Abstract: The effects of facility vegetable cultivation years (three, nine, fourteen or seventeen years) on biodiversity and abundance of soil microorganisms, such as bacteria, ammonia oxidizing bacteria (AOB) and nirK type denitrifying bacteria, in the greenhouse soils in Wuwei of Gansu Province, China were determined by the combined analyses of terminal restriction fragment length polymorphism (T-RFLP) and real-time quantitative PCR. The results showed that the dominant population structure and abundance of bacteria, AOB, nirK type denitrifying bacteria in the soils were significantly different from those in the farmland fields. The dominant population also changed with the cultivation years. With the increase of vegetable cultivation years, the abundance of 16S rRNA and nirK gene in the 0-20 cm soil layer first increased and then decreased, with the maximum values of 9.67×10⁹ and 2.30×10⁷ copies·g^-1 soil at year 14 and year 9, being as 1.51 and 1.52 times of that of the 3year, respectively. However, the abundance of amoA gene showed an opposite trend. The amoA gene copy number in the 14-year sample was 3.28×10⁷ copies·g^-1 soil, which was only 45.7% of that of the 3-year. These results illustrated that the ecological adaptation mechanisms of the different functional microorganisms involved in nitrogen cycling had significant differences in the facility vegetable soils, and provided a base for further researches on exploring and explaining the characteristics and adaptation mechanisms of microorganisms in greenhouse soil.