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Monitoring Prunus Necrotic Ringspot Virus Infection by Hybridization with a CRNA Probe following in Vitro Grafting
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In vitro micrografting was tested as a technique for inoculating peach [Prunus persica (L.) Batsch] with prunus necrotic ringspot virus (PNRSV). Cultured `Suncrest' shoots derived from a naturally infected tree (as indicated by ELISA testing) maintained virus in vitro, with virus concentrations in growing tips and folded leaves being several times those of fully expanded leaves. The infected shoots served as graft bases and the source of virus. Grafted tips were derived from `Suncrest' trees that had tested negative for the virus. Leaf samples were collected from the tips following grafting and analyzed for the presence of virus by slot-blot hybridization with a digoxigenin-labeled cRNA probed derived from PNRSV RNA 3. Rates of successful grafting were 55% and 73% in three trials and PNRSV was found in all tips analyzed. Virus concentrations approximated those found in the source shoots, suggesting that this method should be useful for screening transformed peach shoots for coat protein-mediated resistance to PNRSV.
American Society for Horticultural Science
Title: Monitoring Prunus Necrotic Ringspot Virus Infection by Hybridization with a CRNA Probe following in Vitro Grafting
Description:
In vitro micrografting was tested as a technique for inoculating peach [Prunus persica (L.
) Batsch] with prunus necrotic ringspot virus (PNRSV).
Cultured `Suncrest' shoots derived from a naturally infected tree (as indicated by ELISA testing) maintained virus in vitro, with virus concentrations in growing tips and folded leaves being several times those of fully expanded leaves.
The infected shoots served as graft bases and the source of virus.
Grafted tips were derived from `Suncrest' trees that had tested negative for the virus.
Leaf samples were collected from the tips following grafting and analyzed for the presence of virus by slot-blot hybridization with a digoxigenin-labeled cRNA probed derived from PNRSV RNA 3.
Rates of successful grafting were 55% and 73% in three trials and PNRSV was found in all tips analyzed.
Virus concentrations approximated those found in the source shoots, suggesting that this method should be useful for screening transformed peach shoots for coat protein-mediated resistance to PNRSV.
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