GWAS identified stable loci for TGW and stress tolerance in winter wheat based on two sowing conditions, which will provide opportunities for developing new cultivars with high yield and yield stability. Wheat is an important food crop widely cultivated in the world. Breeding new varieties with high yields and superior adaptability is the main goal of modern wheat breeding program. In order to determine the marker-trait associations (MATs), a set of 688 diverse winter wheat accessions were subjected to genome-wide association study (GWAS) using the wheat 90K array. Field trials under normal-sown (NS) and late-sown (LS) conditions were conducted for thousand grain weight (TGW) and stress susceptibility index (SSI) at three different sites across two consecutive years. A total of 179 (NS) and 158 (LS) MATs corresponded with TGW; of these, 16 and 6 stable MATs for TGWNS and TGWLS were identified on chromosomes 1B, 2B, 3A, 3B, 5A, 5B, 5D, 6B, and 7D across at least three environments. Notably, a QTL hot spot controlling TGW under NS and LS conditions was found on chromosome 5A (140-142 cM). Moreover, 8 of 228 stable MATs on chromosomes 4B, 5A, and 5D for SSI were detected. A haplotype block associated with TGW and SSI was located on chromosome 5A at 91 cM, nearby the vernalization gene VRN-A1. Additionally, analysis of wheat varieties from the different eras revealed that the grain weight and stress tolerance are not improved concurrently. Overall, our results provide promising alleles controlling grain weight and stress tolerance (particularly for thermotolerance) for wheat breeders, which can be used in marker-assisted selection for improving grain yield and yield stability in wheat.