Volume No. 6 Issue No.: 3 Page No.: 452-457 Jan-Mar 2012

 

IN-VITRO PLANT REGENERATION VIA SOMATIC EMBRYO-GENESIS IN RICE-BEAN Vigna umbellate (Thunb.) OHWI AND OHASHI : AN UNDERUTILIZED AND RECALCITRANT GRAIN LEGUME

 

Aparna*, Rajan Chopra and Raman Saini

Department of Biotechnology, Kurukshetra University, Kurukshetra (INDIA) *E-mail : aparna.prabhakar7@gmail.com

 

Received on : October 15, 2011

 

ABSTRACT

 

An efficient in-vitro regeneration protocol for Indian cultivar (RBL-50) of ricebean Vigna umbellate (Thunb.) Ohwi and Ohashi via somatic embryogenesis has been developed. Highly proliferating (98%) calli cultures were initiated from the cotyledonary-node containing both half cotyledons (CN1) on semi-solid MSB medium (MS salts and B5 vitamins) supplemented with 12.5 μM Thidiazuron (TDZ) alone. Type and concentrations of growth regulators influenced the frequency of somatic embryogenesis. TDZ was found responsive for somatic embryogenesis than BAP, 2,4-D and Picloram and the best result (18 somatic embryos per explant) was obtained with 12.5 μM TDZ in combination with 2.5 μM 6-benzylaminopurine (BAP). 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 MS basal medium enabled the embryos to achieve complete maturation and germination. The percentage of germinating embryos increased significantly from 20 to 50 when 1.5 μM BAP and 2.0 μM Gibberellic Acid (GA3) was supplemented to the MS basal media. In vitro-raised plantlets with well-developed roots were successfully hardened in a greenhouse and established in soil. The present study aimed to produce embryos and their subsequent conversion to plants offers potential to use in transformation studies.

 

Keywords : Callus culture, Ricebean, Somatic embryos, Thidiazuron, Vigna umbellate

 

 

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