M-AFLP-based protocol for microsatellite loci isolation in Cynara cardunculus L. (Asteraceae)

Nine microsatellite markers for Cynara cardunculus L. were developed using a two-step 'primer extension' procedure, based on microsatellite-amplified fragment length polymorphism (M-AFLP) technique. In the first step, highly enriched SSR gel profiles were produced and, from the derived sequences of selected bands, forward primers directed towards the microsatellite motif were designed. In the second step, the opposite microsatellite flanking sequence was isolated using a nested approach on a restricted-ligated genomic fraction. Polyp morphism was explored in 24 plants of wild cardoon (Cynara cardunculus L. var. sylvestris) as well as two accessions of both globe artichoke (Cynara cardunculus L. var. scolymus), and cultivated cardoon (Cynara cardunculus L. var. altilis).