Identification of Food and Feed Legumes by RAPD-PCR

Abstract

Sixty-two legume seed samples representing 25 species were selected to investigate the identification of food and feed legumes by random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR). Template DNA was extracted from defatted seed meal with SDS-containing buffer and purified by extraction with chloroform:isoamyl alcohol (24:1, v/v). Amplification was performed with Taq DNA polymerase and six commercially available 10-mer nucleotide primers using standard template dilutions for 12 species and optimum template concentrations ascertained by testing serial dilutions for the remaining species. Identification of 19 legume species, common beans ( Phaseolus vulgaris), runner beans ( P. coccineus), lima beans ( P. lunatus), soybeans ( Glycine max), broadbeans ( Vicia faba), green gram ( Vigna radiata), black gram ( V. mungo), moth beans ( V. aconitifolia), peas ( Pisum sativum), chickpeas ( Cicer arietinum), pigeon peas ( Cajanus cajan), grasspeas ( Lathyrus sativus), broad-leaved peas ( L. latifolius), lentils ( Lens culinaris), blue lupin ( Lupinus angustifolius), European yellow lupin ( L. luteus), white lupin ( L. albus), alfalfa ( Medicago sativa), and common sainfoin ( Onobrychis viciifolia), was achieved using four of the primers. The technique may also be suitable to identify the remaining species studied, jackbeans ( Canavalia ensiformis), swordbeans ( C. gladiata), horsegram ( Macrotyloma uniflorum), Florida velvetbean ( Mucuna deeringiana), sweet trefoil ( Trigonella coerulea), and birdsfoot trefoil ( Lotus corniculatus).