Indirect organogenesis in Phaseolus vulgaris L. cv. CIAP 7247
Journal Title: Biotecnología Vegetal - Year 2008, Vol 8, Issue 2
Abstract
The main difficulty that limits the genetic transformation in Phaseolus vulgaris L. is the low responsiveness for in vitro regeneration. The development of an in vitro regeneration via organogenesis protocol is needed in order to achieve the genetic transformation in one of the most important cultivar of this legume in Cuba. Therefore, the aim of this work was to obtain shoots formation from morphogenetic calli in Phaseolus vulgaris L. cv. CIAP 7247. Matures seeds harvested in greenhouse were used as plant material. Different explants were studied for callus formation (cotyledon, cotyledon with a segment of embryonic axis and segment of embryonic axis with cotyledonal nude). Number of explants with callus formation was evaluated after 28 days. The percentage was also determined. Formed calli were classified according to their consistency and structure in compacts-nodular and soft – spongy. Three concentration of 6-bencylaminopurine (0.5, 1.0 and 2.0 mg.l -1 ) and control treatment without growth regulator were studied to achieve shoot induction and formation from calli. Number of shoots formed on 0.5 g of callus fresh mass, after 45 days of culture, was evaluated. The highest percentage (86%) of explants with callus formation was determinate on cotyledons. Segments of embryonic axis with cotyledonal nude produced 57%, and cotyledons with embryonic axis 38%. However, shoots were only formed on calli grown from cotyledons with embryonic axis. The addition of 6-BAP 2.0 mg.l -1 to shoot formation medium increased in 2.2 the number of shoots observed on 0.5 g of calli. It was demonstrated that the kind of explant had a determinant effect on calli regeneration.
Authors and Affiliations
Raúl Collado , Lourdes R. García , Gert Angenon , Dámaris Torres , Carlos Romero , Idalmis Bermúdez- Caraballoso, Novisel Veitía, Miriam Ramírez
Keywords
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