Agrobacterium tumefaciens-mediated transformation of pigeon pea (Cajanus carjan L. Millsp.) and molecular analysis of regenerated plants

Transformation of pigeon pea (Cajanus cajan L, Millsp.) was achieved using Agrobacterium tumefaciens strain GV2260, containing the construct of isolat ed cowpea protease inhibitor gene (pCPI; Accession-no.: AJ271752), The gene was driven by CaMV 35S promoter containing kanamycin resistance as: plant selection marker. Embryonic axes excised from seeds germinated on MS basal supplemented with BAP (2 mg/l) were used as explants. Transformed calli wer e selected on medium containing kanamycin (50 mug/ml) after 3 weeks of incu bation. Thereafter, the green calli were kept for differentiation. MS basal medium containing IAA (0.2 mg/l) and different concentrations of BAP was t ested for the regeneration frequency of explants (embryo axes and leaves). Rooting ability of differentiated shoots was tested on half MS basal medium containing different concentrations of IBA, Molecular analysis of the tran sformed plants was done by Northern techniques.