Development and Application of Monoclonal Antibodies against the Mycoplasmalike Organism (MLO) Associated with Rice Yellow Dwarf

• Main Authors: Fu-Lin Chang, 張富林
• Other Authors: Chan-Pin Lin
• Format: Others
• Language: zh-TW
• Published: 1993
• Online Access: http://ndltd.ncl.edu.tw/handle/93980832387749070860

碩士 === 國立臺灣大學 === 植物病蟲害學系 === 81 === Eight stable hybridoma cell lines secreting specific monoclonal antibodies against the rice yellow dwarf (RYD)- mycoplasmalike organism (MLO) were produced by employing an immunization scheme for the induction of the immunological tolerance of mice to rice antigens prior to the administration of RYD-MLO immunogens. Neonatal BALB/c mice were injected with nontarget rice antigens present in an immunogen preparation on days 1, 4, 7 and were immunized intrasplenically with RYD-MLO- enriched antigens prepared by Percoll density-gradient fraction 6 wk later. All of these monoclones belonged to the IgG1. With this monoclonal antibody (247B11-2F4), RYD-MLO in diseased rice was specifically identified by indirect ELISA, immunofluorescent staining (IF) and tissue blotting. Antibody titer determinated by indirect ELISA for hybridoma-culture supernatant of 247B11-2 F4 was 5120. Dilution endpoints of RYD-MLO antigens that reacted with the monoclonal antibody from 247B11-2F4 were 1/2048 g/ml. In Western blotting, the antibody from 247B11-2F4 recognized two polypeptides, 15.7kDa and 41 kDa from diseased plants. RYD-MLO was serologically differentiated from the MLOs associated with loofah witches'' broom , paulownia witches'' broom, witches'' broom of Ipomoea obscura, sweet potato witches'' broom, peanut witches'' broom, red-bird cactus witches'' broom, elm yellows and aster yellows (NJ) both in ELISA and IF tests .