Abstract：Abstract Hybridization is a critical pathway to obtain excellent germplasm, however, little publications are referred to hybridization of Pinellia ternata that is an important medicinal plant in China. In this study, different staining methods (iodine-potassium iodide (I-KI), peroxidase, triphenyl tetrazolium chloride (TTC), or acetocarmine method) and in vitro pollen germination were used to detect pollen viability; And benzidine-hydrogen peroxide method and artificial pollination were carried out to test stigma viability; Artificial pollination for different parental combinations that are originated 4 populations of P. ternata were executed to obtain hybrid seeds and traditional chromosome tabletting technique was employed to determine chromosome numbers for parents or hybrids. Results showed that the TTC staining was the most appropriate method to detect pollen viability; Pollen viability was maximum at 9:00~10:00 am in 2~3 day after flowering, Stigma possess high receptivity during 1 d before flowering to 2 d after flowering. The optimal temperature for pollen storage is 4 degree Celsius. Rates of hybrid seed setting (individuals or flower) for Hezhang×Baoding, Hezhang×Pingliang and Xiangyang×Hezhang were 43.75%, 11.11% and 33.33%, respectively whereas rates of hybrid seed setting (ovules) for these 3 combinations were 66.14%, 48.89% and 38.29% respectively. The chromosome number of all hybrids were consistent with its the female parent. These results indicated that new germpalsm of P. ternata could be obtained through hybridization between populations that possess different features. The present study further perfected technical link of hybridization breeding operation of P. ternata, which can be used to guide the practical operation of hybridization breeding of P. ternata.
