Species Identification of Beans, Peas and Other Legumes by RAPD-PCR after DNA Isolation using Membrane Columns

Abstract

Forty legume seed samples representing 11 species were selected to investigate the identification of food legume species by random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR). Template DNA was extracted from seed meal and purified using a commercial membrane column kit. Amplification was performed with commercial RAPD analysis beads and six commercially available decamer nucleotide primers. Electrophoresis of the amplicons on polyacrylamide gels and subsequent silver staining resulted in RAPD profiles from all samples of one species with a given primer that differed from those of the other species studied. Unambiguous identification of six food legume species, common beans ( Phaseolus vulgaris), soybeans ( Glycine max), peas ( Pisum sativum), chickpeas ( Cicer arietinum), lentils ( Lens culinaris), and alfalfa ( Medicago sativa), was obtained applying three of the primers. The technique may also be suitable to identify the remaining five species investigated, scarlet runner beans ( Phaseolus coccineus), lima beans ( P. lunatus), green gram ( Vigna radiata), broadbeans ( Vicia faba), and blue lupin seeds ( Lupinus angustifolius). Using a standard protocol, amplification with Taq DNA polymerase instead of RAPD beads generated RAPD profiles only from soybeans, peas, two of four chickpea samples, green gram, and lupin seeds.