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The C function gene  AGAMOUS  ( AG ) encodes for a MADS-box transcription factor required for floral organ identity and floral meristem (FM) determinacy in angiosperms. Unlike Arabidopsis, most legume plants possess two  AG  homologs arose by an ancient genome duplication event. Recently, two  euAGAMOUS  genes,  MtAGa  and  MtAGb , were characterized and shown to fulfill the C function activity in the model legume  Medicago truncatula . Here, we reported the isolation and characterization of a new  mtaga  allele by screening the  Medicago Tnt1  insertion mutant collection. We found that  MtAGa  was not only required for controlling the stamen and carpel identity but also affected pod and seed development. Genetic analysis indicated that  MtAGa  and  MtAGb  redundantly control  Medicago  floral organ identity, but have minimal distinct functions in regulating stamen and carpel development in a dose-dependent manner. Interestingly, the stamens and carpels are mostly converted to numerous vexillum-like petals in the double mutant of  mtaga mtagb , which is distinguished from Arabidopsis  ag . Further qRT-PCR analysis in different  mtag  mutants revealed that  MtAGa  and  MtAGb  can repress the expression of putative A and B function genes as well as  MtWUS , but promote putative D function genes expression in  M. truncatula . In addition, we found that the abnormal dorsal petal phenotype observed in the  mtaga mtagb  double mutant is associated with the upregulation of  CYCLOIDEA  ( CYC )-like TCP genes. Taken together, our data suggest that the redundant  MtAGa  and  MtAGb  genes of  M. truncatula  employ a conserved mechanism of action similar to Arabidopsis in determining floral organ identity and FM determinacy but may have evolved distinct function in regulating floral symmetry by coordinating with specific floral dorsoventral identity factors.

Functional Specialization of Duplicated AGAMOUS Homologs in Regulating Floral Organ Development of Medicago truncatula