Wheat (Triticum aestivum L.) quality is a major focus of wheat breeding, which is influenced by multiple factors. The Huang-Huai wheat region, one of the main wheat-producing areas in China, provides favourable conditions for cultivating wheat cultivars with strong-gluten and medium-strong-gluten. In this study, a systematic assessment of seven crucial quality traits and important genetic loci (Glu-1 and Sec-1) in 436 wheat cultivars in the Huang-Huai wheat region of China by principal component analysis (PCA) and fuzzy comprehensive evaluation (FCE) methods showed that the stability time (ST), stretch area (SA), and maximum resistance (MAXR) were identified as three key factors, which significantly influenced wheat quality. Glu-1 and Sec-1 primarily impacted these three traits and subsequently influenced wheat quality. Compared to Glu-A1 and Glu-B1, Glu-D1 has a more significant impact on the comprehensive evaluation value D, principal components PC1-PC3, and the main traits ST, SA and MAXR of PC1. Wheat cultivars carrying the high-molecular-weight glutenin subunit (HMW-GS) Dx5+Dy10 exhibited a notable improvement in ST, SA, and MAXR traits compared with those carrying HMW-GS Dx2+Dy12, suggesting that Dx5+Dy10 may enhance wheat quality by improving ST, SA, and MAXR. By combining the results of D value, GYT (genotype by yield×trait) index, and HMW-GS score, 20 high-quality and high yield wheat cultivars were identified, which can be used as elite parents for wheat quality breeding.