The invention relates to the technical field of dendrobium officinale cultivation, in particular to a dendrobium officinale cultivation substrate with a bactericidal effect. The substrate is preparedfrom, by weight, 20 parts of coniferous forest soil, 10 parts of infusorial earth, 8 parts of ceramsite, 5 parts of peat, 18 parts of barks, 15 parts of agar, 6 parts of silkworm excrement, 0.8 part of mineral salt, 6 parts of plant ash, 8 parts of tobacco leaf, 1.8 parts of lysine, 1.0 part of nitrifying bacteria, 12 parts of animal waste and 3 parts of baking soda, after the above components aremixed uniformly, water with dissolved bactericide is added, the dendrobium officinale cultivation substrate has the advantages that the raw materials are cheap and obtained easily, the production cost is low, the air permeability, fertilizer maintenance capacity and water-retaining property are excellent, all nutrient substances needed by the growth of the dendrobium officinale can be provided; when the dendrobium officinale is cultured by using the dendrobium officinale cultivation substrate, tissue culture seedlings grow well, the quality is guaranteed easily, the bactericide is added intothe substrate so as to achieve the effects of sterilization and disinfection, and the survival rate of dendrobium officinale seedlings can be improved.