© 2007 The Authors 342 Journal compilation © 2007 BSPP Blackwell Publishing Ltd NEW DISEASE REPORTS A new tomato leaf curl virus from the Seychelles archipelago P. Lefeuvre a , H. Delatte a , F. Naze a , W. Dogley b , B. Reynaud a and J. M. Lett 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 Ministry of Environment and Natural Resources, PO Box 166, Victoria, Mahé, Seychelles In July 2004, a survey of plants with virus disease symptoms was conducted in the Seychelles archipelago on Mahé Island, located north of Madagascar. A small number of leaf samples showing curling symptoms were collected from bean (Phaseolus vulgaris), pepper (Capsicum annuum) and tomato (Lycopersicon esculentum). Leaf extracts were tested for the presence of begomoviruses by polymerase chain reaction (PCR), using two sets of degenerate primers designed to amplify portions of the coat protein (CP) gene of the DNA A component. The first primer set (AV494 and AC1048) amplified the core region of the CP gene (approxi- mately 550 bp) (Wyatt & Brown, 1996). The second primer set (VD360 and CD1266) amplified another fragment representing more than 90% of the CP gene (approximately 900 bp) (Delatte et al., 2005). No PCR products were obtained from pepper or bean leaf extracts. For two tomato samples, PCR products of the expected size were obtained with both sets of primers. One product, obtained with the primer set VD360-CD1266 (924 bp), was cloned and sequenced (EMBL accession No. AM259286). No PCR products were obtained using degenerate primers designed for begomovirus DNA B or DNA β (Delatte et al., 2005). The most significant nucleotide sequence identities (NCBI BLASTn) were 88% with Tomato leaf curl Mayotte virus from Dembeni (ToLCYTV-[Dem]; AJ865341) and 83% with Pepper yellow vein virus from Mali (AY502935), Tomato leaf curl Madagascar virus isolates (AJ865339 and AJ865338) and a South African cassava mosaic virus isolate (AF155806). Analysis (DNAMAN, Lynnon BioSoft) using the 522bp core CP sequences, previously used to provide provisional identification of begomoviruses (Brown et al., 2001), showed between 88% and 89% nucleotide sequence identity with ToLCYTV isolates (AJ865339 and AJ865340) and between 85 and 86% identity with Tomato yellow leaf curl virus isolates from Réunion Island (TYLCV-[RE], AM234066; and TYLCV-Mld[RE], AJ865337). These results suggest the presence of a new tomato begomovirus in the Seychelles archipelago; genetically related to the ToLCYTV isolates and so to the African tomato monopartite begomoviruses previously identified in the South West Islands of the Indian Ocean (Delatte et al., 2005). The new virus is tentatively named Tomato leaf curl Seychelles virus (ToLCSCV). 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 16 May 2006 at www.bspp.org.uk/ndr where figures relating to this paper can be viewed. Plant Pathology (2007) 56, 342 Doi: 10.1111/j.1365-3059.2006.01471.x Plant Pathology (2007) 56, 342 Doi: 10.1111/j.1365-3059.2006.01472.x First report of Tomato yellow leaf curl China virus infecting kidney bean in China J. H. Dong, Y. Q. Luo, M. Ding, Z. K. Zhang* and C. K. Yang Institute of Biotechnology and Genetic Resources, Yunnan Academy of Agricultural Sciences, Kunming 650223, China In 2004, a virus isolate (Bean-YM) was obtained from kidney bean (Phaseolus vulgaris) showing leaf curl symptoms from Yuanmou, Yunnan province, China. To identify possible begomoviruses, total DNA was extracted from a leaf sample. Degenerate primers PA and PB, designed to amplify part of the intergenic region and coat protein (CP) gene of DNA- A of begomoviruses (Zhou et al., 2003), were used in PCR. A 500 bp DNA fragment was obtained, cloned and sequenced. Sequence comparison showed that the fragment was most closely related to Tomato yellow leaf curl China virus (TYLCCNV) isolate [Y43] (acc. no. AJ781302) with 92% nucleotide sequence identity. Primers WTGs-1 (5′-CGTGCT GCTGTC- CCCACTGT-3′) and WTGs-2 (5′-GTGGAAATGACT ATATCGGC-3′) were then designed to amplify the remaining DNA-A sequences of Bean- YM. The complete DNA-A sequence of Bean-YM was determined to be 2731 nucleotides (DQ256460). Comparisons with other begomoviruses showed that DNA-A of Bean-YM is closely related to TYLCCNYV-[Y43] with 92% nucleotide sequence identity. All previously characterised TYLCCNV isolates have been shown to be associated with DNA β molecules (Cui et al., 2004). To test whether a satel- lite molecule is associated with this isolate, a universal abutting primer pair specific for DNA β was used to amplify the putative DNA β (Briddon et al., 2002). Using this primer set, an amplicon of approximately 1400 bp was obtained. Sequence analysis revealed that DNA β of Bean-YM is 1336 nucleotides long (DQ256459) and most closely related to the DNA β asso- ciated with TYLCCNV-[Y10] (AJ421621; 80·8% nucleotide sequence identity). TYLCCNV has previously been reported to infect tomato, tobacco and various weed spp. This is the first report of TYLCCNV infecting a legume. Acknowledgements This work was supported by NSFC Grant No. 30360005 and NSFYP Grant No. 2005C00122. References Briddon RW, Bull SE, Mansoor S, Amin I, Markham PG, 2002. Universal primers for the PCR-mediated amplification of DNA; a molecule associated with some monopartite begomoviruses. Molecular Biotechnology 20, 315–8. Cui XF, Tao XR, Xie Y, Fauquet CM, Zhou XP, 2004. A DNA, associated with Tomato yellow leaf curl China virus, is required for symptom induction in hosts. Journal of Virology 78, 13966 –74. Zhou XP, Xie Y, Peng Y, Zhang ZK, 2003. Malvastrum yellow vein virus, a new Begomovirus species associated with satellite DNA molecule. Chinese Science Bulletin 48, 2205 –9. *E-mail: zhongkai99@sina.com. Accepted 16 May 2006 at www.bspp.org.uk/ndr where figures relating to this paper can be viewed.