Identification of Oryza Species with the CD Genome Based on RFLP Analysis of Nuclear Ribosomal ITS Sequences

作     者：包颖 葛颂 

作者机构：中国科学院植物研究所系统与进化植物学重点实验室北京100093 山东曲阜师范大学生物系曲阜213765 

出 版 物：《Acta Botanica Sinica》 (Acta Botanica Sinica（植物学报：英文版）)

年 卷 期：2003年第45卷第7期

页      面：762-765页

核心收录：

学科分类：0710[理学-生物学] 07[理学] 09[农学] 071007[理学-遗传学] 0901[农学-作物学] 090102[农学-作物遗传育种] 

基　　金：国家自然科学基金(3002500) 中国科学院知识创新工程重要方向项目(KSCXZSW-101A) 

主　　题：Oryza identification CD genome RFLP ITS 

摘      要：The genus Oryza consists of approximately 24 species with 10 recognized genome types (A, B, C, BC, CD, E, F, G, HJ, HK). The species with the CD genome are endemic to the Central and South America and comprise three species, i.e. O. latifolia Desv., O. alta Swallen and O. grandiglumis (Doell) Prod. Although the three species can be easily distinguished from the species with the other genomes in Oryza by their morphological characteristics, it is very difficult to distinguish them from one another. Recent studies suggested that O. alta and O. grandiglumis should be the same species (O. grandigiumis), whereas O. latifolia retains its taxonomic position unchangeably. In this paper, a total of 77 clones of the nuclear ribosomal internal transcribed spacer (ITS) from 11 samples representing different geographical races were sequenced. Using DNA Strider 1.2 software, the restriction enzyme digestion sites of these clone sequences were analyzed. Based on restriction fragment length polymorphism (RFLP) of ITS sequences, a method to identify the species with CD genome in Oryza was proposed. The method is rapid and convenient and all experimental procedure includes only three steps: (1) to amplify the ITS fragment with the routine primers; (2) to digest PCR products with restriction enzymes Fok I and Dra III or both; (3) to run the digested product on 1% agarose gel and identify the sample based on the restriction profiles.