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以洋葱(Allium cepa L.)品系‘1007’为试材,采用同源基因克隆及RACE-PCR的方法,获得植物开花调控关键基因LEAFY的同源基因的cDNA序列,命名为AcLFY,GenBank登录号为JX275962。序列分析表明,该基因包含1个1119bp的开放阅读框,编码372个氨基酸,该氨基酸序列含有5′-N端脯氨酸富集区,亮氨酸拉链结构和中央酸性区,具有LEAFY(FLO)家族的典型结构特征。同源分析表明,该氨基酸序列与水仙的同源性接近70%,与其他高等植物LEAFY类蛋白的同源性均在50%以上。进化树分析表明AcLFY与单子叶植物的亲缘关系高于双子叶植物。荧光定量结果显示,AcLFY在洋葱抽薹开花的整个过程中都有表达,在抽薹初期的花序分生组织中表达量最高,在花器官中只有微量表达,花器官形成后,主要在叶片中表达。
The cDNA sequence of the homology gene LEAFY, which is the key gene of flowering control, was obtained by homologous cloning and RACE-PCR using the ’Allium cepa L.’ line ’1007’ as test material, and named as AcLFY, GenBank accession number For JX275962. Sequence analysis showed that the gene contains an open reading frame of 1119bp and encodes 372 amino acids. The amino acid sequence contains a 5’-N-terminal proline-rich region, a leucine zipper structure and a central acidic region with LEAFY (FLO The typical structural features of the family. Homology analysis showed that the homology of this amino acid sequence with Narcissus was nearly 70%, and its homology with LEAFY proteins of other higher plants was over 50%. Phylogenetic tree analysis showed that AcLFY is more closely related to monocotyledonous plants than dicotyledons. Fluorescent quantification results showed that AcLFY was expressed throughout the flowering stage of the onion, and was the highest in the inflorescence meristem at the early bolting stage and only slightly expressed in the floral organs. After forming the floral organs, AcLFY was mainly expressed in the leaves.