No abstract

Maize (Zea mays) produces zealexins as phytoalexins, with the inducible production of these antibiotics providing biochemical protection against fungal infection. However, the biosynthesis of these sesquiterpenoids has remained unclear. In particular, it is unclear how the olefinic precursor, (S)-β-macrocarpene produced by the characterized maize sesquiterpene synthases TPS6/11, is further elaborated to form the bioactive zealexins. The first step is likely to be conversion of carbon-15 (C15) from a methyl group to a carboxylic acid by a cytochrome P450 mono-oxygenase (CYP). In this study, CYP71Z18, whose transcription is strongly induced by fungal infection, was found to catalyze oxidation of C15 in (S)-β-macrocarpene, forming zealexin A1. The inducible transcription of CYP71Z18 matches that observed for TPS6/11 and the accumulation of zealexins, which is consistent with a role for CYP71Z18 in sesquiterpenoid phytoalexin production. This completes identification of zealexin A1 biosynthesis, and represents the initial CYP identified for the production of maize terpenoid phytoalexins.

No abstract

CYP701A26 was characterized to exhibit ent-kaurene oxidase activity with substrate promiscuity and might be involved in maize GA biosynthesis, and its homologue CYP701A43 did not show such function and might play roles in stress response.

Maize ( Zea mays ) terpene synthase 7 (ZmTPS7) was characterized as a τ-cadinol synthase, which exhibited constitutive and inducible gene expression patterns, suggesting involvement in stress response. Maize produces a variety of terpenoids involved in defense response. Despite some terpene synthases (TPSs) responsible for these terpenoids have been characterized, biosynthesis of many terpenes, particularly sesquiterpenes, which were produced in response to biotic or abiotic stress, remains largely unknown. Here, we characterized ZmTPS7 biochemically through recombinant expression in Escherichia coli and detected that it catalyzed formation of a blend of sesquiterpenes and sesquiterpenoid alcohols as the sesquiterpene synthase through GC-MS analysis. Subsequently, the major product was purified and identified as τ-cadinol through nuclear magnetic resonance spectroscopy (NMR) analysis, which was also detected in maize tissues infected by pathogen fungus for the first time. ZmTPS7 constitutively expressed in aerial tissues while with trace amount of transcript in roots. Fungus spore inoculation and methyl jasmonate (MeJA) treatment induced gene expression of ZmTPS7 in leaves, while exogenous ABA induced ZmTPS7 dramatically in roots, suggesting that ZmTPS7 might be involved in stress response. τ-cadinol was quantified in infected maize tissues with the concentration of ~200 ng/g fresh weight, however, which was much lower than the inhibitory one on two tested necrotrophic fungi. Such evidences indicate that anti-fungal activity of τ-cadinol is not physiologically relevant, and further investigation is needed to clarify its biological functions in maize. Taken together, ZmTPS7 was characterized as the τ-cadinol synthase and suggested to be involved in stress response, which also increased the diversity of maize terpenoid profile.