Agricultural biodiversity usually leads to greater sustainability in production practices. To understand the environmental implications of the development of village-level multi-cropping in rural China, we compared the environmental impact of a highly diverse vegetable multi-cropping system to a conventional wheat/maize rotation system based on the method of life cycle assessment (LCA). Using household level cultivation data, this study examined the gate-to-gate environmental impacts of on-site cultivation practices relating to the production of 10,000 nutrient equivalent units. Results show that vegetable multi-cropping resulted in decreased average land requirement, and diesel, water and electricity usage by 69.8%, 62.2%, 71.7%, and 63.4%, respectively, while average nitrogen (Total N), phosphorus (P₂O₅), and potassium (K₂O) usage in vegetable multi-cropping systems decreased by 16.3%, 42.1%, and 75.8%, respectively. Additional corresponding effects led to a decrease in the total global warming, eutrophication, and acidification potentials from external inputs by 21.6%, 16.7%, and 16.2% of the entire system, respectively. Moreover, the midpoint human toxicity potential from pesticide usage of the vegetable multi-cropping system was lower than that of the conventional system. However, the midpoint eco-toxicity potential from pesticide usage was higher due to certain highly toxic substances, and both human and eco-toxicity potentials from heavy metals were all higher by a few orders of magnitudes. Thus, to mitigate these detrimental consequences, some related measures are proposed for sustainable practices in the future implementation of multi-cropping systems.