A simple and improved protocol for direct and indirect somatic embryogenesis of peanut (Arachis hypogaea L.)
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Abstract
A simple and efficient protocol for the direct and indirect embryoid induction and plant regeneration is described here. Direct induction of somatic embryos was achieved on proximal end of embryonal axis explant in the medium fortified with MS salts, B5 vitamins, 27.5 mg/L PIC, 30 g/L sucrose and 0.8% agar. After 6 weeks of culture, the highest response to embryoid induction achieved was 97% without an intervening callus phase. About 23.6 ± 0.28 embryoids per explants were induced by the above concentration. Leaf explants were used for the indirect somatic embryogenesis. 500 mg of 30 days old embryogenic callus was used for embryoid induction through suspension culture. High frequency of embryogenic callus induction was observed on the media fortified with MS salts, B5 vitamins, 2 mg/L 2,4-D and 1 mg/L BAP. After 6 weeks, the embryoids were formed on the bottom of the suspension culture. Medium fortified with MS salts, B5 vitamins, 2 mg/L 2,4-D and 1 mg/L KIN showed best response of embryoid induction through suspension culture. The formed matured embryoids on the original medium. The embryoids induced through direct and indirect methods were regenerated on the medium fortified with MS basal salts, B5 vitamins, BAP (0.2 mg/L) and IBA (0.2 mg/l) and 75.8% of plant regeneration was observed at this concentration. The regenerated plantlets showed 100% survival rate in green house conditions. Influences of different carbohydrates, amino acids, and additives were also tested for the high frequency of somatic embryo production.
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