ISHS Acta Horticulturae 521: XXV International Horticultural Congress, Part 11: Application of Biotechnology and Molecular Biology and Breeding - Gene Expression and Molecular Breeding, Genome Analysis.
CONSTRUCTION AND TESTING OF A TRANSFORMATION VECTOR AIMING AT INTRODUCING PDV RESISTANCE IN ALMOND |
| Authors: | M.H. Raquel, C. Miguel, G. Nolasco, M.M. Oliveira |
| Keywords: | Nicotiana tabacum, Almond, Prunus dulcis, genetic transformation, Prune dwarf virus, Virus resistance |
| Abstract:
In South Portugal (Algarve) most almond trees are infected with viruses. It is desirable not only to obtain virus-free plants but also to introduce resistance to the most damaging viruses. To obtain clean plants of portuguese almond genotypes, several methods have been tried including thermotherapy, chemotherapy and meristem-tip culture. Prune Dwarf virus (PDV) was found as one of the most important viruses affecting almond. To introduce resistance to this virus, a transformation vector was constructed using the technique of IC-RT/PCR (immunocapture-reverse transcription/ /polymerase chain reaction) to isolate the coat protein gene. The PDVcp gene was cloned and inserted in pBI121, driven by the 35SCaMV promoter, in substitution of the GUS (beta-glucuronidase) gene. Prior to use in almond transformation, this construction was tested in the transformation of a model plant, Nicotiana tabacum var. Samsun. Genetic transformation was confirmed by PCR and Southern analysis and the protein was shown to express in tobacco as confirmed by ELISA. This construction will be used in almond transformation experiments. | |
Download Adobe Acrobat Reader (free software to read PDF files) | |
