Deng Juan, Huang Xiao, Wang Meng, Li Qian, Gong Ling, Huang Bisheng, Yu Kun
School of Pharmacy, Hubei University of Chinese Medicine, Wuhan, 430065, China.
Hubei Key Laboratory of Resources and Chemistry of Chinese Medicine, Hubei University of Chinese Medicine, Wuhan, 430065, China.
Plant Cell Rep. 2025 May 28;44(6):133. doi: 10.1007/s00299-025-03527-5.
A novel miR164-AlNAC1 module adjusting the expression of genes related to chlorophyll synthesis and ROS synthesis, and regulating leaf senescence in Atractylodes lancea. Leaf senescence is a critical developmental process regulated by complex genetic networks. In this study, a novel miRNA belonging to the MIR164 family was identified as a key regulator of leaf senescence in A. lancea by targeting the transcription factor AlNAC1. Bioinformatics analysis revealed that target gene AlNAC1 was closely related to the senescence-associated gene ANAC092 in Arabidopsis. The qRT-PCR and tobacco transient expression demonstrated nov-miR164 negatively regulated AlNAC1. Subcellular localization and transcriptional activity assays revealed that AlNAC1 localized to the nucleus and functioned as a transcriptional activator. Furthermore, EMSA experiments showed that AlNAC1 bound to the G-box motifs on the promoters of AlNYC, AlPAO, and AlRbohC. With the aging of A. lancea leaves, the expression of nov-miR164 decreased, while AlNAC1 and senescence-associated genes (AlSAG21, AlNYC, AlPAO, and AlRbohC) were significantly upregulated. Agrobacterium-mediated transient expression experiment results demonstrated that nov-miR164 negatively regulated AlNAC1 to suppress chlorophyll degradation and ROS accumulation, thereby delaying leaf senescence. Our findings provide new insights into the molecular mechanisms of leaf senescence in A. lancea and highlight the potential of nov-miR164 as a target for increasing crop yield and improving crop longevity.
一个新型的miR164 - AlNAC1模块调节与叶绿素合成和活性氧合成相关的基因表达，并调控白术叶片衰老。叶片衰老是一个由复杂遗传网络调控的关键发育过程。在本研究中，一个属于MIR164家族的新型miRNA被鉴定为白术叶片衰老的关键调节因子，它靶向转录因子AlNAC1。生物信息学分析表明，靶基因AlNAC1与拟南芥中衰老相关基因ANAC092密切相关。qRT - PCR和烟草瞬时表达表明nov - miR164负调控AlNAC1。亚细胞定位和转录活性分析表明，AlNAC1定位于细胞核并作为转录激活因子发挥作用。此外，EMSA实验表明，AlNAC1与AlNYC、AlPAO和AlRbohC启动子上的G - box基序结合。随着白术叶片的衰老，nov - miR164的表达下降，而AlNAC1和衰老相关基因（AlSAG21、AlNYC、AlPAO和AlRbohC）显著上调。农杆菌介导的瞬时表达实验结果表明，nov - miR164通过负调控AlNAC1来抑制叶绿素降解和活性氧积累，从而延缓叶片衰老。我们的研究结果为白术叶片衰老的分子机制提供了新的见解，并突出了nov - miR164作为提高作物产量和延长作物寿命靶点的潜力。
