A family of auxin conjugate hydrolases from Brassica rapa:: characterization and expression during clubroot disease

During the obligate biotrophic interaction of Plasmodiophora brassicae with members of the Brassicaceae, the host roots show hypertrophy and galls are established. An increased auxin pool appears to correlate with cell expansion and cell division, but the origin of the free auxin is not yet clear. As previous results point to increased IAA-hydrolytic activity in infected roots of Brassica rapa at later time points of infection, we isolated IAA-amidohydrolase-like genes from various tissues. We cloned full-length cDNAs of two genes with high homology to the Arabidopsis IAR3 (Br-IAR3) as well as full-length clones corresponding to the Arabidopsis ILL2/ILL1 (Br-ILL2) and ILL6 (Br-ILL6) hydrolase genes. Using heterologous expression in Escherichia coli, we showed that Br-IAR3 24 and Br-ILL2 possess hydrolytic activity in vitro. Real-time reverse transcription (RT)-PCR revealed that only Br-IAR3 25 and Br-ILL6 are expressed differentially during clubroot disease, but showed a decreased expression at later time point of infection. These findings are discussed with regard to a negative regulation in IAA homeostasis during clubroot disease.