Abstract: The frequency of extreme weather events, such as low and high temperatures, flood and droughts, has increased in recent years, negatively impacting the production, yield, and quality of wheat worldwide. With the rapid socioeconomic development and population growth in China, the demand for wheat is constantly increasing. Therefore, cultivating wheat varieties with a high yield, good quality, and strong resistance is crucial for ensuring national food security. In an attempt to cultivate new varieties of wheat showing adaptability and resistance to adverse weather conditions, the wheat variety ‘Xiaoyan 22’ was radiated with a lethal dose of CO₂ laser to create mutant lines. The resulting mutant lines were screened for their stability, yield, and quality, and assessed against a range of cultivation techniques. ‘Bailu 181’ was selected as the new line most suitable for field environment. The test results showed that: 1) The genetic composition of the mutant strain ‘Bailu181’ from ‘Xiaoyan 22’ was analyzed using the Wheat 660K microarray, identifying 630 517 single nucleotide polymorphisms (SNPs), including 1 indeterminate SNP, 442 859 non-differential SNPs (70.24%), and 187 657 differential SNPs (29.76%). Compared with ‘Xiaoyan 22’, the genetic mutations of ‘Bailu 181’ mainly occurred in the SNPs of AA, CC, GG, and TT (19.23%, 23.50%, 23.40%, and 19.46%, respectively). 2) The 1000-grain weight and yield of ‘Bailu 181’ were significantly greater than those of ‘Xiaoyan 22’, with a mean increase of 9.31% and 5.10%. 3) The contents of 17 amino acids were higher in the seeds of ‘Bailu 181’ than in those of ‘Xiaoyan 22’. Compared with ‘Xiaoyan 22’, the grain bulk density and wet gluten index of ‘Bailu 181’ increased by 7.3% and 15.4%, respectively, while the stability time and maximum tensile resistance increased by 11.0% and 34.8%, respectively. 4) Different fertilization and sowing amounts had varying degrees of impact on the number of grain number per spike, 1000-grain weight, spike number and yield of ‘Bailu 181’. Taking into account both the yield and 1000-grain weight, the economic benefits were maximized under sowing 180 kg of seeds per hectare and applying 750 kg of nitrogen fertilizer per hectare. The results showed that ‘Bailu 181’ was more suitable for planting in the Guanzhong Plain and the Middle Reaches of the Yellow River Region. Approximately 30% of the genes in ‘Bailu 181’ were found to change after irradiation, providing breeding materials for enriching China’s wheat germplasm resources and breeding new varieties of wheat with high yield.