MONOCLONAL-ANTIBODIES TO ANTIGENS OF ANTHERS FROM A DIOECIOUS PLANT - ASPARAGUS-OFFICINALIS-L

Monoclonal antibodies were raised by immunizing mice with a crude extract from anthers of mature male flowers of the dioecious plant Asparagus officinalis L. To isolate anther specific antibodies, hybridoma culture supernatants were screened by enzyme-linked immunosorbent assay (ELISA) against extracts from anthers, leaves, roots of male Asparagus plants. We were able to isolate a set of anther specific monoclonal antibodies recognizing four different epitopes, as shown by ELISA experiments. Western blotting and immunolocalization. Only one of the antibodies is periodate-sensitive, implying that the epitope it recognizes is carbohydrate in nature. Indirect immunofluorescence observations and timing experiments seem to suggest that at least three of the antigens are localized in pollen cells and are present in large amounts only at relatively late times during anther development.