【Objective】In this study, 24 hybrids were made using blueberries of three distinct cultivation cultivar including Vaccinium corymbosum ‘O’Neal’, V. ashei ‘Pink Lemonade’ and V. ashei ‘Bonita’ as the male parent and three ploidy types of blueberries as the female parent. We aimed to investigate the differences in fruit setting and fruit sensory quality after pollinating these crosses. The results may provide some clues for proportion of pollination trees for cultivation and selection of breeding parents in hybrid breeding of blueberry. 【Method】Using the fruits from open pollination of the female parents as the control (CK), sensory quality indices of hybrid fruits from different crosses including fruit appearance parameters, contents of carbohydrates, antioxidant substances, and volatile compounds were evaluated.【Result】Hybridization compatibility between the three male parents and the rabbiteye blueberry cultivars ‘Vernon’ and ‘Briteblue’ was relatively low. Higher fruit set rate was solely found in the hybrid crosses with highbush types using ‘O’Neal’ and ‘Bonita’ as the male parents. After crossing with ‘O’Neal’, the single fruit mass of ‘Primadonna’ was decreased with a significant increase in fruit firmness. In addition, the fruit shape index of ‘Primadonna’ fruits pollinated by ‘Bonita’ increased significantly. Seed observatioin indicated that the hybrid crosses between blueberry ‘Pink Lemonade’ and three southern highbush cultivars all showed the relatively high fruit set rate, and hybrid seeds were not found in the hybrid fruits in the other crosses. Notably, the hybrid crosses between three male parents ‘O’Neal’, ‘Pink Lemonade’ and ‘Bonita’, and the rabbiteye type cultivar ‘Climax’ all revealed no seeds in the fruits. In comparison with each naturally open-pollinated female parent, the contents of sugar substances in the hybrid fruits of highbush types cultivars were significantly increased after the pollination of ‘O’Neal’ and ‘Bonita’. Especially, the solidity-acid ratios of the hybrid fruits of ‘Winsor’ and ‘Primadonna’ were extremely high after the pollination of ‘Bonita’. Further detection of aromatic substances demonstrated that there was a total of 59 volatile compounds in the fruits of all hybrid combinations. Although the types and contents of the volatile substances varied among the combinations, three main common substances, including linalool, caryophyllene oxide, and trans-2-hexenal were detected across almost all hybrid combinations. The content of linalool in the hybrid fruits of ‘Sharpblue’ was both significantly increased after pollination of ‘Bonita’ and ‘O’Neal’, while that of ‘Primadonna’ was significantly increased only after pollination by ‘O’Neal’. Espcecially, the content of trans-2-hexenal in the hybrid fruits of ‘Primadonna’ and ‘Sharpblue’ pollinated by ‘O’Neal’ was significantly higher than those of the naturally open-pollinated fruits. The volatile substances of the rabbiteye blueberry cultivar ‘Climax’ increased significantly after pollination by ‘Pink Lemonade’, and the substances of alkene type covered a relatively higher proportion. The content of total phenols and anthocyanins showed a similar trend in pollination combinations of different ploidy, with the highest content of total phenols and VC in the hybrid fruits of ‘Climax’ pollinated by ‘O’Neal’, and the content of anthocyanins in the fruit of ‘Primadonna’ and ‘Climax’ was relatively higher under natural open-pollination. 【Conclusion】Blueberries of different ploidy levels have a certain cross-compatibility for inter-ploidy hybridization, and the seedless fruit can be obtained in the crosses of the rabbiteye derived cultivar ‘Pink Lemonade’ and three southern highbush varieties. The solidity-acid ratios of hybrid fruits of most of the highbush cultivars were significantly increased after crossing with the rabbiteye cultivar ‘Bonita’. Especially, after crossing with the male parent ‘O’Neal’, the hybrid fruits of rabbiteye cultivar ‘Climax’ bore no seeds with the increase in the accumulation of antioxidant substances.