| Summary: | 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 .
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