细菌性条斑病稻种带菌快速检测方法研究

A QUICK DETECTION FOR RICE BACTERIAL LEAF STRIPE PATHOGENS BORNE IN RICE GRAINS

建立15株分泌抗水稻细菌性条斑病(RBLS)特异性单克隆抗体(McAb)的杂交瘤细胞。对RBLS病菌呈阳性反应,与植物病原细菌5个属的其它检测菌株均无交叉反应。以酶联免疫吸附试验(ELISA)间接法对30份人工接种稻种进行检测,28份呈阳性反应,检出阳性率达93.3%;对46份病田稻种进行检测,36份呈阳性反应,10份呈阴性反应。取检测为阳性反应中的11份病田稻种的浸出液接种稻苗,有7份发病;而10份检测阴性反应的病田稻种浸出液的接种稻苗均无发病。该检测法灵敏度为0.0125—0.00125mg/ml(蛋白浓度)。

Fifteen hybridomas secreting specific monoclonal antibodies (McAb)against the pathogens of rice bacterial leaf stripe (RBLS) were made.The McAb showed the active reaction to the pathogens of RBLS and nocross reaction to the bacterial cultures belonging to different genera res-pectively.28 samples,by the indirect assay of ELISA,were active to theMcAb in the detection of 30 samples infected by the man-made innoculationin the detection rate of 93.3%.Out of 46 samples of grains from diseasedfields,36 samples were positive and 10 were negative to the McAb.11McAb-positive and 10 McAb-negative extracting solutions were respective-ly innoculated to the rice seedlings.There were 7 McAb-positive extrac-ting solutions infected the rice plants tested,while McAb-negative extrac-ting solutions showed no symptoms. The sensitivity of the method was 0.0125mg/ml—0.00125mg/ml (pro-tein concentration).