The putative RNA-dependent RNA polymerase RDR6 acts synergistically with ASYMMETRIC LEAVES1 and 2 to repress BREVIPEDICELLUS and MicroRNA165/166 in Arabidopsis leaf development

The Arabidopsis thaliana ASYMMETRIC LEAVES1 ( AS1) and AS2 genes are important for repressing class I KNOTTED1- like homeobox ( KNOX) genes and specifying leaf adaxial identity in leaf development. RNA- dependent RNA polymerases ( RdRPs) are critical for posttranscriptional and transcriptional gene silencing in eukaryotes; however, very little is known about their functions in plant development. Here, we show that the Arabidopsis RDR6 gene ( also called SDE1 and SGS2) that encodes a putative RdRP, together with AS1 and AS2, regulates leaf development. rdr6 single mutant plants displayed only minor phenotypes, whereas rdr6 as1 and rdr6 as2 double mutants showed dramatically enhanced as1 and as2 phenotypes, with severe defects in the leaf adaxial- abaxial polarity and vascular development. In addition, the double mutant plants produced more lobed leaves than the as1 and as2 single mutants and showed leaf- like structures associated on a proportion of leaf blades. The abnormal leaf morphology of the double mutants was accompanied by an extended ectopic expression of a class I KNOX gene BREVIPEDICELLUS ( BP) and high levels of microRNA165/ 166 that may lead to mRNA degradation of genes in the class III HD- ZIP family. Taken together, our data suggest that the Arabidopsis RDR6- associated epigenetic pathway and the AS1- AS2 pathway synergistically repress BP and MIR165/ 166 for proper plant development.