Plant Pathology (2009) 58, 407 Doi: 10.1111/j.1365-3059.2009.02026.x © 2009 The Authors Journal compilation © 2009 BSPP 407 Blackwell Publishing Ltd Basil (Ocimum basilicum), a new host of Pepino mosaic virus S. Davino c *, G. P. Accotto b , V. Masenga b , L. Torta c and M. Davino a a Dipartimento di Scienze e Tecnologie Fitosanitarie (DISTEF) – sez. di Patologia vegetale, Università degli Studi di Catania, Via S. Sofia, 100, I-95123 Catania; b Institute of Plant Virology, C.N.R., Strada delle Cacce, 73, 10135 Torino; and c Dipartimento di Scienze Entomologiche, Fitopatologiche, Microbiologiche e Zootecniche (SENFIMIZO) – sez. di Patologia vegetale, Università degli Studi di Palermo, Viale delle Scienze, I-90128 Palermo, Italy Pepino mosaic virus (PepMV), a member of the genus Potexvirus, was first described in 1980 on pepino (Solanum muricatum) and was later isolated from different wild species of the genus Solanum (formerly Lycopersicon) (Soler et al., 2002). PepMV has been reported in several European countries and in North America, causing disease in tomato. In July 2008, plants of basil (Ocimum basilicum) showing interveinal chlorosis on young leaves were observed in three greenhouses in Sicily, in the area where tomato plants were found to be infected by PepMV in 2005 (Davino et al., 2006). The disease was transmitted mechanically to Nicotiana benthamiana, producing chlorotic spots and leaf deformation. Electron microscopy of negatively- stained preparations from basil and N. benthamiana leaves, both with symptoms, revealed the presence of flexuous, filamentous virus particles, typical of potexviruses. Leaves from 100 plants with or without symptoms were analysed by double-antibody sandwich (DAS) ELISA using a polyclonal antibody to PepMV (Loewe Biochemica). The presence of the symptoms was correlated with positive ELISA results. Total RNA was extracted from the same samples using an RNeasy Plant Mini Kit (Qiagen) and analysed by RT-PCR using the PepMV-specific primers PepMV-TGB-F and PepMV- UTR-R (Mumford & Metcalfe, 2001). The 840 bp RT-PCR product was sequenced in both directions and the sequence deposited in GenBank (Accession No. EU888637). The sequence had 99% sequence identity with an isolate of PepMV found in Sicily in 2005 (DQ517884). This is the first report of PepMV infecting basil. Although the disease does not appear too severe on basil, the species is frequently cultivated next to tomatoes and PepMV is transmitted mechanically easily; therefore basil may act as a virus reservoir. References Davino S, Bellardi MG, Agosteo GE, Iacono G, Davino M, 2006. Characterization of a strain of Pepino mosaic virus found in Sicily. Journal of Plant Pathology 88 (supplement 3), 40. Mumford RA, Metcalfe EJ, 2001. The partial sequencing of the genomic RNA of a UK isolate of Pepino mosaic virus and the comparison of the coat protein sequence with other isolates from Europe and Peru. Archives of Virology 146, 2455–60. Soler S, Prohens J, Diez MJ, Nuez F, 2002. Natural occurrence of Pepino mosaic virus in Lycopersicon species in Central and Southern Peru. Journal of Phytopathology 150, 49–53. *E-mail: davino@unipa.it. Accepted 19 September 2008 at www.bspp.org.uk/ndr where figures relating to this paper can be viewed. First report of Shallot virus X in shallot in New Zealand Z. Perez-Egusquiza a *, L. I. Ward a , G. R. G. Clover a , J. D. Fletcher b and R. A. A. van der Vlugt c a Plant Health and Environment Laboratory, MAF Biosecurity New Zealand, P.O. Box 2095, Auckland 1140; b Crop & Food Research, Private Bag 4704, Christchurch, New Zealand; and c Plant Research International, P.O. Box 16, 6700 AA Wageningen, The Netherlands In January 2005, mild mosaic and chlorosis were observed on leaves of shallot (Allium cepa var. aggregatum) cv. Red Prisma growing in Marl- borough, New Zealand. Leaves from 100 plants were collected and bulked into groups of ten. Three composite samples tested positive for Shallot mite-borne latent virus (ShMbLV) using polyclonal antiserum (supplied by Dr. E. Barg, Biologische Bundesanstalt, Germany) in an antigen-coated plate enzyme-linked immunosorbent assay (ACP-ELISA). Similar symptoms were seen in shallot cv. Jermor in the same region in December 2007. Leaves from these plants also tested positive for ShMbLV by ACP-ELISA. To confirm the identification of the samples from 2007, dried tissue of the type isolate of ShMbLV was obtained (Van Dijk & van der Vlugt, 1994). RNA was extracted from diseased shallot samples and the ShMbLV type isolate and tested by RT-PCR using primers that amplify a ca. 750 bp fragment between the coat protein (CP) and ORF6 region of allexiviruses (Chen et al., 2004). For both samples, amplicons of the expected size were obtained and sequenced directly. Analysis showed a 93 to 95% nucleotide identity with Shallot virus X (ShVX) (GenBank Accession No. M97264). RT-PCR was then done using specific primers designed to amplify a 912 bp fragment of ShVX including the CP gene (ShVX-CPF: 5′-ATTTAGGG- GTGAAGGTCTGT-3′; ShVX-CPR: 5′-GAGTTTTGAGGTCGTTGG-3′). Amplicons of the correct size were obtained from both diseased shallot samples and the ShMbLV type isolate. Subsequently, one-step immunocap- ture RT-PCR was performed using the ShMbLV antiserum and the ShVX- specific primers, and bands of the correct size were obtained for both samples. The amplicons were cloned and sequenced. A BLAST search showed that the sequence from shallot (EU835197) and that of the type isolate of ShMbLV (EU835196) showed 93 and 95% nucleotide identity, respectively, with ShVX (M97264). According to the criteria demarcating species in the genus (Adams et al., 2004), it is suggested that ShMbLV should be considered a synonym of ShVX as previously proposed (Van Dijk & van der Vlugt, 1994). This is the first report of ShVX in New Zealand and until these findings it was considered a regulated pest. However, no phytosanitary measures will be imposed to eradicate the virus and it is likely to spread in the future, especially as the vector, Aceria tulipae, is present in New Zealand. Allexi- viruses, including ShVX, often infect allium crops in combination with carlaviruses and/or potyviruses and cause significant yield losses (Van Dijk & van der Vlugt, 1994; Chen et al., 2004). References Adams MJ, Antoniw JF, Bar-Joseph M et al., 2004. The new plant virus family Flexiviridae and assessment of molecular criteria for species demarcation. Archives of Virology 149, 1045–60. Chen J, Zheng H-Y, Antoniw JF et al., 2004. Detection and classification of allexiviruses from garlic in China. Archives of Virology 149, 435–45. Van Dijk P, van der Vlugt RAA, 1994. New mite-borne virus isolates from rakkyo, shallot and leek species. European Journal of Plant Pathology 100, 269–77. *E-mail: zoila.perez@maf.govt.nz. Accepted 18 November 2008 at www.bspp.org.uk/ndr where figures relating to this paper can be viewed. Plant Pathology (2009) 58, 407 Doi: 10.1111/j.1365-3059.2009.02031.x