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Photosynthesis plays an extremely important role throughout the life cycle of plants. Improving the photosynthetic rate is a major target for increasing crop productivity. This study was conducted to identify single nucleotide polymorphisms (SNPs) associated with the net photosynthetic rate (Pn), stomatal conductance (Cond), intercellular carbon dioxide concentration (Ci) and transpiration rate (Trmmol) through genome-wide association study (GWAS) and to inspect the relationships among these traits in soybean (Glycine max (L.) Merr.). A population of 219 soybean accessions was used in this research. A total of 12 quantitative trait loci (QTLs) associated with Pn, Cond, Ci and Trmmol were detected and distributed on chromosomes 1, 2, 6, 7, 9, 11, 12, 13, 15, 16, 18, and 19, and some of these QTL overlapped with previously reported QTLs. Furthermore, four candidate genes were identified, and there were significantly different expression levels between the high-light-efficiency accessions and low-light-efficiency accessions. These putative genes may participate in the regulation of photosynthesis through different metabolic pathways. Therefore, the associated novel QTLs and candidate genes detected in this study will provide a theoretical basis for genetic studies of photosynthesis and provide new avenues for crop improvement. [ABSTRACT FROM AUTHOR]