REDBIO México 2010

Folio: 181
Tipo de presentación: Oral
Tema: Metabolómica
Este trabajo se presentará en "Presentaciones Libres 14: Ómicas" el Miercoles dentro del horario 16:40 a 18:40

"Efficient genetic transformation of Digitalis purpurea, an important pharmaceutical crop"

Naivy Pérez-Alonso * (1), Borys Chong-Pérez (1), Alina Capote (1), Anabel Pérez (1), Yovanny Izquierdo (1), Geert Angenon (2), Elio Jiménez (1).
* Autor para correspondencia (Corresponding author) naivy@ibp.co.cu

1- Instituto de Biotecnología de las Plantas. Universidad Central “Marta Abreu” de Las Villas. Carretera a Camajuaní km 5.5, Santa Clara, Villa Clara, Cuba, CP 54 830; 2- Laboratory of Plant Genetics, Department of Applied Biological Sciences, Vrije Universiteit Brussel (VUB) Campus Etterbeek - Gebouw E Pleinlaan 2 B-1050 Brussel Belgium

RESUMEN (ABSTRACT)
"Digitalis purpurea L., contain cardiac glycosides of major interest in pharmaceutical industries. Genetic transformation is a tool of special interest to elucidate genes related with the production of these compounds, exclusively obtained from plants. Besides, it grants the possibility to widen the knowledge on the biosynthesis of these compounds. In this study, an efficient transformation protocol for D. purpurea that would permit the stable expression of transgenes was developed. A regeneration protocol via somatic embryogenesis was obtained from in vitro plant leaf segments. The minimal inhibitory concentration of geneticin was determined for each step of the protocol. Two Agrobacterium strains were used to test the T-DNA transfer ability, EHA 101 and C58C1pMP90, harbouring the binary vector pTJK136. The effectiveness of the method was proven by histochemical, PCR and Southern hybridization. A six days co-cultivation on medium with 1.0 mg.l-1 2,4-D was used to callus induction. The results showed that both leaf explants and callus pieces were sensitive to geneticin 70 mg.l-1. Non differences were found between two A. tumefaciens strain on transient Gus expression. Nevertheless, the strain C58C1pMP90 yielded better results regarding number of transformed plants. GUS histochemical analysis of the putative transgenic tissues further confirmed the transformation event. The transformation efficiency reached 82%. PCR and Southern blot hybridization confirmed the presence of the transgene and their stable integration in the regenerated plants. Till date, only a few papers have been published on studies of genetic transformation of Digitalis species. From these results an Agrobacterium tumefaciens-mediated genetic transformation protocol of D. purpurea was by the first time developed."