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Evolutionary origins and functional diversification of Auxin Response Factors.

Hernández-García, J., et al. · 2024a · Nature Communications   research

doi:10.1038/s41467-024-55278-8   PMID:39738167   PMC11685440

Linked genes (10)  5 core 5 peripheral

Gene ID Name Evidence / Role Function (this paper)
Mp1g12750 MpARF1 experimental subject An A-class ARF studied as the central genetic model for functional conservation. Knock-out Mparf1 mutants were complemented with wild-type and chimeric constructs, and domain-swap, PB1-interaction, and complementation assays defined A-class-specific activity.
Mp4g11820 MpARF2 experimental subject A B-class repressor ARF analyzed for class-specific function. Tested for complementation capacity in Mparf1 mutants (failed like other non-A classes), and PB1-interaction assays showed strong heterotypic interaction between MpARF2 and MpARF3 PB1 domains.
Mp1g07070 MpARF3 experimental subject A C-class ARF used as a model for C-class function. Mparf3 mutants underwent complementation and domain-swap assays for apical growth and gemma formation; non-C classes could not complement, demonstrating C-class specificity.
Mp2g00710 MpBromo4 experimental subject A pre-ARF-related gene investigated as the evolutionary precursor lineage of ARFs. CRISPR/Cas9 Mpphip mutants were generated and showed no genetic or functional resemblance to MpARF1 or MpARF3, and structural alignment of its DD-AD region was compared with ARF.
Mp2g22250 MpRAV experimental subject Studied as a domain-sharing relative used to probe ARF domain function and evolutionary divergence. MpRAV B3 and PB1 domains were swapped into MpARF1/MpARF3 chimeras for complementation tests, and yeast two-hybrid assays showed MpRAV PB1 strongly self-interacted.
Mp3g23400 MpEF1 experimental tool Used as a reference (housekeeping) gene; co-used with MpSAND for qRT-PCR expression normalization.
Mp6g05000 MpIAA experimental tool Aux/IAA repressor used to probe PB1-domain function and auxin regulation; its PB1 domain in MpARF1 chimeras partly restored growth and auxin responsiveness, and it served in yeast two-hybrid interaction profiling.
Mp4g07110 MpSAND missing experimental tool Used as a reference (housekeeping) gene; its expression contributed to the geometric mean for qRT-PCR normalization.
Mp1g09500 MpWIP experimental tool Used as an auxin-responsive marker to validate auxin signaling; its expression was monitored to confirm normal auxin response in mutant plants.
MpWIP1 missing experimental tool Auxin-responsive marker gene measured by qRT-PCR to confirm normal auxin-dependent gene expression in MpphipGE mutants.