Generation of anti-teliospores antibodies for immunolocalization and characterization of antigenic epitopes of teliospores of Karnal bunt (Tilletia indica) of wheat.

ABSTRACT

Polyclonal antibodies raised against intact teliospores of T. indica in New Zealand albino rabbits were used for the development of indirect immunofluorescence tests. Specificity of anti-teliospore antibodies was evaluated by cross reactivity studies on other bunt, smut and related pathogens. The characteristic reactivity pattern indicated that the antibodies reacted with Tilletia species only. Chemical modifications, heat and enzyme treatments followed by indirect immunofluorescence tests were employed to delineate the molecular nature of the surface antigens. There was partial or no loss in immunoreactivity by methanol, periodate, heat or trypsin treatments. Extensive periodate treatment altered the fluorescence pattern due to changes in configuration of carbohydrate antigen present in episporium. Sequential treatment of periodate and trypsin showed diminished fluorescence due to access of proteolytic enzyme into inner site of episporium thereby cleaving peptide epitope(s) after reorientation of carbohydrate moietiesby periodate treatment. It indicated glycoprotein nature or peptide nature of epitopes on the teliospore surface.