Abstract: The effects of 6-BA, NAA, 2.4-D, and IBA on the leave callus of Aloe were studied by using L9(34) orhogonal design. The variance analysis of the experimental results showed that the actions of the four factors were all extremely remarkable in MS medium. Among the four factors, 6-BA was the most important one for the callus induction of Aloe leaves, followed by IBA, then 2.4-D, and NAA was the weakest .The best combination of plant growth substances for callus induction was 6-BA4.0mg/L+NAA0.25mg/L+2.4-D3.0mg/L+IBA0.5mg/L.