REDBIO México 2010

Folio: 134
Tipo de presentación: Oral
Tema: Mejoramiento Genético Vegetal
Este trabajo se presentará en "Presentaciones Libres 11: Biodiversidad, Mejoramiento y Marcadores Moleculares" el Miercoles dentro del horario 14:20 a 16:20

Effect of spermidine on Agrobacterium tumefaciens mediated transformation of banana cv. ‘Dwarf Cavendish’ (Musa spp. AAA group).

Borys Chong-Pérez * (1), Rafael G. Kosky (1), Maritza Reyes (1), Barbara Ocaña (1), Luis Rojas (1), Serge Remy (2), Geert Angenon (3).
* Autor para correspondencia (Corresponding author) boris@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 Tropical Crop Improvement, Katholieke Universiteit Leuven, Kasteelpark Arenberg 13, 3001 Leuven, Belgium ; 3- 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)
"Research in the field of transgenic improvement and functional genomics in banana is constrained by low efficiency transformation systems, and therefore an efficient transformation protocol is crucial for banana genomics and banana improvement initiatives. In this work an study of the effect of spermidine on genetic transformation was done in two different Agrobacterium tumefaciens based protocols, Centrifuge-Assisted Agrobacterium mediated Transformation (CAAT) and six hours Infection period of embryogenic banana cell with A. tumefaciens (6h-IAt). Embryogenic cell suspension culture (ECS) from Dwarf Cavendish (Musa AAA) was established from embryogenic callus obtained from immature male flower. For genetic transformation ECS were co-cultivated with Agrobacterium tumefaciens strain EHA101, harbouring binary vector pFAJ3000, under different conditions. The results indicate major difference between the efficiency of the two transformation protocols and spermidine treated and non treated cells. Significantly higher transient GUS expression was obtained with spemidine treated 6h-IAt protocol (4401±613 blue foci per 50 mg of settled cell volume of ECS) than the other treatments. This result was confirmed on colonies formation and plan regeneration, where 1004±89 independent colonies and 608±104 independent lines were obtained per 50 mg of settled cell volume of ECS after selection procedure on this treatment, two fold higher than the second best treatment (spermidine treated CAAT protocol). Stable expression and integration of the transgenes was confirmed by histochemical GUS assay, PCR and Southern blot analysis. The use of spermidine showed an enhancer effect on Agrobacterium-mediated banana transformation, and gave rise to hundreds of morphologically normal transgenic plants in cv. Dwarf Cavendish."