Plant Molecular Biology 51: 119–133, 2003.
© 2003 Kluwer Academic Publishers. Printed in the Netherlands.
Traumatic resin defense in Norway spruce (Picea abies): Methyl
jasmonate-induced terpene synthase gene expression, and cDNA cloning
and functional characterization of (+)-3-carene synthase
Jenny Fäldt, Diane Martin, Barbara Miller, Suman Rawat and Jörg Bohlmann
Biotechnology Laboratory, Department of Botany and Department of Forest Sciences, University of British
Columbia, 6174 University Boulevard, Vancouver V6T 1Z3, B.C. Canada (
author for correspondence; e-mail:
Received 20 January 2002; accepted in revised form 21 May 2002
Key words: 3-carene, chemical plant defense, conifer biotechnology, methyl jasmonate, monoterpene synthase,
Picea abies (Norway spruce)
Picea abies (L.) Karst. (Norway spruce) employs constitutive and induced resin terpenoids as major chemical
and physical defense-shields against insects and pathogens. In recent work, we showed that a suite of terpenoids,
monoterpenoids and diterpenoids was induced in stems of Norway spruce after treatment of trees with methyl
jasmonate (MeJA) (Martin et al., 2002). Increase of enzyme activities of terpenoid biosynthesis and accumulation
of terpenoids was associated with MeJA-induced de novo differentiation of xylem resin ducts. The formation of
defense-related traumatic resin ducts was also found in Norway spruce after attack by stem boring insects or after
infestation with fungal pathogens. In the present study, we analyzed the traumatic resin response in Norway spruce
further at the molecular genetic level. Treatment of trees with MeJA induced transient transcript accumulation of
monoterpenoid synthases and diterpenoid synthases in stem tissues of Norway spruce. In screening for defense-
related terpenoid synthase (TPS) genes from Norway spruce, a full-length monoterpenoidsynthase cDNA, PaJF67,
was isolated and the recombinant enzyme expressed in E. coli and functionally characterized in vitro. The cloned
PaJF67 cDNA represents a new monoterpenoid synthase gene and the gene product was identiﬁed as 3-carene
synthase. The enzyme encoded by PaJF67 forms stereospeciﬁcally (+)-3-carene (78% of total product) together
with minor acyclic and cyclic monoterpenes, including the mechanistically closely related terpinolene (11% of total
product). (+)-3-Carene is a characteristic monoterpene of constitutive and induced oleoresin defense of Norway
spruce and other members of the Pinaceae.
Abbreviations: 2D-GC, two-dimensional gas chromatography; GC, gas chromatography; GPP, geranyl
diphospahte; GGPP, geranylgeranyl diphospahte; FPP, farnesyl diphosphate; LPP, linalyl diphosphate; MeJA,
methyl jasmonate; MS, mass spectroscopy; TPS, terpenoid synthase.
Conifer resins are composed of a large suite of struc-
turally diverse terpenoid natural products that have
long been recognized for their many functions in the
chemical ecology of conifers and coniferophagus in-
sects (Berryman, 1972; Langenheim, 1994; Bohlmann
and Croteau, 1999; Seybold et al., 2000; Trapp
and Croteau, 2001), as well as for their industrial
value (Dawson, 1994; Schrader and Berger, 2001).
The oleoresin of Norway spruce (Picea abies) con-
sists mainly of monoterpenoids(10-carbon), sesquiter-
penoids (15-carbon), and diterpenoids (20-carbon).
Monoterpenoids and diterpenoids are equally abun-
dant and together account for more than 98% of
resin terpenoids in stems of Norway spruce (Martin
et al., 2002). The turpentine fraction of Norway spruce
resin includes more than 20 different monoterpenoids,