Regulating Interfascicular Fiber Differentiation in Arabidopsis, Encodes a Homeodomain–Leucine Zipper Protein

Arabidopsis inflorescence stems develop extraxylary fibers at specific sites in interfascicular regions. The spatial specification of interfascicular fiber differentiation is regulated by the INTERFASCICULAR FIBERLESS1 ( IFL1 ) gene because mutation of that gene abolishes the formation of normal interfascicular fibers in Arabidopsis stems. To understand further the role of IFL1 in the specification of fiber differentiation, we cloned the IFL1 gene by using a positional cloning strategy. Sequence analysis showed that the IFL1 gene encodes a transcription factor that has the same features as a family of homeodomain–leucine zipper (HD-ZIP) proteins found only in plants. The predicted IFL1 protein is composed of three distinct domains, including a 60–amino acid HD at the N terminus followed by a 28–amino acid ZIP motif and a 724–amino acid C-terminal region. A nuclear targeting assay showed that IFL1 is able to direct a b -glucuronidase fusion protein into the nucleus, which is consistent with IFL1’s presumed function as a transcription factor. Gene expression analysis demonstrated that the IFL1 gene is expressed in the interfascicular regions in which fibers differentiate, which is consistent with its role in the control of interfascicular fiber differentiation. Furthermore, the IFL1 gene was shown to be expressed in the vascular regions, indicating its possible role in the regulation of vascular tissue formation. This possibility is supported by the observation that differentiation of both xylary fibers and vessel elements is altered in the vascular bundles of ifl1 mutants. Our results provide direct evidence that an HD-ZIP protein plays a role in the spatial control of fiber differentiation.