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Coat protein-mediated protection against plum pox virus in herbaceous model plants and transformation of apricot and plum
Authors:Artur da Câmara Machado  Hermann Katinger  Margit Laimer da Câmara Machado
Institution:1. University of Agriculture and Forestry, Institute of Applied Microbiology, Nu?dorfer L?nde 11, 1190, Vienna, Austria
Abstract:Summary The expression of the viral coat protein gene in transgenic plants has been shown to induce tolerance against virus infection (Beachy et al., 1990). Transgenic plants ofNicotiana clevelandii andNicotiana benthamiana- herbaceous host plants for PPV - transformed withAgrobacterium strain LBA 4404 containing the plasmid pBinPPVm, regenerated on selection media containing kanamycin were tested for the expression of the PPV coat protein gene by ELISA and immuno western blot. After rooting and acclimatisation plants were tested for the protection against PPV Following the inoculation plants were investigated for symptom development and virus accumulation. Different lines were identified, according to the different reaction to the mechanical inoculation, ranging from a complete absence to a strong reduction of symptoms. There have not been many reports on transformation of trees in general, and in fruit trees particularly. It is obvious that the major obstacle is the regeneration of transformed plantlets. Attempts to improve crop plants by genetic engineering techniques will always depend very strongly on the availability of reliable protocols for transformation, selection and regeneration (Laimer et al., 1989, 1990). Different systems involving juvenile and adult plant material have been developed allowing the transfer of foreign genes into apricot and plum cultivars. We report the transformation and regeneration ofPrunus armeniaca andPrunus domestica plants withAgrobacterium tumefaciens strain LBA 4404 containing various binary plasmids, pBinGUSint, carrying the marker geneβ-glucuronidase (GUS) and pBinPPVm, carrying the coat protein gene of Plum Pox Virus (PPV), the causal agent of Sharka disease. The marker geneGUS was used for the optical evaluation of the efficiency of different transformation systems involving cotyledons of immature embryos as well as somatic embryos and leaf discs. The coat protein gene of PPV was used to introduce the coat protein mediated resistance against one of the most important pathogens of stone fruit trees in Europe and the whole Mediterranean area.
Keywords:Coat protein-mediated protection  Plum Pox Virus  transformation            Prunus armeniaca                      Prunus domestica            apricot  plum
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