In humid climates, waterlogging from excessive rainfalls can be a major limiting factor for soybean production, particularly during the reproductive stage. However, there is a limited understanding of how soybean growth and physiology respond to waterlogging during this critical stage. Here, we investigated the effects of waterlogging and subsequent reoxygenation on the growth, physiology, yields, and leaf hyperspectral reflectance traits of the soybean cultivar ‘Asgrow AG5332’. The crop was grown to stage R1 (initial flowering) in outdoor pot culture conditions, and then waterlogged for 16 days. The flooded pots were drained and continuously monitored for recovery for an additional 16 days. The results showed that soil oxygen levels declined rapidly to zero in about 5 days after waterlogging, and slowly recovered in about 5–16 days. However, it did not reach the same level as the control plants, which maintained an oxygen concentration of 18%. Increasing waterlogging duration negatively affected leaf chlorophyll index, water potential, and stomatal conductance, with a consequent decline in the photosynthetic rate. Further, decreased photosynthetic rate, leaf area, and mineral nutrients resulted in lower biomass and seed yield. Pod dry weight and leaf number were the most and least sensitive parameters, respectively, decreasing by 81% and 15% after 16 days of waterlogging. Waterlogged plants also had higher reflectance in the PAR, blue, green, and red regions, and lower reflectance in the near-infrared, tissue, and water band regions, indicating changes in chemistry and pigment content. The current study reveals that the soybean crop is susceptible to waterlogging during the reproductive stage, due to poor recovery of soil oxygen levels and physiological parameters. Understanding and integrating the growth, physiology, and hyperspectral reflectance data from this study could be used to develop improved cultivars to ensure the stability of soybean production in waterlogging-prone areas.