Cytoplasmic male sterility (CMS) line, Zhenshan 97A, and its maintainer, Zhenshan 97B, are widely used in hybrid rice (Oryza sativa L.) production in China. Theoretically, these two lines are homologous in genome DNA but different in cytoplasmic genes. The comparison of AFLP maps based on the total DNA, chloroplast DNA and mtDNA between Zhenshan 97A and Zhenshan 97B are herein reported. Eighty-eight different fragments between the CMS line and its maintainer were shown in 32 pairs of amplification primers. Among the differential fragments, A1, produced by primers EcoRⅠ-AAG and MseⅠ-CTC and used total DNA as templates, appeared in the maps of Zhenshan 97A and its hybrid F1 seeds Shanyou 63 and F2 seeds respectively. A2, produced by the same primers showed up in AFLP of Zhenshan 97B and restore line Minghui 63. The fragments A1, A2 also appeared in that of chloroplast DNA. A1, A2 were recovered from the denatured polyacrylamide gel and cloned to pBluescript SK+. Sequence analysis showed that A1 contained one more repeat of 6 basepairs than that of A2 and the repeat was located upstream of the rice chloroplast tRNA-Leu gene.
水稻胞质雄性不育系珍汕97A 及其保持系的AFLP标记及序列分析
侯磊 杨光伟 何光华 唐彬 肖月华 裴炎*
（西南农业大学生物技术研究中心! 农学系，重庆400716）
摘要：用 32对引物组合对水稻 (Oryzasativa L .)珍汕 97不育系与保持系的总DNA进行AFLP分析 ,显示出 88个差异带。其中 ,引物组合EcoRⅠ-AAC×MseⅠ- CTC产生的差异片段——珍汕 97不育系的A1与保持系的A2 ,信号强 ,差异明显。用该引物组合对杂种F1代汕优 6 3及其F2 分离世代单株以及恢复系明恢 6 3进行比较研究 ,结果表明 ,与不育系胞质相同的汕优 6 3和F2 单株均具A1特征带 ;明恢 6 3具有A2 特征带。进而比较了珍汕 97不育系与保持系的叶绿体DNA和线粒体DNAAFLP谱带 ,证明A1和A2 产生自叶绿体基因组 ,而且DNA序列测定结果表明 ,A1比A2 多一个 6碱基的重复片段 :AGAAAA。DNA序列同源性比较也证实了A1和A2 来自于水稻叶绿体基因组。这对差异片段可以作为鉴别珍汕 97不育系与保持系细胞质的分子标记
关键词： 水稻；叶绿体；胞质雄性不育系；AFLP
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