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Efficient in vitro direct shoot organogenesis and regeneration of fertile plants from embryo explants of Bambara groundnut (Vigna subterranea L. Verdc.)

  • Cell Biology and Morphogenesis
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Abstract

An efficient protocol has been developed for direct shoot organogenesis from embryo axes derived from mature seeds of two different landraces of Bambara groundnut. Multiple shoots were initiated on several media containing different concentrations and combinations of benzylaminopurine (BAP) or thidiazuron (TDZ). Efficient regeneration occurred when the embryo axes were first plated for 6 days on a medium containing high concentrations of BAP (1 mg/l) and α-naphthaleneacetic acid (NAA, 1 mg/l) and then cut transversely and transferred onto a medium containing 1.5 mg/l BAP. Shoot regeneration frequency was 100% and from five to eight shoots per explant were obtained. The importance of using embryo explants and cytokinins in the culture media, with respect to controlling the development of a highly organogenic system, was demonstrated. Histological studies revealed that proliferating buds originated directly from the superficial layers of the explants without an intermediate callus phase. The regenerated shoots were rooted on a medium containing 1 mg/l NAA and then transferred to the greenhouse. Flow cytometric analyses and chloroplast counts of guard cells suggested that the regenerants were diploid. All were morphologically normal and fertile. The short duration, high efficiency and low frequency of somaclonal variation of this system make it well suited for wider biotechnological applications of Bambara groundnut—a neglected and under-utilized crop.

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Fig. 1A–L.
Fig. 2.

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Abbreviations

BAP :

N6-Benzylaminopurine

2,4-D :

2,4-Dichlorophenoxyacetic acid

IAA :

Indole-3-acetic acid

NAA :

α-Naphthaleneacetic acid

TDZ :

Thidiazuron

TIBA:

2,3,5-Triiodobenzoicacid

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Acknowledgements

We wish to thank Dr. P. Devaux for his valuable assistance with the flow cytometric analyses and Dr. S. Ochatt for critical reading of this manuscript. This work is part of a FAO/IAEA Coordinated Research Programme "Genetic improvement of under-utilised and neglected crops in LIFDCs through irradiation and related techniques" (1998–2003). We are grateful to Dr. K. Nichterlein of FAO/IAEA Division, Vienna for helpful discussions. Seeds of two different landraces of Bambara groundnut were provided by Dr. H.K. Adu-Dapaah (Crop Research Institute, Kumasi, Ghana).

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  1. B. Lacroix

    Present address: Department of Biochemistry and Cell Biology, State University of New York at Stony Brook, Stony Brook, NY, 11790, USA

Authors and Affiliations

  1. Laboratoire Androgenèse et Biotechnologie, Université de Picardie Jules Verne, 33 Rue Saint-Leu, 80039, Amiens, France

    B. Lacroix, Y. Assoumou & R. S. Sangwan

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  1. B. Lacroix
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  2. Y. Assoumou
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  3. R. S. Sangwan
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Correspondence to R. S. Sangwan.

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Communicated by M.R. Davey

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Lacroix, B., Assoumou, Y. & Sangwan, R.S. Efficient in vitro direct shoot organogenesis and regeneration of fertile plants from embryo explants of Bambara groundnut (Vigna subterranea L. Verdc.). Plant Cell Rep 21, 1153–1158 (2003). https://doi.org/10.1007/s00299-003-0647-0

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