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Evolution of Conifer Diterpene Synthases: Diterpene Resin Acid Biosynthesis in Lodgepole Pine and Jack Pine Involves Monofunctional and Bifunctional Diterpene Synthases Open Access

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Author or creator
Hall, Dawn E.
Zerbe, Philipp
Jancsik, Sharon
Quesada, Alfonso L.
Dullat, Harpreet
Madilao, Lina L.
Yuen, Macaire
Bohlmann, Jörg
Additional contributors
Subject/Keyword
fir abies-grandis
abietadiene synthase
functional-characterization
kaurene synthase
abietadienol/abietadienal oxidase ptao
specialized metabolism
ent-copalyl diphosphate
(+)-copalyl diphosphate
terpene synthases
spruce picea-sitchensis
Type of item
Journal Article (Published)
Language
english
Place
Time
Description
Diterpene resin acids (DRAs) are major components of pine (Pinus spp.) oleoresin. They play critical roles in conifer defense against insects and pathogens and as a renewable resource for industrial bioproducts. The core structures of DRAs are formed in secondary (i.e. specialized) metabolism via cycloisomerization of geranylgeranyl diphosphate (GGPP) by diterpene synthases (diTPSs). Previously described gymnosperm diTPSs of DRA biosynthesis are bifunctional enzymes that catalyze the initial bicyclization of GGPP followed by rearrangement of a (+)-copalyl diphosphate intermediate at two discrete class II and class I active sites. In contrast, similar diterpenes of gibberellin primary (i.e. general) metabolism are produced by the consecutive activity of two monofunctional class II and class I diTPSs. Using high-throughput transcriptome sequencing, we discovered 11 diTPS from jack pine (Pinus banksiana) and lodgepole pine (Pinus contorta). Three of these were orthologous to known conifer bifunctional levopimaradiene/abietadiene synthases. Surprisingly, two sets of orthologous PbdiTPSs and PcdiTPSs were monofunctional class I enzymes that lacked functional class II active sites and converted (+)-copalyl diphosphate, but not GGPP, into isopimaradiene and pimaradiene as major products. Diterpene profiles and transcriptome sequences of lodgepole pine and jack pine are consistent with roles for these diTPSs in DRA biosynthesis. The monofunctional class I diTPSs of DRA biosynthesis form a new clade within the gymnosperm-specific TPS-d3 subfamily that evolved from bifunctional diTPS rather than monofunctional enzymes (TPS-c and TPS-e) of gibberellin metabolism. Homology modeling suggested alterations in the class I active site that may have contributed to their functional specialization relative to other conifer diTPSs.
Date created
2013
DOI
doi:10.7939/R3930NX71
License information
Rights
© 2013 Hall et al. This version of this article is open access and can be downloaded and shared. The original author(s) and source must be cited.
Citation for previous publication
Hall, D.E., Zerbe, P., Jancsik, S., Lara Quesada, A., Dullat, H., Madilao, L.L., Yuen, M., Bohlmann, J. (2013). Evolution of Conifer Diterpene Synthases: Diterpene Resin Acid Biosynthesis in Lodgepole Pine and Jack Pine Involves Monofunctional and Bifunctional Diterpene Synthases. Plant Physiology, 162(2), 1073-1091. doi:10.1104/pp.112.208546.
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