Z-DNA is a left-handed double helix form of DNA that is believed to be involved in various DNA transactions. However, comprehensive investigations aimed at global profiling of Z-DNA landscapes are still missing in both humans and plants. We here report the development of two techniques: anti-Z-DNA antibody-based immunoprecipitation followed by sequencing (ZIP-seq), and cleavage under targets and tagmentation (CUT&TAG) for characterizing Z-DNA in nipponbare rice (Oryza sativa L., Japonica). We found that Z-DNA-IP+ (Z-DNA recognized by the antibody) exhibits distinct genomic features as compared to Z-DNA-IP- (Z-DNA not recognized by the antibody). The concomitant presence of G-quadruplexes (G4s) and i-motifs (iMs) may promote Z-DNA formation. DNA modifications such as DNA-6mA/-4acC generally disfavours Z-DNA formation, while modifications like DNA-5mC (CHH) and 8-oxodG promote it, highlighting the distinct roles of DNA base modifications in modulating Z-DNA formation. Importantly, Z-DNA located at transcription start sites (TSSs) enhances gene expression, whereas Z-DNA in genic regions represses it, underscoring its dual roles in regulating the expression of genes involved in fundamental biological functions and responses to salt stress. Furthermore, Z-DNA may play a role in transcriptional initiation and termination rather than in transcriptional elongation. Finally, the presence of Z-DNA in promoters is correlated with the coevolution of overlapping genes, thereby regulating gene domestication. Consequently, our study represents as a pivotal point and a solid foundation for reliably launching genome-wide investigations of Z-DNA, thereby advancing the understanding of Z-DNA biology in both plants and non-plant systems.
Upland cotton (Gossypium hirsutum) is a principal economic crop and a fundamental raw material for the textile industry. The quality of cotton fibres is significantly influenced by the synthesis of cell wall polysaccharides. This study focuses on GhIRX10, a beta-1,4-xylosyltransferase crucial for xylan backbone synthesis. Overexpression of GhIRX10 enhances xylan synthesis, which impacts fibre elongation and secondary cell wall thickening. GhMYB102, identified as a direct regulator of GhIRX10 expression, was confirmed through comprehensive validation. Overexpression of GhMYB102 resulted in a similar phenotype as OE-GhIRX10: increased cell wall thickness and reduced fibre length. Overexpression of GhMYB102 upregulated the expression of key cell wall synthesis-related genes, including GhCESA4/7/8, GhIRXs, GhCESAs, GhGUXs, GhTBLs, GhXTHs, and GhXXTs. Consequently, the cellulose and hemicellulose contents in OE-GhMYB102 lines were significantly increased. GhMYB102 was also validated as a target gene regulated by GhFSN1 and GhMYB7, with the ability to reciprocally regulate GhFSN1 expression. In summary, we propose a regulatory model where GhMYB102 promotes the expression of GhIRX10 and other cell wall-related genes, thereby affecting fibre quality. This study elucidates the regulatory network of secondary cell wall synthesis in cotton and provides potential targets for improving fibre quality through molecular breeding.
Improving plant architecture and increasing yields are the main goals of rice breeders. However, yield is a complex trait influenced by many yield-related traits. Identifying and characterizing important genes in the coordinated network regulating complex rice traits and their interactions is conducive to cultivating high-yielding rice varieties. In this study, we determined that the interaction between mitogen-activated protein kinase kinase kinase5 (OsMAPKKK5) and brassinosteroid-signalling kinase1-1 (OsBSK1-1) regulates yield-related traits in rice. Specifically, OsMAPKKK5 phosphorylates OsBSK1-1, which enhances the interaction between these two proteins, but adversely affects the OsBSK1-1-OsBRI1 (BR insensitive1) and OsBSK1-1-OsPPKL1 (protein phosphatase with two Kelch-like domains) interactions. Additionally, OsMAPKKK5 disrupts brassinosteroid signal transduction, which prevents OsBZR1 (brassinazole-resistant1) from efficiently entering the nucleus, thereby negatively modulating its function as a transcription factor regulating downstream effector genes, ultimately adversely affecting plant architecture and yield. This study revealed the relationship between the MAPK cascade and the regulatory effects of brassinosteroid on the rice grain yield involves OsMAPKKK5 and OsBSK1-1. The study data may be important for future investigations on the rice yield-regulating molecular network.