Somatic embryogenesis and regeneration of Vigna radiata
2010
Sivakumar, P.,Tamil Nadu Agricultural Univ., Coimbatore (India). Centre for Plant Molecular Biology | Gnanam, R.,Tamil Nadu Agricultural Univ., Coimbatore (India). Centre for Plant Molecular Biology | Ramakrishnan, K.,Centre for Cellular and Molecular Biology, Hyderabad (India) | Manickam, A.,Tamil Nadu Agricultural Univ., Coimbatore (India). Centre for Plant Molecular Biology
An efficient regeneration protocol via somatic embryogenesis was optimized for (Vigna radiata, 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/cubic dm 2,4-dichlorophenoxyacetic acid (2,4-D), 150 mg/cubic dm 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/cubic dm 2,4-D and 50 mg/cubic dm 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/cubic dm 2,4-D, 20 mg/cubic dm Pro, 5 microM abscisic acid, 1,000 mg/cubic dm KNO3, 50 mg/cubic dm polyethylene glycol (PEG 6000) and 30 g/cubic dm D-mannitol. Mature somatic embryos were germinated after dessication for 3 d and complete development of plantlets accomplished in MMS medium containing 30 g/cubic dm maltose, 0.5 mg/cubic dm benzyladenine and 500 mg/cubic dm KNO3. Profuse lateral roots, and regeneration frequency (up to 60 %) were observed in half-strength MMS medium containing 0.5 mg/cubic dm indolebutyric acid (IBA). The regenerated plants were grown to fruiting and were morphologically normal and fertile.
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