Manganese Toxicity Elicits the Degradation of Auxin Transport Carriers to Restrain Arabidopsis Root Growth

Manganese (Mn) is an essential microelement for plant growth but can be toxic when in excessive. Mn toxicity-inhibited root growth possible involves in auxin biosynthesis and transport in plants, but the mechanism remains elusive. Our results have suggested that Mn-inhibited root growth was possible associated with a reduced auxin levels in roots, as shown in experiments with (1) transgenic DII-VENUS and DR5rev::GFP; (2) the application of L-kynurenine (an inhibitor of TAA1 activity), 1-naphthylacetic acid (NAA), 1-naphthylphthalamic acid (NPA) to manipulate auxin levels; (3) using taa1 (defection in auxin biosynthesis) mutants; and (4) determining IAA by enzyme-linked immunosorbent assay. Those results were explained via down-regulated transcriptional levels of auxin biosynthesis-related genes in plants, and reduced abundance of PM-localized auxin transport carriers in roots. Combination of auxin transport-related transgenic pPINs-GFP or pAUX1::AUX1-YFP lines and a real-time in vitro observation of PM-localized PIN2/3/4 carriers using transgenic Dendra2 lines have revealed that excessive Mn induced degradation of these proteins, which might involve in Mn toxicity-elicited up-regulation of partially ubiquitin-, clathrin-meidiated endocytosis (CME)-, and endosomal sorting complex required for transport (ESCRT)-related genes. Overall, our results suggest that Mn toxicity reduces auxin level in the root apex possible via down-regulation of auxin biosynthesis-related genes and post-translational stimulation of the degradation of auxin transport proteins, leading to root growth inhibition caused by Mn toxicity.