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

DNA 标记的开发及其在作物品种育种系和品种分类中的应用

• 批准号: 04556002
• 负责人: SATO Yoichiro
• 金额: $ 4.61万
• 依托单位: FACULTY OF AGRICULTURE,SHIZUOKA UNIVERSITY (1994)National Institute of Genetics (1992-1993)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Developmental Scientific Research (B)
• 财政年份: 1992
• 资助国家: 日本
• 起止时间: 1992 至 1994
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-04556002/
• 关键词: DNA marker; Primers; Crop; Ecotype; Rice; Cucumber; Orange; DNAマーカ; DNAフィンガープリント; 系統進化; DNAマーカー; PCR

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.

我们尝试开发DNA标记及其PCR引物，以建立作物品种和选育系的分类方法。在第一阶段，我们测试了引物的条件，以产生自信和有效的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更有效地产生DNA片段。控制退火温度可能会导致更可靠的电泳带图。该系列引物共12个引物，用于水稻、柑橘和黄瓜的品种和育种系的分类。该系列引物可通过本项目整合的研究人员购买。利用这些引物对栽培稻的籼稻和粳稻以及野生稻的多年生和一年生品系进行了分类。以橙子为例，由形态和生理性状定义的传统生态型得到了部分鉴定。以黄瓜为例，对雌雄幼苗进行了区分。未来，我们将尝试使用基于12-mer随机引物扩增DNA片段的DNA序列设计的大尺寸引物进行STS PCR，以获得更可靠的分类。

项目成果

松浦誠司・斉藤彰・藤田幸雄: "An approach for rapid checking of seed purity by RFLP analysis of nuclear DNA in F1 hybrid of cucumber" J. Japan Soc. Hort. Sci 63:. 63. 379-383 (1994)

Seiji Matsuura、Akira Saito 和 Yukio Fujita：“通过 F1 杂交种黄瓜的 RFLP 分析快速检查种子纯度的方法”J. Japan Soc 63. 379-383 (1994)。

Chen.W.B.,Y.I.Sato,I.Nakamura and H.Nakai: "Indica-Japonica differentiation in Chinese rice landraces" Euphytica. 74. 195-201 (1994)

Chen.W.B.、Y.I.Sato、I.Nakamura 和 H.Nakai：“中国水稻地方品种中的籼粳分化”Euphytica。

Chen,W.B.et at.: "indica-japonica differentiation of Chinese traditional rice cultivars." Japanese Journal of Genetics. (印刷中).

Chen，W.B. 等：“中国传统水稻品种的籼粳分化”。日本遗传学杂志（正在出版）。

CHEN,W.B., Y.I.SATO,I.NAKAMURA and H.NAKAI: "Indica-japonica differentiation in Chinese rice landraces" Euphytica. 74. 195-201 (1994)

CHEN，W.B.，Y.I.SATO，I.NAKAMURA 和 H.NAKAI：“中国水稻地方品种的籼粳分化”Euphytica。

MATSUURA,S., A.SAITO and Y.FUJITA: "An apporach for rapid checking of seed purity by RFLP analysis of nuclear DNA in F1 hybrid of cucumber" J.Japan Soc.Hort.Sci.63. 379-383 (1994)

Matsuura,S.、A.SAITO 和 Y.FUJITA：“通过对黄瓜 F1 杂交种核 DNA 进行 RFLP 分析来快速检查种子纯度的方法”J.Japan Soc.Hort.Sci.63。