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• Expression of 1Dy12 in wheat mutant AS273 was decreased during grain development. • A stop codon caused by base change led to truncated translation of 1Dy12 in AS273. • The absence of 1Dy12 influenced the accumulation of seed storage proteins and GMPs. • The absence of 1Dy12 led to a larger size PBs and an inferior flour-processing quality. Dissecting the functions of high molecular weight glutenin subunits (HMW-GSs) is helpful for improving wheat quality via breeding. In this study, we used a wheat mutant AS273 in which HMW-GS 1Dy12 was silenced to investigate the silencing mechanism of 1Dy12 and its effects on gluten accumulation and flour-processing quality. Results suggested that the expression of 1Dy12 in AS273 was decreased by one fifth during grain development; a stop codon produced by a base mutation (C/T) led to truncated translation; the absence of 1Dy12 stimulated the accumulation of low molecular weight glutenin subunits (LMW-GSs), gliadins, and glutenin macropolymers, and was resulted in larger protein bodies; AS273 had an inferior flour-processing performance. Based on the outputs achieved in this study it is concluded that 1Dy12 makes important contributions to bread, sponge cake and biscuit-processing quality. [ABSTRACT FROM AUTHOR]