Micropropagation, In Vitro Regeneration and Molecular Diversity Study of Moringa Stenopetala (Bakf .) from Ethiopia

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Addis Ababa Universty


The tree species Moringa stenopetala belongs to the family Moringaceae and represented only by a single genus Moringa. M. stenopetala is a native tree in Southern Ethiopia, North Kenya and Eastern Somalia. It plays a vital role for household food security, as source of income, medicine , purification of water, windbreak, fodder, fuel and shade tree all year round. Conventional propagation o/ M. stenopetala involves the use of seeds and cuttings . However, the application of fast and efficient biotechnological methods to propagate and improve different traits of interest is much needed. The objectives of this study were to develop micropropagation and in vitro regeneration protocols for Moringa stenopetala and to analyze its genetic diversity using ISSR molecular marker. Shoot tips obtained from in vitro germinated seedlings were used for culture initiation. All treatments with BAP in combination with NAA showed 100% shoot initiation beginning from the first culture. BAP at lower concentration was found to be the best for shoot multiplication. Of the treatments with BAP, KN, TDZ, IBA and NAA used for shoot multiplication, high quality and maximum mean number (11.16+0.14 and 10.86^0.14 ) of shoot per shoot tip explant were attained on medium containing 0.5 mg/l BAP and 1.0 mg/I BAP with no significant difference. Similarly, application of 1.0 mg/l BAP resulted in the highest number of induced axillary shoots (13.66±0.20) per nodal explant. Half strength MS medium containing NAA combined with IBA was virtually better in promoting root induction than 1/2 MS medium containing IBA or NAA alone. The leaves of in vitro multiplied shoots were used to develop the in vitro regeneration protocol based on using TDZ, 2,4-D and NAA alone or in combination. The use ofTDZ alone at lower concentrations and application of 2,4-D, TDZ and/or NAA together led to very strong callus formation on MS medium. The maximum shoot regeneration percentage (63.66%) was achieved on a medium supplemented with BAP and KN each at 1.0 mg/l concentration. A genetic diversity study based on ISSR was also conducted on a total of 150 samples representing 14 populations from different parts of Ethiopia. The highest level ofpercent polymorphism (53.66%) was obtained from samples of Arba Minch Beto area and the least level of polymorphism (46.34%) was obtained from individuals in Arba Minch Merab Abaya area. The AMOVA results revealed that the genetic variation at population level accounted 21.47%) of the total variation, while the within populations component accounted for 78.53% showing a higher within population variation than among populations. Jaccard’s similarity coefficient for populations ranged from 0.601 to 0.749, with the highest similarity value (0.749) was shown between populations from Konso town and populations from outlying areas of Konso. Findings from this study could be used for mass propagation of disease free planting materials within relatively short period and contribute to further improvement of the plant. In addition, the results obtained here can contribute to food security of both human and cattle, solve the demand of plantlets both in quality and quantity and help to increase source of income for local residents as well as contribute to the genetic improvement of the species. Key words: In vitro regeneration, inter simple sequence repeat (ISSR), micropropagation, Moringa, node, rooting



In vitro regeneration, inter simple sequence repeat (ISSR), Micropropagation, Moringa, Node, Rooting