The invention discloses special bio-fertilizer for blueberries as well as a preparation method and an application of the bio-fertilizer. The bio-fertilizer is prepared from raw materials including 10parts of a Bacillus cereus agent, 5 parts of a bacillus subtilis agent, 5 parts of a trichoderma agent, 30 parts of organic fertilizer, 8 parts of nitrogen fertilizer, 10 parts of phosphate fertilizer, 12 parts of potash fertilizer, 5 parts of trace fertilizer, 4 parts of plant growth promoting substances, 6 parts of humic acid and 5 parts of bentonite; the raw materials are stirred, mixed, sieved, dried and packaged, and the special bio-fertilizer for the blueberries is prepared. The special bio-fertilizer is used for increasing blueberry yield, improving blueberry quality, improving blueberry storing property, induced-enhancing disease resistance of the blueberries, promoting growth of blueberry plants, increasing microbe quantity of blueberry root systems and improving soil enzyme activity, and has the advantages of being remarkable in effect, green, pollution-free, high in fertilizer use ratio, low in cost, easy to popularize and the like, soil is improved, diseases are controlled,and yield and quality are improved.