马铃薯AP2/ERF基因家族鉴定及响应致病疫霉胁迫的表达分析

doi:10.13560/j.cnki.biotech.bull.1985.2025-0798

摘要/Abstract

摘要：

目的对马铃薯全基因组中AP2/ERF转录因子家族基因进行鉴定，分析该家族基因在不同组织及致病疫霉（Phytophthora infestans）胁迫下的表达模式，为阐明AP2/ERF家族基因参与马铃薯响应致病疫霉胁迫提供依据。方法利用生物信息学方法系统鉴定马铃薯AP2/ERF家族成员，并分析其系统发育、染色体定位、基因结构、保守基序及共线性关系。结合转录组数据，分析该家族成员的组织表达特异性和响应致病疫霉胁迫的表达模式。对马铃薯进行致病疫霉不同时间段（0、24、48、72 h）侵染，采用RT-qPCR检测部分家族成员基因的差异表达变化。结果在马铃薯中共鉴定到179个AP2/ERF基因，聚类分为ERF、DREB、AP2、RAV和Soloist五个亚家族，不均匀分布于12条染色体上。不同亚家族基因之间的Motif、内含子和外显子分布差异显著。种内共线性分析表明AP2/ERF基因中有57对共线基因；物种间共线性显示AP2/ERF在水稻和拟南芥中分别存在69对和164对同源基因。组织表达分析显示AP2/ERF基因参与马铃薯营养及生殖生长，部分基因呈组织特异性高表达。致病疫霉胁迫下，大多数AP2/ERF基因被不同程度诱导表达。结论 AP2/ERF转录因子基因家族参与马铃薯生长发育过程，且快速响应致病疫霉胁迫反应。

关键词: 马铃薯, AP2/ERF转录因子, 生物信息学, 致病疫霉胁迫

Abstract:

Objective To identify the AP2/ERF transcription factor family genes in the whole genome of potato (Solanum tuberosum) and analyze their expression patterns in different tissues and under Phytophthora infestans stress, this study aimed to provide a basis for elucidating the involvement of AP2/ERF family genes in potato responses to P. infestans infection. Method Bioinformatics approaches were used to systematically identify AP2/ERF family members in potato, followed by analyses of their phylogeny, chromosomal localization, gene structure, conserved motifs, and collinearity relationships. Combined with previously published transcriptome data, tissue-specific expression profiles and P. infestans-responsive expression patterns of these genes were characterized. Potato plants were inoculated with P. infestans at 0, 24, 48, and 72 h, and RT-qPCR was performed to detect differential expression of selected family members. Result A total of 179 AP2/ERF genes were identified in potato, which were clustered into 5 subfamilies and unevenly distributed across 12 chromosomes. Significant differences in motif, intron, and exon distributions were observed among subfamilies. Intraspecific collinearity analysis revealed 57 pairs of collinear AP2/ERF genes, while interspecific collinearity analysis identified 69 and 164 homologous gene pairs with rice (Oryza sativa) and Arabidopsis respectively. Tissue expression analysis indicated that AP2/ERF genes were involved in both vegetative and reproductive growth, with some showing tissue-specific high expression. Under P. infestans stress, most AP2/ERF genes were induced to varying degrees. Conclusion The AP2/ERF transcription factor family plays roles in potato growth and development, and rapidly responds to P. infestans stress.

Key words: Solanum? tuberosum, AP2/ERF transcription factor, bioinformatics, Phytophthora infestans stress

王江清, 张蝶, 王荟洁, 袁柱东, 付艳鸿, 黄娅楠, 王洪洋. 马铃薯AP2/ERF基因家族鉴定及响应致病疫霉胁迫的表达分析[J]. 生物技术通报, 2026, 42(6): 116-127.

WANG Jiang-qing, ZHANG Die, WANG Hui-jie, YUAN Zhu-dong, FU Yan-hong, HUANG Ya-nan, WANG Hong-yang. Identification of the AP2/ERF Gene Family in Potato and Expression Analysis in Response to Phytophthora infestans Stress[J]. Biotechnology Bulletin, 2026, 42(6): 116-127.

图/表 8

表1 RT-qPCR引物

Table 1 Primers for RT-qPCR

基因名称 Gene name	orwardprimer（5'-3'）上游引物 F	everseprimer（5'-3'）下游引物 R
StERF3	CCCCATCACCACCTGCTAAG	ACTAGCAACGGGCATTTCCA
StERF13	TATGAGATGCCCCGTTTGGG	CCACGGTCGACTCTGTTGAT
StERF101	AGGCACTTCGTGAGGAAACC	CTGAAGGGGAAACTGAGCCC
StERF76	ACGGATGAGGAAGTGGGGTA	AGCGGTATCGTAAGCACGAG
StERF74	GTCGCCACCAAAACCCTCTA	GGGGAAGTTCAGTCGAGCAA
StERF55	GGGGGATCAGACAGAGACCT	TCTTGATGCAGGTGCTGGAG
StERF116	GCTCGAGCTCATGATGTTGC	TAACTGCCGCCTGTCGTATG
StERF32	GGGACTCGGGTTTGGTTAGG	CACCGGCTGTGGAATCTCAA
StERF123	GGCGGAAGCTGAAGAAGAGA	CGAACAGCCAAGGTGAGGAG
StEF1	ATTGGAAACGGATATGCTCCA	TCCTTACCTGAACGCCTGTCA

图1 马铃薯和拟南芥AP2/ERF家族系统进化树

Fig. 1 Phylogenetic tree of the AP2/ERF family in Solanum tuberosum and Arabidopsis thaliana

图2 AP2/ERF 基因家族的系统发育树（A）、保守基序（B）、保守结构域（C）、基因结构（D）D：绿色矩形表示非编码序列，黄色矩形表示外显子，黑线表示内含子

Fig. 2 Phylogenetic tree (A), conserved motif (B), conserved domain (C), and gene structure(D) of AP2/ERF gene familyD: The green rectangles indicate non-coding sequences, the yellow rectangles indicate exons, and the black lines represent introns

图3 马铃薯AP2/ERF 基因在染色体上的分布Chr01-Chr12表示1-12号染色体；黑线表示AP2/ERF 基因位于染色体的位置

Fig. 3 Distribution of potato AP2/ERF genes on chromosomesChr01-Chr12 refer to chromosome 1 to 12. The black lines indicate the chromosomal locations of the AP2/ERF genes

图4 马铃薯AP2/ERF 基因之间的共线性分析（A）以及马铃薯、水稻和拟南芥AP2/ERF基因共线性分析（B）

Fig. 4 Collinearity analysis between AP2/ERF genes of potato (A), and collinearity analysis of AP2/ERF genes in S. tuberosum, O. sativa and A. thaliana (B)

图5 马铃薯 AP2/ERF 家族基因的组织表达热图

Fig. 5 Tissue expression heat map of AP2/ERF family genes in S. tuberosum

图6 致病疫霉感染马铃薯后AP2/ERF家族基因的表达热图Ⅰ：抗性马铃薯感染致病疫霉前表达量；Ⅱ：抗性马铃薯感染致病疫霉后1-2 d的表达量；Ⅲ：易感马铃薯感染致病疫霉前表达量；Ⅳ：易感马铃薯感染致病疫霉后1-2 d的表达量；基因的表达水平经过log₂（FPKM）转换后用于后续的可视化和分析

Fig. 6 Expression heat map of AP2/ERF family genes in S. tuberosum following P. infestans infectionⅠ: Expressions of resistant S. tuberosum before infection with P. infestans. Ⅱ: Expressions of resistant S. tuberosum at 1-2 d after infection with P. infestans. Ⅲ: Expressions of susceptible S. tuberosum before infection with P. infestans. Ⅳ: Expressions of susceptible potato at 1-2 d after infection with P. infestans. The expressions of genes were transformed to log₂(FPKM) values for subsequent visualization and analysis

图7 9个AP2/ERF基因在致病疫霉处理不同时段的表达模式*P0.05，** P0.01（与0 h相比，t检验）

Fig. 7 Expression patterns of nine AP2/ERF genes at different time points after P. infestans infection*P0.05, ** P0.01 (compared with 0 h, Student’s t-test)

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