Narrow leaf 1 (NAL1) regulates leaf shape by affecting cell expansion in rice (Oryza sativa L.)

The narrow leaf1 (nal1) mutant of rice (Oryza sativa L.) exhibits a narrow leaf phenotype. Previous studies have shown that NAL1 modulates leaf size by affecting vein patterning and cell division; however, the underlying mechanism remains unclear. Here, we report that the nal1 mutant shows reduced size of the leaf abaxial epidermal cells and culm parenchyma cells compared with the wild type (WT), indicating that NAL1 also regulates cell expansion. To understand the molecular mechanism of the reduced cell size phenotype, leaves of 40-day-old nal1 mutant and WT seedlings were subjected to RNA-Seq analysis, which has identified 4277 differentially expressed genes (DEGs) between WT and the nal1 mutant. Gene ontology (GO) enrichment analysis revealed a large number of genes down-regulated in the nal1 mutant were involved in cell wall formation. Furthermore, the Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis revealed that NAL1-regulated DEGs, such as ARFs and SAURs, were mapped in auxin signal transduction and auxin-regulated cell expansion pathways. A combination of RNA-Seq analysis and gene expression validation using RT-qPCR suggested that NAL1 is involved in the regulation of auxin-mediated acid growth in rice. These results indicate that, in addition to controlling cell division, NAL1 controls leaf width, at least partially, through its effect on cell expansion, probably via the acid growth mechanism.