Abstract: In order to research the environmental factors coupling effect in fresh-keeping transportation with controlled atmosphere, an experimental platform for environmental control was established. Experiments were carried out to investigate the effects of regulation process such as refrigeration, high-pressure atomizing humidification, liquid nitrogen injection and ventilation on temperature, humidity, O2 concentration, and CO2 concentration of the fresh-keeping environment. Results indicated that liquid nitrogen injection during controlled atmosphere process could rapidly reduce the O2 concentration, meanwhile significantly affecting temperature and humidity in the container. Similarly, refrigeration had a great impact on the humidity when decreasing the temperature. High-pressure atomizing humidification made little effect on temperature when increasing the humidity, whereas the compressor intaking the air outside the container would exerted dramatic effect on the volume fraction of O2 than intaking the air inside. Ventilation had great influence on the volume fraction of O2 and CO2. The results can provide references for integrated control strategy of fresh-keeping transportation with controlled atmosphere.