紫苏ARF转录因子家族全基因组鉴定及其在非生物胁迫响应中的功能分析

doi:10.13560/j.cnki.biotech.bull.1985.2025-0877

生物技术通报 ›› 2026, Vol. 42 ›› Issue (7): 148-162.doi: 10.13560/j.cnki.biotech.bull.1985.2025-0877

• 研究报告 • 上一篇

紫苏ARF转录因子家族全基因组鉴定及其在非生物胁迫响应中的功能分析

黄旭升¹, 周雅莉¹, 贾峥嵘², 沈珊³, 陈锦钗¹, 周广立¹, 李鑫鑫¹, 王计平¹, 李润植¹, 贾小云³()

^1.山西农业大学农学院山西省特用作物遗传和代谢工程研究中心，太谷 030801
^2.山西农业大学高粱研究所高粱遗传与种质创新山西省重点实验室，榆次 030600
^3.山西农业大学生命科学学院山西省特用作物遗传和代谢工程研究中心，太谷 030801

收稿日期:2025-08-12 出版日期:2026-07-26 发布日期:2026-07-20
通讯作者: 贾小云jiaxiaoyun@sxau.edu.cn
作者简介:第一联系人：同等贡献
基金资助:
中央引导地方科技发展资金项目(YDZJSX2024B007);山西省科技合作项目(202304041101009);山西省基础研究计划（自由探索类）项目(202403021212098);山西省基础研究计划（自由探索类）项目(202403021212310);山西省回国留学人员科研资助项目(2021-074);忻州市科学技术局项目(20240212);山西农业大学科技创新提升工程(CXGC2025023);山西农业大学科技创新提升工程(CXGC2023074)

Whole-genome Identification of the Perilla frutescens ARF Transcription Factor Family and Functional Analysis of Its Role in Abiotic Stress Responses

HUANG Xu-sheng¹, ZHOU Ya-li¹, JIA Zheng-rong², SHEN Shan³, CHEN Jin-chai¹, ZHOU Guang-li¹, LI Xin-xin¹, WANG Ji-ping¹, LI Run-zhi¹, JIA Xiao-yun³()

^1.College of Agronomy, Shanxi Agricultural University, Shanxi Engineering Research Center for Genetics and Metabolism of Special Crops, Taigu 030801
^2.Sorghum Research Institute, Shanxi Agricultural University, Shanxi Provincial Key Laboratory of Sorghum Genetics and Germplasm Innovation, Yuci 030600
^3.College of Life Science, Shanxi Agricultural University, Shanxi Engineering Research Center for Genetics and Metabolism of Special Crops, Taigu 030801

Received:2025-08-12 Published:2026-07-26 Online:2026-07-20

摘要/Abstract

摘要：

目的生长素响应因子（auxin response factor, ARF）在植物激素信号转导和非生物胁迫应答过程中发挥关键调控作用，对紫苏PfARF转录因子家族进行全基因组鉴定，分析相关基因在非生物胁迫下的表达特性，为阐明PfARF在紫苏逆境应答中的功能提供理论基础。方法从紫苏基因组中鉴定PfARF成员，利用TBtools、MEGA11、NCBI-CD Search等组学工具分析其染色体定位、系统进化及保守结构等特征。通过RNA-seq和RT-qPCR分析PfARFs基因在紫苏发育种子和幼苗非生物胁迫下的表达情况。应用分子克隆技术获得PfARF22基因，通过本氏烟草叶片瞬时表达检测其编码蛋白亚细胞定位。构建PfARF22转录因子的蛋白互作网络图，利用分子对接技术对其结合位点、结合能等进行分析。结果在紫苏中共鉴定到29个PfARF家族成员，划分为4个亚族，同一亚族PfARF成员的保守基序和基因结构具有相似性，片段重复是其进化和扩增的主要驱动力。PfARF家族成员在紫苏发育种子和幼苗非生物胁迫下差异表达，PfARF22基因在低温和干旱胁迫下显著高表达，亚细胞定位分析表明PfARF22定位于细胞核。分子对接结果表明PfARF22与PfMADS1可能协同调控紫苏生长发育及非生物胁迫应答等过程。结论紫苏PfARF转录因子家族共有29个成员，PfARF22在紫苏生长发育和低温及干旱等胁迫应答过程中具有潜在调控作用。

关键词: 紫苏, 生长素响应因子, 生物信息学分析, 表达特性分析, 非生物胁迫, 生长发育

Abstract:

Objective Auxin response factor (ARF) plays a crucial regulatory role in plant hormone signal transduction and abiotic stress responses. The genome-wide identification of the Perilla frutescens PfARF transcription factor (TF) family and the analysis of the expression characteristics of related genes under abiotic stress provide a theoretical basis for elucidating the function of PfARF in the stress response of perilla. Method The PfARF members were identified from the genome of perilla, and their chromosomal location, phylogenetic evolution and conserved structures were analyzed using omic tools, such as TBtools, MEGA11, NCBI-CD Search, et al. The expressions of PfARF genes in the developing seeds of perilla and in the seedlings under abiotic stress was analyzed through RNA-seq and RT-qPCR. The PfARF22 gene was obtained using molecular cloning technology, and the subcellular localization of its encoded protein was detected by transient transformation in Nicotiana benthamiana leaves. A protein interaction network diagram of the PfARF22 TF was constructed, and its binding sites, binding energies, and other parameters were analyzed using molecular docking technology. Result A total of 29 PfARF family members were identified in perilla, which were classified into four subgroups. PfARF members in the same subgroup exhibited similarities in their conserved motifs and gene structures, and segment duplication was the main driving force for their evolution and expansion. PfARF genes showed differential expression in the developing seeds of perilla and in the abiotic stress conditions of the seedlings. The expression of PfARF22 gene significantly increased under low temperature and drought stress conditions. Subcellular localization analysis revealed that PfARF22 was located in nucleus. The molecular docking results indicated that PfARF22 and PfMADS1 cooperatively regulated biological processes such as the growth and development of perilla and responses to abiotic stress. Conclusion Perilla PfARF TF family consists of 29 members. PfARF22 has potential regulatory effects on the growth and development of perilla as well as responses to stress conditions such as low temperature and drought.

Key words: Perilla frutescens, auxin response factor, bioinformatics analysis, expression characteristic analysis, abiotic stress, groth and development

黄旭升, 周雅莉, 贾峥嵘, 沈珊, 陈锦钗, 周广立, 李鑫鑫, 王计平, 李润植, 贾小云. 紫苏ARF转录因子家族全基因组鉴定及其在非生物胁迫响应中的功能分析[J]. 生物技术通报, 2026, 42(7): 148-162.

HUANG Xu-sheng, ZHOU Ya-li, JIA Zheng-rong, SHEN Shan, CHEN Jin-chai, ZHOU Guang-li, LI Xin-xin, WANG Ji-ping, LI Run-zhi, JIA Xiao-yun. Whole-genome Identification of the Perilla frutescens ARF Transcription Factor Family and Functional Analysis of Its Role in Abiotic Stress Responses[J]. Biotechnology Bulletin, 2026, 42(7): 148-162.

图/表 11

图1 紫苏PfARF基因在染色体上的分布

Fig. 1 Chromosome distribution of PfARF genes in P. frutescens

图2 紫苏ARF蛋白的系统进化树

Fig. 2 Phylogenetic tree of ARF proteins in P. frutescens

图3 紫苏PfARF家族成员的保守motif基序和基因结构A：PfARF成员进化分组；B：功能结构域；C：保守Motif基序；D：基因结构

Fig. 3 Conserved motif pattern and gene structure analysis of PfARF members in P. frutescensA: Evolutionary relationship of PfARF members. B: Protein functional domain. C: Conserved motif pattern. D: Gene structure

图4 紫苏PfARF基因在染色体间的共线性分析

Fig. 4 Synteny analysis of inter-chromosomal relationships of PfARF genes in P. frutescens

图5 紫苏与拟南芥和芝麻ARF基因的同线性关系蓝线为紫苏和拟南芥之间ARF基因的共线性关系；粉线为紫苏和芝麻之间ARF基因的共线性关系

Fig. 5 Collinearity relationship of ARF genes among P. frutescens, A. thaliana and S. indicumThe blue lines indicate the collinearity relationship of ARF genes between P. frutescens and A. thaliana; the pink lines indicate the collinearity relationship of ARF genes between P. frutescens and S. indicum

图6 紫苏PfARF基因启动子顺式作用元件预测

Fig. 6 Cis-acting regulatory elements predicted in the promoters of PfARF genes

图7 紫苏PfARF基因在种子不同发育时期的表达分析10、20和30 DAF分别表示紫苏开花后10、20、30 d种子

Fig. 7 Expression analysis of PfARF genes at different stages of seed development in P. frutescens10, 20, and 30 DAF indicate the seeds of perilla at 10, 20, and 30 d after flowering, respectively

图8 紫苏幼苗低温胁迫下表型（A）及PfARF基因表达（B）分析数据为3次生物学重复的平均值±标准误差。不同小写字母表示样本间差异显著（P<0.05，Tukey多重比较法）。下同

Fig. 8 Analysis of phenotype (A) and PfARF genes expression (B) of P. frutescens seedlings under low temperature stressThe data are the mean values ± standard errors (mean ± SE) of three biological replicates. Different lowercase letters indicate significant differences between samples (P<0.05, Tukey’s multiple comparison method). The same below

图9 紫苏幼苗干旱胁迫下表型（A）及PfARF基因表达（B）分析

Fig. 9 Analysis of phenotype (A) and PfARF genes expression (B) of P. frutescens seedlings under drought stress

图10 紫苏PfARF22基因克隆（A）及PfARF22蛋白的亚细胞定位（B）

Fig. 10 Cloning of PfARF22 gene (A) and subcellular localization of PfARF22 protein (B) in P. frutescens

图11 紫苏PfARF22蛋白互作网络分析（A）及分子对接（B‒F）

Fig. 11 Analysis of PfARF22 protein interaction network (A) and molecular docking (B‒F) of P. frutescens

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紫苏ARF转录因子家族全基因组鉴定及其在非生物胁迫响应中的功能分析

Whole-genome Identification of the Perilla frutescens ARF Transcription Factor Family and Functional Analysis of Its Role in Abiotic Stress Responses

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