In view of the gradual promotion of Single Fuel Concept in the military field and also the rotary engine (RE) which has been widely applied in specialized equipment, it is necessary to study the basic spray characteristics of aviation kerosene under RE operation conditions and optimize the fuel injection strategies. In this research, an optically visible constant volume combustion chamber platform for Mie-scattering and a validated three-dimensional dynamic simulation model of aviation kerosene fueled direct injection rotary engine (DI-RE) were established. The experimental results show that the ambient temperature and pressure affect the spray process differently. There is no linear relationship between ambient temperature and penetration length. The ambient temperature of 600 K is a watershed, when it is below 600 K, the penetration length increases as the temperature rises. When the temperature reaches 600 K, the penetration length decreases rapidly which means a prominent improvement of atomization quality. On the basis of experimental results, the simulation was carried out and the results show that different injection strategies have important effect on distribution of fuel in cylinder, the larger injection angle will promote the atomization of fuel due to the collision between the spray and air flow field. In Case A3, B2 and C2, the fuel mainly distributes near the spark plugs in the center of the combustion chamber, which is supposed to be more conducive to the ignition and diffusion combustion process of the fuel.