Optimization of ISSR reaction system of Liriodendron chinese.

[Objective]A stable ISSR reaction system suitable for Liriodendron chinese was established to provide technical support for ISSR diversity analysis of Liriodendron chinese population. [Method]In the single factor experiment, main factors of ISSR reaction system including MgCl 2 , dNTPs, primer concentration, Taq DNA polymerase, DNA template amount and annealing temperature were optimized and screened. [Result]Using kit to extract DNA in Liriodendron chinese leaves was better than using the improved CTAB method. The optimal PCR reaction system was: 20 μL reaction volume containing 2.0 μL10× buffer, 1.8 mmol/L MgCl 2 , 0.2 mmol/L dNTP, 0.5 μmol/L primer,1U Taq polymerase,60 ng template DNA, and 59.1℃ annealing temperature. [Conclusion]Through the optimized ISSR-PCR reaction system, clear and stable stripes could be attained, thus it’s proper for ISSR diversity analysis of Liriodendron chinese population.