水稻草矮病特异蛋白抗血清的制备及应用

 采用不同pH值沉淀法及超速离心获得提纯的水稻草矮病特异蛋白。提纯的S-蛋白具有典型的蛋白紫外吸收曲线,提纯产量为28 mg/100 g病叶。从SDS-PAGE精提纯的S-蛋白免疫兔子上获得抗血清,经微量沉淀反应、琼脂双扩散测定,其效价分别为1:1024和1:256。获得的S-蛋白抗血清与提纯的S-蛋白及感病植株,在琼脂双扩散反应中,均有明显的沉淀线出现,对照健株及提纯的水稻草矮病毒则无反应。利用制备的S-蛋白抗血清经A-蛋白酶联免疫吸附法检测,结果表明:S-蛋白抗血清与提纯S-蛋白及感病植株有专化性反应,且不同品种、不同部位、接种后不同发病时间的水稻病株中S-蛋白的积累量有差异,这就为RGSV的检测提供了一种快速、简便的方法。

PREPARATION AND APPLICATION OF ANTISERUM OF DISEASE SPECIFIC PROTEIN OF RICE GRASSY STUNT VIRUS

The disease-specific protein of rice grassy stunt virus (RGSV) was purified by differential pH precipitation and ultracentrifugation. The ultraviolet absorbed curve of purified specific protein (SP) was typical protein one. The yield of SP was about 28 mg per 100 g of infected leaves. Antiserum was prepared by immunizing rabbit with RGSV-SP, which was further purified by SDS-PAGE. The produced antiserum had a titre of 1:1024 in microprecipitation and 1:256 in double immunodiffusion test. It reacted strongly with purified SP and infec ted plants in double immunodiffusion test and ELISA, but not reacted with purified RGSV ribonucleoprotein and healthy plants. Results from ELISA showed that the reaction between SP-antiserum and RGSV-SP was strong and specific and the concentration of SP varied with rice varieties, plant parts and disease periods of plant.Therefore, it is a simple and rapid procedure using the antibody against RGSV-SP to detect RGSV in plants.