Phosphorus is an essential macronutrient for plant growth and development, but phosphate resources are limited and rapidly depleting due to massive global agricultural demand. This study identified two genes in the phosphate transporter 2 (PHT2) family of soybean by bioinformatics. The expression patterns of two genes by qRT-PCR at leaves and all were induced by low-phosphate stress. After low-phosphate stress,GmPHT2;2expression was significantly higher thanGmPHT2;1, and the same trend was observed throughout the reproductive period. The result of heterologous expression ofGmPHT2in Arabidopsis knockout mutants ofatpht2;1shows that chloroplasts and whole-plant phosphorus content were significantly higher in plants complementation ofGmPHT2;2than in plants complementation ofGmPHT2;1. This suggests thatGmPHT2;2may play a more important role in plant phosphorus metabolic homeostasis during low-phosphate stress thanGmPHT2;1. In the yeast backfill assay, both genes were able to backfill the ability of the defective yeast to utilize phosphorus.GmPHT2expression was up-regulated by a low-temperature treatment at 4 °C, implying thatGmPHT2;1may play a role in soybean response to low-temperature stress, in addition to being involved in phosphorus transport processes.GmPHT2;1andGmPHT2;2exhibit a cyclic pattern of circadian variation in response to light, with the same pattern of gene expression changes under red, blue, and white light conditions.GmPHT2protein was found in the chloroplast, according to subcellular localization analysis. We conclude thatGmPHT2is a typical phosphate transporter gene that can improve plant acquisition efficiency.