Obtaining calluses with Stevia embryogenic structures rebaudiana Bert. in semi-solid culture media
Journal Title: Biotecnología Vegetal - Year 2008, Vol 8, Issue 2
Abstract
The adjustment of an efficient protocol of somatic embryogenesis in Stevia rebaudiana Bertoni to support programs of genetic breeding by genetic transformation is necessary. Few results are registered up to the moment in this species. The objective of this work was to obtain calli with embryogenic structures from clones of Stevia in vitro plants. Different variables were studied for the formation of calli: explants type, incubation conditions and genotype influences. Culture medium MS with different growth regulators (2,4-D; 6-BAP; TDZ) was used. These were used alone or in combination. Calli were multiplied in the best culture media. Afterwards they were subcultured to culture medium containing 2,4-D or TDZ with 40 and 60 g.l -1 of sucrose to induce proembryogenic mass formation. Calli formation was achieved after 45 days of culture in darkness and in culture media with 2.26 μM 2,4-D and 2.22 μM 6-BAP or 1.13 μM 2,4-D and 0.45 μM of TDZ. Foliar explants of in vitro plants of Stevia were used. Multiplication was achieved in the same culture media for calli formation. The largest amount of calli with development of proembryogenic mass was obtained when subcultures to culture medium with 0.45 μM 2,4-D adding 40 g.l -1 of sucrose was carried out after 45-50 days of culture.
Authors and Affiliations
Mario Kryvenki , Rafael G. Kosky , Diego Guerrero , Martín Dominguez , Maritza Reyes
Keywords
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