bHLH093/NFL and bHLH061 are required for apical meristem function in Arabidopsis thaliana

The basic Helix-Loop-Helix (bHLH) transcription factors SCREAM/ICE1 and SCREAM2 have well-characterized roles in the terminal differentiation of stomatal guard cells in Arabidopsis thaliana. Here we report on the characterization of the functional roles of the remaining members of sub-group IIIB, bHLH093 and bHLH061. The bhlh093/bhlh061 double mutant failed to produce a primary inflorescence shoot and displayed greater phenotypic severity than bhlh093 and bhlh061 single mutants. An ultrastructural investigation revealed structural defects that develop in tissues surrounding the meristem prior to inflorescence emergence. The transition to flowering was restored in bhlh093/bhlh061 with the application of gibberellin to the apex. We also demonstrate that gibberellin application alleviates structural defects that develop in tissues surrounding the meristem and restore meristem activity. Furthermore, the bhlh093/bhlh061 double mutant was affected by delayed flowering, and the severity of the phenotype correlated with photoperiod and light intensity. Our results indicate that bHLH093 and bHLH061 function in the gibberellin-mediated establishment of functional apical meristems during the transition from vegetative to reproductive growth.