水稻条纹病毒外壳蛋白叶绿体离体跨膜运输研究

Studies on in vitro rice stripe virus coat protein import into chloroplasts

【目的】明确离体条件下水稻条纹病毒(RSV)外壳蛋白(CP)能否进入叶绿体,为研究RSV的致病机理提供依据和方法。【方法】参考有关文献的方法提取获得水稻和小麦叶绿体,在离体条件下进行RSV-CP的叶绿体跨膜运输试验,同时研究孵育时间、病毒浓度等对RSV-CP在水稻叶绿体中离体跨膜运输的影响。【结果】在离体条件下5min,RSV-CP即可进入RSV寄主植物水稻、小麦的叶绿体内,其离体跨膜运输的基本条件为RSV浓度58.1μg/mL、孵育时间15min。随着孵育时间的延长,进入叶绿体中的CP量有所增加;反应体系中RSV浓度加大,进入叶绿体中的RSV-CP量随之增加。【结论】离体条件下RSV-CP可进入寄主植物水稻、小麦的叶绿体中,病毒外壳蛋白进入叶绿体可能是其诱发花叶症状的主要原因之一。

[Objective]The present experiment was aimed to find out the possibilities of rice stripe virus coat protein（RSV-CP） import into chloroplasts in vitro in order to provide references for studying the pathogenesis of rice stripe virus.[Method]Rice and wheat chloroplast was obtained by using leaf-cut method.The system of RSV-CP import into chloroplasts in vitro,and the effects of the incubation time and the concentration of RSV on import efficiency were also studied.[Result]RSV-CP was found able to import into isolated chloroplasts of RSV host plants including rice and wheat in 5 min.The lowest concentration of RSV in the import system was 58.1 μg/mL and the best incubation time was 15 min in the current studies.By extending the incubation time or increasing RSV concentration,the amount of RSV-CP import into the chloroplasts was increased.[Conclusion]The study reveals that RSV-CP import into rice and wheat chloroplast in vitro is possible.This may be one of the leading causes of mosaic symtoms.