Genetic transformation by Agrobacterium tumefaciens in papaya hybrid from in vitro plant shoot-tips
Journal Title: Biotecnología Vegetal - Year 2004, Vol 4, Issue 1
Abstract
In papaya (Carica papaya L.) viral infections can decrease production by more than 50%. Genetic transformation consists of an alternative means of obtaining new varieties, which are resistant to this pathogen. The main objective of the present work was to develop a methodology for the transformation of a papaya hybrid via A. tumefaciens. As plant material, shoot-tips (3-5 mm) from in vitro papaya plants were used. The Agrobacterium tumefaciens strains EHA-105 and AT 22-60 were employed, which contained the plasmid pCAMBIA 3301.Co-culture conditions were established to achieve the integration of the foreign gene into the genome of the hybrid. The integration of the transgene sequence was confirmed by the expression of the GUS gene in the tissues of the explants through histo- chemical assay. The greatest transient expression of β --glucuronidase was achieved with the shoot-tips co-cultured at 28 o C during five days, using the strain EHA 105 with an average frequency of 20 blue foci per infected shoot-tip. Complete transgenic plants were obtained 4 subcultures after transformation and these line obtained were resistant to BASTA®. The final results demonstrate that it is possible to obtain plants transformed into hybrids of papaya from its meristematic apexes.
Authors and Affiliations
Jorge Gallardo Colina, Rafael Gómez Kosky, Maritza Reyes, Bárbara Ocaña Díaz, Idalia Herrera O`farri, Marisol Freire Seijo
Keywords
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