This study is focused on the estimation of fuel consumption of the power-shift transmission (PST) tractor based on PTO (power take-off) dynamometer test. The simulation model of PST tractor was developed using the configurations and powertrain of the real PST tractor. The PTO dynamometer was installed to measure the engine load and fuel consumption at various engine load levels (40, 50, 60, 70, 80, and 90%), and verify the simulation model. The axle load was also predicted using tractor’s specifications as an input parameter of the simulation model. The simulation and measured results were analyzed and compared statistically. It was observed that the engine load, as well as fuel consumption, were directly proportional to the engine load levels. However, it was statistically proved that there was no significant difference between the simulation and measured engine torque and fuel consumption at each load level. The regression equations show that there was an exponential relationship between the fuel consumption and engine load levels. However, the specific fuel consumptions (SFC) for both simulation and measured were linear relationships and had no significant difference between them at each engine load level. The results were statistically proved that the simulation and measured SFCs were similar trends. The plow tillage operation could be performed at the gear stage of 7.65 km/h with higher working efficiency at low fuel consumption. The drawback of this study is to use a constant axle load instead of dynamic load. This study can provide useful information for both researchers and manufacturers related to the automated transmission of an agricultural tractor, especially PST tractor for digital farming solutions. Finally, it could contribute to the manufacturers developing a new agricultural tractor with higher fuel efficiency.