Allelic variations at the Glu-1 and Glu-3 glutenin loci play an important role in determining wheat (Triticum aestivum L.) dough properties and bread-making quality. Chromosomal translocations between wheat and rye (Secale cereale L.) have been associated with a decrease in end-use quality in hard winter wheat. To evaluate the effect of glutenin subunit variation and the 1AS.1RS translocation on end-use quality of hard winter wheat, we developed a recombinant inbred line (RIL) population from the cross of TAM107-R7 and Arlin which segregated at several Glu-1 and Glu-3 loci and at 1AS.1RS. The 190 RILs were grown in two Colorado locations, Akron (dryland) and Fort Collins (irrigated) in 2004-05. Gluten strength and dough mixing properties were measured by 10 g Mixograph. Five Mixograph parameters, midline peak time (MPT), midline peak volume (MPV), midline right volume (MRV), middle peak width (MPW), and midline right width (MRW) were used as indicators of dough properties. Results indicated the highly significant (P<0.0001) effect of growing environments on Mixograph traits. 1AS.1RS was highly (P<0.01) associated with the five Mixograph variables in the dryland location, but only MPW, MPV and MRV in the irrigated location. Variation at GluD1 (5+10 vs. 2+12 subunits) had the significant influence (P<0.001) on the five Mixograph variables in irrigated location, but only MRW, MPT and MPV at dryland location. Variation at GluA1 (1 vs. 2* subunits) had a significant influence (P<0.05) on MPV and MRV in both locations, but only MPW in the irrigated location. The GluA3 locus (allele e vs. c) had a significant influence (p<0.05) on MPW, MRV, MPV in both locations, but onl MPT and MRW in dryland location. These results suggested that the influence of 1AS.1RS translocation and Glu-1 and Glu-3 loci on wheat dough mixing properties largely depending on the environment in which the wheat is grown.