Experience with the diagnostics and purification of plum pox virus

Abstract

Some diagnostic methods devised for the demonstration of the presence of plum pox virus in plum (Prunus domestica L.) and apricot (Armeniaca vulgarisLam.) leaves were examined. The method of radial diffusion in agar can be recommended as the simplest and the least time consuming method which can be used during the entire vegetation period. In order to obtain antisera, some preparation methods of PPV antigen were verified. The best preparation method was a modification of Van Oosten’s method in which HEPES buffer pH 6.7 was used for the homogenization-of leaves from infectedNicotiana clevelandiiGray plants and for the solution of sediments after ultracentrifugation. In this way, antigens with titre up to 1: 2048 and during the immunization of rabbit antisera with titre 1: 512 to 1: 1024 were obtained. After the saturation of antisera according to Uyemoto, the titre of the antisera was 1: 64 to 1: 256. Antisera were used for agar preparation in transparent plastic boxes. 0.2 to 1 g portions of leaf material were homogenized with 0.05 M Tris-HCl buffer pH 7.2, 3% pyrrolidine and 1 % polyvinylpyrrolidone in the ratio of 1: 3 for the determination of PPV in plum and apricot leaves.