Globularia nudicaulis (family Globulariaceae ) is a perennial, found naturally on European mountains at altitudes between 900 and 2000 m. In June 2004, G. nudicaulis plants, with a yellow mosaic and/or variegation on malformed leaves, were noted among plant species cultivated in the Botanical Garden at Bologna University, Italy. No elongated virus-like particles were observed in affected-leaf extracts by transmission electron microscopy using a leaf dip method. By applying a protein A sandwich enzyme-linked immunosorbent assay (PAS-ELISA) technique (Edwards & Cooper, 1985), Cucumber mosaic virus (CMV) was detected using a polyclonal antiserum (PVAS 30, American Type Culture Collection, Manassas, VA, USA). CMV from G . nudicaulis leaves was mechanically transmitted to 10 of 14 species tested. Local lesions were observed in Chenopodium murale and C. quinoa ; systemic symptoms were observed in Nicotiana tabacum , N. benthamiana , N. glutinosa , N. clevelandii and Capsicum annuum , and Cucumis sativus and C. melo . Reverse transcription-polymerase chain reaction (RT-PCR) and single-strand conformation polymorphism (SSCP) were employed to characterize the CMV isolate. Total RNA was extracted from affected G. nudicaulis leaf samples with a RNeasy Plant Minikit (Qiagen) according to the manufacturer’s instructions. RT-PCR was carried out using specific primers for the movement protein gene of RNA3 of CMV (forward MP + CATGGCTTTCCAAGGTACCAG, genomic position 118nt to 138nt, and reverse CTAAAGACCGTTAACCACCTGC, genomic position 938nt to 959nt; Lin et al ., 2004). All samples yielded DNA fragments of the expected size: 841 bp. PCR products were then analysed by SSCP to identify specific sequence variants and compare genetic relationships with other CMV isolates from the Botanical Garden (MGB & SD, unpublished results). The results showed a sequence variant that was different from other local CMV isolates, indicating that CMV isolate G in G. nudicaulis is a new accession in this location. This is the first time that CMV has been isolated from G. nudicaulis.
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