Optimization of DNA amplification fingerprinting techniques to study genetic relationships of white lupin germplasm

DNA amplification fingerprinting (DAF) techniques were used to evaluate genetic relationships among 24 randomly selected white lupin (Lupinus albus L.) accessions originating from four endemic regions. Parameters affecting DAF were optimized to obtain high resolution and reproducible banding patterns. DNA amplification was tolerant to a wide range of both template and primer concentrations but sensitive to Mg 2+ concentration, and annealing temperature and time. Pure template DNA was found to be essential for obtaining reproducible banding patterns. Primer annealing occurred at multiple sites, permitting detection of polymorphism using only a single primer. Of 56 octamer primers tested, 22 yielded highly polymorphic DAF bands that could be used to distinguish closely related accessions. Genetic similarity coefficients were calculated and a dendrogram, using the unweighted pair group method with arithmetic average (UPGMA), was constructed. A high level of genome diversity was found among the accessions, but several from the Middle East and West Africa tended to cluster together. The results support the future use of DAF markers for the characterization and identification of white lupin germplasm.