马铃薯Y病毒SYRI株系外壳蛋白抗血清制备及应用OA

[Objective]Potato virus Y(PVY)is the main pathogen of tobacco and potato virus diseases.SYRI strain is the dominant strain in China.Preparation of coat protein(CP)antiserum of SYRI strain can reduce the missed detection rate of PVY.[Method]The PVY CPSYRI gene sequence was amplified and ligated into the prokaryotic expression vector pEHISTEV to obtain the recombinant plasmid pEHISTEV-CPSYRI.This recombinant plasmid was then transformed into Escherichia coli Rosetta,and after prokaryotic expression,New Zealand white rabbits were immunized to obtain polyclonal antibodies.[Result]Western blot results showed that the prepared antiserum could detect PVY CPSYRI protein diluted 100 000-fold,and it exhibited no cross-reaction with the CP protein of Turnip mosaic virus(TuMV),which belonged to the same genus.Tests on potato and tobacco samples in the field showed that the antiserum prepared was effective in detecting the diseased plants in the field.[Conclusion]The antiserum has showed good sensitivity and specificity,and could be used to detect PVY in crops in the field,which has laid a foundation for the monitoring and early warning of PVY epidemic.