Multiple shoot regeneration of plumular apices of chickpea

Abstract

Chickpea (Cicer arietinum L.) is an important grain legume used almost all over the world. Development of new repeatable and reliable tissue culture protocols is very important for improvement of breeding and genetic transformation studies. It is a highly recalcitrant plant and there is an urgent need to develop a regeneration protocol that can ensure easy multiple and qualitative superior shoots that could be rooted and yield fertile plants. Unconditioned plumular apices and plumular apices preconditioned with 10 mg L-1 benzylaminopurine (BA) for 10 days were cultured on Murashige and Skoog (MS) medium supplemented with 0.25-2.00 mg L-1 BA with or without 0.25 mg L-1 naphthalene acetic acid (NAA). Comparing the 2 culture conditions, the preconditioned explants had 2-fold to 5-fold more regeneration and shoots per explant compared to the unconditioned plumular apices. The presence of NAA in the culture medium positively increased the number of shoots per preconditioned explant at the lower concentrations of BA. In contrast, NAA inhibited the number of shoots per explants and the mean shoot length of the unconditioned explants at all concentrations of BA. An increased sucrose concentration of 45 and 60 g L-1 (R1) with 1.00 mg L-1 indole-3 butyric acid resulted in 50% rooting with multiple and earlier hardening of shoots. Subculturing of multiple shoots on MS medium containing 45-60 g L-1 sucrose (R2) enhanced the rooting frequency by 60%-100%. The rooted plantlets were successfully acclimatized. © TϋBİTAK.