In vitro regeneration of some Ethiopian maize (Zea mays L.) lines and varieties from immature embryos through callus induction

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Research Paper 01/06/2022
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In vitro regeneration of some Ethiopian maize (Zea mays L.) lines and varieties from immature embryos through callus induction

Tarekegn Gebreyesus Abisso, Lemma Abayneh Tumebo
Int. J. Biosci.20( 6), 140-154, June 2022.
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Maize is an important food crop in Ethiopia. Success in genetic transformation requires efficient in vitro regeneration protocols. However, inadequate information is available on in vitro regeneration of maize varieties/lines developed for the Ethiopian climate. Therefore, this study was initiated to optimize in vitro regeneration protocols for some inbred lines and open-pollinated maize varieties from immature embryos. Immature embryos evaluated for their ability to form callus were cultured in N6 medium and incubated at room temperature in the dark to initiate callus. Embryogenic calli were transferred from callus maintenance medium to embryo maturation medium supplemented with 2 mg/l glycine, 1 mg/l NAA and different levels of sucrose (55, 60, 65, or 70 g/l). Matured somatic embryos were subcultured in a shoot regeneration medium consisting of MS medium supplemented with 2 mg/l glycine, 2% sucrose and different levels (0, 0.1, 0.2 and 0.3 mg/l) of BAP. Roots were induced by subculturing individual shoots on half-strength MS medium supplemented with 2 mg/l glycine, 2% sucrose and different levels (0, 0.1, 0.2 and 0.3 mg/l) of NAA. Immature embryos were harvested between 16-20 days after pollination, depending on the variety for average callus induction. Better plant regeneration was obtained at basal (hormone-free) MS medium. Better root formation was at 0.1 mg/l NAA with an average of 2.82- 4.50 roots per shoot. Regenerated plantlets were successfully acclimatized in greenhouse and field conditions with survival rates of 83.7% and 75.6%, respectively. This study established a regeneration scheme for maize lines/ varieties via somatic embryogenesis from immature embryos.


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