Somatic embryogenesis and regeneration of Vigna radiata

Abstract

An efficient regeneration protocol via somatic embryogenesis was optimized for mung bean [Vigna radiata (L.) Wilczek; cv. Vamban 1]. Primary leaf explants were used for embryogenic callus induction in MMS medium (Murashige and Skoog salts with B5 vitamins) containing 2.0 mg dm-3 2,4-dichlorophenoxyacetic acid (2,4-D), 150 mg dm-3 glutamine and 3 % sucrose. Fast growing, highly embryogenic cell suspensions were established from 21-d-old calli in MMS medium supplemented with 0.5 mg dm-3 2,4-D and 50 mg dm-3 proline (Pro), and maximum recovery of globular (39.0 %), heart-shaped (26.3 %) and torpedo-stage (21.0 %) somatic embryos were observed in this medium. Mature cotyledonary-stage somatic embryos were cultured for 5 d in half strength B5 liquid medium containing 0.05 mg dm-3 2,4-D, 20 mg dm-3 Pro, 5 μM abscisic acid, 1000 mg dm-3 KNO3, 50 mg dm-3 polyethylene glycol (PEG 6000) and 30 g dm-3 D-mannitol. Mature somatic embryos were germinated after dessication for 3 d and complete development of plantlets accomplished in MMS medium containing 30 g dm-3 maltose, 0.5 mg dm-3 benzyladenine and 500 mg dm-3 KNO3. Profuse lateral roots, and regeneration frequency (up to 60 %) were observed in half-strength MMS medium containing 0.5 mg dm-3 indolebutyric acid (IBA). The regenerated plants were grown to fruiting and were morphologically normal and fertile.

Keywords

• cell suspension cultures
• mung bean
• plant growth regulators
• primary leaf explant