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Transgenic plums (Prunus domestica L.) express the plum pox virus coat protein gene
Authors:Ralph Scorza  Michel Ravelonandro  Ann M Callahan  John M Cordts  Marc Fuchs  Jean Dunez  Dennis Gonsalves
Institution:(1) USDA-ARS Appalachian Fruit Research Station, 45 Wiltshire Rd., 25430 Kearneysville, WV, USA;(2) Station de Pathologie Vegetale, INRA, BP 81, 33 883 Villenave d'Ornon, France;(3) Department of Plant Pathology, Cornell University, 14456 Geneva, NY, USA
Abstract:Summary Plum hypocotyl slices were transformed with the coat protein (CP) gene of plum pox virus (PPV-CP) following cocultivation with Agrobacterium tumefaciens containing the plasmid pGA482GG/PPVCP-33. This binary vector carries the PPV-CP gene construct, as well as the chimeric neomycin phosphotransferase and beta-glucuronidase genes. Integration and expression of the transferred genes into regenerated plum plants was verified through kan resistance, GUS assays, and PCR amplification of the PPV-CP gene. Twenty-two transgenic clones were identified from approximately 1800 hypocotyl slices. DNA, mRNA, and protein analyses of five transgenic plants confirmed the integration of the engineered CP gene, the accumulation of CP mRNA and of PPV-CP-immunoreactive protein. CP mRNA levels ranged from high to undetectable levels, apparently correlated with gene structure, as indicated by DNA blot analysis. Western analysis showed that transgenic plants produced amounts of CP which generally correlated with amounts of detected mRNA.
Keywords:Agrobacterium tumefaciens  Cross protection  Sharka  Transformation  Virus resistance
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