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

本研究尝试开发DNA标记及其PCR引物，建立农作物品种和育种系的分类方法。在第一阶段中，我们测试了引物的条件，以产生用于该目的的可靠和有效的DNA片段。我们发现，具有特定DNA序列的12-mer随机引物比完全随机设计的引物有效地产生DNA片段。控制退火温度可能使电泳带型更可靠。合成的12-mer引物可用于水稻、橙子和黄瓜的品种和育种系的分类。该系列引物可通过本项目的研究人员购买。利用这些引物对栽培稻的籼、粳稻和野生稻的多年生、一年生品系进行了分类。在橙子的情况下，传统的生态类型定义的形态和生理特征的部分确定。在黄瓜的情况下，区分雄性和雌性幼苗。在未来的研究中，我们将尝试利用12-mer随机引物扩增的DNA片段的DNA序列设计大引物，以获得更可靠的分类。

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.