Paper Details

PJB-2021-312

Sustainable plantlet regeneration from hypocotyl segments and zygotic embryo of Securidaca longipedunculata Fresen. (Polygalaceae) - an endangered medicinal plant

Uche Cyprian Okafor, Adaugo Ozioma Nwakuche, Nneka Sheily Egonu, Felix Ifeanyi Nwafor, Chinelo Chinyere Onyekwuluje, Hyacinth Chikwado Obayi, Hannah Ifunanya Iwuoha, Chinedum Raphael Anajemba and Nwanneka Maryann Okafor
DOI:10.30848/PJB2023-4(5)


Abstract

Plant regeneration was achieved by culturing sterile hypocotyl segments, and mature zygotic embryos (MZE) of S. longipedunculata Fresen excised from sterile seeds. In this study, de-coated and sterilized seeds were first grown into seedlings In vitro, disjointed into root, hypocotyl, and leaf, segments. The hypocotyl segments (0.5cm) were cultured in root induction media containing MS basal medium + sucrose (30 g l-1) fortified with 0-5 mg l-1 NAA and 0-5 mg l-1 IBA either alone or in combination to produce roots. Rooted hypocotyl segments were cultured in shoot induction media containing 0-1mg l-1 IAA, 0-1 mg l-1 KIN, and 0-1 mg l-1 GA for shoot induction. Second, mature zygotic embryos (MZEs) excised from seeds were cultured based on 3×4×5 factorial – Control, MS and B5 basal media, three carbon sources (sucrose, glucose, and fructose), and control at five concentrations (0-4%) in a completely randomized design at p≤0.05 and repeated two independent times for reproducibility of results. Results of the observed parameters revealed that after 45 days, the highest number of roots (11.66±0.23), most increased root length (3.43±0.23), and highest percent root generation (86.66±3.33) were produced in hypocotyl explants in 0 mg l-1 NAA+5.0 mg l-1 IBA. More so, the earliest induction (35.66±0.66) was seen in 0.5 mg l-1 NAA+0.5 mg l-1 IBA. After 72 days of culture, the highest number of new shoots (2.0 per explant), the highest number of leaves (5.0±1.00), the highest number of shoots (3.80±0.11), highest percent regeneration (88.33±1.60), and highest leaf area (2.06±0.23) were produced in rooted hypocotyl explants at 1mg l-1 each for IAA, KIN, and GA relative to other treatments. In MZE culture, 3% sucrose in MS medium was the most preferred for zygotic embryo culture and differed significantly from other treatments at p≤0.05. These young plantlets, In vitro, were hardened and raised in the field. These regenerated plantlets would ensure the conservation of this endangered plant as a primary source of medicine

To Cite this article: Okafor, U.C., A.O. Nwakuche, N.S. Egonu, F.I. Nwafor, C.C. Onyekwuluje, H.C. Obayi, H.I. Iwuoha, C.R. Anajemba and N.M. Okafor. 2023. Sustainable plantlet regeneration from hypocotyl segments and zygotic embryo of Securidaca longipedunculata Fresen. (Polygalaceae) - an endangered medicinal plant. Pak. J. Bot., 55(4): DOI: http://dx.doi.org/10.30848/PJB2023-4(5)  
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