Articles
PRODUCTION OF TRANSGENIC POINSETTIA WITH RESISTANCE AGAINST POINSETTIA MOSAIC VIRUS (PNMV) USING AGROBACTERIUM-MEDIATED TRANSFORMATION
Article number
901_10
Pages
87 – 93
Language
English
Abstract
Genetic engineering plays a significant role for the improvement of ornamental crops.
Using an Agrobacterium-mediated transformation approach, we have produced transgenic poinsettia conferring resistance against Poinsettia mosaic virus (PnMV). A. tumefaciens harbouring three hairpin (hp) RNA gene constructs to generate silencing-based resistance to PnMV was introduced into internode stem explants of poinsettia cultivar Millenium.
Stable integration of transgenes into the poinsettia nuclear genome was confirmed by PCR and Southern blot analysis.
Double antibody sandwich enzyme-linked immunosorbent assays (DAS-ELISA) revealed that resistance to mechanical inoculation of PnMV was found among the transgenic poinsettia lines, whereas the controls were susceptible.
Grafting technique was used to re-introduce the free branching factor, phytoplasma, back to the selected desirable transgenic poinsettia lines.
Subsequently, fresh cuttings derived from these lines with phytoplasma and branches were further investigated for the stability of PnMV resistance and their morphological performance compared with non-transformed controls.
Currently, these lines are undergoing evaluation at the growers facility for future commercialization.
Using an Agrobacterium-mediated transformation approach, we have produced transgenic poinsettia conferring resistance against Poinsettia mosaic virus (PnMV). A. tumefaciens harbouring three hairpin (hp) RNA gene constructs to generate silencing-based resistance to PnMV was introduced into internode stem explants of poinsettia cultivar Millenium.
Stable integration of transgenes into the poinsettia nuclear genome was confirmed by PCR and Southern blot analysis.
Double antibody sandwich enzyme-linked immunosorbent assays (DAS-ELISA) revealed that resistance to mechanical inoculation of PnMV was found among the transgenic poinsettia lines, whereas the controls were susceptible.
Grafting technique was used to re-introduce the free branching factor, phytoplasma, back to the selected desirable transgenic poinsettia lines.
Subsequently, fresh cuttings derived from these lines with phytoplasma and branches were further investigated for the stability of PnMV resistance and their morphological performance compared with non-transformed controls.
Currently, these lines are undergoing evaluation at the growers facility for future commercialization.
Authors
J.L. Clarke, C. Spetz, S. Haugslien, M.W. Dees, R. Moe, D.-R. Blystad
Keywords
Euphorbia pulcherrima, Poinsettia, transformation, Agrobacterium tumefaciens, Poinsettia mosaic virus
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