Interactions among proteins of floral MADS-box genes in basal eudicots: implications for evolution of the regulatory...

Floral MADS-box genes encode transcription factors that play critical roles in the development and evolution of the flower. Proteins of floral MADS-box genes regulate the expression of their downstream genes by forming various homodimers/heterodimers and quaternary complexes. Interactions among proteins of floral MADS-box genes have been documented in several model species, yet the information accumulated so far is still not sufficient to draw a general picture of the evolution of the interactions. We have characterized 28 putative floral MADS-box genes from three representative basal eudicots (i.e., Euptelea pleiospermum, Akebia trifoliata, and Pachysandra terminalis) and investigated the protein-protein interactions (PPIs) among the proteins encoded by these genes using yeast two-hybrid assays. We found that, although the PPIs in basal eudicots are largely consistent with those in core eudicots and monocots, there are lineage-specific features that have not been observed elsewhere. We also reconstructed the evolutionary histories of the PPIs among members of seven MADS-box gene lineages (i.e., AP1, AP3, PI, AG, STK, AGL2, and AGL9) in angiosperms. We revealed that the PPIs were extremely conserved in nine (or 32.1%) of the 28 possible combinations, whereas considerable variations existed in seven (25.0%) of them; in the remaining 12 (or 42.9%) combinations, however, no interaction was observed. Notably, most of the PPIs required for the formation of quaternary complexes, as suggested by the "quartet model," were highly conserved. This suggested that the evolutionarily conservative PPIs may have played critical roles in the establishment of the basic structure (or architecture) of the flower and experienced coevolution to maintain their functions. The evolutionarily variable PPIs, however, seem to have played subsidiary roles in flower development and have contributed to the variation in floral traits.