Document Type : Research Paper
Authors
1 Agriculture and Natural Resources of Tehran University, Iran
2 Department of Agriculture, Tehran University, Iran
3 Agricultural and Natural Resources Research Center of Esfahan, Iran
Abstract
Seeds were cultured on Whatman paper by sterile water in cube, solid MS and solid MS. Seeds were not germinated on medium but 80% of seeds were germinated on Whatman paper, so this method is used as a basic method. Seeds were germinated after four days and after two weeks of culture they had normal roots, shoots and leaves. Cultures were incubated at 25◦ ±2◦C and exposed to 16 hours light per day. Explants were cultured on MS medium supplemented with 3% sucrose and solidified with 0.8% (w/v) agar. The pH of the medium was adjusted to 5.8 before autoclaving. Different types of explants were used for this experiment; root, crown, apical meristem, hypocotyls and leaf. Between different kinds of explants leaf didn’t response to callogenesis. The effects of different combinations of 2,4-D (2,4-Diclorophenoxy acetic acid) and BAP (6-Benzylaminopurine) were studied. Subculture was done every 3 weeks. In order to determine regeneration ability, the initiated callus were transferred to a regeneration medium which was composed of macronutrient, micronutrient and organic components of MS, 2,4-D (0.2 mgL-1), BAP (0.5, 2, 4, 15, 20 and 25 mgL-1) and MS without hormones, 0/3% sucrose, pH 5.8 for 4 weeks. In the presence of 2,4-D (2 and 4 mgL-1) and BAP (0.25 and 0.5 mgL-1) in the callus induction medium, high callus production percentage was reported. The hypocotyls, in contrast to the primary leaf explants, and apical meristem segments were more responsive to the tested combinations of 2,4-D and BAP. The callus from all explants was soft, watery and loose friable. During subculture period, hypocotyls and apical meristem were proliferated more on medium with the addition of (0.25 and 0.5 mgL-1) BAP and (2 and 4 mgL-1) 2,4-D than the medium contain BAP (1 mgL-1) and 2,4-D (8 mgL-1). The present study, in F. vulgare MS media without any hormone was sufficient to regenerate the plantlet from the hypocotyls, roots and apical meristems explants. In MS medium supplemented with BAP (0.5, 2 and 4 mgL-1) and 2,4-D (0.2 mgL-1) shoots were formed earlier when the number of subculture was increased 4 times.
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