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Bread wheat (Triticum aestivum L.) is one of the important cereals for human life. As people pursue higher quality of life, the requirements of improving the quality of wheat flour also increase accordingly. Global warming exhibits asymmetrically, that is, the warming rate during the night is significantly greater than that during the day. However, the effect of moderate warming on the quality formation of wheat is still unclear. In this study, night warming treatment was carried out at the grain filling stage to clarify the effects of night warming on the accumulation of UPP% (percentage of SDS-unextractable polymeric protein) and micro-structure of wheat gluten, with a strong gluten wheat cultivar Xinong979 and a plain gluten wheat cultivar Tam107 as materials. Results showed that night warming treatment significantly increased the protein content in the grains of the two varieties, accelerated the accumulation rate of UPP%, made the micro-structure of wheat dough more compact and dense and increased the flour processing quality. Quantitative analysis of protein networks showed night warming increased junction density, total protein length and lacunarity of micro-structure of dough in both varieties. Night warming treatment in this study helped the two varieties demonstrate better quality traits in both wheat grain and dough.