The Possibility of In Vitro Haploid Production in Chickpea (Cicer arietinum L.)

In order to investigate the androgenic response of chickpea cultivars, two Iranian Chickpea cultivars, Pirooz and Karaj 12-60-31, were used in this study. After 7 to 10 days of cold pretreatment of flower buds, anther containing uninucleate stage of microspores were placed aseptically on MS medium supplemented with various combinations of growth regulators of 2,4-D (1,2 and 3 mg/l) and kinetin (0.1, 0.2 and 0.5 mg/l). Callus regeneration achieved using MS and modified Blayd’s media with various hormones and different sucrose concentrations. The results indicated that the callus initiation was significantly affected by 2,4-D and kinetin concentrations, and that increasing these hormones reduced callus induction. The best response obtained on media with the lowest concentration levels of 2,4-D and kinetin (1 and 0.2 mg/l, respectively). A highly significant genotypic effect and a genotype  2,4-D interaction were detected, which proved that Pirooz response was the best. Callus differentiation and organogensis occurred in MS medium supplmented with NAA, BA and 3% sucrose. Mature embryos also obtained in modified Blayd’s plus 0.5 mg/l kinetin and 10% sucrose. Cytological studies revealed the presence of haploid cells with chromosome variation in the anther derived callus. Therefore, optimizing the hormone levels of different basal media with a particular sucrose cocentration may improve haploid regeneration in chickpea. It seems a further study should be carried out to characterize calli from induction to regeneration and to determine the effect of cold pretreatment, the results of which could be used to improve anther culture response of chickpea.