PDK1 has a pleiotropic PINOID-independent role in Arabidopsis development

Abstract

The 3-Phosphoinositide-Dependent Protein Kinase 1 (PDK1) is a conserved and important master regulator of AGC kinases in eukaryotic organisms. pdk1 loss-of-function causes a lethal phenotype in animals and yeast. In contrast, only very mild phenotypic defects have been reported for the pdk1 loss-of-function mutant of the model plant Arabidopsis thaliana (Arabidopsis). The Arabidopsis genome contains two PDK1 genes, hereafter called PDK1 and PDK2. Here we show that the previously reported Arabidopsis pdk1 T-DNA insertion alleles are not true loss-of-function mutants. By using CRISPR/Cas9 technology, we created true loss-of-function pdk1 alleles, and pdk1 pdk2 double mutants carrying these alleles showed multiple growth and development defect, including fused cotyledons, a short primary root, dwarf stature, late flowering, and reduced seed production caused by defects in male fertility. Surprisingly, pdk1 pdk2 mutants did not phenocopy pid mutants, and together with the observations that PDK1 overexpression does not phenocopy the effect of PID overexpression, and that pdk1 pdk2 loss-of-function does not change PID subcellular localization, we conclude that PDK1 is not essential for PID membrane localization or functionality in planta. Nonetheless, most pdk1 pdk2 phenotypes could be correlated with impaired auxin transport. PDK1 is highly expressed in vascular tissues and YFP:PDK1 is relatively abundant at the basal/rootward side of root stele cells, where it colocalizes with PIN auxin efflux carriers, and the AGC1 kinases PAX and D6PK/D6PKLs. Our genetic and phenotypic analysis suggests that PDK1 is likely to control auxin transport as master regulator of these AGC1 kinases in Arabidopsis.