棉花大丽轮枝菌(Verticillium dahliae)血清学检测——抗原处理的比较  

Production and characterization of polyclonal antibodies prepared against different antigens of Verticillium dahliae from cotton

作　　者：李亚宁[1] 魏艳敏[1] 刘大群[1] 杨文香[1] 张汀[1] 

机构地区：[1]河北农业大学植物病理系河北省农作物病虫害生物防治工程技术中心,河北保定071001

出　　处：《河北农业大学学报》2005年第2期79-84,共6页Journal of Hebei Agricultural University

基　　金：国家863项目(2003AA241130)

摘　　要：分别以棉花黄萎病菌V20的菌丝研磨物、菌丝蛋白、超氧化物歧化酶酶带Ⅰ、超氧化物歧化酶酶带Ⅱ为抗原免疫家兔,制备了抗血清As1、As2、As3、As4。琼脂双扩散法(ADD)和间接ELISA方法测定其特异性,发现这4种抗血清既可与同种的抗原发生阳性反应,也可与不同种的抗原发生交叉反应。用异种抗原黑白轮枝菌和尖孢镰刀菌吸附这4种抗血清,排除其非特异性成分后,抗血清专化性明显提高,消除了交叉反应。特异性测定表明:经吸附后的4种抗血清可以准确地将大丽轮枝菌鉴定到种的水平,但仍不能区分其生理型和致病类型。另外,当分别使用菌丝研磨物或菌丝蛋白为特异性测定抗原时,ADD检测的结果虽然一致,但前者阳性反应沉淀线粗糙,轮廓不清晰,后者的沉淀线较细,且清晰、明显。因此适宜选择菌丝蛋白为抗原进行抗血清的特异性测定试验。The antisera As1, As2, As3, As4 with different specificity prepared against different antigens, induding mycelium, protein, band I of SOD isozyme, and band II of SOD isozyme of V20, a nondefoliating strain of Verticillium dahliae, were prepared. The specificities of four different antisera were tested by agar dimension diffusion (ADD) and indirect enzyme-linked immunosorbent assay (ELISA), respectively. All antisera showed nonspecific reaction with homogeneous and heterogeneous antigens. Antibodies specific for V. dahliae were purified from polyclonal antibodies by mixing them with the antigen V. albo-atrum and Fusarium oxysporum. After purified, the nonspecific polyclonal antibodies reacted with V. dahliae but not with other fungi commonly found in cotton diseases, although the antisera still can not determinate physiological type and pathotype of V. dahliae. The results from ADD were the same with it from ELISA. However, as the results of ADD, to a certain degree, the specialty of the antisera prepared against antigens of the proteins of V. dahliae was improved more than the antisera prepared against mycelial preparations, which was shown by the different quality of deposit lines of ADD. Therefore, we chose protein of mycelial as the antigen to do the specificity test.

关 键 词：大丽轮枝菌 血清学检测 超氧化物歧化酶 e抗原 间接ELISA方法 棉花黄萎病菌 菌丝蛋白 交叉反应 抗血清 尖孢镰刀菌 抗原免疫 阳性反应 非特异性 致病类型 反应沉淀 测定试验 V20 扩散法 专化性 生理型 ADD 研磨 酶带 吸附 

分 类 号：S435.621.2[农业科学—农业昆虫与害虫防治] S858.28[农业科学—植物保护]