Micropropagation of an Endangered Ethiopian Medicinal Plant, Kebericho (Echinops Kebericho Mesfin)
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Date
2014-01-01
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Addis Ababa University
Abstract
Echinops kebericho Mesfin, endemic to Ethiopia, is a critically endangered medicinal plant. It is among the most important medicinal plants of the country. The commercial harvesting and sale of roots of E. kebericho is widespread in the growing regions of the country. This involves uprooting of the whole plant or removal of the root, which may seriously damage the root system or the whole plant. This unsustainable harvesting of the plant reduces regeneration and thus, threatens local populations. In vitro mass propagation of this species could offer large number of cloned plants for rehabilitation of the threatened population, for commercialization, conservation and sustainable utilization of the species. The aim of this study was to develop micropropagation protocol for E. kebericho using shoot tips as explant. The study was first started with seed germination test using seeds stored for different months. The shoot tip explants excised from in vitro germinated seedlings was cultured on shoot initiation MS media supplemented with different concentrations of BAP (0, 0.1, 0.5, 1.0, 1.5, 2.0 mg/l) or KN (0, 0.1, 0.5, 1.0, 1.5, 2.0 mg/l) alone. Explants from initiated shoots were cultured on shoot proliferation MS medium supplemented with various concentrations (0.0, 0.5, 1.0, 2.5, 3.0, 5.0 mg/l) of cytokinins [(Kinetin, 6-benzylaminopurine (BAP), and thidiazuron (TDZ)] either alone or in combination with different concentrations (0.0, 0.1, 0.25, 0.5 mg/l) of α-Naphthaleneacetic acid (NAA) in a completely randomized design (CRD). For root induction, full, half and 1/3 strength MS media supplemented with various concentrations (0, 0.05, 0.1, 0.5, 1.0, 1.5, 2.5 mg/l) of auxins [indole-3-butyric acid (IBA) and NAA] were used. In all cases, growth regulator free MS medium was used as a control. Based on the study results, seed germination percentage gradually decreased with increased storage time. The best germination (100%) was obtained with fresh seeds. Mean germination percentage dropped as low as 65.18 and 22.3% with seeds stored for 3 and 5 months respectively. MS medium containing 0.5 mg/l KN gave 100% shoot induction. On shoot multiplication MS media, medium fortified with 0.5 mg/l KN + 0.1 mg/l NAA showed maximum shoot proliferation with 11.07 shoots per explant and 4.82 cm shoot length. Among different salt strengths of MS medium (full, half and 1/3) containing IBA and NAA, best rooting (100%) was obtained on 1/3 strength medium containing 1.5 mg/l NAA with maximum of 8.23 roots/shoot and 4.82 cm root length and well established under greenhouse conditions with 83% survival rate. Generally, the micropropagation protocol reported here was characterized with a rapid proliferation of shoots, easy rooting of the micro-shoots and the plantlets were acclimatized to the greenhouse environment with 83% survival rate and undergoing normal physiological development.
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Keywords
Echinops Kebericho, Micropropagation, Shoot Tips, Plant Growth Regulator