The tribe triticeae represents a potential gene pool for improvement of crops such as wheat, rye and barley. Roegneria shandongensis is a tetraploid species widely distributing in the Eastern part of China. The species contains resistance to wheat yellow dwarf disease. However, the molecular markers used to investigate the genetic diversity of R. shandongensis were poorly studied. In this paper, a SSR-PCR system of R. shandongensis was optimized and tested. PCR system was optimized in five factors (Mg 2+, dNTP, primer, DNA template and Taq DNA polymerase) at five levels respectively. To discover the economic, rapid, and stable PCR system to screen SSR primers of R. shandongensis and detect the generality of the established system, one SSR primer Xgwm43-7B was used to screen the best PCR reaction system, and nine pairs of SSR primers (Xgwm18-1B, Xgwm32-3A, Xgwm6-4B, Xgwm60-7A, Xgwm67-5B, Xgwm77-3B, Xgwm88-6B, Xgwm95-2A and Xgwm99-1A) were used to test the generality. As a result, a satisfactory SSR-PCR reaction system for R. shandongensis with desirable repeatability and polymorphic bands was established. 20 L SSR-PCR system contained 1 * buffer, 2.875 mmol/L Mg 2+, 200 mol/L dNTP, 3 pmol primer, 45 ng template DNA, 1.5 U Taq polymerase, and ddH 2O then added up to terminal volume of 20 L. The generality test of PCR optimized system of R. shandongensis was carried out. The test result indicated that this system is also suitable for the amplification done by other SSR primers.
