Tobacco yellow crinkle virus, a new bipartite begomovirus infecting tobacco and pepper in Cuba
*yamila@censa.edu.cu
1 Department of Microbiology, Faculty of Biology, University of Havana, Cuba
2 Laboratory of Plant Molecular Virology, National Centre for Animal and Plant Health (CENSA), Cuba
3 Department of Plant Genetic Engineering, CINVESTAV-IPN/Unidad Irapuato, Mexico
Accepted: 24 Feb 2009
Tobacco (Nicotiana tabacum) is an important crop in Cuba. Recently two bipartite begomoviruses have been reported infecting tobacco in the country, Tobacco leaf rugose virus (TbLRV) and Tobacco leaf curl Cuba virus (TbLCuCUV) (Domínguez et al., 2002; Morán et al., 2006). In January 2007, foliar yellowing and crinkling symptoms appeared on tobacco plants in eastern Cuba. DNA was isolated from seven leaf samples showing symptoms. PCR using Tomato yellow leaf curl virus (TYLCV)-specific primers was negative, demonstrating the absence of TYLCV, the most wide-spread begomovirus in the region (Martínez et al., 2003). All seven samples showed amplification with generic primers for genome components DNA A and B (amplicons of approximately 1.1 and 0.6 kb, respectively); this demonstrated the presence of bipartite begomovirus (Rojas et al., 1993). The same samples showed strong hybridisation signals in Southern blots using av1 and bc1/bv1 genes from Pepper golden mosaic virus as probes, under low stringency washing conditions. The positive samples were amplified by rollingcircle amplification and digested with several restriction enzymes. Digestion with EcoRI or PstI yielded DNA fragments of ~2.8kb, which were cloned. Four clones were sequenced and showed 100% identity (GenBank Accession No. FJ213931). The genome organization was typical of that of the DNA-A components of bipartite begomoviruses originating from the New World. The sequence was compared with those of other begomoviruses using the program CLUSTAL V and showed less than 88% nucleotide sequence identity to all previously reported begomoviruses. The highest levels of identity were with Cabbage leaf curl Jamaica virus (81.9%; DQ178614), TbLCuCUV (74%; AM050143) and with TbLRV (73.9%;AJ488768).
At the same time foliar yellow mottle symptoms were observed on sweet pepper plants (Capsicum annuum) in eastern Cuba and the presence of a bipartite begomovirus was demonstrated. The DNA-A component of the virus from one sample was cloned and sequenced (FJ222587). This showed 99.3% nucleotide sequence identity to the virus from tobacco. Thus the virus identified in both tobacco and pepper is a new begomovirus species for which the name Tobacco yellow crinkle virus (TbYCV) is proposed.
Acknowledgements
The authors would like to thank Red Latinoamericana de Botánica for a fellowship (RLB08-P10) to E. Fiallo-Olivé.
References
- Dominguez M, Ramos PL, Echemendía AL, Peral R, Crespo J, Andino V, Pujol M, Borroto C, 2002. Molecular characterization of Tobacco leaf rugose virus, a new begomovirus infecting tobacco in Cuba. Plant Disease 86, 1050.
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- Rojas MR, Gilbertson RL, Russell DR, Maxwell DP, 1993. Use of degenerate primers in the polymerase chain reaction to detect whitefly-transmitted geminiviruses. Plant Disease 77, 340-347.
This report was formally published in Plant Pathology
©2009 The Authors