The cloning and nucleotide sequence of a new bipartite geminivirus found in Cuba is described. DNA A (2620 nt) and DNA B (2586 nt) presented a genomic structure resembling that of other geminiviruses transmitted by Bemisia tabaci. Both components had a common region of 168 nt with a 95% identity. Typical elements involved in replication and transcription were found in this region, though group-characteristic arrangement of iterons was not conserved in this virus. Sequence was compared with geminivirus sequences deposited in the GenBank. Interestingly, when total DNAs or individual ORFs and deduced amino acid sequences were compared, the highest scores were for different viruses. It showed to be most closely related to tomato mottle virus (81.9% and 65.5% similarity with DNAs A and B, respectively) and a member of the abutilon mosaic cluster of New World Begomoviruses. When clones A and B were co-agroinoculated they resulted highly infectious and induced symptoms in Nicotiana benthamiana plants. The A component alone was infectious but induced only mild symptoms, while the B component was not infectious. The presence of viral DNA in N. benthamiana plants was confirmed by dot-blot hybridization using specific probes. These data show that the cuban isolate is a new geminivirus for which the name of Havana tomato virus is proposed.
Beans with yellow mosaic and/or leaf crumple symptoms were collected in three fields in the southern area of the province of Havana, Cuba in December 2001 and February 2002. DNA was extracted from the fresh bean leaves of 25 samples (1). Dot blot hybridization was performed at high stringency with a specific probe for Tomato yellow leaf curl virus (TYLCV). The specific probe was prepared by alkaline phosphatase labeling of the polymerase chain reaction (PCR) fragment amplified with primer pair, PTYIRv21/PTYIRc287, containing the intergenic region (IR) of TYLCV, and chemiluminescent hybridization was completed as described by the manufacturer (AlkPhos Direct Labeling and Detection Systems, Amersham Pharmacia Biotech Inc., Piscataway, NJ). Four of the samples had positive hybridization signals. PCR was performed with overlapping primers for TYLCV (2) with the DNA extract from sample 01–44, which gave a positive hybridization signal with the TYLCV probe, and a 2.8-kb fragment was obtained. This fragment was cloned in pGem T-Easy (pBeTY44) and partially sequenced. Greater than 96% nt identity was obtained for the 591 nt of the IR and 504 nt of the N-terminus of the Rep gene with TYLCV (GenBank Accession No. AF260331). Also, PCR was completed on 11 of the 25 samples with the degenerate primer pair PAL1v1978/PAR1c715 for DNA-A (3). Eight samples gave fragment sizes of 1.4 kb and one sample gave a fragment of 1.3 kb. The 1.3-kb fragment from sample number 01–50 was cloned in pGem T-Easy (pBeBG50) and partially sequenced. Pairwise nucleotide comparisons with Bean golden yellow mosaic virus (BGYMV, GenBank Accession No. M91604) were 95% for 719 nt of the N-terminus of the Rep gene. These results are consistent with the association of both TYLCV and BGYMV in beans and have important implications for future disease management strategies. References: (1) G. P. Accotto et al. Eur. J. Plant. Pathol. 106:179, 2000. (2) M. K. Nakhla et al. Plant Dis. 78:926, 1994. (3) M. Rojas et al. Plant Dis. 77:340, 1993.
Os begomovírus causam doenças de grande importância econômica em diversas culturas, principalmente em regiões tropicais e subtropicais. Juntamente com outras famílias de vírus, os begomovírus têm causado grande prejuízo para os produtores de tomate in natura e para processamento industrial. O objetivo deste trabalho foi avaliar o comportamento de 11 genótipos resistentes ao Tomato yellow leaf curl virus (TYLCV) frente à infecção pelos begomovírus Tomato yellow spot virus (ToYSV) e Tomato severe rugose virus (ToSRV) em condições de casa-vegetação. A inoculação das plantas foi realizada via biobalística no estádio de duas folhas verdadeiras. A infecção viral confirmou-se pelo desenvolvimento dos sintomas e pela técnica de hibridização dot blot. Selecionaram-se como promissores os genótipos STY2, STY5, STY6 e L7, por não apresentarem sintomas e por terem concentrações virais muito baixas para os dois vírus. O espectro de resistência dos genes Ty-1 e Ty-2 não resultaram efetivos ante as espécies virais empregadas no estudo. As linhagens TY52, H24 e CLN2116B, portadoras destes genes, foram suscetíveis aos vírus ToYSV e ToSRV.
Tomato plants displaying symptoms of yellowing and leaf curling were analysed for the presence of geminiviruses. Two distinct geminiviruses were present in the plants studied. One had a genome size and coat protein gene sequence similar to the Israeli strain of tomato yellow leaf curl virus (TYLCV), while the other had a smaller genome size than TYLCV that could not be amplified using primers specific for Israeli TYLCV. The presence of the Israeli strain of TYLCV has been reported in other Caribbean islands, but not in Southern Florida (USA) which is close to those islands where TYLCV has been detected. This suggests that the introduction of the Israeli strain of TYLCV to the Caribbean area may have occurred within recent times.
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