Abstract:

To establish and optimize AFLP reaction system for Asparagus and lay foundation to analy its genetic diversity, the single-factor was applied for optimizing four factors in the reaction system including restriction enzyme amount, time of digestion, preamplification system and final amplification. The DNA was completely digested by 5 U enzyme EcoRⅠand MseⅠin 37℃ water-baths for 3 h. The digested fragments were legated with 2.0 pmol EcoRⅠ, 2.0 pmol MseⅠadaptors and 1 U T4-DNA ligase at 16℃ to stand overnight. The Final amplification system included 1.0 U Taq polymerase, 0.4 μL EcoR I and Mse I primers. On this basis, 8 primers were screened with stable amplification and rich polymorphism. It was proved that this system was stable and credible. This optimized AFLP system would provide the basis for the genetic analysis of Asparagus.