Somatic embryogenesis and in vitro plant regeneration in moth bean [Vigna aconitifolia (Jacq.) Marechal]: a recalcitrant grain legume |
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Authors: | Kailash Choudhary M. Singh M. S. Rathore N. S. Shekhawat |
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Affiliation: | (1) Department of Biotechnology, Lachoo Memorial College of Science and Technology, J.N.V. University, Jodhpur, 342001, Rajasthan, India;(2) Biotechnology Laboratory, FASCL, Mody Institute of Technology and Science, Lakshmangarh, Sikar, 332311, Rajasthan, India;(3) Department of Botany, Biotechnology Center, J.N.V. University, Jodhpur, 342001, Rajasthan, India |
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Abstract: | An efficient in vitro regeneration protocol for moth bean [Vigna aconitifolia (Jacq.) Marechal] via somatic embryogenesis has been developed. Embryogenic callus cultures were established from the cotyledonary node as explant on semi-solid Murashige and Skoog (MS) medium supplemented with 0.75 mg l−1 2,4-dichlorophenoxyacetic acid (2,4-D) and 1.5 mg l−1 6-benzylaminopurine (BA) and with various additives (50 mg l−1 ascorbic acid and 25 mg l−1 each of adenine sulphate, citric acid and l-arginine). Numerous somatic embryos differentiated on MS basal nutrient medium supplemented with 0.25 mg l−1 2,4-D and 0.5 mg l−1 of kinetin (Kin). Sustained cell division resulted in the formation of cell aggregates, which progressed to the globular- and heart-shaped somatic embryos and then, if they differentiated properly, to the torpedo shape and cotyledonary stages. The transfer of embryos onto fresh MS basal medium containing 0.2 mg l−1 BA and 2.0 mg l−1 gibberellic acid enabled the embryos to achieve complete maturation and germination. More than 80% of somatic embryos were converted into true-to-type fertile plants. In vitro-regenerated plantlets with well-developed roots were successfully hardened in a greenhouse and established in soil. |
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Keywords: | Embryogenic callus Germination Heart and torpedo shape True-to-type |
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