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Cell suspension culture and plant regeneration in the latex-producing plant, Calotropis gigantea (linn.) R. Br
journal contributionposted on 2023-05-16, 12:41 authored by Roy, A, Anthony KoutoulisAnthony Koutoulis, De, DN
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 Î¼M Î±-naphthaleneacetic acid and 4.44 Î¼M 6-benzyl-aminopurine. Friable calluses were transferred to modified Murashige and Skoog liquid medium containing 500 mg 1-1 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 Î¼M 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 Î¼M 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 Î¼M 6-benzyl-aminopurine and 0.45 Î¼M 2,4-dichlorophenoxyacetic acid. Shoots were rooted using Bonner's solution containing 0.49 Î¼M indole-3-butyric acid, and the plants successfully transferred to soil.
Publication titlePlant Cell Tissue & rgan Culture
Department/SchoolSchool of Natural Sciences
PublisherKluwer Academic Publishing Group
Place of publicationNetherlands