In order to understand the effect of preharvest spray with organic calcium on the quality of blueberry, the appearance, physical and chemical indexes, nutrition components and antioxidant ability in vitro of blueberry fruit were analyzed after spraying 3 times organic calcium in blueberry cultivars. The results showed that spraying organic calcium had different effects on the quality of different cultivars, it can significantly increase fruit size and color of the Baldwin, also increase color and anthocyanin content of Tifblue. It can significantly reduce the polyphenols of blueberry, reduce the pH, juice yield, DPPH, and FRAP of Baldwin, reduce the juice yield, DPPH, FRAP, and ABTS of Tifblue, reduce the sugar, anthocyanin, and ABTS of Powderblue. It can significantly change the species and content of organic acids in blueberry, the formic acid and quinic acid of the tested blueberry was decreased significantly, the acetic acid of the tested blueberry, the malic acid and lactic acid of Powderblue, and the lactic acid of Tifblue were increased significantly.