Two hundred and forty diverse set of wheat cultivars released in India during the last several decades were evaluated for HMW and LMW glutenin alleles, for assessing their diversity and effect on sedimentation volume and mixograph parameters. Both SDS-PAGE and PCR based markers were employed in identifying alleles encoded at Glu-1 and Glu-3 loci. Extensive allelic variation was observed at both the Glu-1 and Glu-3 loci. There was prevalence of Glu-A1b, Glu-B1i, Glu-D1a, Glu-A3c, Glu-B3b, Glu-B3g and Glu-D3b. The alleles Glu-A1b, Glu-B1i, Glu-D1d, Glu-A3b, Glu-B3g/h and Glu-D3b exhibited high SDS-sedimentation volume. Glu-B1i and Glu-D1d showed highly significant positive effect (p < 0.001) on sedimentation volume and also had additive effects. However, surprisingly overall there was decline in the frequency of Glu-B1i allele during last two decades in Indian wheat breeding and not a single 1B/1R translocation cultivar possessed this allele. Glu-A1b showed significant positive effect on mixograph peak time, peak slope and peak width. Glu-B3g exhibited significantly higher mixograph peak time and width at 8 and Glu-B3h showed higher dough stability. Glu-B3j (1B/1R translocation) exhibited highest peak slope indicating the negative effect on dough strength. This information can be useful in designing breeding program for the improvement of Indian bread wheat quality.