Initiation of embryogenic cell suspensions of taro (Colocasia esculenta var. esculenta) and plant regeneration
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Embryogenic callus was initiated by culturing in vitro taro corm slices on agar-solidified half-strength MS medium containing 2.0 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) for 20 days followed by transfer to 1.0 mg/L thidiazuron (TDZ). Callus was subsequently proliferated on solid medium containing 1.0 mg/L TDZ, 0.5 mg/L 2,4-D and 800 mg/L glutamine before transfer to liquid medium containing the same components but with reduced glutamine (100 mg/L). After 3 months in liquid culture on an orbital shaker, cytoplasmically dense cell aggregates began to form. Somatic embryogenesis was induced by plating suspension cells onto solid media containing reduced levels of hormones (0.1 mg/L TDZ, 0.05 mg/L 2,4-D), high concentrations of sucrose (40–50 g/L) and biotin (1.0 mg/L). Embryo maturation and germination was then induced on media containing 0.05 mg/L benzyladenine (BA) and 0.1 mg/L indole-3-acetic acid (IAA). Histological studies of the developing embryos revealed the presence of typical shoot and root poles suggesting that these structures were true somatic embryos. The rate of somatic embryos formation was 500–3,000 per mL settled cell volume while approximately 60% of the embryos regenerated into plants.
KeywordsSomatic embryogenesis Callus Cell suspension Taro Colocasia esculenta var. esculenta
The authors wish to thank New Zealand’s International Aid and Development Agency and The University of the South Pacific for their financial support, and the Centre for Tropical Crops and Biocommodities (Queensland University of Technology) and the Secretariat of the Pacific Community for their provision of facilities and technical support during this project. PCD was a PhD candidate at The University of the South Pacific.
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