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<p>MicroProteins (miPs) represent a small but functionally relevant class of regulatory proteins involved in transcriptional and developmental processes in plants. Here, we used an integrative computational and experimental approach to identify and functionally characterize three candidate miPs (AT4G32105, AT2G32765, and AT1G16640) in <i>Arabidopsis thaliana</i>. Transcriptomic profiling and co-expression network analyses highlighted their expression during embryogenesis and their association with lipid-related pathways. Interaction predictions from STRING, conserved motif analysis, and docking simulations indicated potential protein–protein interaction compatibility for AT2G32765 and AT1G16640 with selected transcription factors, whereas AT4G32105 showed weaker predicted connectivity. Phenotypic characterization of the corresponding T-DNA insertion mutants revealed clear, genotype-specific developmental differences, including alterations in flowering time, shoot and root architecture, trichome density, and seed size. Fatty acid profiling of mature seeds further demonstrated that AT2G32765 and AT1G16640 mutants exhibit shifts in major fatty acid species, including increased proportions of C18:0 and C18:1 in AT2G32765 and altered C16:0 and C18:3 levels in AT1G16640. These experimentally measurable changes support a functional contribution of the candidate miPs to seed traits and fatty acid composition. Overall, this work expands the repertoire of putative plant microProteins and provides a systematic framework for identifying and prioritizing candidates with potential roles in seed development and oil-related traits. The findings establish a basis for future mechanistic studies and highlight candidate miPs as promising genetic entry points for metabolic and developmental modulation in crops.</p>

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Functional characterization of candidate microproteins involved in seed oil biosynthesis in Arabidopsis thaliana

  • 期刊论文

  • 发表日期：

    2026年5月2日

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  • Khadijeh Shokri,
  • Naser Farrokhi,
  • Mahdi Safaeizadeh,
  • Asadollah Ahmadikhah

摘要

MicroProteins (miPs) represent a small but functionally relevant class of regulatory proteins involved in transcriptional and developmental processes in plants. Here, we used an integrative computational and experimental approach to identify and functionally characterize three candidate miPs (AT4G32105, AT2G32765, and AT1G16640) in Arabidopsis thaliana. Transcriptomic profiling and co-expression network analyses highlighted their expression during embryogenesis and their association with lipid-related pathways. Interaction predictions from STRING, conserved motif analysis, and docking simulations indicated potential protein–protein interaction compatibility for AT2G32765 and AT1G16640 with selected transcription factors, whereas AT4G32105 showed weaker predicted connectivity. Phenotypic characterization of the corresponding T-DNA insertion mutants revealed clear, genotype-specific developmental differences, including alterations in flowering time, shoot and root architecture, trichome density, and seed size. Fatty acid profiling of mature seeds further demonstrated that AT2G32765 and AT1G16640 mutants exhibit shifts in major fatty acid species, including increased proportions of C18:0 and C18:1 in AT2G32765 and altered C16:0 and C18:3 levels in AT1G16640. These experimentally measurable changes support a functional contribution of the candidate miPs to seed traits and fatty acid composition. Overall, this work expands the repertoire of putative plant microProteins and provides a systematic framework for identifying and prioritizing candidates with potential roles in seed development and oil-related traits. The findings establish a basis for future mechanistic studies and highlight candidate miPs as promising genetic entry points for metabolic and developmental modulation in crops.