目的 通过对滇龙胆Gentiana rigescen中的生长素响应因子（auxin response factors，ARFs）家族成员进行全基因组鉴定和功能分析，探究其在滇龙胆花冠感温开闭运动中的潜在功能。方法 基于全基因组数据和转录组数据，利用生物信息学方法分析滇龙胆ARFs（Gentiana rigescens ARFs，GrARFs）基因家族的理化性质、保守基序和基因结构、顺式作用元件以及进化分析，采取实时荧光定量（RT-qPCR）技术分析其在滇龙胆不同开闭状态下的表达模式，并通过亚细胞定位实验验证GrARF9的蛋白定位，初步分析了GrARFs基因家族的生物学功能。结果 在滇龙胆基因组中共鉴定到21条GrARFs，编码612～1 065个氨基酸，相对分子质量在68 082.29～118 117.97，等电点范围为5.52～8.43，二级结构以无规则卷曲为主。系统进化树分析将滇龙胆中的ARFs分为4个亚群，即Group A、Group B、Group C、Group D；大多数GrARFs含有Auxin-resp和B3 DNA结合结构域；GrARFs启动子顺式作用元件含有丰富的激素响应与光响应原件；21条GrARFs不均匀分布在8条染色体上；共线性分析显示滇龙胆中的GrARFs在双子叶植物中比较保守，且与已经报道过功能的月季RhARFs有较多的共线性对；21个GrARFs基因在滇龙胆花冠感温过程中均存在差异表达；亚细胞定位实验结果显示GrARF9蛋白定位于细胞核，具有转录因子的核定位特征。结论 初步阐明了GrARFs在滇龙胆感温运动中的表达模式，为进一步研究GrARFs在滇龙胆花冠运动中的功能奠定了基础。

Objective By conducting a whole-genome identification and functional analysis of auxin response factors (ARFs) family members in Gentiana rigescens, this study investigate their potential roles in the thermosensitive corolla opening-closing movement of its corolla. Methods Based on whole-genome data and transcriptome data, bioinformatics methods were used to analyze the physicochemical properties, conserved motifs, gene structure, cis-acting elements, and evolutionary relationships of the G. rigescens ARFs (GrARFs) gene family. Real-time quantitative reverse transcription PCR (RT-qPCR) technology was employed to detect the expression patterns of GrARFs genes under different opening and closing states. Additionally, subcellular localization experiments were conducted to verify the protein localization of GrARF9, so as to preliminarily analyze the biological functions of the GrARFs gene family. Results A total of 21 GrARFs were identified in the G. rigescens genome, encoding 612—1 065 amino acids. Their relative molecular weights ranged from 68 082.29 to 118 117.97, and their isoelectric points spanned 5.52—8.43. The secondary structure was dominated by random coils. Phylogenetic tree analysis classified the ARFs in G. rigescens into four subgroups, namely Group A, Group B, Group C, and Group D. Most GrARFs contained Auxin-resp and B3 DNA-binding domains. The promoter cis-acting elements of GrARFs were rich in hormone-responsive and light-responsive elements. The 21 GrARFs were unevenly distributed across eight chromosomes. Collinearity analysis revealed that the GrARFs in G. rigescens were relatively conserved among dicotyledonous plants and shared a considerable number of collinear pairs with the functionally characterized ARFs from Rosa hybrida. All 21 GrARFs genes exhibited differential expression during the thermosensitive process of G. rigescens corollas. Results of subcellular localization experiments showed that the GrARF9 protein was localized in the nucleus, displaying the nuclear localization characteristic of transcription factors. Conclusion The results of this study preliminarily clarify the expression pattern of GrARFs in the thermosensitive movement of G. rigescens, and lay a foundation for further research on the function of GrARFs in the corolla movement of G. rigescens.

R282.12

云南省科技厅创新引导与科技型企业培育计划（202404BT090021）；云南省生物医药和大健康产业推进中心—重大科技专项计划（202402AA310040）；兴滇人才计划—产业创新人才（XDYC-CYCX-2022-0032）；迪庆州科技计划项目（2025ZXXN01）