Abstract
To develop a DNA fingerprinting technique suitable for edible tissues of vegetables, the reliable DNA extraction methods followed by fluorescent AFLP were proposed for each material. DNA was extracted from edible tissues of 33 vegetables using the miniprep DNA isolation method. In all materials, the DNA concentration and purity (A260/280 ratios of more than 1.8, A230/260 ratios of less than 2.95) estimated by UV spectrophotometry were thought to be sufficient for the next PCR step, but the browned DNA solution extracted from leaf lettuce or edible burdock inhibited PCR. The subsequent PCR was successful after purification of the crude DNA solutions, including 2-Mercaptethanol at a final concentration of 10mM, using Nucleon PhytoPure Kit (Amersham Biosciences Corp.). A total of 65 to 249 peaks were obtained by fluorescent AFLP analysis using six primer combinations in 33 vegetables. The technique developed in this study seems to have potential for cultivar identification in various vegetables. Furthermore, a similar AFLP analysis using bulked DNA samples from 16 plants each in three cultivars and 13 strains of Japanese bunching onion (Allium fistulosum L.) was conducted to examine the possibility of cultivar identification in outcrossing species. The DNA fingerprint obtained in this study allowed these cultivars and strains to be distinguished from each other.
