Ying
Affiliation:
Department of Ornamental Horticulture and Landscape Architecture, China Agriculture University, Beijing 100094, China
Zhao Liang-Jun*
Affiliation:
Department of Ornamental Horticulture and Landscape Architecture, China Agriculture University, Beijing 100094, China
Tian Ying-Chuan
Affiliation:
Institute of Microbiology, Chinese Academy of Sciences, Beijing 100080, China
*
*Corresponding author. E-mail: [email protected]

Abstract

Using hypocotyls as explants, the adventitious shoots of Euonymus fortunei var. radicans were differentiated directly from basal Murashige and Skoog (MS) medium supplemented with different plant growth regulators. The highest regeneration frequency was obtained with MS medium containing 0.5 mg/l 6-benzylaminopurine (BAP) and 0.01 mg/l α-naphthalene acetic acid (NAA). A regeneration frequency of 92% and 4.2 shoots per explant were obtained after 30 days of culture. The binary vector pBCGm, containing Galanthus nivalis agglutinin (GNA) gene, was introduced into Agrobacterium tumefaciens LBA4404. Hypocotyl segments of E. fortunei var. radicans were infected through A. tumefaciens-mediated transformation. Polymerase chain reaction (PCR) and PCR–Southern blot analysis results confirmed that the GNA gene was integrated into the genome of transgenic plants. The highest transformation frequency was obtained with un-precultured explants infected for 30 min with OD600=0.6 Agrobacterium tumefaciens, and co-cultivated for 3 days.

Type
Research Papers
Copyright
Copyright © China Agricultural University 2008

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Footnotes

First published in Journal of Agricultural Biotechnology 2008, 16(1): 121–126

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