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Investigating Interactions Between Histone Modifying Enzymes and Transcription Factors in vivo by Fluorescence Resonance Energy Transfer
Published on: October 14, 2022
LAX PANICLE1 acts as a canonical bHLH transcription factor to regulate inflorescence axillary meristem formation in
Debao Fu1, Xiaohu Xiong1, Minghua Wei2
1National Key Laboratory of Crop Genetic Improvement, Hubei Hongshan Laboratory, College of Bio-X, Huazhong Agricultural University, Wuhan 430070, China.
Abstract:
The development of axillary meristems (AMs) is critical for tillering and panicle branching in rice, processes that ultimately determine grain yield. LAX PANICLE1 (LAX1) encodes a basic helix-loop-helix (bHLH) transcription factor essential for AM formation during panicle development. However, because LAX1 possesses a non-canonical basic region that contains fewer than five basic amino acids, it has long been presumed to lack intrinsic DNA-binding capacity, leaving its molecular mechanism of action unclear. Here, we demonstrate that LAX1 localizes to the nucleus and exhibits transcriptional activation activity. Using systematic evolution of ligands by exponential enrichment (SELEX) assays, we found that LAX1 possesses intrinsic DNA-binding capability in vitro. Furthermore, chromatin immunoprecipitation sequencing (ChIP-seq) analysis of young rice panicles identified direct LAX1 target genes; peaks were significantly enriched for the motif TTTTGC(T/A/C)NNN(G/A/T), which is directly recognized by the LAX1 bHLH domain. Subsequent experiments showed that LAX1 directly binds to the promoters of OsPID, OsIAA7, LOG, and OsTPS8. Molecular and genetic analyses provided evidence supporting a functional role for the LAX1-OsPID module in the regulation of inflorescence architecture. Comparative transcriptome analysis of lax1 mutants and wild-type plants further indicated that LAX1 participates in multiple biological processes. Interestingly, we found that LAX1 and its orthologs likely regulate downstream gene expression by recognizing a common set of cis-regulatory elements across diverse cereal crops. Collectively, our findings redefine LAX1 and its orthologs as canonical bHLH transcription factors, elucidate the regulatory network through which they control panicle branching, and provide a theoretical foundation for the cross-species application of LAX1 in crop yield improvement.
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