Callus Induction and Plant Regeneration from Embryo Culture in GF677 Rootstock

In this research, complete micropropagation protocol of GF677 has been described by indirect organogenesis from embryo culture. In order to break the chilling requirement, mature seeds after disinfection were treated with 250 mg-l of sterile solution of GA3 for different periods of time (0, 12, 24, 48, 96 h). Then, embryo explants separated carefully from the seeds and for callus induction were cultured in factorial experiment on Murashige and Skoog medium (MS) supplemented with different growth regulator treatments. The results showed, only in the presence of 2,4-D and BA, GA3 treatment increased callus induction of explants. Calluses were transferred on medium containing BA (0.0, 1.0, 2.0 and 4.0 mg-l) with 0.25 mg-l NAA and 0.25 mg-l IBA. The highest percentage rate of regeneration (90%) and the highest shoot number, (16.60) per explant were obtained in the presence of 2.0 mg-l BA. Seeds treatment with GA3 at this stage reduced regeneration of shoots from callus. For rooting of regenerated shoots, the effect of three types of auxin IBA, NAA and IAA in different concentrations (0.0, 0.5, 1.0, 2.0, 4.0 and 8.0 mg-l) on 1/2MS medium were studied. The highest rooting index was observed in the presence of IBA. Rooted plantlets after transferred to the soil mixture (peat, perlite and sand, 2: 2: 1) acclimatized with 85% success in greenhouse condition.