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<p>In angiosperms, pollen tube growth is crucial for double fertilization and directly affects crop yield. Rapid alkalization factor (RALF) peptides play essential roles in fertilization by interacting with receptor-like kinases in the apoplast. However, the function of atypical types of RALFs remains largely unknown. We identified the pollen-specific <i>Oryza sativa</i> RALF gene, <i>OsRALF5</i>, and investigated its role in reproduction. OsRALF5 belongs to a distinct clade lacking the conserved motifs of typical RALFs, as revealed by phylogenetic analysis. <i>OsRALF5</i> was highly expressed in pollen, as shown using reverse transcription quantitative polymerase chain reaction (RT-qPCR) and reporter assays using native promoter-derived β-glucuronidase (GUS)-and green fluorescent protein (GFP)-tagging transgenic lines. Subcellular localization in tobacco and rice indicated that OsRALF5 was predominantly cytoplasmic and could be secreted into plasma membrane and apoplasts. Clustered regularly interspaced palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9)-generated <i>osralf5</i> knockout mutants displayed normal pollen viability but showed significantly reduced pollen germination rates and seed sets. Our findings demonstrated that OsRALF5, which is an atypical pollen-preferential RALF, positively regulates pollen germination and fertilization in rice.</p>

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Pollen-Specific OsRALF5 Regulates Pollen Tube Growth in Oryza sativa

  • 期刊论文

  • 发表日期：

    2025年12月16日

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  • Hyo-Jeong Kim,
  • Ji-Hyun Kim,
  • Eui-Jung Kim,
  • Ye-Jin Son,
  • Woo-Jong Hong,
  • Da Eun Kim,
  • Byoung Il Je,
  • Ki-Hong Jung,
  • Yu-Jin Kim

摘要

In angiosperms, pollen tube growth is crucial for double fertilization and directly affects crop yield. Rapid alkalization factor (RALF) peptides play essential roles in fertilization by interacting with receptor-like kinases in the apoplast. However, the function of atypical types of RALFs remains largely unknown. We identified the pollen-specific Oryza sativa RALF gene, OsRALF5, and investigated its role in reproduction. OsRALF5 belongs to a distinct clade lacking the conserved motifs of typical RALFs, as revealed by phylogenetic analysis. OsRALF5 was highly expressed in pollen, as shown using reverse transcription quantitative polymerase chain reaction (RT-qPCR) and reporter assays using native promoter-derived β-glucuronidase (GUS)-and green fluorescent protein (GFP)-tagging transgenic lines. Subcellular localization in tobacco and rice indicated that OsRALF5 was predominantly cytoplasmic and could be secreted into plasma membrane and apoplasts. Clustered regularly interspaced palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9)-generated osralf5 knockout mutants displayed normal pollen viability but showed significantly reduced pollen germination rates and seed sets. Our findings demonstrated that OsRALF5, which is an atypical pollen-preferential RALF, positively regulates pollen germination and fertilization in rice.