Abstract: In this paper,3α-HSD transgenic rice rhizosphere microorganism of the soil were cultured and counted to reveal the effects of rhizosphere microbial secretions on the number of microorganisms at different times. And 2 strains that utilized cholesterol as sole carbon and energy sources were isolated from the soil by the enrichment culture method. The results showed that, the quantities of3α-HSD transgenic rice rhizosphere microorganism were greater than that of conventional rice. Two strains were selected for degradation of cholesterol. In the culture conditions of 30℃ and 180 r/min, the degradation rate to cholesterol of 2 strains were 38.75% and 64.64%, with 0.1 mg/mL concentrations within four days. 2 strains were identified to be Klebsiella pneumoniae by morphological characteristics and the similarity analysis of the 16S rDNA sequence.