FINGERPRINTING OF ALFALFA MEIOTIC MUTANTS USING RAPD MARKERS

Polymerase chain reaction (PCR), with single ten-base-long nucleotide primers, was used to amplify random regions of genomic DNA from eleven diploid meiotic mutants (2n egg, 2n pollen and jumbo pollen producers ) of the Medicago sativa-coerulea-falcata complex. Electrophoretic pat terns of the resulting random amplified polymorphic DNA (RAPD) fragmen ts were evaluated to develop a graphical representation of amplificati on products from which conserved and individual-specific markers could be readily identified. Oligonucleotide primers differed significantly in their capacity of detecting polymorphism. Jumbo pollen (jp) mutant s showed higher levels of similarity, as proportion of shared amplific ation products, than 2n pollen mutants. A diploid plant of alfalfa, te sted for the occurrence of two mechanisms of unreduced egg formation ( second division restitution and apomeiosis) was analyzed to obtain gen ome specific fingerprints. This PCR-based characterization could be us ed to determine whether the apomeiotic 2n eggs develop through parthen ogenesis. The application of RAPD markers for germplasm evaluation and alfalfa improvement programs is also discussed.