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稻温病菌的分生孢子、芽管、附着胞的混合物作为抗原免疫BALB/c小鼠 ,取免疫小鼠的脾细胞与SP2 /0骨髓瘤细胞在 50 %PEG下融合成杂交瘤细胞 ,用间接ELISA筛选阳性孔 ,获 1 1株单克隆抗体。间接免疫荧光试验表明其中 4株单克隆抗体 2B4、4A1 、1D1 和 2H4分别与孢子、芽管或附着胞有特异性结合 ;Westernblotting分析发现 2B4、4A1 、1D1 单克隆抗体分别与孢子、芽管表面的提取物有不同的结合带 ;此四株单克隆抗体均干扰稻温病菌附着胞形成 ,并抑制稻温病菌在叶表的致病性。
BALB / c mice were immunized with a mixture of conidia, germ tube and adherent cells of T. grisea, and splenocytes from immunized mice and SP2 / 0 myeloma cells were fused into hybridoma cells under 50% PEG. Indirect ELISA screening of positive wells, 11 monoclonal antibodies were obtained. Indirect immunofluorescence assay showed that the four monoclonal antibodies 2B4, 4A1, 1D1 and 2H4 were specifically associated with spores, germ tubes or appressorium, respectively. Western blotting analysis showed that 2B4, 4A1 and 1D1 monoclonal antibodies were specific to spores, Of the extracts have different binding bands; the four monoclonal antibodies interfere with the formation of Appressorium thermophila appressorium, and inhibit the pathogenicity of rice warm bacteria in the leaf table.