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以毛白杨为材料,采用同源基因克隆法从毛白杨中分离了FT(FLOWERING LOCUS T)同源基因PtFT1和PtFT2编码区序列。测序结果表明PtFT1和PtFT2编码区长度均为525bp,可编码174个氨基酸。蛋白序列比对发现这两个基因与拟南芥、葡萄等物种中FT同源基因所编码的氨基酸同源性达到75%以上,PtFT1和PtFT2所推测的氨基酸序列包含FT类蛋白保守基序(LGRQTVYAPGWRQN)和两个关键性氨基酸残基Tyr84(Y),Gln139(Q)。系统进化分析进一步表明PtFT1和PtFT2属于FT亚家族成员。采用Real-time qRT-PCR技术检测PtFT1和PtFT2在各个组织部位中的表达模式,结果表明PtFT1和PtFT2在各个组织部位均有表达,但这两个基因的表达水平存在差异;两个基因在早期雌雄花芽(7月5日)表达量明显高于成熟雌雄花芽(翌年3月10日)表达量,进而推测在毛白杨中PtFT1和PtFT2的表达响应日照长短,长日照条件促进这两个基因的表达,它们可能在光周期调控的开花途径中促进花芽分化和开花发挥特定作用。这些研究对于阐明PtFT1和PtFT2在光周期开花调控途径中的作用机制具有重要意义,为进一步开展毛白杨开花调控基因工程研究奠定了工作基础。
Using Populus tomentosa as a material, the homologous genes PtFT1 and PtFT2 of FT (FLOWERING LOCUS T) were isolated from Populus tomentosa by homologous cloning method. Sequencing results showed that both PtFT1 and PtFT2 coding regions were 525 bp in length and encoded 174 amino acids. Protein sequence alignment revealed that the homologies of these two genes to the homologous genes of FT homology in Arabidopsis and grape reached more than 75%. The putative amino acid sequence of PtFT1 and PtFT2 contained the conserved motifs of FT protein LGRQTVYAPGWRQN) and two key amino acid residues Tyr84 (Y), Gln139 (Q). Phylogenetic analysis further indicates that PtFT1 and PtFT2 belong to members of the FT subfamily. Real-time qRT-PCR was used to detect the expression pattern of PtFT1 and PtFT2 in various tissues. The results showed that PtFT1 and PtFT2 were expressed in all tissues and tissues, but the expression levels of these two genes were different. The expression levels of male and female flower buds (July 5) were significantly higher than those of mature male and female flower buds (March 10 the following year), and then the expressions of PtFT1 and PtFT2 in Populus tomentosa were estimated to be in response to the length of daylight and long-day conditions to promote the expression of these two genes Expression, they may play a specific role in promoting flower bud differentiation and flowering in the photoperiod-controlling flowering pathway. These studies are of great significance for clarifying the mechanism of action of PtFT1 and PtFT2 in the photoperiod-flowering regulatory pathway, which laid the foundation of further research on genetic engineering of Flowering Populus tomentosa.