花生ZF-HD基因家族的鉴定和非生物胁迫响应分析

doi:10.13560/j.cnki.biotech.bull.1985.2025-0806

摘要/Abstract

摘要：

目的分析花生ZF-HD基因家族的基本特性及其在多种非生物胁迫下的表达模式，为揭示ZF-HD在调控花生非生物胁迫中的作用奠定基础。方法利用生物信息学方法筛选花生ZF-HD基因家族成员，并对其理化性质、系统进化关系、保守基序、保守结构域、基因结构、启动子顺式作用元件、染色体定位、基因共线性和蛋白互作等进行分析，基于转录组数据分析AhZHDs基因在花生不同组织器官和不同非生物胁迫中的表达模式，以及利用实时荧光定量PCR技术分析该家族部分成员在不同非生物胁迫处理后的表达情况。结果在花生Tifrunner基因组中共鉴定到40个花生ZF-HD基因，根据系统发育关系将其分为ZHD Ⅰ、ZHD Ⅱ、ZHD Ⅲ、ZHD Ⅳ、ZHD Ⅴ和MIF 6个亚族，同一亚家族中的AhZHDs基因具有相似的结构特征，且大部分AhZHDs基因没有内含子。顺式作用元件分析表明，花生ZF-HD基因家族成员广泛参与激素响应、生长发育响应、非生物胁迫和光响应。染色体定位和共线性分析可知，40个AhZHDs基因不均匀地分布在16条染色体上，全基因组复制（WGD）事件或片段复制可能是AhZHDs基因进化的主要动力。蛋白互作分析表明大部分AhZHD蛋白之间存在复杂的互作关系，可能通过协同调控的方式参与花生发育和胁迫反应。转录组分析及荧光定量PCR验证实验表明，AhZHD5/9/10/17/23/29/30/32/40等9个AhZHD基因在花生不同组织器官或胁迫下表达模式丰富，其中大部分AhZHD基因在非生物胁迫下表达水平显著变化，推测该基因家族在花生应对非生物胁迫中可能发挥着重要作用；亚细胞定位实验表明，AhZHD5/17/29/32在细胞核中发挥作用。结论 40个AhZHD基因在结构和特性上具有差异，AhZHDs不仅广泛参与花生生长发育和激素信号转导，还在非生物胁迫中发挥重要调控作用。

关键词: 花生, ZF-HD基因, 基因家族, 生物信息学, 组织特异性, 非生物胁迫, 表达分析, 亚细胞定位

Abstract:

Objective To analyze the basic characteristics of the ZF-HD gene family in peanut (A. hypogaea) and its expression patterns under various abiotic stresses, laying a foundation for revealing the role of ZF-HD in regulating abiotic stress in peanut. Method Bioinformatics methods were used to screen the peanut ZF-HD gene family members and analyze 1) their physicochemical properties, phylogenetic relationships, conserved motifs, conserved structural domains, gene structures, promoter cis-acting elements, chromosomal localization and gene covariance; 2) the expression patterns of the AhZHDs genes in different tissues and organs of peanut and in different abiotic stresses based on the transcriptome data; 3) the expressions of some members of this family in peanut following different abiotic stress treatments using RT-qPCR. The expression patterns of AhZHDs genes in different tissues and organs of peanut under different abiotic stresses were analyzed based on transcriptome data, and the expression of some members of this family in different abiotic stresses was analyzed by real-time fluorescence PCR. Result A total of 40 peanut ZF-HD genes were identified in the peanut Tifrunner genome, which were classified into six subfamilies, ZHD Ⅰ, ZHD Ⅱ, ZHD Ⅲ, ZHD Ⅳ, ZHD Ⅴ and MIF, according to their phylogenetic relationships. AhZHDs genes in the same subfamily had similar structural features, and most of them had no introns. Cis-acting element analysis indicated that members of the peanut ZF-HD gene family were widely involved in hormone response, growth and development response, abiotic stress and light response. Chromosomal localization and covariance analysis showed that the 40 AhZHDs genes were unevenly distributed on 16 chromosomes, and that the whole genome duplication (WGD) event or fragment duplication might be the main driving force for the evolution of the AhZHDs genes. Protein interactions analysis showed that most AhZHD proteins had complex interactions with each other and might be involved in peanut development and stress response through co-regulation. Transcriptome analysis and qRT-PCR validation indicated that nine AhZHD genes (AhZHD5/9/10/17/23/29/30/32/40) showed diverse expression patterns across various peanut tissues and under stress conditions, with the majority showing significantly altered expression levels in response to abiotic stress, subcellular localization experiments showed that AhZHD5/17/29/32 function in the nucleus. Conclusion The 40 AhZHD genes differ in structure and characterization, and AhZHDs are widely involved not only in peanut growth and development and hormone signaling, but also play important regulatory roles in abiotic stresses.

Key words: peanut (A. hypogaea), ZF-HD gene, gene family, bioinformatics, tissue-specific expression, abiotic stress, expression analysis, subcellular localization

陈登科, 兰刚, 夏芝, 侯保国, 杨六六, 曹彩荣, 李朋波, 吴翠翠. 花生ZF-HD基因家族的鉴定和非生物胁迫响应分析[J]. 生物技术通报, 2026, 42(4): 114-128.

CHEN Deng-ke, LAN Gang, XIA Zhi, HOU Bao-guo, YANG Liu-liu, CAO Cai-rong, LI Peng-bo, WU Cui-cui. Identification of ZF-HD Gene Family in Arachis hypogaea and Analysis in Response to Abiotic Stress[J]. Biotechnology Bulletin, 2026, 42(4): 114-128.

图/表 12

图1 花生、拟南芥和水稻ZF-HD基因家族系统进化树

Fig. 1 Phylogenetic tree of ZF-HD gene family in A. hypogaea, A. thaliana and O. sativa

图2 AhZF-HD家族成员进化关系（A）、保守基序（B）、保守结构域（C）及基因结构分析（D）

Fig. 2 Phylogenetic relationship (A), conserved motif (B), conserved domain (C), and gene structure analysis (D) of AhZF-HD family members

图3 花生ZF-HD基因启动子区顺式作用元件分析A：AhZHD家族成员顺式作用元件（a：AhZHD家族成员进化关系；b：AhZHD家族顺式元件；c：AhZHD家族顺式元件热图）；B：AhZHD家族中顺式调控元件的功能分类（a：植物激素响应类元件；b：生长发育相关元件；c：非生物胁迫响应类元件；d：光响应类元件）。图中百分比数值代表了各类特定元件在其所属功能类别中的比例

Fig. 3 Analysis of cis-acting elements in the promoter regions of ZF-HD genes in A. hypogaeaA: Cis-acting elements in AhZHD family members (a: Phylogenetic relationship of AhZHD family members. b: Cis-acting elements in the AhZHD family. c: Heatmap of cis-acting elements in the AhZHD family). B: Functional categorization of cis-regulatory elements in the AhZHD family (a: Phytohormone-responsive elements. b: Elements involved in growth and development. c: Abiotic stress-responsive elements. d: Light-responsive elements). The proportion of each specific element type within the respective functional category is indicated as a percentage

图4 AhZHDs基因在染色体上的分布

Fig. 4 Distribution of AhZHDs genes on chromosomes

图5 花生ZF-HD基因家族物种内共线性分析

Fig. 5 Collinearity analysis of the ZF-HD gene family in A. hypogaea L.

图6 栽培种花生与其他6个代表性物种之间ZF-HD基因的共线性分析

Fig. 6 Collinearity analysis of ZF-HD genes between cultivated A. hypogaea and six other representative species

图7 花生ZF-HD蛋白互作网络分析

Fig. 7 Interaction network analysis of A. hypogaea ZF-HD proteins

图8 花生ZF-HD基因在不同组织中的表达分析

Fig. 8 Expression analysis of A. hypogaea ZF-HD genes in different tissues

图9 花生ZF-HD基因在高温、低温、干旱和盐处理后的表达分析

Fig. 9 Expression analysis of ZF-HD genes in A. hypogaea under high temperature, low temperature, drought, and salt stress treatments

图10 基于RT-qPCR的不同组织器官中9个AhZHDs基因的表达差异*，**分别表示在0.05、0.01水平上差异显著，下同

Fig. 10 Expression differences of 9 AhZHDs genes under different tissues and organs based on RT-qPCR* and ** indicate significant differences at the level of 0.05, 0.01 respectively. The same below

图11 基于RT-qPCR的不同胁迫处理下9个AhZHDs基因的表达差异

Fig. 11 Expression differences of 9 AhZHDs genes under different stress treatments based on RT-qPCR

图12 AhZHD5、AhZHD17、AhZHD29和AhZHD32的亚细胞定位

Fig. 12 Subcellular localization of AhZHD5, AhZHD17, AhZHD29 and AhZHD32 (Bar=50 μm)

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