Pelargonium chlorotic ring pattern virus: first report in Spain

A recent survey of symptomless ornamental geraniums ( Pelargonium zonale , P. peltatum and P. grandiflorum ) in Spain (2000–2003) revealed viral infections in 52·4% of the tested plants (Alonso & Borja, 2005). Most infected plants contained tombus-like viruses such as Pelargonium line pattern virus (PLPV), either alone or in combination with other viral pathogens. Several little-studied PLPV-related viruses have been reported in Pelargonium spp. but their distribution and importance to geranium growers is unknown since they are not usually included in phytosanitary programmes. One of these viruses, Pelargonium chlorotic ring pattern virus (PCRPV), was recently sequenced (GenBank Acc. No. AY038069) and found to be related to PLPV in nucleotide sequence and genome organization (Kinard & Jordan, 2002; Castano & Hernandez, 2005). To assess the occurrence of PCRPV in Spain 5 samples from the previous survey and 3 gathered in 2006 were tested. Total RNA was extracted using Plant RNA Reagent (Invitrogen) and reverse transcribed with Superscript II-RT (Invitrogen) and primer CH149 (5 ′ -CCGATTCTCGAGCTGCCAATC-3 ′ ), complementary to positions 1543–1523 of PCRPV. PCR used primers CH149 and CH148 (5 ′ -CGGTTTGAGCACTCTGCGTATC-3 ′ ), homologous to positions 1277–1298 of PCRPV. An amplicon of the expected size (267 bp comprising a region of the viral polymerase gene) was generated in five samples from different geographical origins (Zaragoza, Zamora, Aranjuez and Cordoba) and collecting times (2000, 2003 and 2006). The amplicons derived from two separate samples were sequenced (GenBank Acc. Nos. EF635383 and EF635384) and showed 95% nucleotide and 99–100% amino acid sequence identity with the PCRPV reference sequence. An Italian isolate of PCRPV was recovered from P. zonale with vein banding and leaf distortion (Lisa et al ., 1996; Kinard & Jordan, 2002); however from our results, PCRPV infection can also be asymptomatic. This is the first report of PCRPV in Spain.