Abstract: In order to establish and optimize the SSR-PCR reaction system of Hydrangea macrophylla,and the orthogonal design L₂₅(5⁶)was applied to optimize the five main factors(Mg²⁺,dNTPs,primers,DNA template,and Taq enzyme)at five levels of SSR-PCR reaction system.The PCR result was analyzed to select the best levels of five factors and established the suitable SSR reaction system for Hydrangea macrophylla.The results showed that the optimal SSR-PCR system of 20 μL volumes consists of 60 ng DNA template,1.5 mmol/L Mg²⁺,0.3 mmol/L dNTPs,0.4 μmol/L primer,and 0.8 U Taq polymerase.PCR amplification procedures were pre-denaturation for 5 min at 94℃;followed by 33 cycles of denaturation for 1 min at 94℃,annealing at a suitable for primers for 40 s,extension for 1 min at 72℃;and a final extension for 10 min at 72℃,then stored at 4℃.The optimal SSR-PCR reaction system was used to verify by 10 cultivars of Hydrangea macrophylla,and the results showed that it was excellent stability and repeatability for the optimal reaction system.The establishment of this reaction system provides a theoretical basis and technical references for further research on the genus Hydrangea by SSR markers.

Key words: Hydrangea macrophylla, SSR-PCR, Orthogonal design, Reaction system

CLC Number: 

• S68.03