The invention discloses a pot vegetable substrate and a preparation method thereof. The raw material of the pot vegetable substrate is composed of mushroom material, corncob, chicken manure, cinder, fine sand and cake fertilizer, the organic material content is 30%, and the density is 55kg/dm<3>. The mushroom material, the corncob, the chicken manure and the cake fertilizer are respectively fermented, the fermentation products are mixed, the mixed bacteria of Trichoderma harzianum and lactobacillus plantarum is added to the mixture for further fermentation, finally, the fine sand is added, and the pot vegetable substrate is prepared; bifidobacterium, saccharomycetes, lactic acid bacteria, lactobacillus plantarum and Trichoderma harzianum, which are used in the fermentation process, survive in the prepared pot vegetable substrate, the strains can perform long-term synergistic effect to regulate the nutritional ingredients of the pot vegetable substrate, thus, the vegetable grows better. The pot vegetable substrate is harmless to human beings and livestock during long-term storage, is odor-free and green, and can be safely used for a long time.