New Polish Isolate of Pepino mosaic virus Highly Distinct from European Tomato, Peruvian, and US2 Strains

Abstract

Pepino mosaic virus (PepMV, genus Potexvirus) was first described on pepino (Solanum muricatum) in Peru during 1980. Since 1999, the virus was reported in several European countries and in North and South America as an agent of viral disease of tomato crops. In Poland in 2002, the PepMV-SW isolate that was genetically similar to European isolates (approximately 99% identity) was identified (3). In November 2005, in the western part of the Wielkopolska Region, a virus with flexuous filamentous particles approximately 500 nm long was isolated from tomato fruits exhibiting symptoms of discoloration. Crude sap from Nicotiana benthamiana leaves was used for mechanical inoculation of indicator plants. The virus caused symptoms on N. benthamiana, N. clevelandii, Datura inoxia, and Lycopersicon esculentum. Symptomless infection on N. tabacum cv. Xanthi nc, N. tabacum cv. White Burley, and N. debneyi was confirmed by back-inoculation on N. benthamiana. The virus did not infect N. glutinosa, Physalis floridana, Petunia hybrida, Capsicum annuum, Chenopodium quinoa, Cucumis sativus, or Phaseolus vulgaris. The virus was initially identified using double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) with polyclonal antiserum against PepMV (DSMZ, Braunschweig, Germany). Positive serological reactions were obtained with sap from inoculated N. benthamiana, L. esculentum, and N. clevelandii plants. The serological identification was confirmed using a reverse transcription-polymerase chain reaction (RT-PCR) with primers generated from a sequence of the RNA polymerase region of an isolate of PepMV reported in the United Kingdom (1). Sequence information obtained from the amplified fragment of the virus designated PepMV-PK (GenBank Accession No. DQ387870), showed only 81% nt identity and 89% amino acid identity with PepMV-SW (GenBank Accession No. DQ387869). PepMV isolates can be divided into three strains including European tomato, Peruvian, and US2 based on their genetic diversity (2). The PepMV-PK isolate resulted in nucleotide identities ranging from 79 to 81% with isolates of the European tomato strain (GenBank Accession Nos. AJ438767, AF340024, AF484251, AJ271991, AJ606359, and AJ290424), 81% with the Peruvian strain (GenBank Accession Nos. AM109896 and AJ606361), and 78% identity with each of the U.S. isolates US1 (GenBank Accession No. AY509926) and US2 (GenBank Accession No. AY509927). These results show that the new Polish isolate is distinct from all other PepMV isolates reported to date.