目的 Dof（DNA binding with one finger）家族是高等植物中特有的一类转录因子家族，参与植物中光、激素、非生物胁迫等多种胁迫响应调控。本研究基于全基因组数据对黄花蒿Dof（AaDof）转录因子家族进行鉴定及表达模式分析，探究Dof家族基因在青蒿素合成调控中的作用。方法 经PFAM数据库鉴定获得AaDof序列，通过生物信息学软件分析其理化性质、亚细胞定位、基因结构、蛋白保守结构以及启动子序列结合元件等，并基于赤霉素（Gibberellic acid， GA）、紫外线B（UV-B）及二者协同胁迫下黄花蒿转录组数据对其表达模式进行分析。结果 本研究从全基因组水平共鉴定出51个AaDof基因，均含有保守的C₂-C₂单锌指结构，依据系统发育分析分为8个亚族，同一亚族内基因结构与蛋白保守结构域相对保守。亚细胞定位预测显示12个AaDof蛋白定位在细胞外，其余均定位在细胞核。启动子元件分析发现AaDof家族基因启动子区富含光、激素等多种响应元件。对AaDof在GA、UV-B和GA+UV-B处理下的表达模式分析发现，AaDof基因对GA胁迫处理响应较弱，仅有少量基因敏感，其表达主要受到UV-B胁迫影响。C1及C2.1亚族大部分基因在UV-B胁迫下上调表达，而A亚族大部分基因在UV-B胁迫下下调表达。qRT-PCR验证表明AaDof1、AaDof17和AaDof44在GA和UV-B处理下表达量显著上调，推测其可能通过参与GA和UV-B调控网络，正向调控青蒿素生物合成。结论 本研究系统鉴定了黄花蒿AaDof家族基因并筛选了3个可能正向调控青蒿素生物合成的候选AaDof基因，为黄花蒿Dof家族基因功能研究及其在青蒿素生物合成中的调控机制解析奠定基础。

Objective Dof (DNA binding with one finger) family is a plant-specific transcription factor family, which is involved in the regulation of light, hormones, abiotic stress and other stress responses. To explore the Dof family role on the regulation of artemisinin biosynthesis, based on identifying the Dof transcription family in Artemisia annua from the whole genome data and analyzing their expression patterns.Methods The AaDof sequences were identified by Pfam database. The physical and chemical properties, subcellular localization, gene structure, protein conserved structure and promoter sequence binding elements were analyzed by bioinformatics software, and their expression patterns were analyzed based on the transcriptome data of A. annua under gibberellin (GA), ultraviolet B (UV-B) and GA+UV-B treatment.Results A total of 51 AaDof genes were identified at the whole genome level, and their proteins contained conserved C₂-C₂ single zinc finger structure. According to phylogenetic analysis, they were divided into 8 subfamilies. The gene structure and protein conserved domains in the same subfamily were relatively conserved. Subcellular localization prediction showed that 12 AaDof proteins were located in the extracellular matrix, while others were located in the nucleus. Promoter element analysis indicated that the light, hormones and other stress-related elements were highly rich in promoter regions of AaDof family genes. The expression heatmap of AaDof genes under GA, UV-B and GA+UV-B treatment indicated the response of AaDofs to GA treatment were weak, only a few genes were sensitive. Their expression were mainly affected by UV-B stress. Most genes of C1 and C2.1 subfamilies were up-regulated under UV-B stress, while most genes of A subfamily were down-regulated under UV-B stress. qRT-PCR showed that the expressions of AaDof1, AaDof17 and AaDof44 were significantly up-regulated under GA and UV-B treatments, suggesting that they might play a positive regulatory role in artemisinin biosynthesis by participating in GA and UV-B regulatory networks.Conclusion This study systematically identify the Dof gene family of A.annua and screened three candidate AaDof genes that may positively regulate artemisinin biosynthesis. The results will lay a foundation for the functional study of Dof family genes in A.annua and the understanding of their regulatory mechanisms in artemisinin biosynthesis.

中国中医科学院科研管理处中国中医科学院科技创新工程项目（CI2021A05103）：基于青蒿药材的兽用产品开发，负责人：向丽；中国中医科学院科研管理处优秀青年科技人才（创新类）培养专项（ZZ13-YQ-101）：应用CRISPR/Cas9基因编辑技术培育高青蒿素含量青蒿品种的研究，负责人：师玉华。