The bitter taste character of Ningxia wild bitter wolfberry was studied, using methods of botany classification, conventional hybridization, PCR-SSR molecular marker detection, Ultra performance liquid chromatography high resolution mass spectrometry?［UPLC-MS(HRMS), and other in this paper. The results showed that, comparing with other Chinese wolfberry germplasm, the length of filament and style, seed size and color etc. of Ningxia wild bitter wolfberry had larger differences. Its stamen filaments and pistil style of flower buds were curly shape. Most berries clustered. PCR-SSR molecular marker detection results showed that DNA of Ningxia wild bitter wolfberry was amplified a band, which hasn ‘t found it in cultivativars that Ningqi 1, 2, 3, 4, 5, 7, etc..Menwhile, betwenm 111bp and 147bp, Ningxia wild bitter wolfberry DNA amplified a band which was slightly higher than other Chinese wolfberry germplasm resources, It means that the molecular weight of the site was higher than other germplasm resources of Chinese Lycium barbarum L. It had a significant difference in genetic material. As a female parent, Ningqi 7, a major cultivar in Ningxia, crossed by Ningxia wild bitter wolfberry, the mature berries of hybrid F1 generation had a proportion of bitterness and sweetness separation, which closed to 1:1. Meanwhile, the separation ratio of bitter to sweet berries had 1:4 in F1, when Ningqi 7, as female parent, crossed by wild semi bitter wolfberry. However, all of F1 fruits were bitter from the reverse cross. Obviously, bitter genes may be heterozygous of wild bitter wolfberries in rural environment.However, it indicated that bitter traits might be controlled by multiple dominant genes with additive genetic effects, and it might relate to the interaction between nuclear and plastid genetic material. The sweet trait might be controlled by recessive genes. According to preliminary determination, the main source of bitter substance might be flavonoids with Taxifolin structure.