ABSTRACT
To date, no begomovirus has been fully characterized from Euphorbia heterophylla, a widely distributed weed, in Brazil. Here, we show the occurrence of a new begomovirus on E. heterophylla plants showing bright yellow mosaic. The bipartite viral genome was cloned from 10 samples, and all clones are almost identical to each other (95.6-98.8% nucleotide sequence identity). The DNA-A sequences shared a maximum nucleotide sequence identity of 87.3% with euphorbia mosaic Peru virus (EuMPV) and thus were classified as belonging to a novel begomovirus species, tentatively named Euphorbia yellow mosaic virus (EuYMV). The EuYMV DNA-B sequences share a maximum nucleotide sequence identity of 56.2% with a euphorbia mosaic virus (EuMV) isolate from Mexico. Phylogenetic analysis demonstrated that this new virus belongs to a different lineage than EuMV isolates from Central America.
Subject(s)
Begomovirus/genetics , Begomovirus/isolation & purification , Euphorbia/virology , Plant Diseases/virology , Begomovirus/classification , Brazil , Molecular Sequence Data , PhylogenyABSTRACT
The bacteriophage phiDNA polymerase amplifies circular DNA in a rolling circle amplification mechanism. This characteristic was applied to amplify and clone the complete circular DNA genome of a begomovirus. Total DNA extracted from infected tissue was used as the template of an amplification reaction using the commercial kit TempliPhi (Amersham Biosciences). The amplified DNA could be used for direct sequencing and was cloned after digestion with a single cutting restriction endonuclease. The use of this enzyme simplified the cloning steps and increased the cloning efficiency of the complete genome of a circular plant DNA virus.
