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为了发掘小麦育性基因,采用同源克隆的方法,从小麦花药中克隆了3个与拟南芥AtMS2和水稻OsMS2基因同源的cDNA序列。TaMSR-1、TaMSR-2和TaMSR-3分别具有1 842,1 824,1 830 bp的开放阅读框,各自编码613,607,609个氨基酸。推导的蛋白质序列中包含2个保守区:NAD_binding和Sterile保守功能域。氨基酸序列分析发现,TaMSR与水稻OsMSR2和拟南芥AtMSR2具有较高的相似性。采用基因特异定位引物PCR技术对中国春缺失系材料进行扩增,将TaMSR-1、TaMSR-2和TaMSR-3基因分别定位于4AS、4DL和4BL染色体上。半定量RT-PCR分析表明,TaMSR基因是花药组织特异表达的基因。这些结果说明,TaMSR基因可能在小麦花药发育过程中起重要作用。
In order to explore the wheat fertility genes, three cDNA sequences homologous to AtMS2 and OsMS2 genes of Arabidopsis were cloned from wheat anthers using homologous cloning method. TaMSR-1, TaMSR-2 and TaMSR-3 have open reading frames of 1 842, 1 824 and 1 830 bp, respectively, encoding 613,607,609 amino acids. The deduced protein sequence contains two conserved regions: NAD_binding and Sterile conserved domains. Amino acid sequence analysis showed that TaMSR had higher similarity with rice OsMSR2 and Arabidopsis AtMSR2. Genomic DNA was used to amplify the Chinese spring deletion lines. The TaMSR-1, TaMSR-2 and TaMSR-3 genes were located on 4AS, 4DL and 4BL chromosomes respectively. Semi-quantitative RT-PCR analysis showed that TaMSR gene is an anther tissue-specific gene. These results indicate that the TaMSR gene may play an important role in the development of wheat anthers.