水稻Ds插入纯合体的筛选和鉴定

Selection and Identification of Ds Insertion Homozygotes in Rice (Oryza sativa L.)

采用Basta抗性鉴定、潮霉素抗性鉴定和PCR检测相结合的方法筛选和鉴定了水稻Ds插入纯合体。在T1代 2 36个转化株系中 ,有 16个株系的全部植株表现出对Basta的敏感 ,其余 2 2 0个株系的植株表现出对Basta的抗性。经过 3代的纯合筛选 ,共鉴定出Ds插入纯合体 2 0 3个 ,这些Ds插入纯合体可用于构建Ac/Ds系统和对Ds插入突变体进行筛选和鉴定 ,为水稻功能基因组学研究提供了材料。

Transposon tagging is a useful approach for gene isolation in higher plant.Its first step is the development of Ac/Ds system. In this paper, the identification of the homozygotes with Ds insertion in rice ( Oryza sativa L.) by a method combining Basta resistance, hygromycin resistance and PCR polymorphism (Fig.1) is described. A total of 236 transformed lines were subjected to Ds homozygote selection.In T 1 generation, 220 lines were found to be resistant to Basta and 16 lines were susceptible (Table 1). After two generations of selfing of the 220 Basta-resistant lines, 203 Ds homozygotes were identified and the other 17 lines were still segregating (Tables 2, 3). The Ds homozygotes can be used to develop Ac/Ds system and to detect mutants for rice functional genomic study.