Generation of diagnostic anti-teliospore antibodies for development of specific immunodiagnostic format for detection of Karnal bunt (Tilletia indica) of wheat

Abstract

Rabbit antisera were generated against four preparations of teliospores of Tilletia indica the fungus responsible for Karnal bunt in wheat having titre ranging from 1:2500 to 1:10,000. The suitability of four solubilising agents (I–IV) used for extraction of teliospores/spore proteins was assessed, highest protein extractability was observed with solubilising agent I (0.5% sodium dodecyl sulphate). The maximum immunoreactivity in enzyme linked immunosorbent assay (ELISA) was observed with the teliosporic protein(s) extracted using solubilising agent I followed by agent II, IV and III. Further, the results of western blotting with proteins from different fungal pathogens using anti-intact teliospores antibody showed one unique immunoreactive band of 28 kDa with teliosporic protein of T. indica only. The potential diagnostic antibody was generated against the characterised protein (28 kDa) of teliospore wall's antigen of T. indica. The developed diagnostic antibody and solubilising agent I could therefore be employed for development of specific and rapid immunodiagnostic format for detection of Karnal bunt.