Efficient and rapid in vitro plant regeneration system for Indian cultivars of chickpea (Cicer arietinum L.)

Lacking of an efficient regeneration protocol for the recalcitrant crop chickpea is a limiting factor for adapting genetic engineering approaches for its improvement. The present study describes a rapid and efficient method for multiple shoot regeneration for three Indian cultivars, B115, C235, ICCV89314, using single cotyledons with half embryos as explant. Modified MS medium with 1.5 mg l(-1) 6-benzyladenine (BA) and 0.04 mg l(-1) alpha-naphthaleneacetic acid (NAA) induced a maximum of 26 shoots from a single explant after 20 days of culture. When cultured in modified MS medium containing 0.2 mg l(-1) indole-3-acetic acid (IAA), 80% of the shoots from each regenerating explant elongated in another 20-25 days. Following a root-grafting protocol, 90-95% of the elongated shoots survived in soil which subsequently produced seeds. The regeneration process from explant preparation to complete plants took 55-60 days. The presently optimized rapid regeneration method holds promise for facilitating the deployment of agronomically important components through genetic transformation for betterment of this important food crop.