Magnesium (Mg) is an essential macronutrient for plant growth and development; however, the adaptive mechanisms of Mg deficiency to underlying changes in Mg translocation, subcellular distribution and chemical forms in citrus plants are unknown. In this study, we conducted a sand culture experiment with 0 (Mg-deficiency) or 2 (Mg-sufficiency) mmoll-1Mg2+ treatments to investigate the responses underlying Mg adaptability, as well as the resulting growth and Mg transport features in citrus seedlings [Citrus sinensis (L.) Osbeck cv. 'Xuegan']. We found that Mg-deficiency significantly depressed biomass by 39% in the whole plant and by 66% in branch organs compared with Mg-sufficient conditions, which further resulted in a subsequent decrease in Mg concentration and accumulation with changes in its distribution in different organs and a reduction in root growth. Under Mg-sufficiency, >50% of Mg was sequestered in the soluble fraction and this was reduced by 30% under Mg-deficiency. Furthermore, >70% of Mg existed as inorganic (42%) and water-soluble (31%) forms with high mobility across treatments and organs. Under Mg-deficiency, the proportion of water-soluble Mg was reduced in leaf and increased in root, whereas the proportion of inorganic Mg increased in main stem leaves and decreased in branch leaves and root. However, under Mg-deficiency, the proportion of Mg forms with low mobility, including pectates and proteins, phosphates, oxalates and residues, was increased in leaf and root organs, with the exception of pectate and protein Mg, which was decreased in root. The Mg transfer factor showed that Mg-deficiency improved Mg transport from parent to branch organs, which was related to Mg subcellular distribution and chemical forms. Taken together, our study establishes a defined process to clarify the mechanisms of Mg absorption and translocation and reveals a possible strategy to effectively improve Mg mobility and availability in citrus plants.