Modified AFLP technique for rapid genetic characterization in plants

The standard amplified fragment-length polymorphism (AFLP) technique was mo dified to develop a conveniens and reliable technique for rapid genetic cha racterization of plants. Modifications included (i) using one restriction e nzyme, one adapter molecule and primer (ii) incorporating formamide to gene rate more intense and uniform bands and (iii) using agarose gel electrophor esis. Sea oats (Uniola paniculata L.), pickerel-weed (Pontederia cordata L. ), Bermudagrass (Cynodon dactylon L.) and Penstemon heterophyllus Lindl. we re used to determine the ability to generate adequate resolution power with both self and cross-pollinated plant species including cultivars, ecotypes and individuals within populations. Reproducibility of bands was higher in all the AFLP experiments compared to random amplified polymorphic DNA (RAP D). Formamide with or without bovine serum albumin improved band intensitie s compared to dimethyl sulfoxide and the standard reaction mixture with no organic solvents. Comparison between RAPD and modified AFLP using sea-oats population samples proved that modified AFLP exhibits (i) a low number of f aint bands with increased specificity of amplified bands, (ii) a significan tly higher number of polymorphic loci per primer (iii) less primer screenin g time, (iv) easy scoring associated with fewer faint bands and (v) greatly enhanced reproducibility. The technique described here can be applied with a high degree of accuracy for plant genetic characterization.