Plant Physiol. Drug Metab Dispos
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Limonene Synthase, the Enzyme Responsible for Monoterpene Biosynthesis in Peppermint, Is Localized to Leucoplasts of Oil Gland Secretory Cells1

Glenn Turner, Jonathan Gershenzon2, Erik E. Nielson, John E. Froehlich, and Rodney Croteau*

Institute of Biological Chemistry, and Department of Biochemistry and Biophysics, Washington State University, Pullman, Washington 99164-6340 (G.T., J.G., R.C.); and Department of Energy Plant Research Laboratory, Michigan State University, East Lansing, Michigan 48823 (E.E.N., J.E.F.)

Circumstantial evidence based on ultrastructural correlation, specific labeling, and subcellular fractionation studies indicates that at least the early steps of monoterpene biosynthesis occur in plastids. (4S)-Limonene synthase, which is responsible for the first dedicated step of monoterpene biosynthesis in mint species, appears to be translated as a preprotein bearing a long plastidial transit peptide. Immunogold labeling using polyclonal antibodies raised to the native enzyme demonstrated the specific localization of limonene synthase to the leucoplasts of peppermint (Mentha × piperita) oil gland secretory cells during the period of essential oil production. Labeling was shown to be absent from all other plastid types examined, including the basal and stalk cell plastids of the secretory phase glandular trichomes. Furthermore, in vitro translation of the preprotein and import experiments with isolated pea chloroplasts were consistent in demonstrating import of the nascent protein to the plastid stroma and proteolytic processing to the mature enzyme at this site. These experiments confirm that the leucoplastidome of the oil gland secretory cells is the exclusive location of limonene synthase, and almost certainly the preceding steps of monoterpene biosynthesis, in peppermint leaves. However, succeeding steps of monoterpene metabolism in mint appear to occur outside the leucoplasts of oil gland cells.

1   This work was supported in part by the U.S. Department of Energy, Division of Energy Biosciences, the National Science Foundation Cell Biology Program, the Mint Industry Research Council, and Project 0268 from the Agricultural Research Center, Washington State University.
2   Present address: Max-Planck-Institut für Chemische Ökologie, D-07745 Jena, Germany.
*   Corresponding author; e-mail croteau{at}mail.wsu.edu; fax 1-509-335-7643.

Plant Physiol. (1999) 120: 879-886
Copyright Clearance Center:   0032-0889/99/120//08
© 1999 American Society of Plant Physiologists




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