The effects of long-term fertilization on soil microbial communities of tea plantation were investigated by using BIOLOG techniques. Based on AWCD and community diversity index, the treatments with fertilizers resulted in increasing the abundance and diversity of soil microorganisms compared to the non-fertilizing treatment, the combination of inorganic and organic fertilizers showed the greatest effect followed by the application of organic matter while inorganic fertilizer had less effect. Long-term fertilization exhibited little influence on the common species of soil microorganisms according to the analysis of multiple indexes; however, application of organic fertilizer caused unevenly distribution of microbial communities in soil. After analyzing the principle components, it was found that the differentiation of soil microbial communities was mainly caused by carbohydrate and carboxylic acid. The improvement of soil fertility of tea plantation enhanced the microbial activity.