百合LiLFY1基因的克隆和表达分析

Isolation of LiLFY1 and Its Expression in Lily (Lilium longiflorum Thunb.)

【目的】分离百合的LFY类基因,检测百合中LFY类基因的拷贝数并分析其表达模式。【方法】利用RT-PCR结合RACE的策略克隆百合的LFY类基因,通过Southern杂交检测百合中LFY类基因的拷贝数,通过RT-PCR分析LiLFY1基因的表达模式。【结果】获得了包括全长开放阅读框(ORF)的LiLFY1基因的cDNA序列。与已克隆的LFY类蛋白的同源性分析表明,LiLFY1编码的蛋白与单子叶植物水稻和玉米的LFY类蛋白具有更高的同源性。Southern杂交结果表明,二倍体百合中有2个拷贝的LFY类基因。LiLFY1基因在百合的幼嫩花芽和顶端分生组织中表达,而在根、茎、成熟的叶片和成熟花的各轮器官中不表达。【结论】百合的LiLFY1基因与已克隆的其它作物的LFY类基因高度同源,在百合的顶端分生组织和幼嫩花芽中表达,LiLFY1基因的克隆将有助于调整百合开花时间的基因工程研究。

[Objective] The aim of this study is to isolate the LFY family genes in lily and analyze the copies number and the expression of the LFY family genes isolated. [Method] LiLFY1 was isolated with the strategy of RT-PCR combining RACE. The copies number of the LFY family genes was analyzed by Southern blotting and the expression of LiLFY1 was analyzed by RT-PCR. [Result] The cDNA of LiLFY1 containing the full-length open reading frame （ORF） was isolated from lily （Lilium longiflorum Thunb.） by the RT-PCR and RACE strategy. The alignment analysis of the deduced LiLFY1 protein with other known LFY family proteins indicated that LiLFY1 is highly homologous with rice RFL and maize FLL. The result of Southern hybridization showed that there are two copies of LFY family genes in lily. LiLFY1 is expressed in young flower buds and shoot apical meristem （SAM） but not in roots, shoots, mature leaves and mature floral organs. [ Conclusion] The LiLFY1 isolated from lily are highly homologous with the other LFY family genes isolated from other species. LiLFYI is expressed in SAM and young flower buds. The cloning of LiLFY1 gene may be applied to the genetic engineering aiming for regulating the flowering time in lily.