检测棉花黄萎病菌的间接ELISA法建立与应用

棉花黄萎病菌（大丽轮枝菌－ＶＤ８）培养液中产生的毒素经纯化后获得电泳纯毒素ＰＬＰｓ，制备ＰＬＰｓ的抗血清，建立了检测棉花黄萎病菌毒素的间接ＥＬＩＳＡ方法。研究结果显示，ＰＬＰｓ抗血清的效价为１：１０２４００，最低检测为１．５６ｎｇ。间接ＥＬＩＳＡ法检测显示，ＰＬＰｓ的免疫抗血清可以与多种棉花黄萎病菌菌株体外培养液，带菌棉籽的培养液，以及病棉的叶脉、叶柄及茎杆的榨取液发生特异性的反应，而与其他致病菌

Indirect ELISA for detection of verticillium wilt of cotton and its application

The Vitro toxin(PLPs)produced by a strain of Verticillium dahliae (VD 8)pathogenic on cotton was purified from filtered culture fluids by ion chromatography and polyarcylamide gel electrophoresis(PAGE).Polyclonal antiserum was prepared against PLPs,and the specificity of the antiserum was tested by using indirect ELISA,and no cross reactivity was observed among the eight isolates representing other species of fungi.The pathogen in the veins and stems of infected cotton and cottonseeds could be deduced by detecting the toxin in them with ELISA.The PLPs could be detected as low aas 1 56 ng at 490 nm.