**5. Conclusions**

In the present study, a new set of 5RKu dissection lines was identified, and 111 new PCR-based and 5RKu-specific markers were developed. The 111 markers were combined with the 52 5RKu-specific markers reported previously, and 65 *S. cereale* Lo7 scaffolds were physically mapped to six regions of the 5RKu chromosome. Additionally, the 5RLKu arm carried stripe rust resistance gene(s) and it was mapped to region L2 of the 5RKu chromosome, the same region where 22 5RKu-specific markers and 11 *S. cereale* Lo7 scaffolds were mapped. The stripe rust resistance gene(s) located in the 5RLKu arm enriches the resistance source of stripe rust for wheat breeding programs, and the markers and the 11 *S. cereale* Lo7 scaffolds that were mapped to the L2 region of the 5RKu chromosome facilitates the utilization of the stripe rust resistance gene(s) in the improvement of wheat cultivars.

**Supplementary Materials:** The following are available online at http://www.mdpi.com/2073-4395/9/9/498/s1, Table S1: The information of 5RKu-specific markers. Table S2: Location of 5RKu-specific markers on six regions of 5RKu chromosome using 5RKu dissection lines.

**Author Contributions:** Conceived and designed the study, S.F. and Z.T.; performed the experiments, W.X. and J.L.; data analysis, W.X.; writing-original draft preparation, Z.T.; writing-review and editing, S.F.

**Funding:** This manuscript is provided by the National Key Research and Development Program of China (No. 2016YFD0102001) and National Natural Science Foundation of China (No. 31770373).

**Acknowledgments:** We are thankful to Yanling Duan, Ruiying Liao and Shuyao Tang at the College of Agronomy, Sichuan Agricultural University for their helpful work on nucleotide blast searching and data submission to NCBI.

**Conflicts of Interest:** The authors declare no conflict of interest.
