Transgenic almond (Prunus dulcis Mill.) plants obtained by Agrobacterium-mediated transformation of leaf explants |
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Authors: | C M Miguel M M Oliveira |
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Institution: | (1) Instituto de Biologia Experimental e Tecnológica, Apartado 12, P-2780 Oeiras, Portugal e-mail: cmiguel@itqb.unl.pt Fax: +351-1-4421161, PT;(2) Departamento de Biologia Vegetal, Faculdade de Ciências de Lisboa, Bloco C2, Campo Grande, P-1700 Lisboa, Portugal, PT |
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Abstract: | Almond (Prunus dulcis Mill.) leaves were transformed with the marker genes gusA (β-glucuronidase) and nptII (neomycin phosphotransferase II) via Agrobacterium-mediated transformation. Bacterial strains and preculture of explants affected efficiency of gene transfer evaluated by transient
expression assays. Following transformation, shoots were induced from primary explants on medium without kanamycin and exposed
to selection 20 days after cocultivation. From 1419 original leaves, four shoots (A, B, C and D) were obtained that showed
amplification of the predicted DNA fragments by polymerase chain reaction (PCR). After micropropagation of these shoots, only
those cloned from shoot D gave consistently positive results in histochemical GUS detection and PCR amplification. Southern
blot hybridisation confirmed stable transgene integration in clone D, which was also negative in PCR amplification of an Agrobacterium gene. Additional molecular analysis suggested that the remaining three shoots (A, B and C) were chimeric.
Received: 28 March 1998 / Revision received: 18 April 1998 / Accepted: 12 May 1998 |
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Keywords: | Almond Prunus Transformation Agrobacterium Adventitious regeneration |
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