Preparation and application of antiserum to soybean stay-green associated virus coat protein
Soybean stay-green associated virus(SoSGV)is a newly discovered plant virus that can cause soy-bean stay-green.Soybean stay-green is widespread in the soybean producing area of China and seriously threatens the development of soybean industry.Therefore,it is of great importance to establish a rapid and effective SoS-GV detection method for the monitoring and control of soybean stay-green.In this study,SoSGV CP primers were designed according to the nucleotide sequence of the SoSGV coat protein(CP)gene,CP gene of 786 bp length was amplified from infected soybean,and cloned into prokaryotic expression vector pET-28a to obtain the recombinant plasmid pET-28a-CP.The recombinant vector pET-28a-CP was transformed into Escherichia coli strain Rosseta.After induction with IPTG,a 30 kDa protein was obtained,which was consistent with the molec-ular weight of SoSGV CP.SoSGV CP antiserum was prepared by immunizing mice with recombinant SoSGV CP protein.The In-ELISA test results showed that the serum titer of the SoSGV CP antiserum was ≥3.2×104.West-ern blot analysis showed that the SoSGV CP antiserum could specifically recognize the SoSGV CP protein,which was 30 kDa,in the SoSGV-infected tobacco and soybean plants.These results indicate that the SoSGV CP antiserum provides favorable conditions for rapid detection of soybean stay-green in soybeans-infected by SoSGV.
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