Both phytohormones and non-coding microRNAs (miRNAs) play important role in root development in Arabidopsis thaliana. Mature miR166/165 s, which are derived from precursor transcripts of concerned genes, regulate developmental processes, including leaf and root patterning, by targeting Class III HOMEODOMAIN LEUCINE-ZIPPER (HD-ZIP III) transcription factors (TFs). However, their regulation through hormones remained poorly understood. Here, we show that several phytohormones dynamically regulate the spatio-temporal expression pattern of miR166/165 and target HD-ZIP IIIs in developing roots. Hormone signaling pathway mutants show differential expression pattern of miR166/165, providing further genetic evidence for multilayered regulation of these genes through phytohormones. We further show that a crosstalk of at least six different phytohormones regulate the miR166/165, their target HD-ZIP IIIs, and KANADI (KANs). Our results suggest that HD-ZIP IIIs mediated root development is modulated both transcriptionally through phytohormones and KANs, and post-transcriptionally by miR166/165 that in turn are also regulated by the phytohormonal crosstalk.

The main root and continuously emerging lateral roots constitute the root architecture of an adult plant during its postembryonic development. Epigenetic modifications like methylation or deacetylation of histones have been suggested to regulate root development. SWP1/LDL1, a component of plant specific corepressor complex, has been implicated in the induction of flowers and root through histone modifications in Arabidopsis. However, molecular role of SWP1 in regulating the lateral root development remained unexplored. Here we show that SWP1 regulates lateral root initiation and elongation in Arabidopsis. Mutation in SWP1 increases both the density and length of lateral roots. SWP1 negatively regulates lateral root initiation through direct/indirect transcriptional repression of lateral root promoting factors, such as AUXIN RESPONSE FACTORS (ARFs) and GATA23.