DEVELOPMENT OF DNA MARKER AND ITS UTILIZATION FOR CLASSIFYING BREEDING LINES AND CULTIVARS OF CROP SPECIES

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We attempted develop DNA markers and their PCR primers to establish the methodology for classification of the varieties and breeding lines of crop species. In the first stage, we tested conditions of primers for producing confident and effect DNA fragments for the purpose. We found that 12-mer random primers having particular DNA sequences effectively produced DNA fragments than fully randomly designed primers. It was probable that controlling annealing temperature may result in more reliable banding pattern of electrophoresis. Such series of 12-mer primers are synthesized for the classification of cultivars and breeding lines of rice, orange and cucumber. The series of primers can be purchased through the researchers integrated in this project. By using these primers, indica and japonica of cultivated rice, and perennial and annual strains of wild rice were classified. In the case of oranges, traditional ecotypes defined by morphological and physiological traits were partly identified. In case of cucumber, male and female seedlings were distinguished. In the future, we attempt STS PCR that uses big-size primers designed based on the DNA sequences of the DNA fragments amplified by 12-mer random primers for more reliable classification.