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Development of Plum pox virus resistance in plum via expression of an intron-spliced hairpin HC-Pro transgene.

Tian, L.-N., Svircev, A.M., Sanfaçon, H., Brown, D.C.W., Schneider, K.E., Sibbald, S., and Malutan, T. (2010). "Development of Plum pox virus resistance in plum via expression of an intron-spliced hairpin HC-Pro transgene.", Genes, Genomes & Genomics, 4(Suppl. 1), pp. 28-31.

Abstract

Plum pox virus (PPV) is a serious viral disease of Prunus species. Few sources of natural resistance to PPV are available. In this light, biotechnology is a strong option to develop PPV resistance in plants. We have previously shown that a transgene consisting of a region of the PPV HC-Pro sequence in an intron-spliced hairpin conformation could provide high level of resistance to PPV in Nicotiana benthamiana, a herbaceous model host. We have now generated transgenic plum lines expressing the same transgene. Plants were inoculated with PPV virus using a chip bud grafting method and were subjected to three cycles of dormant/growth periods. Plants were analyzed for PPV resistance after each cold treatment (dormant period) using a real-time polymerase chain reaction (PCR) assay. After the first cold treatment, transgenic plum plants remained PPV-free whereas all the control plants were PPV positive. After three cycles of cold treatment, many transgenic plants still exhibited strong resistance to PPV. Presence of siRNA molecules corresponding to the HC-Pro transgene were confirmed in transgenic lines that were highly resistant to PPV. This study shows that induction of PPV-specific RNA silencing via expression of a intron-spliced hairpin HC-Pro transgene is an effective approach for high level and stable resistance to plum pox virus in plum.

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