Cell suspension culture and plant regeneration in the latex-producing plant,Calotropis gigantea (Linn.) R. Br. |
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Authors: | Roy Ashis T. Koutoulis Anthony De Deepesh N. |
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Affiliation: | (1) Applied Botany Section, Agricultural Engineering Department, Indian Institute of Technology, Kharagpur, West Bengal, 721 302, India;(2) Cell Biology Group, School of Plant Science, University of Tasmania, GPO Box 252-55, Hobart, TAS 7001, Australia |
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Abstract: | A procedure is outlined for the establishment of a proliferating cell suspension culture and subsequent plant regeneration of the latex-producing plant,Calotropis gigantea (Linn.) R. Br. Friable calluses were obtained by culturing hypocotyl explants on modified Murashige and Skoog medium with 2.69 μM α-naphthaleneacetic acid and 4.44 μM 6-benzyl-aminopurine. Friable calluses were transferred to modified Murashige and Skoog liquid medium containing 500 mg l−1 casein hydrolysate, 5% (v/v) coconut water and 5% (w/v) sucrose to initiate suspension cultures. Suspensions were subcultured every 10–12 days and supplemented with 13.56 μM 2,4-dichlorophenoxyacetic acid (2,4-D). After 3–4 subcultures, suspensions consisted of small, highly cytoplasmic cell clumps and large vacuolate cells. Plating the suspension clumps on medium containing 4.52 μM 2,4-dichlorophenoxyacetic acid and culturing in darkness produced macroscopic calluses, which subsequently produced a high number of shoots when placed in light and supplemented with 2.22 μM 6-benzyl-aminopurine and 0.45 μM 2,4-dichlorophenoxyacetic acid. Shoots were rooted using Bonner's solution containing 0.49 μM indole-3-butyric acid, and the plants successfully transferred to soil. This revised version was published online in June 2006 with corrections to the Cover Date. |
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Keywords: | Calotropis gigantea cell suspension culture latex organogenesis |
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