ISHS Acta Horticulturae 235: XIV International Symposium on Fruit Tree Virus Diseases
SEROLOGICAL DISCRIMINATION AMONG MYCOPLASMA-LIKE ORGANISMS USING POLYCLONAL AND MONOCLONAL ANTIBODIES |
| Authors: | M.F. Clark, D.L. Davies, S.L. Buss, A. Morton |
| Abstract:
Preparations of mycoplasma antigens partially purified from infected Catharanthus roseus L.plants were used to prepare polyclonal and monoclonal antibodies. The antibodies were compared for their specificity for detecting and discriminating among MLOs by indirect F(ab')2 ELISA and by fluoresence microscopy using fluorescein isothyocyanate (FITC) labelled antibodies. In ELISA tests with extracts of infected plants the different polyclonal antibodies clearly distinguished between MLO-associated antigens (MLO-ag) of peach yellow leaf roll (PYLR), Molieres disease of cherry (Mol), clover phyllody (CP), European aster yellows (EAY) and Spiroplasma citri (SC). Strong cross reactions were obtained with PYLR antiserum and extracts of C.roseus infected with green valley X (GVX) MLO, and also in reciprocal tests between PY-ag and EAY-ag and their respective antisera. A monoclonal antibody developed against PY-ag did not distinguish between PY and EAY either by ELISA or by fluorescence microscopy when labelled with FITC, indicating a close serological relationship between these two MLOs. Double staining of cryo-sections with DAPI and FITC-labelled antibodies was particularly useful for rapid confirmation of MLO identity. | |
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