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Research Article Open access CC BY 4.0

Regeneration Procedure for Three Arachis hypogaea L. Botanicals in Uganda through Embryogenesis

D. K. Okello, L. B. Akello, P. Tukamuhabwa, S. M. Ochwo, T. L. Odong, J. Adriko, C. Mwami, C. M. Deom

Biotechnology Journal International · pp. 122–133 · Published 6 May 2015

10.9734/BBJ/2015/17418

Abstract

Aims: A procedure was developed for embryogenesis from embryo explants derived from mature seeds of freshly harvested Serenut 4T, Serenut 1R and Acholi-white groundnut cultivars representing the three broad groundnut botanical classifications. Methodology: This study explored the use of mature embryo axes as explants for somatic embryogenesis, and determined the factors that affect regeneration of three Ugandan groundnut cultivars. Freshly harvested mature seeds of the three groundnut cultivars were collected and the embryo explants were initiated on 3 media namely; Murashige and Skoog (MS) basal media with varying concentrations of the growth regulator 2,4-Dichlorophenoxy acetic acid (2,4-D); Chu N6 basal medium with vitamins (N6); and Callus Induction Medium (CIM). The shoot formation and elongation medium contained MS basal medium supplemented with indolebutyric acid (IBA) and 6-Benzylamminopurine (BAP) in isolation, and BAP in combination with a-naphthaleneacetic acid (NAA) and indoleacetic acid (IAA). For root induction, elongated shoots were transferred to MS medium supplemented with various combinations of NAA with IBA, BAP and a combination of IBA and Kinetin. Results and Conclusion: Different concentrations of 2,4-D elicited different callogenesis responses in the cultivars with Acholi white (Valencia botanical) and Serenut 4T (Spanish botanical) giving the optimal response at 5mg/l whereas Serenut 1R (Virginia botanical) showed best response at a concentration of 30 mg/l. N6 and CIM supported callogenesis in Acholi white (AW) and Serenut 4T only. In all cultivars, maximum root production was gained when using MS medium supplemented with NAA- 1 mg/l and IBA -2.0 mg/l. On the other hand, for Serenut 1R and Serenut 4T, BAP 2.5 mg/l; NAA 0.5 mg/l combination yielded higher shoot regeneration percentage whereas for AW BAP 3 mg/l; NAA 0.5 mg/l supported maximum shoot production.This is the first ever report of successful regeneration of the three groundnuts botanicals in Uganda. These results are likely to facilitate genetic transformation of three preferred Ugandan groundnut varieties.

Groundnuts cultivars callus induction 2,4-D, N6 callus induction medium in vitro morphogenesis

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