菊芋WRKY转录因子家族基因的鉴定及分析

doi:10.13560/j.cnki.biotech.bull.1985.2022-0551

摘要/Abstract

摘要：

菊芋属抗旱性物种，通过对菊芋转录组数据进行分析，筛选与菊芋响应干旱胁迫密切相关的WRKY转录因子基因，为明晰菊芋WRKY家族基因的功能奠定理论基础。采用生物信息学方法，对菊芋WRKY基因家族成员的理化性质、保守结构域、蛋白质三级结构及其与其他物种的系统进化关系进行分析，利用RT-qPCR方法检测干旱胁迫下，菊芋WRKY基因在种子、根、茎、叶、花瓣及块茎的相对表达量，以及菊芋WRKY基因家族成员的转录水平。结果表明，在菊芋叶片中共鉴定出74个WRKY基因，序列长度为510-2 040 bp，检测到的motif基序的长度为25-50个碱基，系统进化树分析显示，大部分HtWRKY基因能够与其他WRKY基因聚为一类；组织特异性表达分析结果显示，HtWRKY基因在花瓣和块茎的相对表达量较高，干旱诱导表达分析显示，分别有20个和19个HtWRKY基因在菊芋叶片和根中呈现出诱导后上升表达的趋势。HtWRKY参与了菊芋抵御干旱胁迫的过程。

关键词: 菊芋, WRKY, 转录因子, 鉴定分析

Abstract:

Jerusalem artichoke（JA）（Helianthus tuberosus L.）is a drought-resistant species. Through analyzing the transcriptomic data of JA, WRKY transcription factor genes closely related to drought stress response of JA were screened and analyzed, laying a theoretical foundation for clarifying the function of WRKY family genes of JA. Bioinformatics methods were applied to analyze the physicochemical properties, conserved domains, tertiary protein structures of WRKY gene family members in JA and their phylogenetic relationships with other species. RT-qPCR was used to detect the relative expression levels of WRKY genes in the seeds, roots, stems, leaves, petals and tubers under drought stress, and the transcription levels of WRKY gene family members in JA. A total of 74 WRKY genes were identified in JA leaves, with the sequence length ranging from 510 to 2 040 bp. The detected motif lengths ranged from 25 to 50 bases. Phylogenetic tree analysis showed that the most of the HtWRKY genes clustered with other WRKY genes. The results of tissue-specific expression analysis showed that the relative expressions of HtWRKY genes were higher in petals and tubers, and the drought-induced expression analysis showed that 20 and 19 HtWRKY genes were up-regulated in the leaves and roots of JA, respectively. HtWRKY is involved in the process against drought stress.

Key words: Helianthus tuberosus L., WRKY, transcription factors, identification and analysis

赵孟良, 郭怡婷, 任延靖. 菊芋WRKY转录因子家族基因的鉴定及分析[J]. 生物技术通报, 2023, 39(2): 116-125.

ZHAO Meng-liang, GUO Yi-ting, REN Yan-jing. Identification and Analysis of WRKY Transcription Factor Family Genes in Helianthus tuberosus[J]. Biotechnology Bulletin, 2023, 39(2): 116-125.

图/表 6

图1 HtWRKY基因的保守基序标识

Fig. 1 Conversed motif logo of HtWRKY genes

图2 HtWRKY基因的聚类分析图及基序位置

Fig. 2 Phylogenetic relationship and motif locations of HtWRKY genes

图3 4个物种WRKY转录因子的系统进化树

Fig. 3 Phylogenetic tree of WRKY transcription factors in four species

图4 HtWRKY基因在PEG-6000模拟干旱胁迫下的组织特异性表达水平热图数字表示2-ΔΔCT值

Fig. 4 Tissue-specificity expression heatmap of HtWRKY genes under PEG-6000 -simulated drought stresses Number indicates 2-ΔΔCT value

图5 74个HtWRKY基因在PEG-6000模拟干旱胁迫下的菊芋叶片中的表达水平热图数字表示log2PFKM值

Fig. 5 Expression heatmap of 74 HtWRKY genes under PEG-6000-simulated drought stress in H. tuberosus leaves Number indicates log2 PFKM value

图6 19个HtWRKY基因在干旱胁迫下菊芋根中的表达情况

Fig. 6 Expressions of 19 HtWRKY genes in the Jerusalem artichoke roots under drought stress H. tuberosus

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