Wang Jiemin, Rong Zhengpu, Shi Weihong, Zhang Ye, Wang Wenshuai, Zheng Yuguang, Han Xiaowei, Sun Huigai
School of Pharmacy, Hebei University of Chinese Medicine, Shijiazhuang, Hebei, 050200, China.
Traditional Chinese Medicine Processing Technology Inheritance Base of the State Administration of Traditional Chinese Medicine (Hebei), Shijiazhuang, Hebei, 050200, China.
BMC Plant Biol. 2025 May 6;25(1):593. doi: 10.1186/s12870-025-06592-w.
Drought and other abiotic stressors exert significant impacts on plant growth, development, and yield. WRKY transcription factors (TFs) are pivotal in regulating plant responses to such stresses. This study conducted a genome-wide identification of the Astragalus membranaceus (A. membranaceus) WRKY TF family, aiming at providing essential stress-tolerance genes for molecular breeding in A. membranaceus. Using bioinformatics approaches, including phylogenetic analysis, gene structure analysis, protein conserved motif prediction, and promoter cis-acting element examination, we identified 76 WRKY TF family members unevenly distributed across eight chromosomes. Phylogenetic analysis categorized the 76 AmWRKY proteins into three major classes: 14 in Class I, 52 in Class II, and 10 in Class III. Analysis of conserved structural domains revealed that similarities among domains of the same class AmWRKYs, with a predominance of members containing the conserved "WRKYGQK" heptapeptide domain and zinc finger structure, despite a few members had conserved domain variants. Gene structure analysis demonstrated that AmWRKY genes include 1-6 exons and 1-5 introns. GO and KEGG analyses revealed the differentially expressed WRKY genes are primarily transcription regulators involved in stress response pathways, with significant enrichment in DNA-binding activities, nuclear localization, and plant-pathogen/MAPK signaling. Transcriptome and qRT-PCR analyses indicated that several WRKY TFs, including AmWRKY8, were involved in the response to drought stress. Furthermore, it was confirmed that AmWRKY8 is localized exclusively in the nucleus and possesses transcriptional activation activity. Protein interaction analysis revealed that AmWRKY8 coordinates drought stress responses by interacting with key regulators of hormone signaling (AtWRKY70, ERF6), mitochondrial stress (NAC017), and developmental processes (HBI1). These findings are useful in further investigations into the regulatory role of this this TF in abiotic stress responses.
干旱和其他非生物胁迫因子对植物的生长、发育和产量产生重大影响。WRKY转录因子在调节植物对这类胁迫的反应中起着关键作用。本研究对膜荚黄芪WRKY转录因子家族进行了全基因组鉴定，旨在为膜荚黄芪分子育种提供重要的耐胁迫基因。利用系统发育分析、基因结构分析、蛋白质保守基序预测和启动子顺式作用元件检测等生物信息学方法，我们鉴定出76个WRKY转录因子家族成员，它们不均匀地分布在8条染色体上。系统发育分析将76个膜荚黄芪WRKY蛋白分为三大类：I类14个，II类52个，III类10个。保守结构域分析表明，同一类膜荚黄芪WRKY结构域之间存在相似性，大多数成员含有保守的“WRKYGQK”七肽结构域和锌指结构，尽管有少数成员具有保守结构域变体。基因结构分析表明，膜荚黄芪WRKY基因包含1 - 6个外显子和1 - 5个内含子。GO和KEGG分析表明，差异表达的WRKY基因主要是参与胁迫反应途径的转录调节因子，在DNA结合活性、核定位以及植物 - 病原体/MAPK信号传导方面显著富集。转录组和qRT - PCR分析表明，包括膜荚黄芪WRKY8在内的几个WRKY转录因子参与了对干旱胁迫的响应。此外，证实膜荚黄芪WRKY8仅定位于细胞核并具有转录激活活性。蛋白质相互作用分析表明，膜荚黄芪WRKY8通过与激素信号传导（拟南芥WRKY70、ERF6）、线粒体胁迫（NAC017）和发育过程（HBI1）的关键调节因子相互作用来协调干旱胁迫反应。这些发现有助于进一步研究该转录因子在非生物胁迫反应中的调控作用。
