The methods of traditional microbiology, molecular biology and biochemistry were combined to study the differences of rhizosphere soil physical and chemical properties, soil enzyme activities and the culturable microorganism diversities between healthy and diseased cherry trees in the White Deer Prairie Shaanxi for the prevention and control of ‘Black tumor’ disease. The results showed that the contents of total phosphorus, available potassium, available phosphorus and available nitrogen of healthy rhizosphere soil were significantly higher than those of diseased one, organic carbon content of healthy rhizosphere soil was significantly lower than that of diseased one. Total nitrogen, total potassium contents and pH had no significant difference between healthy and diseased rhizosphere soils. The activities of protease, urease, invertase and PPO enzymes of healthy rhizosphere soil were much higher than those of diseased one, but the activities of DHA and CAT enzymes of healthy rhizosphere soil were lower. The population of culturable bacteria and actinomyces of healthy rhizosphere soil were significantly more than diseased one, particularly of dominant bacteria, the mean population of streptomyces of healthy rhizosphere soil were lower than those of diseased one but without significant difference. The decrease of the number of culturable bacteria and actinomycetes, the imbalance proportion of dominant bacteria in the rhizosphere soil, the decrease of available fertilizer and enzyme activities may be the causes of Black tumor disease.