ISHS Acta Horticulturae 386: XVI International Symposium on Fruit Tree Virus diseases
3’ NON-CODING REGION RT-PCR DETECTION AND MOLECULAR HYBRIDIZATION OF PLUM POX VIRUS IN ANTHERS OF INFECTED STONE FRUIT |
| Authors: | L. Levy, A. Hadidi, M. Kolber, G. Tokes, M. Nemeth |
| Abstract:
The international movement of germplasm may require movement of stone fruit anthers for germplasm collection, breeding programs, or other purposes. To investigate the feasibilty of dissemination of plum pox virus (PPV) through the exchange of Prunus anthers collected from PPV-infected trees, we assayed over 120 trees of various cultivars of peach, plum, and apricot grown in a PPV-infected experimental cultivar evaluation orchard in Cegled, Hungary. Individual trees were verified to be PPV-infected by either visual symptoms on leaves or fruit, and/or by indirect ELISA in Hungary prior to these experiments. We evaluated PPV detection using RT-PCR from dessicated anthers after storage at 4°C for 1–2 years. Anther tissues were prepared with Gene Releaser™ polymeric matrix, and then RT-PCR assayed using DNA primers for the 3’ non-coding region (NCR) of PPV. Polyacrylamide gel electrophoresis and dot-blot hybridization of the amplified PPV product from anthers using a 3’ NCR digoxigenin-labeled PPV-D cRNA probe showed high levels of hybridization signal. Our results suggest that stone fruit anthers could be a source of PPV dissemination during international movement of Prunus germplasm. | |
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