Abstract: 【Objective】This study aimed to investigate the effects of different pollinating varieties on fruit quality of O’Neal blueberry and comprehensive evaluation，and to provide reference for selecting suitable pollinated varieties to improve the fruit quality and economic benefits of O’Neal blueberry.【Method】O’Neal blueberry was used as the main variety，with selfpollination as the control. Six other varieties of blueberries，including Emerald，Primadonna，Eureka， Biloxi，Millennia，and Legacy，were used as pollinating varieties for O’Neal. Pollen viability of each variety was determined by germination method. The fruit setting rate of O’Neal blueberry under different pollinating varieties was statistically analyzed. The physiological indexes of O’Neal blueberry pollinated by different varieties were measured，including fruit appearance quality indexes of transverse diameter，longitudinal diameter，single fruit weight and fruit shape index， and taste quality indexes of fruit hardness，soluble sugar content，total soluble solids（TSS）content，titratable acid（TA） content and TTS/TA value. O’Neal blueberry fruit related components contents under different pollinating varieties such as the contents of vitamin C，anthocyanins，flavonoids and total phenols. Finally，principal component analysis was used to comprehensively evaluate the fruit quality of O’Neal blueberry under different pollinating varieties.【Result】The pollen viability of the seven blueberry varieties was fine，ranging from 92.49% to 97.04%. There was no significant difference in fruit setting rate of O’Neal blueberries among different pollinating varieties（P>0.05）. Among them，the blueberries pollinated by Emerald had the highest fruit setting rate of 74.04%. Except for Emerald and Eureka，the transverse diameter， longitudinal diameter and single fruit weight of other cross-pollinated O’Neal blueberries were significantly higher than those of self-pollinated fruits（P<0.05，the same below）. The fruit shape index of Legacy-pollinated O’Neal blueberry fruit was significantly smaller than that of self-pollinated fruits. The soluble sugar content in O’Neal blueberry pollinated by Primadonna was significantly higher than that of self-pollination fruits. The TSS content of O’Neal blueberries pollinated by Legacy was significantly increased by 124.57% compared with that of self-pollination fruits. Except Biloxi，the vitamin C content of O’Neal blueberry fruits pollinated by other varieties was significantly higher than that of selfpollination fruits，and among them，the vitamin C content increase of fruits pollinated by Legacy was the highest. Compared with self-pollination fruits，the flavonoid content of O’Neal blueberry fruits pollinated by Biloxi，Legacy and Emerald significantly increased by 160.94%，155.83% and 154.91% respectively. The total phenol content increase of O’Neal blueburry fruit pollinated by Biloxi was the largest，and increased by 141.81% compared with that of selfpollination fruits. The cumulative contribution rate of the first principal component（PC1）and the second principal component（PC2） reached 79.052%. The comprehensive scores of the seven varieties were ranked as follows：Legacy > Primadonna > Biloxi > Millennia > Eureka > Emerald > O’Neal.【Conclusion】Pollinating varieties have great influence on the fruit quality of O’Neal blueberry. Xenia effect can be used to improve the fruit quality of O’Neal blueberry. Legacy and Primadonna produce the best comprehensive effect on improving the fruit quality of O’Neal blueberry as pollinating varieties.