Abstract: The different effects such as plant growth regulators, light conditions, and genotypes on the maintenance of the three species of Acacia were compared in the experiment, and the results showed that the medium supplemented with 0.2-1 mg昄-12,4-D was beneficial to the maintenance of calli. Moreover, the calli could be kept loose and soft under the weak light by comparing the effects of different light conditions. And the superior inoculation amount was 0.3 g calli in each bottle. By the cytohistological observation on different calli in acacia, it showed that there were different Morphological cells and calli in different species of Acacia, which also changed during the culture.