Computer-aided RAPD fingerprinting of accessions from the ryegrass–fescue complex

Abstract

RAPD (Randomly Amplified Polymorphic DNA) assay of 32 cultivar accessions from the ryegrass–fescue (Lolium–Festuca) complex was accomplished using ten decamer primers to assess (i) the power of RAPD technology to discriminate between individual commercial accessions and to produce cultivar fingerprinting, (ii) the degree of relatedness of accessions based on RAPD profiles in comparison with other existing classifications, and (iii) the possibility of automation of RAPD technology.The variation of the correlation coefficient r as the primary output from the automated RAPD-profile processing summarizes variability derived from DNA isolation, the RAPD reaction, and final computer-image processing of RAPD profiles. The AII (Accession Identity Interval) of r for accession Festuca arundinacea cv. Lekora was determined experimentally and the value obtained was accepted as a valid interval for all the other accessions studied. In order to evaluate the discrimination potential of all ten primers together, a pooled-similarity matrix was computed. Employing this approach, we achieved 100% discrimination between all 35 accessions when using all ten primers. A dendrogram for all 35 accessions was obtained using average linkage cluster analysis (UPGMA – Unweighted Pair Group Method with Arithmetic Means). This procedure successfully produced smaller groups of higher taxonomic homogeneity. The relationships between the Lolium–Festuca accessions were also revealed by principal coordinate analysis (PCO) based on absorbance profiles from the RAPD assay. Again, all accessions were well separated, recognising even subspecies relationships. In general, PCO analysis confirmed the inferences made from the UPGMA method.We successfully applied the computer-aided system of RAPD assay, based on an IBM PC computer, for discrimination of cultivars as well as for description of DNA-based relationships of accessions from various taxonomic groups of the Lolium–Festuca complex.