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, ν/ν). 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). © 2002 Elsevier Science Ltd. All rights reserved.