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用 32对引物组合对水稻 (OryzasativaL .)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。
AFLP analysis of the total DNA of rice (Oryzasativa L.) Zhenshan97 male sterile line and its maintainer using 32 pairs of primers showed 88 differential bands. Among them, the difference between the fragment of EcoRⅠ_AAC × MseⅠ_CTC, A1 of maintainer line of Zhenshan 97 and A2 of maintainer line was strong, and the difference was obvious. The primer combination was used to compare the single F1 generation Shanyou 6 3 and its F2 segregation generation and restorer line Minghui 6 3. The results showed that both Shanyou 6 3 and F2 single plants with the same cytoplasm With A1 characteristic band; Ming Hui 6 3 has A2 characteristic band. Furthermore, chloroplast DNA and mitochondrial DNAAFLP bands of CMS lines and maintainer lines of Zhenshan 97 were compared to prove that A1 and A2 were derived from the chloroplast genome. The results of DNA sequencing showed that A1 had a 6-base repeat fragment more than A2: AGAAAA . DNA sequence homology also confirmed that A1 and A2 are from the rice chloroplast genome. This pair of differential fragments can be used as a molecular marker to identify the cytoplasm of Zhenshan97 male sterile line and maintainer line.