These findings establish OsDPMS1 as a critical regulator of pistil development and female fertility in rice and highlight its potential application in hybrid rice breeding programs.
Abstract
The pistil, a key female reproductive organ in flowering plants, plays a critical role in sexual reproduction. Although pistil function has been well studied in some species, the molecular mechanisms governing its development in rice (Oryza sativa) remain unclear. Here, we isolated and characterized a female-sterile rice mutant, Osdpms1, that exhibits pleiotropic reproductive defects, including greater stigma number and aberrant gametophyte development, while maintaining normal vegetative growth and male fertility. Using map-based cloning, genetic complementation, and CRISPR-Cas9 knockout experiments, we determined that LOC_Os07g03160 (named OsDPMS1), encoding a protein containing a WD40 repeat domain, is the causal gene underlying the mutant phenotype. OsDPMS1 was predominantly expressed during the flowering stage, and subcellular localization experiments revealed that OsDPMS1 localizes to both the nucleus and the cytoplasm. Using yeast two-hybrid, bimolecular fluorescence complementation, and GST-pulldown assays, we demonstrated that OsDPMS1 physically interacts with three components of the 26S proteasome pathway: OsUBQ, OsCDC48, and OsCDC48E. Furthermore, we showed that the female sterility of Osdpms1 can be efficiently maintained via seed production technology. When we combined Osdpms1 with male-sterile lines for hybrid seed production in a mixed planting manner, we achieved yield potentials comparable to those using conventional row-planting methods. Our findings establish OsDPMS1 as a critical regulator of pistil development and female fertility in rice and highlight its potential application in hybrid rice breeding programs.
Rice blast, caused by Magnaporthe oryzae (M.oryzae), is one of the three most devastating rice diseases, significantly reducing both yield and grain quality. The Elongator complex, first characterized in Arabidopsis thaliana, regulates growth, development, and innate immunity by tightly associating with hyperphosphorylated RNA polymerase II. However, its immunological role in rice remains unexplored. Here, we demonstrate that Elongator plays an essential role in rice blast resistance. The OsELP3 subunit mutant exhibited susceptibility to M.oryzae, while its overexpression increased resistance. OsELP3 interacts with its homologous subunit OsELP4 in the nucleus, and that OsELP4 also positively regulates rice blast resistance. RNA-seq and histone acetylation analyses demonstrated that OsELP3/ELP4 enhances transcriptional activation of key rice blast resistance genes by modulating histone acetylation levels in both the jasmonic acid (JA) signaling pathway (OsAOS1, OsLOX6, OsPROPEP3) and lignin biosynthesis pathway (OsMYB30, OsMYB55, OsMYB110), thereby strengthening plant defense mechanisms against M. oryzae. These findings reveal that OsELP3/ELP4 regulates histone acetylation-mediated defense responses against M.oryzae, providing both theoretical foundations and genetic resources for developing blast-resistant rice cultivars.
Yanfei Wu, Zhensheng Qiao, Yuquan Fu et al.· Plant Physiology· 0 citations
Auxin is an important regulator of reproductive development. The families of the AUXIN RESPONSE FACTOR (ARF) genes, important components of the auxin signal, have expanded in flowering plants. Tomato (Solanum lycopersicum) has two copies of ARF8, which are negatively regulated by miR167. Here, we investigated the involvement of this expanded SlmiR167-SlARF8 module in flower development. Mutants were generated in 5 SlmiR167 and 2 SlARF8 genes, and flower development and fruit set were characterized in mutant combinations. We show that the SlmiR167-SlARF8 module regulates flower-organ development and fruit set. The two SlARF8 paralogs are found to have both overlapping and distinct roles. They additively affect stigma position by oppositely affecting stamen and pistil growth, while SlARF8B has evolved a unique role in the inhibition of anther dehiscence. Loss of SlARF8B in the Slmir167a background uncovers a unique reproductive syndrome of fast, early, fertilization-independent fruit set and increased yield under cold stress, mediated by the deregulation of SlARF8A. These results demonstrate how gene duplication within the miR167-ARF8 module enables fine-tuning of auxin signaling to achieve coordinated, robust reproductive development, with implications for the control of fruit set in tomato.
Nave Man, Naama Teboul, Amir Bahu et al.· New Phytologist· 0 citations
Chaperones are essential for Clp protease that governs protein quality control in chloroplast. However, it remains poorly understood whether and how individual ClpC proteins play distinct roles in plant growth and development. In this study, we isolated a new allele of clpc2, designated sot11, from genetic suppressor screening of the thf1 leaf variegation phenotype. DNA sequencing combined with protein structural modeling revealed that the sot11 mutation causes a substitution of Thr to Ile at position 868 located within a loop region of ClpC2, leading to partial loss-of-function of ClpC2. Genetic and phenotypic assays demonstrated that sot11 suppresses not only thf1-mediated but also var2/ftsh2-mediated leaf variegation. Notably, sot11 fails to restore high-light hypersensitivity and the reduced FtsH protease levels observed in thf1. In addition, CRISPR-Cas9-generated complete knockout mutants of clpc2 exhibit severe growth retardation, markedly shortened roots, and pronounced anthocyanin accumulation, compared to the wild type (WT) plants, revealing a dosage-dependent relationship between ClpC2 activity and plant fitness. Taken together, these findings suggest that ClpC2 plays an important role in both shoot and root growth, opening a new avenue for investigating the organ-specific molecular mechanisms underlying ClpC2 function.
Xiao-Mei Chen, Ting Guo, Danni Lin et al.· Biochemical and Biophysical...· 0 citations
Eukaryotic translation initiation factor 5A (eIF5A) is a highly conserved protein family unique to eukaryotes, yet its functional characterization in woody plants remains limited. In this study, we identified four eIF5A genes (PtoeIF5A1–PtoeIF5A4) from the genome of Populus tomentosa, a fast-growing tree species indigenous to China, and characterized their expression patterns and functional roles through bioinformatics analysis, quantitative real-time PCR, stable overexpression in Arabidopsis thaliana, and transient expression in Nicotiana benthamiana leaves. Our results demonstrated that all PtoeIF5A proteins contain a conserved OB-fold domain and multiple phosphorylation sites, with PtoeIF5A1 showing predominant expression in roots and secondary xylem. Functional assays revealed that PtoeIF5A1 overexpression accelerated inflorescence stem elongation and early flowering in Arabidopsis, induced visible chlorosis and programmed cell death (PCD) in tobacco leaves, and significantly enhanced salt tolerance under NaCl treatment. Collectively, these findings establish PtoeIF5A1 in poplar as a pleiotropic regulator integrating developmental cues, programmed cell death, and stress responses; and as a valuable genetic resource for breeding stress-resilient woody plants.
Dan Zhu, Y. Guan, Feng Feng et al.· Plants· 0 citations
Chloroplasts drive photosynthesis and regulate cellular signaling and phytohormone biosynthesis processes that greatly influence crop yield and quality traits. In this study, we identified a recessive chlorophyll-deficient mutant, reduced chlorophyll mutant 1t (rcm1t), from an advanced-generation tomato inbred line. Genetic and phenotypic characterization confirmed that chloroplast degradation accelerates rapidly and chlorosis occurs during late leaf development in the rcm1t mutant. Subsequently, we fine-mapped rcm1t to an approximately 60 kb region on chromosome 8. This interval encompasses Lutescent (L1)/SlRCM1, a well-characterized gene essential for chloroplast biogenesis and developmental maintenance. An allelism test between rcm1t and the previously characterized ethyl methanesulfonate (EMS)-induced rcm1 mutant revealed non-complementation. Moreover, transgenic complementation with the wild-type SlRCM1 coding sequence rescued the chloroplast premature senescence phenotype in rcm1t, providing definitive genetic evidence that rcm1t is a loss-of-function allele of SlRCM1. Resequencing identified a ~4.8 kb retrotransposon insertion in the first exon of the gene, which disrupts normal transcription and results in a premature stop codon. Quantification of abscisic acid (ABA) revealed significantly higher ABA levels in the rcm1t mutant than in the wild type. Protein–protein interaction analyses demonstrate that SlRCM1 interacts with SlNCED2. Osmotic stress assays showed that the rcm1t near-isogenic line exhibited enhanced tolerance under osmotic stress. This study successfully mapped the natural allele rcm1t of SlRCM1, preliminarily explored its potential application in enhancing plant tolerance to osmotic stress, and provided valuable insights for further elucidation of SlRCM1 function.
Zhuo-Yao Qiu, Chunyang Pan, Dong-Ling Lan et al.· International Journal of Mol...· 0 citations
Evidence is provided suggesting that StFKF1 influences the timing of flowering and tuber initiation, as well as the expression patterns of key regulatory genes, under long-day conditions, thereby contributing to a theoretical foundation for understanding developmental transitions in this specific environmental context.
Zefeng Zhai, Yongguang Liu, Haicai Li et al.· Molecular breeding· 0 citations
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