In vitro differentiation of tracheary elements is induced by suppression of Arabidopsis phytoglobins.

Differentiation of tracheary elements (TEs) in vitro was affected by the expression level of the Arabidopsis thaliana Col-0 phytoglobins (Pgbs). Over-expression of Pgb1 or Pgb2 (35S:Pgb1 or 35S:Pgb2 lines) reduced the differentiation process while suppression of either Pgb (Pgb1-RNAi or pgb2 lines) enhanced the production of TEs. The inductive effect of Pgb suppression on TE differentiation was linked to the reduced expression of the transcription factor MYC2. Suppression of this gene, observed under conditions of high NO levels or low Pgb expression, was sufficient to promote TE differentiation, while its over-expression abolished the promotive effect of Pgb suppression on the differentiation process. Cells in which MYC2 was mutated accumulated ethylene which induced the expression of the homeodomain-leucine zipper (HD-Zip) III ATHB8. Production of ethylene was reduced in cells over-expressing MYC2 in a WT or a pgb mutant background. While stabilizing procambial cell specification, ATHB8 in known to activate downstream components triggering programmed cell death (PCD) and modifications of cell wall components, required steps of the TE differentiation process. Collectively, we provide evidence that in addition to their recognised participation in stress responses, Pgbs may play a key role in the specification of cell fate during development.