Pepino mosaic virus (PepMV) was detected for the first time in 1974 in Peru on pepino (Solanum muricatum). It infected tomato (Lycopersicon esculentum) latently and did not pose any threat to this plant. The presence of the PepMV in Europe was confirmed in 1999, but the virus performed as a very dangerous pathogen for tomato. The virus was also detected in tomato crops in North and South America. PepMV is under official control in Europe and in many other parts of the world and it has been placed in the EPPO alert list. The isolates of PepMV belong to different strains: 1)Peruvian 2)European tomato 3)American US1, and 4) American US2. In Poland in 2002 and 2005 two different isolates of Pepino mosaic virus signed PepMV-SW and PepMV-PK were obtained. The studies showed that Polish isolates differ from each other and belong to two strains: PepMV-SW to European and PepMV-PK to US2 strain. A rapid and highly sensitive detection and quantification method for PepMV was develop using SYBR Green chemistry. We designed specific primers which amplified both strains without non-specific products. The polymerase chain reaction mixture contained a fluorescent dye SYBR Green which exhibits fluorescent enhancement upon biding to double strand DNA. A linear relationship was observed between input cDNA and cycle threshold (Ct) values for 10⁶ down to a single copy of both strains. We obtained different Tm values for PepMV-PK and PepMV-SW according to differences in nucleotide composition of amplified products. To control for the variation in sample processing and in PCR among samples, ß -actin gene was amplified in parallel with the viral cDNA. The specificity of PepMV and ß- actin amplifications was confirmed by analyzing the dissociation curve of the target amplicon. The Ct values of PepMV samples were normalized against ß-actin values for determining the absolute copy number from the standard curve of the corresponding virus. The method describe above is highly robust and specific and it is a useful tool for diagnostics, epidemiological and genetic studies of PepMV .

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1. Phylogenetic analysis of Polish two types of Beet necrotic yellow vein virus
2. Comparative analysis of 2a polymerase protein sequence of chosen Polish strains of Peanut stunt virus, PSV.