Salt and alkali stress are two of the main environmental factors limiting rice production. Thus, understanding the mechanisms of salinity and alkali stress tolerance is necessary to modify rice to increase its resistance to salinity and alkaline stress. MicroRNAs (miRNAs) are approximately 21-nucleotide RNAs that are ubiquitous regulators of gene expression in eukaryotic organisms. In plants, miRNAs constitute one of five classes of small RNAs that function primarily as negative regulators for gene expression at the posttranscriptional level. Several plant miRNAs, such as miR396, play vital roles in plant growth, development and resistance to stresses. In this study, we identified osa-MIR396c, which shows dramatic transcript change under salt and alkali stress conditions in Oryza sativa. We designed an experiment to detect miRNA-target interaction and demonstrated that several transcription factors related to growth, development, and stress tolerance are targeted by osa-MIR396c. Transgenic rice and Arabidopsis thaliana plants constitutively over-expressing osa-MIR396c showed reduced salt and alkali stress tolerance compared to that of wild-type plants. Overall, this study further established a link between salt and alkali stress and osa-MIR396c in rice.

Constitutive androstane receptor (CAR) induces xenobiotic, bilirubin, and thyroid hormone metabolism as a heterodimer with the retinoid X receptor (RXR). Unlike ligand-dependent nuclear receptors, CAR is constitutively active. Here, we report the heterodimeric structure of the CAR and RXR ligand binding domains (LBDs), which reveals an unusually large dimerization interface and a small CAR ligand binding pocket. Constitutive CAR activity appears to be mediated by the compact nature of the CAR LBD that displays several unique features including a shortened AF2 helix and helix H10, which are linked by a two-turn helix that normally adopts an extended loop in other receptors, and an extended helix H2 that stabilizes the canonical LBD fold by packing tightly against helix H3. These structural observations provide a molecular framework for understanding the atypical transcriptional activation properties of CAR.

The major root system of cereals consists of crown roots (or adventitious roots), which are important for anchoring plants in the soil and for water and nutrient uptake. However, the molecular basis of crown root formation is largely unknown. Here, we isolated a rice (Oryza sativa) mutant with fewer crown roots, named lower crown root number1 (lcrn1). Map-based cloning revealed that lcrn1 is caused by a mutation of a putative transcription factor-coding gene, O. sativa SQUAMOSA PROMOTER BINDING PROTEIN-LIKE3 (OsSPL3). We demonstrate that the point mutation in lcrn1 perturbs theO. sativa microRNA156 (OsmiR156)-directed cleavage of OsSPL3 transcripts, resulting in the mutant phenotype. Chromatin immunoprecipitation sequencing assays of OsSPL3 binding sites and RNA sequencing of differentially expressed transcripts in lcrn1 further identified potential direct targets of OsSPL3 in basal nodes, including a MADS-box transcription factor, OsMADS50. OsMADS50-overexpressing plants produced fewer crown roots, phenocopying lcrn1, while knocking out OsMADS50 in the lcrn1 background reversed this phenotype. We also show that OsSPL12, another OsmiR156 target gene, regulates OsMADS50 and crown root development. Taken together, our findings suggest a novel regulatory pathway in which the OsmiR156-OsSPL3/OsSPL12 module directly activates OsMADS50 in the node to regulate crown root development in rice.

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