Plant material and shoot regeneration from semi-solid cultures

Young leaves of Urginea altissima (L.f.) Baker were collected from the Botanical Garden, University of KwaZulu-Natal, Pietermaritzburg, South Africa. Explants were decontaminated using 0.2% (w/v) aqueous HgCl₂ for 10 min followed by five rinses with sterile distilled water. Disinfected leaf explants (approximately, 15 × 10 mm) were used for direct and indirect shoot regeneration. The explants were cultured in solidified (8 g L⁻¹ agar) MS (Murashige and Skoog 1962) medium (semi-solid MS) containing 30 g L⁻¹ sucrose and supplemented with 10 µM benzyladenine (BA), meta-topolin (mT) and thidiazuron (TDZ) alone or in combination with 2 µM indole-3-acetic acid (IAA) or indole-3-butyric acid (IBA) or naphthaleneacetic acid (NAA) or phloroglucinol (PG) for direct shoot regeneration. White compact callus obtained with semi-solid MS medium containing 10 µM 2,4-D and 2 µM TDZ was established onto semi-solid MS medium containing 10 µM BA or mT or TDZ and in combinations with 2 µM picloram (Pic) or 2,4-dichlorophenoxyacetic acid (2,4-D) or NAA or IAA or BA or mT or TDZ for indirect shoot regeneration.