目的 从红花Carthamus tinctorius基因组中克隆CtFD1与CtFD2基因，对其进行生物信息学、表达特异性和亚细胞定位分析。方法 基于红花基因组筛选并克隆CtFD1与CtFD2基因，利用生物信息学在线网站进行理化性质、保守结构域等分析；通过实时荧光定量PCR（quantitative real-time polymerase chain reaction，qRT-PCR）检测其组织表达特性；利用烟草瞬时表达系统检测观察亚细胞定位情况；通过优化诱导剂浓度及温度使蛋白为可溶性表达。结果 CtFD1与CtFD2基因CDS大小分别为726 bp和540 bp，编码241和179个氨基酸。二者均包含典型的bZIP转录因子结合域，为亲水性蛋白，定位于细胞核。系统进化树显示与黄花蒿Artemisia annua亲缘关系较近。组织表达分析表明，CtFD1与CtFD2基因在茎尖分生组织（shoot apical meristem，SAM）中表达量最高。原核表达分析结果显示在16 ℃、IPTG浓度为0.2 mmol/L时，蛋白可在上清中大量表达。结论 CtFD1与CtFD2基因属于bZIP家族成员，在红花茎尖分生组织中表达量最高，表明这2个基因对红花开花调控可能发挥重要作用，为进一步研究基因功能及开花机制解析提供了科学依据。

Objective To clone the CtFD1 and CtFD2 genes from the genome of Carthamus tinctorius and perform bioinformatics, expression specificity, and subcellular localization analyses. Methods The CtFD1 and CtFD2 genes were screened and cloned based on the C. tinctorius genome. Bioinformatics online tools were used to analyze physicochemical properties, conserved domains, and other features. Quantitative real-time polymerase chain reaction (qRT-PCR) was employed to examine tissue-specific expression patterns. Subcellular localization was observed using a tobacco transient expression system. Soluble expression of the proteins was achieved by optimizing inducer concentration and temperature. Results The coding sequences (CDS) of CtFD1 and CtFD2 were 726 bp and 540 bp in length, encoding 241 and 179 amino acids, respectively. Both proteins contained a typical bZIP transcription factor binding domain, which were hydrophilic, and localized to the nucleus. Phylogenetic analysis indicated a close evolutionary relationship with Artemisia annua. Tissue expression analysis showed that CtFD1 and CtFD2 were most highly expressed in shoot apical meristem (SAM). Prokaryotic expression analysis revealed that both proteins were abundantly expressed in the supernatant at 16  ℃ with an IPTG concentration of 0.2 mmol/L. Conclusion The CtFD1 and CtFD2 genes belong to the bZIP family and exhibit the highest expression levels in the SAM of C. tinctorius, suggesting that they may play important roles in regulating flowering. This study provides a scientific basis for further functional characterization of these genes and elucidation of the flowering mechanism in C. tinctorius.

R282.12

兵团自然科学支持计划重点项目（2025DA003）；新疆院士人才发展基金