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Terpenoid Secondary Metabolism in Arabidopsis thaliana: cDNA Cloning, Characterization, and Functional Expression of a Myrcene/(E)-β-Ocimene Synthase

https://doi.org/10.1006/abbi.1999.1669Get rights and content

Abstract

The Arabidopsis genome project has recently reported sequences with similarity to members of the terpene synthase (TPS) gene family of higher plants. Surprisingly, several Arabidopsis terpene synthase-like sequences (AtTPS) share the most identity with TPS genes that participate in secondary metabolism in terpenoid-accumulating plant species. Expression of a putative Arabidopsis terpene synthase gene, designated AtTPS03, was demonstrated by amplification of a 392-bp cDNA fragment using primers designed to conserved regions of plant terpene synthases. Using the AtTPS03 fragment as a hybridization probe, a second AtTPS cDNA, designated AtTPS10, was isolated from a jasmonate-induced cDNA library. The partial AtTPS10 cDNA clone contained an open reading frame of 1665 bp encoding a protein of 555 amino acids. Functional expression of AtTPS10 in Escherichia coli yielded an active monoterpene synthase enzyme, which converted geranyl diphosphate (C10) into the acyclic monoterpenes β-myrcene and (E)-β-ocimene and small amounts of cyclic monoterpenes. Based on sequence relatedness, AtTPS10 was classified as a member of the TPSb subfamily of angiosperm monoterpene synthases. Sequence comparison of AtTPS10 with previously cloned monoterpene synthases suggests independent events of functional specialization of terpene synthases during the evolution of terpenoid secondary metabolism in gymnosperms and angiosperms. Functional characterization of the AtTPS10 gene was prompted by the availability of Arabidopsis genome sequences. Although Arabidoposis has not been reported to form terpenoid secondary metabolites, the unexpected expression of TPS genes belonging to the TPSb subfamily in this species strongly suggests that terpenoid secondary metabolism is active in the model system Arabidopsis.

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      They also reported not only that photoperiod could indirectly affect terpene profile but also that temperature had considerable effects on respiratory substrate concentrations because of its effect on modifying the critical day length (Burbott and Loomis, 1967). Our finding indicating the coproduction of β-myrcene/ocimene and α-pinene/β-pinene confirms those reported elsewhere suggesting that a single multiproduct monoterpene synthase could be possibly involved in the biosynthesis of various types of terpenes (Bohlmann et al., 2000; Croteau et al., 1978; Dehal and Croteau, 1987; Martin et al., 2010). On the other hand, it has been suggested that the coordinated formation and emission of monoterpene compounds could be due to the involvement of separate enzymes that use similar substrates (Dudareva et al., 2003).

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    The nucleotide sequence reported in this paper will appear in GenBank under Accession Nos. AF178535 and AF180366.

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