Tomato (Lycopersicon esculentum Mill.) plants showing severe chlorotic and necrotic ringspots, line patterns on leaves, and concentric chlorotic ringspots on stems and fruits were observed in plastic greenhouse-grown tomato crops cv. Royesta during the spring of 1996 in Zaragoza province, Northeast Spain. Symptoms were similar to those associated with Pelargonium zonate spot virus (PZSV) infection on tomato in Italy (1,2). The causal agent was mechanically transmitted from leaf, fruit, and stem samples to several indicator species. The following host reactions were recorded: chlorotic local lesions on Chenopodium amaranticolor, C. quinoa, Cucumis sativus, and Cucurbita pepo, and systemic reactions, sometimes associated with localized reactions, on Capsicum annuum 'Doux des Landes' and 'Yolo Wonder', Datura stramonium, Gomphrena globosa, Nicotiana clevelandii, N. glutinosa, N. megalosiphon, N. rustica, N. sylvestris, N. tabacum 'Paraguay', 'Samsun', and 'Xanthi nc', Ocimum basilicum, Petunia hybrida, Physalis floridana, Solanum melongena, and Vigna unguiculata. Symptoms obtained in indicator species were erratic. During the spring of 1999, naturally occurring symptoms appeared again on tomato plants, cultivars Royesta and Bond, growing in greenhouses in the same area. Positive serological reactions with the enzyme-linked immunosorbent assay (ELISA) using a commercial PZSV antiserum (Agdia Inc.), developed against an Italian isolate of PZSV, were obtained with extracts from leaves, stems, and fruits of tomato plants naturally infected (1999) and from systemically infected indicator species mechanically inoculated with sap from tomato samples (1996 and 1999). Serological results were confirmed by molecular hybridization analysis using a PZSV-specific riboprobe (D. Gallitelli, personal communication). Some of the weeds growing around the greenhouses (Capsella bursa-pastoris, Diplotaxis erucoides, Picris echioides, and Sonchus oleraceus) also tested positive for PZSV (A405nm values greater than three times that of healthy plants). However, other weed species such as Anacyclus tomentosus, Beta maritima, Cardaria draba, Malva sylvestris, Medicago sp., Polygonum aviculare, Rumex sp., and Sisymbrium irio tested negative, while results from tests on Borago officinalis, Bromus rigidus, and Convolvulus arvensis were inconclusive. Symptoms like those of naturally infected tomato plants were reproduced by mechanically inoculating tomato seedlings with sap from PZSV-infected tobacco (Nicotiana glutinosa and N. tabacum 'Paraguay') or from Physalis floridana plants.
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