Aegilops comosa (2n = 2x = 14, MM) contains many excellent genes/traits for wheat breeding. Wheat–Ae. comosa introgression lines have potential value in the genetic improvement of wheat quality. A disomic 1M (1B) Triticum aestivum–Ae. comosa substitution line NAL-35 was identified by fluorescence in situ hybridization and genomic in situ hybridization analysis from a hybridization cross between a disomic 1M (1D) substitution line NB 4-8-5-9 with CS N1BT1D. The observation of pollen mother cells showed that NAL-35 had normal chromosome pairing, suggesting that NAL-35 could be used for the quality test. NAL-35 with alien Mx and My subunits showed positive effects on some protein-related parameters including high protein content and high ratios of high-molecular-weight glutenin subunits (HMW-GSs)/glutenin and HMW-GS/low-molecular-weight glutenin subunits. The changes in gluten composition improved the rheological properties of the dough of NAL-35, resulting in a tighter and more uniform microstructure. NAL-35 is a potential material for wheat quality improvement that transferred quality-related genes from Ae. comosa.