Amplified fragment length polymorphism analysis of vandaceous orchids

We investigated the application of the PCR-based fingerprinting technique, amplified fragment length polymorphism (AFLP), in orchids. The optimal AFLP patterns have been determined using primer combinations of EcoRI + 4 and M seI + 3 selective nucleotides. The same reproducible AFLP patterns were dem onstrated in genomic DNAs isolated both from: (1) different orchid tissues, e.g. leaves and flowers; and (2) orchid flowers collected at different tim es. Genomic variations among different cultivars of vandaceous orchid hybri ds were successfully determined by AFLP analysis. More than 10% of the AFLP bands were polymorphic DNA when siblings, derived from the same original c rosses (two cultivars of Aranda Christine, five cultivars of Mokara Willie How), were used. Only 0.3-0.7% of the AFLP patterns were shown to be polymo rphic when different cultivars, originating from somatic mutations during m eristem culture for massive propagation, were used (two cultivars of AI. Ch ristine, foul cultivars of M. Chark Kuan). (C) 1999 Elsevier Science Irelan d Ltd. All rights reserved.