Plant Pathology (2006) 55, 817 New Disease Report 50 Blackwell Oxford, PPA Plant 0032-0862 3 Pathology Pathology, UK Publishing 2006Ltd
Doi: 10.1111/j.1365-3059.2006.01473.x NEW N XXXX S.ew K.Disease Raj DISEASE et alReport . REPORTS 3 50
First report of Tomato leaf curl Karnatka virus infecting soybean in India S. K. Raja*, M. S. Khana, S. K. Snehia, S. Srivastavab and H. B. Singhb a
Molecular Virology and bPlant Pathology, National Botanical Research Institute, Lucknow-226001, U.P., India
During a survey in September 2005, a leaf crumple disease of soybean (Glycine max) was observed in Lucknow with a disease incidence of more than 80%. The diseased plants exhibited severe yellowing, crumpling and distortion of leaves. The infected plants remained dwarf with fewer flowers which resulted in a very low yield. The causal pathogen was suspected to be a begomovirus due to the large population of whitefly (Bemisia tabaci; the vector of begomoviruses) observed on the crop. A begomovirus was detected by PCR using begomovirus coat protein gene-specific primers (Singh, 2005) and total DNA isolated from infected leaf tissues. An amplicon of the expected size (∼800 bp) was obtained only from leaf samples with symptoms. The PCR product was cloned, sequenced and the 771 bp sequence data was submitted to GenBank (accession number DQ343284). Analysis of nucleotide sequence data by Blast search revealed the highest nucleotide sequence identity (96–98%) with Tomato leaf curl Karnataka virus (ToLCKV) [TLU38239, AJ810342, AY753203, AJ810370 and AJ810347]. Mungbean yellow mosaic India
virus (Usahrani et al., 2004) and Pepper golden mosaic virus (MendezLozano et al., 2006) have been reported on soybean from India and Mexico respectively. However, ToLCKV infection of soybean in India has not been reported previously. References Mendez-Lozano J, Quintero-Zamora E, Barbossa-Jasso MP, Leyva-Lopez NE, 2006. A begomovirus associated with leaf curling and chlorosis of soybean in Sinaloa, Mexico is related to Pepper golden mosaic virus. Plant Disease 90, 109. Singh R, 2005. Molecular characterization of a virus causing yellow mosaic disease in Cucurbita maxima and development of diagnostics for detection of the virus. Lucknow, India: University of Lucknow, PhD thesis. Usharani KS, Surendranath B, Haq QMR, Malathi VG, 2004. Yellow mosaic virus infecting soybean in Northern India is distinct from the species infecting soybean in southern and western India. Current Science 86, 845 – 50.
*E-mail:
[email protected]. Accepted 9 March 2006 at www.bspp.org.uk/ndr where figures relating to this paper can be viewed.
© 2006 The Authors Journal compilation © 2006 BSPP
Plant Pathology (2006) 55, 817
Doi: 10.1111/j.1365-3059.2006.01474.x
A new begomovirus infecting pepper plants in Cuba
Blackwell Publishing Ltd
Y. Martinez Zubiaur*, Y. Muñiz Martín and M. Quiñones Pantoja Plant Pathology Group, National Centre for Animal and Plant Health (CENSA). P.O. Box 10, San José De las lajas, La Habana, Cuba Diseases caused by begomoviruses are a major threat to vegetable production for countries in the Caribbean and Central America. In 1996, an isolate of Tomato yellow leaf curl virus (TYLCV) was reported to be affecting tomato plants (Lycopersicon esculentum) throughout Cuba (Martínez et al., 1996). In 2002, the same virus was detected in pepper (Capsicum annuum) in Camagüey, Sancti Spíritus and Havana provinces and in bean (Phaseolus vulgaris) plants in Havana province (Martínez et al., 2002; Quiñones et al., 2002). During a survey in 2005, in the pepper production areas of eastern Cuban, about 70% of pepper plants growing at an Experimental Station in Holguin province, showed symptoms that differed from those previously described for TYLCV in pepper. Symptoms consisted of plant stunting and a severe leaf yellow mosaic with crinkling and curling. Ten representative plants with symptoms were collected and leaf samples analyzed by transmission electron microscopy. Total nucleic acid was extracted and assayed by non-radioactive nucleic acid hybridisation, using a TYLCV intergenic region probe under low stringency conditions (Quiñones et al., 2002) and by polymerase chain reaction (PCR). All leaf samples showed the presence of typical geminivirus particles, whereas none exhibited hybridisation signals. Using PCR with degenerate primers that specifically amplify the coat protein region from begomoviruses (Wyatt & Brown, 1996), all samples produced amplicons of the expected size (576 bp). PCR amplicons from three samples were cloned in pGEM T-Easy vectors (Promega). The clones were sequenced and BLAST analysis showed the highest identity (98%) within clones; between 93% and 94% identity with Cabbage leaf curl virus (U65529 and AF065815, respec-
tively) and between 87% and 90% identity with Pepper yellow vein virus (AF 063139), Pepper golden mosaic virus (AY368336) and Texas pepper mosaic virus (AF077025). However there was only 68% identity with TYLCV (AJ223505). One of the sequences was deposited in GenBank (DQ207808) as representative of the three clones. This report shows the occurrence of a new begomovirus in pepper plants in Cuba. Acknowledgement This work was support by an INCO BETOCARIB project (PL ICA42000-10328) and the Cuban Agriculture Biotechnology programs. References Martínez-Zubiaur Y, Zabalgogeazcoa I, de Blas C, Sánchez F, Peralta EL, Romero J, Ponz FM, 1996. Geminiviruses associated with diseased tomatoes in Cuba. Journal of Phytopathology 144, 277 – 9. Martínez Y, Quiñones M, Fonseca D, Potter J, Maxwell DP, 2002. First report of Tomato yellow leaf curl virus infecting bean (Phaseolus vulgaris) in Cuba. Plant Disease 86, 814. Quiñones M, Fonseca D, Acotto GP, Martínez Y, 2002. Viral infection associated with the presence of Begomoviruses in pepper plants in Cuba. Plant Disease 86, 73. 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:
[email protected]. Accepted 5 April 2006 at www.bspp.org.uk/ndr where figures relating to this paper can be viewed.
© 2006 The Authors Journal compilation © 2006 BSPP
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