Partial Characterization of Two Whitefly-Transmitted Geminiviruses Infecting Tomatoes in Venezuela.

Whitefly-transmitted geminiviruses can cause significant yield losses on tomatoes (Lycopersicon esculentum Mill.) in Venezuela. To identify the geminivirus(es) infecting tomatoes in Venezuela, 20 tomato samples from commercial tomato fields in four states and one weed (Euphorbia heterophylla L.) sample were examined for geminivirus infection. All samples showed symptoms generally associated with geminivirus infection, including golden or yellow mosaic, mottling, crumpling and/or distortion of leaves, and, in some cases, stunted and distorted growth. Through the use of squash blot hybridization analysis and a general probe for Western Hemisphere whitefly-transmitted geminiviruses (4), geminivirus nucleic acids were detected in 19 of 20 tomato samples and the weed sample. No samples were infected with tomato yellow leaf curl virus (TYLCV), based on squash blot hybridization analysis with a TYLCV-specific probe. With polymerase chain reaction (PCR) and degenerate primers for whitefly-transmitted geminiviruses (PAL1v1978 and PAR1c496) (4), an approximately 1.2-kb DNA-A fragment was amplified from the 19 squash blot-positive tomato samples and from the weed sample. No DNA fragment was amplified from any samples when TYLCV-specific primers (PTYC1v2406 and PTYIRc287) (3) were used. PCR-amplified DNA-A fragments from four samples representing four different states [Monagas (5L), Guarico (3M), Aragua (3R), and Portuguesa (2U)] were cloned and sequenced. Partial AC1, AV1, and complete common region (CR) sequences of the 5L, 3M, and 2U DNA-A fragments were 92 to 93, 93, and 95 to 97% identical, respectively, indicating that these were DNA-A clones of the same virus. Furthermore, these sequences were 91 to 92, 92 to 95, and 93 to 95% identical, respectively, to sequences of homologous regions of potato yellow mosaic virus (PYMV), indicating that these tomato-infecting geminiviruses are isolates or strains of PYMV. The partial AC1, AV1, and complete CR sequences of the 3R DNA-A fragment were 79, 95, and 77% identical to those of 5L, 3M, and 2U clones, respectively, suggesting that this is a different geminivirus. These sequences were 75 to 87, 82 to 88, and 73 to 81% identical, respectively, to sequences of homologous regions of other tomato geminiviruses, including tomato golden mosaic from Brazil, tomato mottle from Florida, and tomato leaf crumple from Mexico. The bipartite nature of the geminiviruses that were present in the 5L, 3M, 3R, and 2U samples was suggested by the amplification of a DNA-B fragment with degenerate DNA-B primers (PBL1v2040 and PCRc1) (4). These results suggest at least two distinct bipartite Western Hemisphere whitefly-transmitted geminiviruses are associated with tomato virus diseases in Venezuela, and that one of these (sample 3R) may be an undescribed geminivirus. The sequence of the DNA-A fragment from the weed sample was not closely related to the tomato-infecting geminiviruses and, therefore, this weed was not an alternate host of these viruses. Furthermore, because PYMV has been shown to infect tomatoes and cause yellow mosaic symptoms (1), it would be of interest to determine the relationship of PYMV and tomato yellow mosaic geminivirus (ToYMV), which has been reported infecting tomatoes in Venezuela (2), but has not been characterized on the molecular level. References: (1) A. K. Buragohain et al. J. Gen. Virol. 75:2857, 1994. (2) R. C. de Uzcátegui and R. Lastra. Phytopathology 68:985, 1978. (3) M. K. Nakhla et al. Phy-topathol. Mediterr. 32:163, 1993. (4) M. R. Rojas et al. Plant Dis. 77:340, 1993.