The invention belongs to the technical field of ecological agriculture, and discloses a preparation method of a biological light carbon mixture. The biological light carbon mixture comprises the following components in percentage by mass: 13-25% of microalgae, 5-7% of an adsorbent, 1-1.5% of a colloid agent, 2-3% of a water absorbent, 1-2% of a complexing agent, 1-2% of an activating agent, 8-12%of yeast sugar and 55-65% of purified water, and is prepared by the following steps: (1) respectively taking microalgae, an adsorbent, a colloid agent, a water absorbent, a complexing agent, an activating agent, yeast sugar and purified water for later use; and (2) respectively weighing the raw materials for later use in the step (1) according to the mass percent of the preparation amount: 13-25%of microalgae, 5-7% of an adsorbent, 1-1.5% of a colloid agent, 2-3% of a water absorbent, 1-2% of a complexing agent, 1-2% of an activating agent, 8-12% of yeast sugar and 55-65% of purified water, fully stirring in a reaction kettle at normal temperature and normal pressure, and carrying out a reaction for 3 h to obtain the biological light carbon mixture, wherein the biological light carbon mixture is used for activating and improving soil, enhancing the photosynthetic action of plants, resisting diseases, expelling parasites, storing water, retaining fertilizers and improving the comprehensive functions of crops.