微小RNA(miRNA)是一类非编码内源性小分子,单链RNA,参与植物转录后基因表达调控,在植物生长发育中起重要作用。其中,miRNA156调控靶SPL基因家族成员,参与植物生长发育,荷尔蒙反应和逆境压力。然而,在人参中没有报道。在这项研究中,基于先前对SPL基因家族的分析,在根中获得了与年龄相关且稳定表达的SPL基因PgSPL24-09。使用靶基因预测工具分析miRNA156与该基因的结合位点,通过双荧光素酶报告基因测定和RT-qPCR验证了miRNA156与PgSPL24-09基因之间的相互作用。同时,构建miRNA156沉默载体和过表达载体,转化人参不定根和拟南芥,分析miRNA156-SPL模块调控人参不定根生长的分子机制。本研究为深入研究miRNAs在人参生长中的分子作用提供了理论依据,同时也为研究miRNA156-SPL模块在人参生长发育调控中的作用奠定了基础。
MicroRNA (miRNA) is a class of non-coding endogenous small-molecule, single-stranded RNAs, and it is involved in post-transcriptional gene expression regulation in plants and plays an important role in plant growth and development. Among them, miRNA156 regulates members of target SPL gene family and thus participates in plant growth and development, hormonal response and adversity stress. However, it has not been reported in ginseng. In this study, based on the previous analysis of the SPL gene family, the age-related and stably expressed SPL gene PgSPL24-09 was obtained in roots. The binding site of miRNA156 to this gene was analyzed using target gene prediction tools, and the interactions between miRNA156 and PgSPL24-09 gene were verified by dual luciferase reporter gene assay and RT-qPCR. At the same time, miRNA156 silencing vector and overexpression vector were constructed and transformed into ginseng adventitious roots and Arabidopsis thaliana to analyze the molecular mechanism of miRNA156-SPL module in regulating the growth of ginseng adventitious roots. This study provides a theoretical basis for the in-depth study of the molecular role of miRNAs in ginseng growth, and also lays the foundation for the study of the role of miRNA156-SPL module in regulating the growth and development of ginseng.