1华北作物改良与调控国家重点实验室/河北省作物种质资源实验室/河北农业大学农学院,保定 071000;2河北省藜麦产业技术研究院, 张家口 075000;3南洋理工大学生物科学学院,新加坡 639798;4西北农林科技大学农学院,陕西杨凌 712100;5保定理工学院教育学院,河北保定 071000
研究方向为种质资源的创制与利用,E-mail: 2749354952@qq.com
国家重点研发计划“政府间国际创新合作”专项(SQ2026YFE0103026);河北农业大学“太行山农业创新驿站建设”项目(311718002)
1North China State Key Laboratory of Crop Improvement and Regulation/Hebei Provincial Laboratory of Crop Germplasm Resources/College of Agronomy, Hebei Agricultural University, Baoding 071000;2The Quinoa Industrial Technology Research Institute of Hebei Province, Zhangjiakou 075000;3School of Biological Sciences, Nanyang Technological University, Singapore 639798;4College of Agronomy, Northwest A&F University, Yangling 712100, Shaanxi;5School of Education, Baoding University of Technology, Baoding 071000, Hebei
Special Project of International Innovation Cooperation of the National Key Research and Development Program of China (SQ2026YFE0103026);Project of “Taihang Mountain Agricultural Innovation Station Construction” of Hebei Agricultural University (311718002)
藜麦属于全营养假谷物,含有人类所需的全部必需氨基酸。为探究藜麦发芽过程中γ-氨基丁酸(GABA)的积累机制,以藜麦M059为材料,对其发芽12 h和24 h的GABA含量进行测定,并比较其转录组差异。结果显示,发芽24 h时,GABA含量达峰值(24.47 μg/g),较12 h提升2.27倍。RNA-seq分析共鉴定到1146个差异表达基因(DEGs),其中上调基因913个,下调基因233个。GO富集分析发现,有19个与发芽过程中GABA合成密切相关的GO条目;KEGG通路分析表明,与发芽过程中GABA合成密切相关的代谢途径为6条,其中丙氨酸、天冬氨酸和谷氨酸代谢(ko00250)以及精氨酸和脯氨酸代谢(ko00330)为核心通路,24 h时关键基因CqGAD(LOC110718614)和CqALDH3H1(LOC110709762)的表达量较12 h时分别上调1.9倍和1.1倍。转录因子分析鉴定到关键基因的潜在转录调控关系表明,转录因子CqSPL8(r=0.77)、CqWRKY24(r=0.91)、CqWRKY6(r=0.97)正调控CqGAD,CqHSFC1(r=-0.97)负调控CqGAD;转录因子CqERF091(r=0.89)、CqWRKY24(r=0.88)、CqTGA9(r=0.91)正调控CqALDH3H1,CqABI4(r=-0.91)负调控CqALDH3H1。其中,CqWRKY24作为共同调控因子,同时参与CqGAD和CqALDH3H1的正调控。qRT-PCR结果表明,CqSPL8(LOC110714232)、CqWRKY6(LOC110720931)、CqHSFC1(LOC110691034)、CqERF091(LOC110693708)、CqWRKY24(LOC110730454)、CqTGA9(LOC110684250)、CqABI4(LOC110699305)与转录组表达一致。本研究表明藜麦发芽过程中GABA的高效积累是谷氨酸脱羧酶(GAD)催化的谷氨酸途径与醛脱氢酶(ALDH)介导的多胺降解途径协同作用的结果,WRKY家族转录因子在其中发挥关键调控作用。本研究结果为藜麦发芽过程中GABA关键代谢通路及其分子机制提供了参考。
Quinoa (Chenopodium quinoa Willd.) is a nutritionally complete pseudocereal containing all the essential amino acids required by humans. To investigate the accumulation mechanism of γ-aminobutyric acid (GABA) during quinoa germination, we used quinoa cultivar M059 as experimental material, determined GABA contents at 12 h and 24 h of germination, and compared transcriptomic differences between these two time points.The results showed that the GABA content reached a peak of 24.47 μg/g at 24 h of germination, which was 2.27-fold higher than that at 12 h. Transcriptome analysis identified 1146 differentially expressed genes (DEGs), including 913 up-regulated genes and 233 down-regulated genes. GO enrichment analysis indicated 19 GO pathways closely related to GABA synthesis during germination. KEGG pathway analysis identified six metabolic pathways associated with GABA synthesis, among which the alanine, aspartate and glutamate metabolism (ko00250) and arginine and proline metabolism (ko00330) pathways were highlighted as the core metabolic pathways.The expression levels of the key genes CqGAD (LOC110718614) and CqALDH3H1 (LOC110709762) were up-regulated by 1.9-fold and 1.1-fold, respectively. Transcription factor analysis identified potential transcriptional regulatory relationships of key genes: CqSPL8 (r=0.77), CqWRKY24 (r=0.91), and CqWRKY6 (r=0.97) positively regulate CqGAD, while CqHSFC1 (r=-0.97) negatively regulates CqGAD. For CqALDH3H1, CqERF091 (r=0.89), CqWRKY24 (r=0.88), and CqTGA9 (r=0.91) positively regulate its expression, while CqABI4 (r=-0.91) negatively regulates it. Notably, CqWRKY24 serves as a common regulatory factor that positively regulates both CqGAD and CqALDH3H1. qRT-PCR results confirmed that the expression levels of CqSPL8(LOC110714232), CqWRKY6(LOC110720931), CqHSFC1(LOC110691034), CqERF091(LOC110693708), CqWRKY24(LOC110730454), CqTGA9(LOC110684250) and CqABI4(LOC110699305) were consistent with the transcriptome data.This study demonstrated that the efficient accumulation of GABA during quinoa germination is the result of the synergistic effect of the GAD-catalyzed glutamate pathway and the ALDH-mediated polyamine degradation pathway, and WRKY family transcription factors play a key regulatory role. The findings provide a valuable reference for elucidating the key metabolic pathways and molecular mechanisms underlying GABA accumulation during quinoa germination.
魏博翔,刘阳,张敏,等.藜麦发芽过程中
