Navjot kaur2017-02-202017-02-202012http://krishikosh.egranth.ac.in/handle/1/5810001820Root knot nematodes (RKNs) of genus Meloidogyne are among the most widely spread parasitic nematodes which attack a wide range of crops and limit their production. The egg masses of Meloidogyne nematodes were cultured on the tomato plants and were identified on the basis of perineal pattern of females by Scanning Electron Microscopy. Only single species of Meloidogyne i.e. M. incognita was identified, indicating that the maintained culture was pure. The crude homogenate/antigen was prepared from the egg masses of established pure culture by homogenization, sonication and centrifugation. Quantitative analysis of prepared homogenate showed an average protein content (mg/gm, wet weight of nematodes) of 128.43 ± 11.10. Protein profile of this crude protein extract by SDS-PAGE indicated a total of seven bands (five major and two minor) observed within a range of molecular weight 27 kDa to 118 kDa. Major band was observed between 50-90 kDa followed by a band between 36-50 kDa. Two minor bands were also observed between 27-36 kDa and 50-90 kDa. Healthy rabbits were immunized with crude protein/antigen of M. incognita egg masses and hyperimmune serum showed significantly higher levels (% of total protein) of Ig from 2.37±0.32 (0 DPI) to 6.98±1.06 (60 DPI), and CIC from 3.48±0.80 (0 DPI) to 12.13±0.56 (60 DPI). Double immunodiffusion technique indicated the production of antibodies, as precipitation line was observed due to the formation of lattices of antigen-antibody complexes. Western blotting resulted in the transfer of single band (lying between 50-90 kDa) from the protein profile of crude antigenic extract of M. incognita analysed on SDS-PAGE on the nitrocellulose membrane probed with anti-Meloidogyne antibodies, thus indicating the antigenic specificity of this proteinennullThesis