Integrated Transcriptome-metabolome Analysis of the Molecular Network Underpinning Reproductive Defects in the Rice (Oryza sativa) OsmiR159 Mutant
HE Yu-Xuan*, YAN Wei*, DONG Li-Ming, YIN Yue-Jia, XIE Yan-Bo, MA Yue, YANG Fan, LONG Li-Kun**, LI Fei-Wu**
Institute of Agricultural Quality Standards and Testing Technology, Jilin Academy of Agricultural Sciences (Northeast Agricultural Research Center of China), Changchun 130033, China
Abstract：MicroRNA159 (miR159) is a conserved plant microRNA that regulates anther development and fertility maintenance. Its roles in rice (Oryza sativa) reproductive development are still unclear. In this study, the CRISPR/Cas9 system was employed to generate multi-locus loss-of-function mutants of the OsmiR159 family in rice. These mutants exhibited a distinct phenotype characterized by a failure to transition from vegetative to reproductive growth, resulting in complete absence of flowering and seed set. Integrated transcriptomic and metabolomic analyses revealed that OsmiR159 deficiency disrupts normal rice development by perturbing key biological processes, including hormone signaling, energy metabolism, and cell wall biosynthesis. KEGG pathway enrichment analysis demonstrated that differentially expressed genes and metabolites were significantly enriched in core metabolic pathways, such as starch and sucrose metabolism, phenylpropanoid biosynthesis, and the TCA cycle. Coordinated dysregulation of these pathways led to imbalanced energy supply, aberrant carbon skeleton allocation, and disturbances in secondary metabolism, collectively impairing the developmental phase transition in rice. By integrating multi-omics datasets, we systematically dissected the regulatory interactions underlying the OsmiR159-mediated network in reproductive development, thereby establishing a robust theoretical framework and providing valuable genetic resources for crop improvement.
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