PRODUCTION OF MONOSPECIFIC POLYCLONAL ANTIBODIES AGAINST ASTER YELLOWS MYCOPLASMALIKE ORGANISM-ASSOCIATED ANTIGEN

Proteins of mycoplasmalike organisms (MLO) associated with aster yellows were detected by Western blotting in partially purified preparations from aster yellows-infected plants. Polyclonal antiserum produced against aster yellows (AY) MLO recognized a 23-kDa protein in AY-infected plants that was not found in healthy controls. In addition, this antiserum recognized proteins common to AY-infected and healthy samples. Cross-absorption of the PA with healthy plant extracts did not completely eliminate the nonspecific antibodies. Antibodies specific for AY MLO-associated protein were purified from polyclonal antibodies by trapping them on AY MLO protein obtained by electrophoresis of infected plant extracts and transferred onto nitrocellulose. The resultant monospecific antibodies, eluted from the membrane, reacted with specific AY MLO-associated protein. The monospecific polyclonal antibodies reacted with AY isolates from Oklahoma carrot and lettuce but not with other MLO and spiroplasma isolates tested.