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ISHS Acta Horticulturae 235: XIV International Symposium on Fruit Tree Virus Diseases

VIRUS AND RNA-SPECIFIC MOLECULAR HYBRIDIZATION PROBES FOR TWO NEPOVIRUSES

Authors:   C. Bretout, T. Candresse, O. Le Gall, V. Brault, M. Ravelonandro, J. Dunez
Abstract:
Working with two related nepoviruses, tomato black ring virus (TBRV) and Hungarian latent grapevine chrome mosaic virus (GCMV), we have obtained a set of four probes which can be used in molecular hybridization experiments to specifically detect the two genomic RNAs of the two viruses. The probes are derived from the 3' region of each of the RNAs and are 700 to 1500 bp long. The two genomic RNAs of TBRV and of GCMV have identical 3' non coding ends, and these are about 73 % homologous between the two viruses. So, it is necessary to delete these 3' non coding regions in order to obtain RNA-specific probes. When using cDNA probes labeled by random priming under optimized conditions, it is possible to specifically detect as little as 3 pg of viral RNA in a simple dot blot hybridization assay.

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