Genetic tranformation of cotyledon explants of cowpea (Vigna unguiculata L. Walp) using Agrobacterium tumefaciens |
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Authors: | B Muthukumar M Mariamma K Veluthambi A Gnanam |
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Institution: | (1) Department of Plant Science, School of Biological Sciences, Madurai Kamaraj University, 625021 Madurai, India;(2) School of Biotechnology, Madurai Kamaraj University, 625021 Madurai, India;(3) Pondicherry University, 605 014 Pondicherry, India |
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Abstract: | 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× 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.Abbreviations BAP
6-Benzylaminopurine
- 2,4-D
2,4-dichlorophenoxyacetic acid
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hpt
hygromycin phosphotransferase
- IAA
Indole-3-acetic acid
- NAA
1-naphthaleneacetic acid |
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Keywords: | De-embryonated cotyledons shoot regeneration co-cultivation Vigna unguiculata |
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