Rosa rugosa leaf distortion virus(RrLDV)belongs to the genus Pelarspovirus in the family Tombusviridae,which can infect roses and cause symptoms such as leaf deformity,yellowing and premature aging.In order to establish a rapid detection method for detecting RrLDV,total RNA of susceptible rose leaves was extracted by hot borate method.The gene which encodes coat protein of the virus was amplified by RT-PCR and constructed to the prokaryotic expression vector pDB-His-MBP.The vector was transformed into Escherichia coli BL21 and the expression of the recombinant protein was induced by IPTG.The recombinant protein with a concentration of 8 mg/mL was obtained and the polyclonal anti-rabbit antiserum was produced.Western blotting showed the titer of the antiserum was 1∶2 048 000 and its sensitivity was 1∶1 024 when the antiserum was diluted to 1∶1 000.The serological detection result was consistent with RT-PCR,indicating that the antiserum can be used to detect the occurrence and distribution of RrLDV.
rosa rugosa leaf distortion viruscoat proteinprokaryotic expressionantiserum preparation
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