Molecular and immunoelectron microscopic identification of Pepino mosaic potexvirus in tomato fruits imported from Spain [Molekulinis ir imunoelektroninis mikroskopinis pepino mozaikos poteksviruso identifikavimas importuotų iš Ispanijos pomidorų vaisiuose]. – Botanica Lithuanica, 10(1): 89–98
In Lithuania Pepino mosaic potexvirus (PepMV) causing disease in tomato (Lycopersicon esculentum Mill.) crops has been isolated from imported tomato fruits and identified on the basis of determination of host range, symptom expression, morphological, and serological properties of the virus isolates. Data on the antigen purification and preparation of specific diagnostic antiserum against PepMV are presented. Antiserum to PepMV was produced by immunization of rabbits. The serological properties of PepMV antisera were determined by microprecipitin tests under mineral oil. The homologous titres of produced antisera were 1:512 and 1:1024. Using immunosorbent electron microscopy (ISEM) potexvirus particles of PepMV were decorated using immunoglobulin (IgG) prepared against PepMV. The primers designed on the basis of published sequences were applied for amplification of PepMV RNA fragments in reverse transcription-polymerase chain reaction (RT-PCR) using naturally and experimentaly PepMV affected tomato fruits of infected host plants. The detection of PepMV in tomato fruits imported from Spain and inoculated test plants was confirmed by RT-PCR. Analysis of PCR products in gel electrophoresis revealed amplification of 844 bp (base pair) fragments, which were in agreement with size of the fragment expected from the sequence data.
Keywords: tomato, Pepino mosaic virus, identification, purification, antiserum, immunoglobulin, ISEM, RT-PCR.