Biological and molecular characterization of begomoviruses associated with tobacco leaf curl disease from Karnataka by sequencing the complete genome
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Authors
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Gurudevi V Navali
College of Horticulture, Hiriyuru, KSN University of Agricultural and Horticultural Sciences, Shivamogga 577 598
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K T Rangaswamy
College of Agriculture, University of Agricultural Sciences, GKVK, Bengaluru 560 065
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K S Shankarappa
College of Horticulture, UHS Campus GKVK, Bengaluru 560 065
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S Basavaraj
National Institute of Plant Health Management, Hyderabad- 500 030
Keywords:
Tobacco, leaf curl, isolates, DNA-A, molecular characterization.Abstract
Biological characterization of begomoviruses associated tobacco leaf curl disease of different isolates were collected during 2016-2018 viz., TbLCV-MYS, TbLCV-MAN, TbLCV-CHN, TbLCV-SHG, TbLCV-DVG
and TbLCV-BEL were successfully transmitted to tobacco, tomato, chilli, datura, goat weed, sunflower,
sesamum, and milkweed, whereas TbLCV-MAN and TbLCV-SHG isolates able to infect jatropha and
cassava in addition to above plant species. The complete genome characterization of TbLCV-MYS, TbLCVMAN, TbLCV-CHN, TbLCV-SHG, TbLCV-DVG and TbLCV-BEL isolates were performed by PCR for
complete genome of TbLCV by using three pairs of specific overlapping primers viz., GEMA, MK and
OY. Six TbLCV isolates and positive control yielded amplicons of 1.1kb, 1.4kb and 1.2kb for GEMA, MK
and OY, respectively. The amplified products were cloned, sequenced and aligned. The nBLAST analysis
revealed that, TbLCV-MYS and TbLCV-DVG isolates shared maximum identity with Tomato leaf curl
Karnataka virus whereas TbLCV-CHN and TbLCV-BEL isolates shared maximum identity with Tobacco
leaf curl Yunnan virus and Tomato leaf curl Kerala virus, respectively. The TbLCV-SHG and TbLCV-MAN
isolates shared maximum sequence similarly of less than 87 percent in nBLAST analysis indicated their
distinctness from other isolates. Hence, they renamed as Tobacco leaf curl Shivamogga virus (TbLCSV)
and Tobacco leaf curl Mandya virus (TbLCMV). The phylogenetic analysis revealed that TbLCV-MYS
and TbLCV-DVG isolates clustered with Tomato leaf curl Karnataka virus. Similarly, TbLCV-BEL isolate
clustered with Tomato leaf curl Kerala virus, TbLCV-CHN isolate clustered with Tobacco leaf curl Yunnan virus, whereas TbLCV-SHG and TbLCV-MAN isolates clustered with the Chilli leaf curl Multan virus and SriLankan cassava mosaic virus, respectively.
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Manuscript will be accepted on the understanding that their content is original and that permission has been received in writing wherever necessary to produce previously published material (including quotations, data and illustrations) and that the manuscript has not been submitted/ accepted for publication elsewhere. Copyright resides with the Plant Protection Association of India.