Mapping cytosine methylation on BYVMV genome

Geminiviruses are molecular pathogens, contains small circular single stranded DNA genome which is prone for Transcriptional gene silencing (TGS). To counteract this TGS mediated host defence, Geminiviruses encodes silencing suppressor proteins. Bhendi Yellow vein mosaic virus is a monopartite geminivirus contains DNA A genome and assiciated DNA Beta. To find out whether BYVMV interferes in TGS Bisulfite sequencing using Next Generation Sequencing (NGS) was performed on wild type BYVMV viral DNA isolated from infiltrated N. benthamiana plants and the base level cytosine methylation was identified. further to know the involvement of DNA A genes in TGS suppression, C2 and C4 genes of BYVMV was mutated by introducing stop codons in the N-terminal of C2 and C4 ORFs seperately. C2 mutant and C4 mutant BYVMV A DNA was infiltrated alone or along with betasatellite in N. benthamiana. the viral DNA was isolated from infiltrated plants at 16 dpi and Bisulfite sequencing was performed by using NGS. the NGS Data was analysed for the identification of transcriptional silencing suppressor. Through this study the role of BYVMV A DNA genes in TGS suppression can be identified.