目的 以药用植物红花Carthamus tinctorius为研究对象，克隆光信号途径关键转录因子HY5基因，并对其进行生物信息学和表达模式分析，为红花HY5基因的功能研究提供参考。方法 以红花转录数据为参考，设计引物，采用PCR扩增方法从红花中克隆得到HY5的全长cDNA和DNA序列。运用生物信息学方法对该基因进行分析，预测编码蛋白的结构与功能，并通过荧光定量PCR方法检测CtHY5基因在红花不同组织及花发育不同时期的表达情况。结果 CtHY5基因的cDNA全长为462 bp，编码153个氨基酸，DNA全长为1941 bp，包含4个外显子和3个内含子。生物信息学分析表明，CtHY5为亲水性蛋白，定位于细胞核中。系统进化树及模体结构分析结果表明CtHY5与来自菊科的刺菜蓟、黄花蒿、薇甘菊、向日葵、莴苣中的HY5进化关系较近。定量分析表明，在白色红花和红色红花中，CtHY5基因均在花中表达量最高，其次为茎和苞片，在根中表达量最低。此外，除了根外，CtHY5基因在白色红花各组织中的表达量要明显高于红色红花。结论 首次从红花中克隆得到了光信号途径关键转录因子CtHY5基因，并研究了该基因在不同花色红花品系中的表达模式，为进一步研究其生物学功能奠定了基础。

Objective HY5 (ELONGATED HYPOCOTYL5) is a key transcription factor in light signaling pathway. In this study, the CtHY5 gene was cloned from Carthamus tinctorius, and the bioinformatics and expression profile analysis of CtHY5 were performed with bioinformatics and quantitative Real-time PCR (qRT-PCR) methods to provide a reference for the functional study of the HY5 gene. Methods Based on the transcription data of safflower, primers were designed and the full-length cDNA of HY5 were cloned from safflower to obtain DNA sequences. The characteristics of physiochemical properties, structures and function of the deduced CtHY5 protein were determined using a series of bioinformatics tools. The expression of CtHY5 in different tissues and flower development stages of safflower was detected by fluorescence quantitative PCR. Results The full length cDNA sequence of CtHY5 gene was 462 bp, encoding 153 amino acids, while the full length DNA sequences of CtHY5 gene were 1 941 bp, containing four exons and three introns. Bioinformatics analysis showed that CtHY5 was hydrophilic protein and was located in the nucleus. Phylogenetic tree and motif structure analysis revealed that CtHY5 was more closely related to HY5 homologous protein in Cynara cardunculus, Artemisia annua, Mikania micrantha, Helianthus annuus and Lactuca sativa, which all of them belonged to Asteraceae. The results of qRT-PCR analysis showed that the expression of CtHY5 gene was the highest in the flowers, followed by stems and bracts, and the lowest in roots. And the expression of CtHY5 gene in all tissues of safflower was significantly higher in white safflower line than that in red safflower line. Conclusion CtHY5, the key transcription factor of light signaling pathway, was first isolated from safflower in this study. The expression pattern of CtHY5 in different lines of safflower was studied, which laid a foundation for further study on its biological function.

R286.12

财政部和农业农村部：国家现代农业产业技术体系资助（CARS-21）；河南省中央引导地方科技发展专项自由探索类项目（YDZX20214100001804）；河南省农科院新兴学科发展专项（2022XK03）；河南省科技攻关项目（222102110379、222102110466）；河南省农科院自主创新专项基金（2022ZC64）