© 2006 The Authors Journal compilation © 2006 BSPP 567 Blackwell Publishing Ltd NEW DISEASE REPORTS First report of Tomato chlorosis virus on tomato crops in Cyprus L. C. Papayiannis a *, N. Ioannou a , C. I. Dovas b , V. I. Maliogka c and N. I. Katis c a Agricultural Research Institute, P.O.B. 22016, 1516 Nicosia, Cyprus; b National Agricultural Research Foundation (NAGREF), Plant Protection Institute of Thessaloniki, P.O.B. 324, Thermi 570 01, Thessaloniki; and c Aristotle University of Thessaloniki, Faculty of Agriculture, Plant Pathology Laboratory, P.O.B. 269, 54 124 Thessaloniki, Greece In the summer of 2004, yellowing symptoms similar to those caused by nitrogen and/or magnesium deficiency were observed in field- and glasshouse-grown tomatoes (Lycopersicon esculentum), in the Parekklisia area of Cyprus. Initially, lower leaves showed extensive interveinal yellowing with necrotic flecks, brittleness and occasional upward leaf rolling, before finally the whole plant turned yellow. Similar symptoms were observed during 2005 in glasshouse tomatoes grown in areas located on the south- west coastal region of the island. The abundance of whiteflies on the affected plants suggested the involvement of the whitefly-transmitted Tomato chlorosis virus (ToCV) and/or Tomato infectious chlorosis virus (TICV), both of the genus Crinivirus (Wisler et al., 1998). Leaves of 18 affected plants were collected, total RNA was isolated and RT-PCR was performed in a single tube using primers HS-11 and HS-12, followed by a multiplex nested-PCR with primers TIC-3/TIC-4 and ToC- 5/ToC-6, for the detection of TICV and ToCV, respectively (Dovas et al., 2002). A PCR product of 463 bp, corresponding to the HSP 70 gene of ToCV, was amplified for all tested samples. The sequences of four cloned PCR products were identical (EMBL accession number AM158958) and showed 99% nucleotide identity to a ToCV isolate from Florida (accession number AY903448). ToCV is vectored by Bemisia tabaci (biotypes A and B), Trialeurodes vaporariorum and T. abutilonea. Although there have been no systematic studies on whitefly incidence and distribution in Cyprus, it seems that B. tabaci is the predominant species present, as Tomato yellow leaf curl virus (Ioannou, 1985) and Cucurbit yellow stunting disorder virus (Papayiannis et al., 2005), vectored by this species, are prevalent in tomatoes and cucurbit crops, respectively. On the other hand, the incidence of Beet pseudo-yellows virus (transmitted by T. vaporariorum) is much lower. This is the first report of ToCV in Cyprus. References Dovas CI, Katis NI, Avgelis AD, 2002. Multiplex detection of criniviruses associated with epidemics of a yellowing disease of tomato in Greece. Plant Disease 86, 1345–9. Ioannou N, 1985. Yellow leaf curl and other virus diseases of tomato in Cyprus. Plant Pathology 34, 428–34. Papayiannis LC, Ioannou N, Boubourakas IN, Katis NI, Falk BW, 2005. Incidence of viruses infecting cucurbits in Cyprus. Journal of Phytopathology 153, 530–5. Wisler GC, Li HY, Lowry DS, Duffus JE, 1998. Tomato chlorosis virus: a new whitefly-transmitted, phloem-limited, bipartite closterovirus of tomato. Phytopathology 88, 402–9. *E-mail: L.Papayiannis@arinet.ari.gov.cy. Accepted 16 December 2005 at www.bspp.org.uk/ndr where figures relating to this paper can be viewed. Plant Pathology (2006) 55, 567 Doi: 10.1111/j.1365-3059.2006.01423.x Plant Pathology (2006) 55, 567 Doi: 10.1111/j.1365-3059.2006.01421.x Blackwell Publishing Ltd First report of Tobacco leaf curl Zimbabwe virus affecting tobacco in the Comoros Archipelago J. M. Lett a *, F. Lefeuvre a , F. Naze a , H. Delatte a , Y. Mohamed-Ali b and B. Reynaud a a CIRAD, UMR PVBMT CIRAD-Université de La Réunion, Pôle de Protection des Plantes, Ligne Paradis, 97410 Saint-Pierre, La Réunion, France; and b INRAPE-DECVAS, BP1406, Moroni, Grande-Comore, Union des Comores In March 2004, within the Regional Program of Plant Protection, an inventory of plant pathogens was organized in the Comoros Archipelago, located in the northern part of the Mozambique Channel. Symptoms of leaf curling and yellowing were observed on tobacco plants in the Grande- Comore Island, the most northern island of the archipelago and the closest to the African continent. Leaf samples from tobacco plants presenting the most severe symptoms were collected from three different locations: Fou- mboudzivouni (eastern region), Foumbouni (southeast region) and Sim- boussa (southern region). Samples were conserved by dehydration and tested for the presence of begomoviruses using PCR assays with two sets of degenerate primers designed to amplify part of the coat protein (CP) gene of the DNA A component. The first primer set used (AV494 and AC1048) amplified the approximately 550 bp core region of the CP gene (Wyatt & Brown, 1996). The second primer set used (VD360 and CD1266) amplified an approximately 800 bp fragment, representing more than 90% of the CP gene (Delatte et al., 2005). PCR products of the expected size were obtained with both sets of primers. No PCR products were obtained with degenerate primers designed for begomovirus DNA B or DNA β. PCR products obtained with primers VD360 and CD1266 from one sample each originating from Foumboudzivouni (EMBL acces- sion no. AM156758), Foumbouni (AM156760) and Simboussa (AM156759) were cloned and sequenced. The three sequences showed 97% nucleotide sequence identity (DNAMAN, Lynnon BioSoft). The most significant sequence alignments (NCBI, BLASTn) were 95–96% with Tobacco leaf curl Zimbabwe virus (TbLCZV; AF350330) and 83–85% with Chayotte yellow mosaic virus (ChaYMV; AJ223191). The 552 bp core CP sequences, which are sometimes used to provide provisional iden- tification of begomoviruses (Brown et al., 2001), showed 95–97% nucle- otide sequence identity with TbLCZV, and 83–84% with ChaYMV. These results demonstrate the presence of TbLCZV on tobacco in the Grande- Comore Island of the Comoros Archipelago. Previous reports have identi- fied the presence of distinct indigenous begomoviruses on tomato in the southwest islands of the Indian Ocean (Delatte et al., 2005), but this is the first report of TbLCZV. Acknowledgements This study was initiated by the Regional Program of Plant Protection and funded by the European Union and the Conseil Régional de La Réunion. References Brown JK, Idris AM, Torres-Jerez I, Banks GK, Wyatt SD, 2001. The core region of the coat protein gene is highly useful for establishing the provisional identification and classification of begomoviruses. Archives of Virology 146, 1581–98. Delatte H, Martin DP, Naze F, Goldbach R, Reynaud B, Peterschmitt M, Lett JM, 2005. South West Indian Ocean islands tomato begomovirus populations represent a new major monopartite begomovirus group. Journal of General Virology 86, 1533–42. Wyatt SD, Brown JK, 1996. Detection of subgroup III geminivirus isolates in leaf extracts by degenerate primers and polymerase chain reaction. Phytopathology 86, 1288–93. *E-mail: lett@cirad.fr. Accepted 6 January 2006 at www.bspp.org.uk/ndr where figures relating to this paper can be viewed.