Sensitive and high-throughput polyclonal antibody-based serological methods for rice stripe virus detection in both rice and small brown planthopper

Abstract

Rice stripe virus (RSV) is a plant virus, and its infection leads to a devastating disease that often causes serious yield losses in rice-producing regions. In this study, the capsid protein (CP) gene of the RSV was cloned and expressed in E. coli (Rosetta), and the purified recombinant His-CPRSV protein was used to immunize New Zealand white rabbits. After six weeks, a polyclonal antibody against the His-CPRSV protein (PAb-CPRSV) was obtained from the serum of the immunized rabbits. Using the prepared PAb-CPRSV, three serology-based detection methods were developed, which could specifically test for RSV-infected rice plants or small brown planthoppers (SBPHs). Methods that combine dot-blot with enzyme-linked immunosorbent assay to high-throughput methods were also developed to detect RSV in rice seedlings and SBPHs simultaneously in wild fields. The field surveys indicated that the incidence of RSV in rice plants and SBPHs in the suburbs of Yangzhou City was much lower than the value ten years ago. This research presents multiple serology-based RSV detection methods, using the prepared PAb-CPRSV, which can simultaneously identify RSV-infected rice plants and SBPHs in wild field conditions. Our study will benefit RSV control in rice planting regions.