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Plant Physiol, January 2000, Vol. 122, pp. 215-224
Developmental Regulation of Monoterpene Biosynthesis in the
Glandular Trichomes of Peppermint1
Marie E.
McConkey,
Jonathan
Gershenzon,2 and
Rodney B.
Croteau*
Institute of Biological Chemistry, and Department of Biochemistry
and Biophysics, Washington State University, Pullman, Washington
99164-6340.
Monoterpene
production in peppermint (Mentha × piperita L.) glandular trichomes is determined by the
rate of biosynthesis, as determined by 14CO2
incorporation, and is restricted to leaves 12 to 20 d of age. Using oil glands isolated from peppermint leaves of different ages, in
vitro assay of the eight sequential enzymes responsible for the
biosynthesis of the principal monoterpene ( )-menthol indicated that
all but one biosynthetic enzyme had a very similar developmental
profile. Activities were highest in leaves 12 to 20 d of age, with
a sharp peak centered at 15 d. The exception, ()-menthone
reductase, the last enzyme of the pathway, exhibited a later peak of
activity, which was centered at approximately 21 d. The
correlation between in vitro enzyme activity and the rate of
biosynthesis measured in vivo suggests that monoterpene formation is
controlled mainly by the coordinately regulated activity of the
relevant biosynthetic enzymes. Developmental immunoblotting of limonene
synthase, which catalyzes the committed step of the pathway,
demonstrated a direct correlation between enzyme activity and enzyme
protein, suggesting that the dynamic time course for the remaining
pathway enzyme activities also reflects the corresponding protein
levels. RNA-blot analyses indicated that the genes encoding enzymes of
the early pathway steps are transcriptionally activated in a
coordinated fashion, with a time course superimposible with activity
measurements and immunoblot data. These results demonstrating coincidental temporal changes in enzyme activities, enzyme protein level, and steady-state transcript abundances indicate that most of the
monoterpene biosynthetic enzymes in peppermint are developmentally regulated at the level of gene expression.
1
This work was supported in part by the U.S.
Department of Energy Division of Energy Biosciences, the Mint Industry
Research Council, and project no. 0268 from the Agricultural Research
Center, Washington State University.
2
Present address: Max Planck Institut für
Chemische Ökologie, Tatzendpromenade 1a, D-07745 Jena, Germany.
*
Corresponding author; e-mail croteau{at}mail.wsu.edu; fax
509-335-7643.
© 2000 American Society of Plant Physiologists
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