Two lines, L-19-613 and L-19-626, were produced from the common wheat cultivar Longmai 19 (L-19) by six consecutive backcrosses using biochemical marker-assisted selection. L-19 (Glu-D1a, Glu-A3c/Gli-A1?; Gli-A1? is a gene coding for unnamed gliadin) and L-19-613 (Glu-D1d, Glu-A3c/Gli-A1?) formed a set of near-isogenic lines (NILs) for HMW-GS, while L-19-613 and L-19-626 (Glu-D1d, Glu-A3e/Gli-A1m) constituted another set of NILs for the LMW-GS/gliadins. The three L-19 NILs were grown in the wheat breeding nursery in 2007 and 2008. The field experiments were designed using the three-column contrast arrangement method with four replicates. The three lines were ranked as follows for measurements of gluten strength, which was determined by the gluten index, Zeleny sedimentation, the stability and breakdown time of the farinogram, the maximum resistance and area of the extensogram, and the P andWvalues of the alveogram: L-19-613 > L-19-626 > L-19. The parameters listed above were significantly different between lines at the 0.05 or 0.01 level. The Glu-D1 and Glu-A3/Gli-A1 loci had additive effects on the gluten index, Zeleny sedimentation, stability, breakdown time, maximum resistance, area, P and W values. Although genetic variation at the Glu-A3/Gli-A1 locus had a great influence on wheat quality, the genetic difference between Glu-D1d and Glu-D1a at the Glu-D1 locus was much larger than that of Glu-A3c/Gli-A1? and Glu-A3e/Gli-A1m at the Glu-A3/Gli-A1 locus. Glu-D1d had negative effects on the extensibility and the L value compared with Glu-D1a. In contrast, Glu-A3c/Gli-A1? had a positive effect on these traits compared with Glu-A3e/Gli-A1m.