Abstract
Fast-growing normal root culture of Vernonia amygdalina, a potent African medicinal plant was established from leaf explants of in vitro raised shoot induced from the stem nodal segments on murashige and skoog (MS) medium containing 0.5 mg l-1 6-benzylaminopurine (BA) in combination with 0.5 mg l-1 naphthalene acetic acid (NAA). In vitro raised plantlets were maintained on MS agar medium and sub cultured at 4 weeks interval and used as leaf explant source. Explants were cultured on half-strength MS medium supplemented with different concentrations of Indole-3-acetic acid (IAA), indole-3-butyric acid (IBA) and NAA. Basal medium supplemented with IBA at 0.25 and 2.0 mg l-1 and under 16 photoperiod condition favoured induction of the longest root (2.7 ± 1.1 cm) and highest number of roots/explant (38.3 ± 1.1) respectively. After 6 weeks well established roots were separated. Fresh root tissue, in amount of a 100 mg were cultured in 50 ml full-strength MS liquid medium supplemented with 2.0 mg l-1 IBA and under continuous agitation (80 rpm). The biomass of root culture was increased to 2.1949 g after 5 weeks of culture. The root culture was maintained up to 6 weeks. The protocol developed in this study provides a basis for adventitious root induction and for further investigation of medicinally active constituents of this elite medicinal plant. Key words: Vernonia amygdalina, nodal segment, leaf explant, root culture, medicinal plant, suspension culture.
Highlights
Vernonia amygdalina is a small tree between 1 to 3 m in height (Igile et al, 1995) grows predominantly in the tropical parts of Africa and well known for its medicinal and nutritional values (Farombi, 2003)
Fast-growing normal root culture of Vernonia amygdalina, a potent African medicinal plant was established from leaf explants of in vitro raised shoot induced from the stem nodal segments on murashige and skoog (MS) medium containing 0.5 mg l-1 6-benzylaminopurine (BA) in combination with 0.5 mg l-1 naphthalene acetic acid (NAA)
Under a laminar flow cabinet leaf explants were inoculated on half-strength MS (Murashige and Skoog, 1962) medium supplemented with varied concentration of different auxins; indole-3-acetic acid (IAA), indole-3- butyric acid (IBA) or αnaphthalene acetic acid (NAA)
Summary
Vernonia amygdalina (compositae) is a small tree between 1 to 3 m in height (Igile et al, 1995) grows predominantly in the tropical parts of Africa and well known for its medicinal and nutritional values (Farombi, 2003). All parts of the plant are pharmacologically useful (Ojiako and Nwanjo, 2006), roots are the principle material used in the traditional medicine. In view of the increasing demand for V. amygdalina for use as herbal medicine and the fact that roots are the principle material for herbal preparation which involve destructive harvesting. Development of biotechnological methods such as micropropagation, cell/ root and hairy root culture is one of the major solutions to circumvent these problems. On this line, in vitro micropropagation protocol using nodal explant recently has been developed in our laboratory (Khalafalla et al, 2007). Root cultures can be used in many ways including studies of carbohydrate metabolism, mineral nutrient requirements, essentiality of vitamins and other growth regulators, differentiation of the root apex and gravitropism
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