摘要： 为探究FT基因在大麻光周期途径中调控开花的作用，从工业大麻品种庆麻1号（Q1）中使用PCR技术克隆了FT同源基因cDNA序列，命名为CsHd3a。利用生物信息方法对该基因序列特征进行了分析，并使用实时荧光定量（qRT-PCR）技术研究其组织特异性表达模式。选取了晚花品种云麻7号（Y7）与早花品种Q1使用qRT-PCR技术进行了长、短日照下的表达谱分析，并分别克隆了2个品种的CsHd3a基因，对其氨基酸序列进行了差异分析。结果表明，CsHd3a基因CDS全长为543 bp，编码180个氨基酸，包含PEBP家族蛋白结构域。系统进化分析表明，CsHd3a蛋白与棉花的GhFT1蛋白亲缘关系最近。组织特异性分析表明，大麻CsHd3a基因在叶片中高表达，在根和茎中几乎不表达。qRT-PCR分析结果表明，Q1在短日照（SD）处理下表现出节律性变化，且在8：00达到最高，而在长日照（LD）处理下几乎不表达。SD条件下，早花品种Q1 CsHd3a基因表达量显著高于晚熟品种Y7。此外，CsHd3a氨基酸序列在Q1和Y7存在5处氨基酸差异，其差异是否因其保守结构域PEBP功能的变化还有待进一步研究。综上，获得了CsHd3a基因的CDS序列，初步探究了早、晚花品种Q1和Y7 CsHd3a基因的时空表达模式，为探究大麻开花光周期途径的机理奠定了研究基础。

关键词: 大麻, CsHd3a, 基因克隆, 生物信息学分析, 表达谱分析

Abstract: In order to explore the role of the FT gene in the regulation of flowering by the photoperiod pathway of cannabis, the FT homologous gene cDNA sequence was cloned from the industrial cannabis variety Qingma 1 hao (Q1) using PCR technology and named CsHd3a. The sequence characteristics of the gene using biological information were analyzed, and the Real-time fluorescence quantification (qRT-PCR) technology was used to study its tissue-specific expression pattern. The late-flowering variety Yunma 7 hao (Y7) and the early-flowering variety Q1 were selected for long-day and short-day expression profiling using qRT-PCR technology, and the CsHd3a genes of the two varieties were cloned, and the amino acid sequence difference analysis was carried out. The results showed that the CDS sequence of CsHd3a gene was 543 bp in length, encoded 180 amino acids, and contained the protein domain of the PEBP family. Phylogenetic analysis showed that the CsHd3a protein was closely related to the GhFT1 protein of cotton. Tissue-specific analysis showed that the gene was highly expressed in leaves and almost not expressed in roots and stems. The results of qRT-PCR analysis showed that Q1 showed a rhythmic change under short-day (SD) treatment, and reached the highest at 8:00, while it was almost not expressed under long-day (LD) treatment. Under SD conditions, the CsHd3a gene expression of early-flowering variety Q1 was significantly higher than that of late-maturing variety Y7. In addition, the amino acid sequence of CsHd3a had 5 amino acid differences between Q1 and Y7. Whether the difference was due to the change in the function of its conserved domain PEBP remains to be further studied. In summary, this study obtained the CDS sequence of the CsHd3a gene, and initially explored the spatiotemporal expression patterns of CsHd3a genes in the early and late flowering varieties Q1 and Y7, which laid a research foundation for exploring the mechanism of the flowering photoperiod pathway of cannabis.

Key words: Cannabis, CsHd3a, Gene cloning, Bioinformatics analysis, Expression profile analysis

中图分类号: 

• Q78

• S563.03