Conferring High IAA Productivity on Low-IAA-Producing Organisms with PonAAS2, an Aromatic Aldehyde Synthase of a Galling Sawfly, and Identification of Its Inhibitor.

Gall-inducing insects often contain high concentrations of phytohormones, such as auxin and cytokinin, which are suggested to be involved in gall induction, but no conclusive evidence has yet been obtained. There are two possible approaches to investigating the importance of phytohormones in gall induction: demonstrating either that high phytohormone productivity can induce gall-inducing ability in non-gall-inducing insects or that the gall-inducing ability is inhibited when phytohormone productivity in galling insects is suppressed. In this study, we show that the overexpression of PonAAS2, which encodes an aromatic aldehyde synthase (AAS) responsible for the rate-limiting step in indoleacetic acid (IAA) biosynthesis in a galling sawfly (Pontania sp.) that contains high levels of endogenous IAA, conferred high IAA productivity on Caenorhabditis elegans, as the model system. This result strongly suggests that PonAAS2 can also confer high IAA productivity on low-IAA-producing insects. We also successfully identified an inhibitor of PonAAS2 in a chemical library. This highly selective inhibitor showed stronger inhibitory activity against AAS than against aromatic amino acid decarboxylase, which belongs to the same superfamily as AAS. We also confirm that this inhibitor clearly inhibited IAA productivity in the high-IAA-producing C. elegans engineered here.

Identification of an aromatic aldehyde synthase involved in indole-3-acetic acid biosynthesis in the galling sawfly (Pontania sp.) and screening of an inhibitor.

Indole-3-acetic acid (IAA), a phytohormone auxin, may be involved in insect gall induction. We previously proposed that the IAA biosynthetic pathway is Trp → indole-3-acetaldoxime → indole-3-acetaldehyde (IAAld) → IAA or Trp → IAAld → IAA. In this study, we surveyed galling sawfly enzymes responsible for the rate-limiting steps using a heterologous protein expression system and identified PonAAS2, an aromatic aldehyde synthase, that catalyzed the conversion of Trp to IAAld. The PonAAS2 gene was highly expressed in early- and mid-stage larvae that contained high concentrations of IAA, but the expression level was almost negligible in larvae that had escaped from their gall in autumn and contained very low concentrations of IAA. An inhibitor of PonAAS2, obtained by screening a chemical library, inhibited IAA production in sawfly enzyme solution by 80%, suggesting the important role of this enzyme in IAA biosynthesis in sawfly.