Distant hybridization is one of the important ways for wheat improvement, but a number of sister lines were usually derived in the process of obtaining stable novel germplasm. Thus how to scientifically deal with these sister lines is an urgent problem to be solved. In order to provide theoretical guidance to the effective utilization of different innovated sister lines, 205 sister lines "Pubing2011" obtained by wheat-Agropyron cristatum distant hybridization, were analyzed through agronomic characters evaluation of the conventional breeding, identification of powdery mildew and stripe rust resistance and HMW-GS composition, detection of vernalization and photoperiod genes. The results showed that 205 sister lines were similar with the genetic similarity coefficient of 0.9854-0.9999, however, they showed some differences that the coefficient of variation of different characters was in the range of 0.54%-20.21%. 205 lines conferred adult-plant resistance to powdery mildew, 204 of them conferred adult-plant resistance to stripe rust, two of them conferred seedling resistance to stripe rust. 112 sister lines carried high quality subunits 5 10, including 4 lines with both 2* and 5 10. The composition of vernalization genes of Pubing2011-2 was (vrn-A1, vrn-B1, Vrn-D1, vrn-B3), the others were (vrn-A1, vrn-B1, vrn-D1, vrn-B3), the photoperiod gene at the Ppd-D1 locus of the 205 lines was Ppd-D1a with non-sensitive character. Further analysis showed that three high and stable yield and lodging resistant sister lines could be used directly in the production practice, eight outstanding sister lines could be used as parents to improve the yielding ability, quality and disease resistance of common wheat.