OPTIMIZATION OF DNA AMPLIFICATION FINGERPRINTING TECHNIQUES TO STUDY GENETIC-RELATIONSHIPS OF WHITE LUPIN GERMPLASM

DNA amplification fingerprinting (DAF) techniques were used to evaluat e genetic relationships among 24 randomly selected white lupin (Lupinu s albus L.) accessions originating from four endemic regions. Paramete rs affecting DAF were optimized to obtain high resolution and reproduc ible banding patterns. DNA amplification was tolerant to a wide range of both template and primer concentrations but sensitive to Mg2+ conce ntration, and annealing temperature and time. Pure template DNA was fo und to be essential for obtaining reproducible banding patterns. Prime r annealing occurred at multiple sites, permitting detection of polymo rphism using only a single primer. Of 56 octamer primers tested, 22 yi elded highly polymorphic DAF bands that could be used to distinguish c losely related accessions. Genetic similarity coefficients were calcul ated and a dendrogram, using the unweighted pair group method with ari thmetic average (UPGMA), was constructed. A high level of genome diver sity was found among the accessions, but several from the Middle East and West Africa tended to cluster together. The results support the fu ture use of DAF markers for the characterization and identification of white lupin germplasm.