title: Identification of genes in Thuja plicata foliar terpenoid defenses.
abstract: Thuja plicata (western redcedar) is a long-lived conifer species whose foliage is rarely affected by disease or insect pests, but can be severely damaged by ungulate browsing. Deterrence to browsing correlates with high foliar levels of terpenoids, in particular the monoterpenoid α-thujone. Here, we set out to identify genes whose products may be involved in the production of α-thujone and other terpenoids in this species. First, we generated a foliar transcriptome database from which to draw candidate genes. Second, we mapped the storage of thujones and other terpenoids to foliar glands. Third, we used global expression profiling to identify more than 600 genes that are expressed at high levels in foliage with glands, but can either not be detected or are expressed at low levels in a natural variant lacking foliar glands. Fourth, we used in situ RNA hybridization to map the expression of a putative monoterpene synthase to the epithelium of glands and used enzyme assays with recombinant protein of the same gene to show that it produces sabinene, the monoterpene precursor of α-thujone. Finally, we identified candidate genes with predicted enzymatic functions for the conversion of sabinene to α-thujone. Taken together, this approach generated both general resources and detailed functional characterization in the identification of genes of foliar terpenoid biosynthesis in T. plicata.
authors: Department of Biological Sciences, Simon Fraser University, Burnaby, British Columbia, Canada V5A 1S6.
authors: Adam J
authors: AJ
authors: Foster
authors: Dawn E
authors: DE
authors: Hall
authors: Leanne
authors: L
authors: Mortimer
authors: Shelley
authors: S
authors: Abercromby
authors: Regine
authors: R
authors: Gries
authors: Gerhard
authors: G
authors: Gries
authors: Jörg
authors: J
authors: Bohlmann
authors: John
authors: J
authors: Russell
authors: Jim
authors: J
authors: Mattsson
mesh_terms: D000081008:Bicyclic Monoterpenes; D053898:Biosynthetic Pathways / Q000235:genetics; D020869:Gene Expression Profiling; D018506:Gene Expression Regulation, Plant; D017343:Genes, Plant / Q000235:genetics*; D056726:Genetic Association Studies; D005838:Genotype; D019753:Intramolecular Lyases / Q000378:metabolism; D008969:Molecular Sequence Data; D039821:Monoterpenes / Q000378:metabolism; D010802:Phylogeny; D018515:Plant Leaves / Q000201:enzymology / Q000235:genetics* / Q000276:immunology*; D060888:Real-Time Polymerase Chain Reaction; D015203:Reproducibility of Results; D012116:Resins, Plant / Q000378:metabolism; D013729:Terpenes / Q000378:metabolism*; D029783:Thuja / Q000033:anatomy & histology / Q000201:enzymology / Q000235:genetics* / Q000276:immunology*; D059467:Transcriptome / Q000235:genetics; D015854:Up-Regulation / Q000235:genetics
chemical_list: D000081008:Bicyclic Monoterpenes; D039821:Monoterpenes; D012116:Resins, Plant; D013729:Terpenes; C005790:beta-thujone; D019753:Intramolecular Lyases; C040265:pinene cyclase I
references: Nat Prod Rep. 2008 Feb;25(1):118-38
references: 18250899
references: Plant Cell Rep. 2009 Jan;28(1):123-33
references: 18854999
references: Nucleic Acids Res. 2008 Jun;36(10):3420-35
references: 18445632
references: Curr Biol. 2006 Oct 10;16(19):1911-7
references: 17027487
references: Plant J. 2001 Jan;25(1):43-53
references: 11169181
references: BMC Genomics. 2010 Dec 01;11:681
references: 21122097
references: Nat Protoc. 2006;1(3):1462-7
references: 17406436
references: J Chem Ecol. 2002 Dec;28(12):2411-27
references: 12564790
references: J Integr Plant Biol. 2010 Jan;52(1):86-97
references: 20074143
references: Plant Physiol. 2005 Jan;137(1):369-82
references: 15618433
references: Biochem Soc Trans. 2006 Dec;34(Pt 6):1209-14
references: 17073787
references: Genomics. 2008 Nov;92(5):255-64
references: 18703132
references: Proc Natl Acad Sci U S A. 2008 Jan 22;105(3):1085-90
references: 18198275
references: Int J Plant Genomics. 2008;2008:619832
references: 18483572
references: BMC Plant Biol. 2011 Mar 07;11:43
references: 21385377
references: J Ultrastruct Res. 1965 Oct;13(3):233-44
references: 5320627
references: Arch Biochem Biophys. 1987 Oct;258(1):287-91
references: 3310901
references: BMC Plant Biol. 2010 Jun 12;10:106
references: 20540781
references: J Chromatogr Sci. 2007 May-Jun;45(5):281-5
references: 17555638
references: Appl Microbiol Biotechnol. 2010 Jan;85(4):1105-14
references: 19820924
references: New Phytol. 2006;170(4):657-75
references: 16684230
references: Naturwissenschaften. 2005 Dec;92(12):562-77
references: 16292524
references: Biotechniques. 2004 May;36(5):821-4
references: 15152602
references: J Chem Ecol. 2002 Jan;28(1):211-28
references: 11868675
references: Arch Biochem Biophys. 1987 Jul;256(1):179-93
references: 3111374
references: Anal Biochem. 2004 Dec 15;335(2):210-7
references: 15556559
references: J Chromatogr. 1963 Aug;11:463-71
references: 14062605
references: Plant J. 2011 Mar;65(6):936-48
references: 21323772
references: Plant Physiol. 2004 Aug;135(4):1908-27
references: 15310829
references: Proc Natl Acad Sci U S A. 2005 May 31;102(22):8060-5
references: 15911762
references: Oecologia. 2001 Jan;126(1):84-93
references: 28547441
references: Trends Plant Sci. 2004 May;9(5):229-35
references: 15130548
references: Crit Rev Food Sci Nutr. 2006;46(5):365-77
references: 16891209
references: J Chromatogr Sci. 2005 May-Jun;43(5):253-8
references: 15975244
references: Genomics. 2010 Dec;96(6):369-76
references: 20828606
references: Plant J. 2009 Dec;60(6):1015-30
references: 19754519
references: Plant Physiol. 2011 Dec;157(4):1677-95
references: 21994349
references: Plant J. 2011 Apr;66(1):212-29
references: 21443633
references: New Phytol. 2006;172(1):47-62
references: 16945088
references: Plant Physiol. 2006 Mar;140(3):1009-21
references: 16415217
references: J Hered. 2002 Sep-Oct;93(5):370-6
references: 12547927