In Vitro Plant Regeneration from Coleoptilar Node of Maize Seedling: a New Tool to Bioengineer the Maize Rapidly
In Vitro Plant Regeneration from Coleoptilar Node of Maize Seedling: a New Tool to Bioengineer the Maize Rapidly
Zaheer Abbas*, Shaukat Ali, Jalal-Ud-Din and Ghulam M. Ali*
ABSTRACT
Efficient and reproducible regeneration system from coleoptilar node of elite maize inbred line was developed. Regeneration from coleoptilar node circumvented tedious procedures of raising the immature embryos as a starting material for genetic transformation. Unlike immature embryos, the development of coleoptilar node is seasonal free and green house independent which would accelerate the work relevant to genetic transformation of maize. MS medium fortified with 3 mgL-1 BAP and 5 mgL-1 picloram produced maize seedlings with expanded coleoptilar nodes. Longitudinal cut sections of expanded coleoptilar nodes produced maximum primary calli on callus induction medium containing both 0.5mgL-1 2, 4-D and 1 mgL-1 picloram. Eighty percent of embryogenic calli were obtained on MS medium containing only single auxin while combination of auxin and cytokine in MS and Chu’s N6 media produced 53.33 and 35.56% embryogenic calli, respectively. Inclusion of 5 mgL-1 silver nitrate in embryogenic medium enhanced the production of type II embryogenic calli by 54.90% compared to control treatment. Maximum regeneration (58.33%) was observed when embryogenic calli were placed on N6 medium supplemented with 60 gL-1 sucrose and 1 mgL-1 NAA for one week initially in dark and then three weeks on MS medium augmented with 2 gL-1 myo-inositol and 20 gL-1 sucrose under 16/8 hours light/dark photoperiod. The study confirmed that embryogenic type II calli obtained from half of coleoptilar node of maize seedling is fully capable to regenerate into whole plant under specific chemical and environmental conditions.
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