High-efficiency Agrobacterium-mediated transformation of citrus via sonication and vacuum infiltration.
Plant Cell Rep
; 28(3): 387-95, 2009 Mar.
Article
en En
| MEDLINE
| ID: mdl-19048258
ABSTRACT
An improved method for the Agrobacterium infiltration of epicotyl segments of 'Pineapple' sweet orange [Citrus sinensis (L.) Osbeck] and 'Swingle' citrumelo [Citrus paradisi Macf. X Poncirus trifoliata (L.) Raf.] was developed in order to increase transformation frequency. Sonication-assisted Agrobacterium-mediated transformation (SAAT), vacuum infiltration, and a combination of the two procedures were compared with conventional Agrobacterium-mediated inoculation method ('dipping' method). It was observed that the morphogenic potential of the epicotyl segments decreased as the duration of SAAT and vacuum treatments increased. Transient GUS expression was not affected by the different SAAT treatments, but it was significantly enhanced by the vacuum infiltration treatments. The highest transformation efficiencies were obtained when the explants were subjected to a combination of SAAT for 2 s followed by 10 min of vacuum infiltration. PCR and Southern blot analysis of the uidA gene were used to confirm the integration of the transgenes. The transformation frequencies achieved in this study (8.4% for 'Pineapple' sweet orange and 11.2% for 'Swingle' citrumelo) are the highest ones reported for both cultivars.
Texto completo:
1
Colección:
01-internacional
Banco de datos:
MEDLINE
Asunto principal:
Citrus
/
Plantas Modificadas Genéticamente
/
Técnicas de Transferencia de Gen
Idioma:
En
Revista:
Plant Cell Rep
Asunto de la revista:
BOTANICA
Año:
2009
Tipo del documento:
Article
País de afiliación:
Brasil