Abstract:

The aim was to establish SRAP-PCR reaction system and provide a new tool for future research on Aechmea germplasm. A orthogonal design of L16(45) was used to optimize SRAP-PCR reaction system for Aechmea species with five factors, namely Mg2+, dNTPs, primers, Taq DNA polymerase and template DNA, and SRAP primer combinations were screened for polymorphism. The result showed that a suitable SRAP-PCR reaction system for Aechmea species was 1.50 mmol/L Mg2+, 400 μmol?/L dNTPs, 1.5 U Taq DNA polymerase, 15 μmol/L primer combination, 30 ng template DNA and 1× PCR buffer. In addition, each factor in SRAP-PCR reaction system had different effects on the amplified patterns and the descending order was DNA > Taq DNA polymerase > dNTPs > primer > Mg2+. A total of 51 polymorphic SRAP primer combinations were screened out from 56 SRAP primer combinations, with the polymorphic primer ratio more than 90%. The optimized SRAP-PCR reaction system was identified by different genomic DNAs of Aechmea species and different SRAP primer combinations; as a result, the amplified pattern with rich polymorphism and clear bands was obtained. It proved that the established SRAP-PCR reaction system for Aechmea species was steady and reliable.