As an excellent biological resource, Chlorella has wide applications for production of biofuel, bioactive substances and water environment restoration. Therefore, it is very important to understand the photosynthetic physiology characteristics of Chlorella. Magnesium ions play an important role in the growth of microalgae, not only the central atom of chlorophyll, but also the cofactor of some key enzyme in the metabolic pathway. A laboratory study was conducted to evaluate the effects of magnesium deficiency on several photosynthetic and physiological parameters and the triacylglyceride (TAG) accumulation of the green alga, Chlorella vulgaris, in the photoautotrophic culture process. Chlorella vulgaris biomass, protein, chlorophyll a and chlorophyll b contents decreased by 20%, 43.96%, 27.52% and 28.07% in response to magnesium deficiency, while the total oil content increased by 19.60%. Moreover, magnesium deficiency decreased the maximal photochemical efficiency F(v)/F(m) by 22.54%, but increased the non-photochemical quenching parameters qN. Our results indicated the decline of chlorophyll caused by magnesium, which affected the photosynthesis efficiency, lead to the growth inhibition of Chlorella vulgaris and affected the protein synthesis and increased the triacylglyceride (TAG) accumulation.