Identification and Expression Analysis of the DOF Gene Family in Huperzia serrata
LI Hai-Bo1, XIE Li-Bo1, ZHANG Kai1, SHI Ji-Dong2, CAO Yu3, LI Mu-Zi3, HUA Yang-Guang3, WANG De-Kai3,*
1 Yuyao City Seed and Seedling Management Station, Ningbo 315499, China; 2 Agricultural and Rural Affairs Office of Luting Township, Ningbo 315434, China; 3 College of Life Sciences and Medicine, Zhejiang Sci-Tech University, Hangzhou 310018, China
摘要蛇足石杉(Huperzia serrata)为石松科植物,其含有的石杉碱甲(huperzine A, Hup A)是一种高选择性的乙酰胆碱酯酶抑制剂,用于阿尔海默茨病(Alzheimer's disease, AD)的治疗和症状改善。DOF (DNA binding with one zine finger)家族是一类植物特有的转录因子,广泛参与植物的生长发育和非生物胁迫反应。本研究基于蛇足石杉三代全长转录组数据,鉴定HsDOF基因家族,分析其蛋白的理化性质、保守结构域、亚细胞定位、二级与三级结构、调控网络,利用RNA-seq数据分析HsDOF家族基因在不同组织及高温、干旱胁迫下的表达模式。结果显示,共鉴定出19个HsDOF基因;其编码蛋白质的氨基酸数目在213~574之间,分子量约为23.89~60.50 kD,等电点为5.15~9.93;亚细胞定位预测结果显示,全部HsDOF定位于细胞核;二级结构显示主要为α-螺旋和无规则卷曲,19个HsDOF蛋白中所用的三级结构模型有7种。系统发育分析显示,拟南芥(Arabidopsis thaliana)和蛇足石杉DOF可分为9个亚组,HsDOF分布于其中的2个亚组。表达模式分析显示,HsDOFs基因在不同的组织中呈现差异表达;在高温胁迫下,6个HsDOF基因在1个或多个时间点上调或下调2倍以上;在干旱胁迫下,有6个HsDOFs基因在1个或多个时间点上调或下调2倍以上,其中3个上调;2个下调。本研究为深入探究HsDOF基因家族的生物学功能和分子育种提供了理论依据。
Abstract：Huperzia serrata, a member of the Lycopodiaceae family, contains huperzine A (Hup A), a highly selective acetylcholinesterase inhibitor used to treat and alleviate symptoms in Alzheimer's disease (AD). The DOF family, a transcription factor family unique to plants, plays a significant role in plant growth, development, and responses to abiotic stress. Based on the full-length transcriptome data of H. serrata, this study identified the DOF gene family (HsFOF), analyzed its protein physicochemical properties, conserved domains, subcellular localization, secondary and tertiary structures, and regulatory networks. By utilizing RNA-seq data, the expression patterns of HsDOF were analyzed across various tissues and under conditions of high temperature and drought stress. The results showed that a total of 19 HsDOF genes were identified; The number of amino acids encoded the protein ranges from 213 to 574, with a molecular weight of approximately 23.89 to 60.50 kD and an isoelectric point of 5.15 to 9.93; The subcellular localization prediction results showed that all HsDOFs were located to nucleus. The secondary structure was mainly displayed as α-helix and irregular curl, with 7 types of tertiary structure models used in the 19 HsDOF proteins. Phylogenetic analysis showed that the DOF protein of Arabidopsis thaliana and H. serrata could be divided into 9 subgroups, with HsDOF distributed in 2 of them. Expression pattern analysis showed that the HsDOF genes showed differential expression in different tissues. Under high temperature stress, 6 HsDOF genes were upregulated or downregulated by more than 2 times at 1 or more time points. Under drought stress, 6 HsDOF genes were upregulated or downregulated by more than 2 times at 1 or more time points, of which 3 were upregulated; 2 downregulated. The research results provide a theoretical basis for further exploring the biological functions and molecular breeding of the HsDOF gene family.
李海波, 谢锂波, 张恺, 史济东, 曹禹, 李沐子, 化阳光, 汪得凯. 蛇足石杉DOF基因家族的鉴定和表达分析[J]. 农业生物技术学报, 2025, 33(3): 594-607.
LI Hai-Bo, XIE Li-Bo, ZHANG Kai, SHI Ji-Dong, CAO Yu, LI Mu-Zi, HUA Yang-Guang, WANG De-Kai. Identification and Expression Analysis of the DOF Gene Family in Huperzia serrata. 农业生物技术学报, 2025, 33(3): 594-607.
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