Genetic transformation of cotyledon explants of cowpea (Vigna unguiculata L. Walp) using Agrobacterium tumefaciens

Mature de-embryonated cotyledons with intact proximal end of Vigna unguiculata were cultured on B5 basal medium containing varying concentrations of BAP. Thirty-six percent of the explants produced shoots on B5 medium supplemented with 8 X 10(-6) M BAP. Cotyledon explants were pre-incubated for 24 h...

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Bibliographic Details
Published in:Plant cell reports 1996-09, Vol.15 (12), p.980-985
Main Authors: Muthukumar, B, Mariamma, M, Veluthambi, K, Gnanam, A
Format: Article
Language:English
Subjects:
2,4-D
Agrobacterium tumefaciens
b5 basal medium
b5 medium
benzyladenine
Biological and medical sciences
Biotechnology
cotyledons
culture media
dose response
enzyme activity
explants
Fundamental and applied biological sciences. Psychology
gene expression
gene transfer
Genetic engineering
Genetic technics
genetic techniques and protocols
in vitro culture
indole acetic acid
Methods. Procedures. Technologies
naphthaleneacetic acid
phosphotransferases (phosphomutases)
regenerative ability
shoot induction medium
transformation
Transgenic animals and transgenic plants
Transgenic plants
Vigna unguiculata
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Summary:Mature de-embryonated cotyledons with intact proximal end of Vigna unguiculata were cultured on B5 basal medium containing varying concentrations of BAP. Thirty-six percent of the explants produced shoots on B5 medium supplemented with 8 X 10(-6) M BAP. Cotyledon explants were pre-incubated for 24 h, inoculated with A. tumefaciens pUCD2614 carrying pUCD2340, co-cultivated for 48 h and transferred to hygromycin-B (25 mg/l) containing shoot induction medium. Approximately 15-19% of the explants produced shoots on the selection medium. The elongated shoots were subsequently rooted on B5 basal medium containing hygromycin. The transgenic plants were later established in pots. The presence of hpt gene in the transgenic plants was confirmed by Southern blot hybridization.
ISSN:0721-7714
1432-203X
DOI:10.1007/BF00231600