Callus induction and organogenesis from leaf explants of Tectonagrandis | Abstract
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Callus induction and organogenesis from leaf explants of Tectonagrandis

Author(s): M. LW. O. M. Karunaratne, S. E. Peries and W. C. P. Egodawatta

Organogenesis and embryogenesis of Tectonagrandis was attempted in this study with leaves collected from stumps and mature trees as explants. Tissue contamination and browning were the main problems in in vitro establishment of stumps. Leaf explants from stumps were sterilized using different concentrations of Clorox™, fungicide and adding alcohol. The sterilization based on dipping in 0.05% Bavistin™ as well as adding it in 0.05% concentration to culture medium was more efficient as it produced 98% of vital explants. Keep cultures in dark condition for 3days produced 96% browning free cultures. Different Cytokinins (6-benzyladenine and kinetin) and Auxines (NAA)at a range of various concentrations in Murashige and Skoog (MS) medium have been investigated for organogenesis. Medium supplemented with 0.5 mg/l NAA and 1.5 BAP mg/l produced compact and fibrous callus under light condition after two weeks. Callus initiation was not observed in MS medium supplemented with BAP, Kinetin and NAA. The explants which were obtained from L2growth stage (second leaf from the tip) produced 96% of yellow colour globular shapes callus. Callus proliferated from 2, 4–D was introduced to no hormone medium, GA3and liquid MS medium and observed different type of callus formation. This study provides callus induction protocol derived from T.grandis leaf explants as a basis for future studies