Immunodetection of Teliospores of Karnal Bunt (Tilletia indica) of Wheat using Fluorescent Staining Tests

Polyclonal antibodies were raised against intact teliospores in Newzealand white rabbits for the immunodetection of karnal bunt (KB) teliospores.The indirect immunofluoresence (IIF) test was developed using anti-teliospores serum and binding was monitored by goat anti-rabbit antibody conjugated to fluorescein (FITC) lable. The standardization of IIF test was carried out by optimization of dilutions of anti-teliospore antibodies, fluorescent probe and exposure time. The teliospores of T. indica showed bright green, patchy and ring shaped fluorescence around the teliospore. The spore exhibited uniform distribution in discrete regions of spore surface probably in spore episporium. Similar fluorescence pattern in the teliospores of KB isolated from infected wheat seeds of cultivars HD 2328, UP 2338, PBW 393, WH 542, as well as RR 21 (susceptible cultivars) respectively, is an indication of the presence of similar antigenic configuration of teliospores. Again, they did not exhibit variation in the expression of teliospore associated molecular pattern (TAMP) during previous and subsequent years of infection.