High percentage of regeneration and transformation in chickpea

Abstract

Efficient tissue culture and gene transfer systems of Cicer arietinum L. cvs. Red chickpea, Canitez 87 and MB-10 varieties were established. The explants were taken from the shoot primordium part of the mature embryo. For callus induction MS+0.1 mg/l NAA+ 0.5 mg/l 2,4-D and for plant regeneration MS+0.2 mg/l IAA+ 0.5 mg/l Kn+ 0.2 mg/l 6-BAP containing media were used. Best regeneration response was for the MB-10 variety (85%). Young and wounded regenerated shoots were co-cultivated with A. tumefaciens 4404 and A. rhizogenes 9402 both containing pBI-121 plasmids. Explants cocultivated with A. tumefaciens produced crown gall and plantlets on selective media. However, applications with A. rhizogenes produced hairy roots. The existence of GUS and NPT II genes in the DNA isolated from plantlets and hairy roots was confirmed with Southern blot and digoxigenin labeling. The best transformation response was in Canitez 87 (12.7%) with A. tumefaciens and MB-10 variety (10.4%) with A. rhizogenes.