Introduction: Psyllium (Plantago ovata Forsk) is medicinally used mainly for its mucilage content. Objective: In the present study, an attempt was made to improve mucilage yield under in vitro callus culture using different genotypes, explants and culture media. Methods: The effects of a range of concentrations of plant growth regulators including 2,4-dichlorophenoxyacetic acid (2,4-D) and kinetin (Kin) were evaluated on mucilage synthesis under in vitro culture using cotyledon, hypocotyl and seed explants. Fourteen genotypes originating from different geographical regions of Iran were used to evaluate their response to in vitro mucilage synthesis. Results: The highest rate of callus induction (76%) and callus growth rate CGR (0.38 mm/day) were induced on MS medium supplemented with 0.5 mg/l 2,4-D and 1 mg/l Kin and the hypocotyl explant. The results of analysis of variance showed significant genotypic differences for callus induction, CGR and mucilage content of callus and seeds. The mucilage content ranged from 0.38 to 0.08 (g/g DW) and 0.13 to 0.042 (g/g DW) for callus and seed, respectively. The superior callus induction (73%), CGR (0.45 mm/day) and mucilage content of callus (0.38 g/g DW) was denoted to Po1 genotype. The callus produced nearly three times more mucilage than the seeds using superior genotype (Po1). Conclusion: The results of this study revealed that high efficiency of callus culture of P. ovata using hypocotyl explant accompanied by the exploration of genetic diversity are important to improve the yield of mucilage synthesis by in vitro callus culture.
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