Abstract Regulation of the homeodomain transcription factor WUSCHEL concentration is critical for stem cell homeostasis inArabidopsisshoot apical meristems. WUSCHEL regulates the transcription ofCLAVATA3through a concentration-dependent activation-repression switch.CLAVATA3, a secreted peptide, activates receptor kinase signaling to repressWUSCHELtranscription. Considering the revised regulation,CLAVATA3mediated repression ofWUSCHELtranscription alone will lead to an unstable system. Here we show thatCLAVATA3signaling regulates nuclear-cytoplasmic partitioning ofWUSCHELto control nuclear levels and its diffusion into adjacent cells. Our work also reveals that WUSCHEL directly interacts with EXPORTINS via EAR-like domain which is also required for destabilizing WUSCHEL in the cytoplasm. We develop a combined experimental and computational modeling approach that integratesCLAVATA3-mediated transcriptional repression ofWUSCHELand post-translational control of nuclear levels with the WUSCHEL concentration-dependent regulation ofCLAVATA3. We show that the dual control by the same signal forms a seamless connection between de novo WUSCHEL synthesis and sub-cellular partitioning in providing robustness to the WUSCHEL gradient.
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Large-scale single-cell profiling of stem cells uncovers redundant regulators of shoot development and yield trait variation
SUMMARY Stem cells in plant shoots are a rare population of cells that produce leaves, fruits and seeds, vital sources for food and bioethanol. Uncovering regulators expressed in these stem cells will inform crop engineering to boost productivity. Single-cell analysis is a powerful tool for identifying regulators expressed in specific groups of cells. However, accessing plant shoot stem cells is challenging. Recent single-cell analyses of plant shoots have not captured these cells, and failed to detect stem cell regulators likeCLAVATA3andWUSCHEL. In this study, we finely dissected stem cell-enriched shoot tissues from both maize and arabidopsis for single-cell RNA-seq profiling. We optimized protocols to efficiently recover thousands ofCLAVATA3andWUSCHELexpressed cells. A cross-species comparison identified conserved stem cell regulators between maize and arabidopsis. We also performed single-cell RNA-seq on maize stem cell overproliferation mutants to find additional candidate regulators. Expression of candidate stem cell genes was validated using spatial transcriptomics, and we functionally confirmed roles in shoot development. These candidates include a family of ribosome-associated RNA-binding proteins, and two families of sugar kinase genes related to hypoxia signaling and cytokinin hormone homeostasis. These large-scale single-cell profiling of stem cells provide a resource for mining stem cell regulators, which show significant association with yield traits. Overall, our discoveries advance the understanding of shoot development and open avenues for manipulating diverse crops to enhance food and energy security.
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- Award ID(s):
- 2026561
- PAR ID:
- 10531696
- Publisher / Repository:
- bioRxiv
- Date Published:
- Format(s):
- Medium: X
- Institution:
- bioRxiv
- Sponsoring Org:
- National Science Foundation
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