Co-Authors:
Rotem, N., The R.H. Smith Institute of Plant Sciences and Genetics in Agriculture, R.H. Smith Faculty of Agriculture, Food and Environment, The Hebrew University of Jerusalem, Rehovot, Israel
Shtein, C., The R.H. Smith Institute of Plant Sciences and Genetics in Agriculture, R.H. Smith Faculty of Agriculture, Food and Environment, The Hebrew University of Jerusalem, Rehovot, Israel
Rosner, A., The Department of Virology, Institute of Plant Protection, The Volcani Center, ARO, Bet Dagan, Israel
Levy, D., The R.H. Smith Institute of Plant Sciences and Genetics in Agriculture, R.H. Smith Faculty of Agriculture, Food and Environment, The Hebrew University of Jerusalem, Rehovot, Israel
Rabinowitch, H.D., The R.H. Smith Institute of Plant Sciences and Genetics in Agriculture, R.H. Smith Faculty of Agriculture, Food and Environment, The Hebrew University of Jerusalem, Rehovot, Israel
Abstract:
A novel simple, fast qualitative method for detection and differentiation of PVY strains in potatoes by the LightCycler technology is described. Fluorescent-labeled probe designed to contain variable degree of homology with a ‘target sequence’ of several known reference PVY strains was annealed to PCR products of these viruses followed by a graded melting analysis. The specific characteristics of the melting curves enable the detection, distinction and differentiation of each of following four known PVY strains O-FL, O-RB, N and NTN, in a single reaction obviating the need for size or nucleotide sequence analyses. In addition, it was demonstrated that virus extracts from plants infected with more than one strain can be resolved using this procedure. The melting-curves of extracts from 55 market size tubers harvested randomly from commercial fields and compared with those of the reference virus strains, revealed the presence of O-FL, O-RB and NTN strains of PVY. © 2016, The Potato Association of America.