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Cloning and characterization of a monoterpene synthase gene from flowers of Camelina sativa

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A Correction to this article was published on 13 November 2017

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Abstract

Main conclusion

CsTPS1 encodes for a monoterpene synthase that contributes to the emission of a blend of volatile compounds emitted from flowers of Camelina sativa.

The work describes the in vitro characterization of a monoterpene synthase and its regulatory region that we cloned from Camelina sativa (Camelina). Here, we named this gene as C. sativa terpene synthase 1 (CsTPS1). In vitro experiments performed with the CsTPS1 protein after expression and purification from Escherichia coli (E. coli) showed production of a blend of monoterpene volatile organic compounds, of which the emission was also detected in the floral bouquet of wild-type Camelina plants. Quantitative-PCR measurements revealed a high abundance of CsTPS1 transcripts in flowers and experiments performed with the GUS reporter showed high CsTPS1 expression in the pistil, in the cells of the wall of the ovary and in the stigma. Subcellular localization of the CsTPS1 protein was investigated with a GFP reporter construct that showed expression in plastids. The CsTPS1 gene identified in this study belongs to a mid-size family of 60 genes putatively codifying for TPS enzymes. This enlarged family of TPS genes suggests that Camelina has the structural framework for the production of terpenes and other secondary metabolites of relevance for the consumers.

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  • 13 November 2017

    In the original publication, the order of figures and citations was incorrect. The corrections are listed below:

Abbreviations

GPP:

Geranyl pyrophosphate

VOCs:

Volatiles organic compounds

TPS:

Terpene synthase

PTP:

Plastid targeting peptide

MVA:

Mevalonate

MEP:

2-C-Methyl-d-erythritol 4-phosphate

References

  • Aharoni A, Jongsma MA, Bouwmeester HJ (2005) Volatile science? Metabolic engineering of terpenoids in plants. Trends Plant Sci 10(12):594–602

    Article  CAS  PubMed  Google Scholar 

  • Aleklett K, Hart M, Shade A (2014) The microbial ecology of flowers: an emerging frontier in phyllosphere research. Botany 92(4):253–266

    Article  Google Scholar 

  • Aubourg S, Lecharny A, Bohlmann J (2002) Genomic analysis of the terpenoid synthase (AtTPS) gene family of Arabidopsis thaliana. Mol Genet Genom 267(6):730–745. doi:10.1007/s00438-002-0709-y

    Article  CAS  Google Scholar 

  • Augustin JM, Higashi Y, Feng X, Kutchan TM (2015) Production of mono- and sesquiterpenes in Camelina sativa oilseed. Planta 242(3):693–708. doi:10.1007/s00425-015-2367-4

    Article  CAS  PubMed  Google Scholar 

  • Blande JD, Glinwood R (2016) Deciphering chemical language of plant communication. Springer International Publishing, Switzerland

    Book  Google Scholar 

  • Boachon B, Junker RR, Miesch L, Bassard J-E, Höfer R, Caillieaudeaux R, Seidel DE, Lesot A, Heinrich C, Ginglinger J-F (2015) CYP76C1 (Cytochrome P450)-mediated linalool metabolism and the formation of volatile and soluble linalool oxides in Arabidopsis flowers: a strategy for defense against floral antagonists. Plant Cell 27(10):2972–2990

    CAS  PubMed  PubMed Central  Google Scholar 

  • Bohlmann J, Martin D, Oldham NJ, Gershenzon J (2000) Terpenoid secondary metabolism in Arabidopsis thaliana: cDNA cloning, characterization, and functional expression of a myrcene/(E)-beta-ocimene synthase. Arch Biochem Biophys 375(2):261–269. doi:10.1006/abbi.1999.1669

    Article  CAS  PubMed  Google Scholar 

  • Borghi M, Xie DY (2016) Tissue-specific production of limonene in Camelina sativa with the Arabidopsis promoters of genes BANYULS and FRUITFULL. Planta 243(2):549–561. doi:10.1007/s00425-015-2425-y

    Article  CAS  PubMed  Google Scholar 

  • Borghi M, Fernie AR, Schiestl FP, Bouwmeester HJ (2017) The sexual advantage of looking, smelling, and tasting good: the metabolic network that produces signals for pollinators. Trends Plant Sci 22(4):338–350

    Article  CAS  PubMed  Google Scholar 

  • Chen F, Tholl D, D’Auria JC, Farooq A, Pichersky E, Gershenzon J (2003) Biosynthesis and emission of terpenoid volatiles from Arabidopsis flowers. Plant Cell 15(2):481–494

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Chen F, Ro D-K, Petri J, Gershenzon J, Bohlmann J, Pichersky E, Tholl D (2004) Characterization of a root-specific Arabidopsis terpene synthase responsible for the formation of the volatile monoterpene 1, 8-cineole. Plant Physiol 135(4):1956–1966

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Chen F, Tholl D, Bohlmann J, Pichersky E (2011) The family of terpene synthases in plants: a mid-size family of genes for specialized metabolism that is highly diversified throughout the kingdom. Plant J Cell Mol Biol 66(1):212–229. doi:10.1111/j.1365-313X.2011.04520.x

    Article  CAS  Google Scholar 

  • Corpet F (1988) Multiple sequence alignment with hierarchical clustering. Nucleic Acids Res 16(22):10881–10890

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Dalal J, Lopez H, Vasani NB, Hu Z, Swift JE, Yalamanchili R, Dvora M, Lin X, Xie D, Qu R (2015) A photorespiratory bypass increases plant growth and seed yield in biofuel crop Camelina sativa. Biotechnol Biofuels 8(1):175

    Article  PubMed  PubMed Central  Google Scholar 

  • Eberle CA, Thom MD, Nemec KT, Forcella F, Lundgren JG, Gesch RW, Riedell WE, Papiernik SK, Wagner A, Peterson DH (2015) Using pennycress, camelina, and canola cash cover crops to provision pollinators. Ind Crops Prod 75:20–25

    Article  Google Scholar 

  • Edgar RC (2004) MUSCLE: multiple sequence alignment with high accuracy and high throughput. Nucleic Acids Res 32(5):1792–1797

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Emanuelsson O, Nielsen H, Von Heijne G (1999) ChloroP, a neural network-based method for predicting chloroplast transit peptides and their cleavage sites. Protein Sci 8(5):978–984

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Falara V, Akhtar TA, Nguyen TT, Spyropoulou EA, Bleeker PM, Schauvinhold I, Matsuba Y, Bonini ME, Schilmiller AL, Last RL, Schuurink RC, Pichersky E (2011) The tomato terpene synthase gene family. Plant Physiol 157(2):770–789. doi:10.1104/pp.111.179648

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Faure J-D, Tepfer M (2016) Camelina, a Swiss knife for plant lipid biotechnology. OCL 23(5):D503

    Article  Google Scholar 

  • Groeneveld JH, Klein AM (2014) Pollination of two oil producing plant species: Camelina (Camelina sativa L. Crantz) and pennycress (Thlaspi arvense L.) double cropping in Germany. Gcb Bioenergy 6(3):242–251

    Article  Google Scholar 

  • Gutensohn M, Nagegowda DA, Dudareva N (2012) Involvement of compartmentalization in monoterpene and sesquiterpene biosynthesis in plants. Isoprenoid synthesis in plants and microorganisms. Springer, Berlin, pp 155–169

    Chapter  Google Scholar 

  • Heilmann M, Iven T, Ahmann K, Hornung E, Stymne S, Feussner I (2012) Production of wax esters in plant seed oils by oleosomal cotargeting of biosynthetic enzymes. J Lipid Res 53(10):2153–2161. doi:10.1194/jlr.M029512

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Horton P, Park K-J, Obayashi T, Fujita N, Harada H, Adams-Collier C, Nakai K (2007) WoLF PSORT: protein localization predictor. Nucleic acids Res 35(suppl_2):W585–W587

    Article  PubMed  PubMed Central  Google Scholar 

  • Hutcheon JA, Chiolero A, Hanley JA (2010) Random measurement error and regression dilution bias. BMJ 340:c2289

    Article  PubMed  Google Scholar 

  • Jia Q, Li G, Köllner TG, Fu J, Chen X, Xiong W, Crandall-Stotler BJ, Bowman JL, Weston DJ, Zhang Y (2016) Microbial-type terpene synthase genes occur widely in nonseed land plants, but not in seed plants. Proc Natl Acad Sci 113(43):12328–12333

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ju S, Go YS, Choi HJ, Park JM, Suh MC (2017) DEWAX transcription factor is involved in resistance to Botrytis cinerea in Arabidopsis thaliana and Camelina sativa. Front Plant Sci 8:1210

    Article  PubMed  PubMed Central  Google Scholar 

  • Junker RR, Tholl D (2013) Volatile organic compound mediated interactions at the plant-microbe interface. J Chem Ecol 39(7):810–825

    Article  CAS  PubMed  Google Scholar 

  • Kagale S, Koh C, Nixon J, Bollina V, Clarke WE, Tuteja R, Spillane C, Robinson SJ, Links MG, Clarke C (2014) The emerging biofuel crop Camelina sativa retains a highly undifferentiated hexaploid genome structure. Nat Commun 5:3706

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Källberg M, Wang H, Wang S, Peng J, Wang Z, Lu H, Xu J (2012) Template-based protein structure modeling using the RaptorX web server. Nat Protoc 7(8):1511–1522

    Article  PubMed  PubMed Central  Google Scholar 

  • Keeling CI, Weisshaar S, Ralph SG, Jancsik S, Hamberger B, Dullat HK, Bohlmann J (2011) Transcriptome mining, functional characterization, and phylogeny of a large terpene synthase gene family in spruce (Picea spp.). BMC Plant Biol 11:43. doi:10.1186/1471-2229-11-43

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Korbie DJ, Mattick JS (2008) Touchdown PCR for increased specificity and sensitivity in PCR amplification. Nat Protoc 3(9):1452–1456

    Article  CAS  PubMed  Google Scholar 

  • Kram BW, Carter CJ (2009) Arabidopsis thaliana as a model for functional nectary analysis. Sex Plant Reprod 22(4):235–246

    Article  PubMed  Google Scholar 

  • Kumar S, Nei M, Dudley J, Tamura K (2008) MEGA: a biologist-centric software for evolutionary analysis of DNA and protein sequences. Brief Bioinform 9(4):299–306

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Lardizabal KD, Metz JG, Sakamoto T, Hutton WC, Pollard MR, Lassner MW (2000) Purification of a jojoba embryo wax synthase, cloning of its cDNA, and production of high levels of wax in seeds of transgenic arabidopsis. Plant Physiol 122(3):645–655

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Lee S, Chappell J (2008) Biochemical and genomic characterization of terpene synthases in Magnolia grandiflora. Plant Physiol 147(3):1017–1033

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Li G, Köllner TG, Yin Y, Jiang Y, Chen H, Xu Y, Gershenzon J, Pichersky E, Chen F (2012) Nonseed plant Selaginella moellendorffii has both seed plant and microbial types of terpene synthases. Proc Natl Acad Sci 109(36):14711–14715

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Liu YG, Mitsukawa N, Oosumi T, Whittier RF (1995) Efficient isolation and mapping of Arabidopsis thaliana T-DNA insert junctions by thermal asymmetric interlaced PCR. Plant J 8(3):457–463

    Article  CAS  PubMed  Google Scholar 

  • Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods 25(4):402–408

    Article  CAS  PubMed  Google Scholar 

  • Lu C, Kang J (2008) Generation of transgenic plants of a potential oilseed crop Camelina sativa by Agrobacterium-mediated transformation. Plant Cell Rep 27(2):273–278

    Article  CAS  PubMed  Google Scholar 

  • Lücker J, Schwab W, van Hautum B, Blaas J, van der Plas LH, Bouwmeester HJ, Verhoeven HA (2004) Increased and altered fragrance of tobacco plants after metabolic engineering using three monoterpene synthases from lemon. Plant Physiol 134(1):510–519

    Article  PubMed  PubMed Central  Google Scholar 

  • Magnard JL, Roccia A, Caissard JC, Vergne P, Sun P, Hecquet R, Dubois A, Hibrand-Saint Oyant L, Jullien F, Nicole F, Raymond O, Huguet S, Baltenweck R, Meyer S, Claudel P, Jeauffre J, Rohmer M, Foucher F, Hugueney P, Bendahmane M, Baudino S (2015) PLANT VOLATILES. Biosynthesis of monoterpene scent compounds in roses. Science 349(6243):81–83. doi:10.1126/science.aab0696

    Article  CAS  PubMed  Google Scholar 

  • Malik MR, Yang W, Patterson N, Tang J, Wellinghoff RL, Preuss ML, Burkitt C, Sharma N, Ji Y, Jez JM, Peoples OP, Jaworski JG, Cahoon EB, Snell KD (2015) Production of high levels of poly-3-hydroxybutyrate in plastids of Camelina sativa seeds. Plant Biotechnol J 13(5):675–688. doi:10.1111/pbi.12290

    Article  CAS  PubMed  Google Scholar 

  • Martin DM, Aubourg S, Schouwey MB, Daviet L, Schalk M, Toub O, Lund ST, Bohlmann J (2010) Functional annotation, genome organization and phylogeny of the grapevine (Vitis vinifera) terpene synthase gene family based on genome assembly, FLcDNA cloning, and enzyme assays. BMC Plant Biol 10:226. doi:10.1186/1471-2229-10-226

    Article  PubMed  PubMed Central  Google Scholar 

  • Nguyen HT, Park H, Koster KL, Cahoon RE, Nguyen HT, Shanklin J, Clemente TE, Cahoon EB (2015) Redirection of metabolic flux for high levels of omega-7 monounsaturated fatty acid accumulation in camelina seeds. Plant Biotechnol J 13(1):38–50. doi:10.1111/pbi.12233

    Article  CAS  PubMed  Google Scholar 

  • Nützmann HW, Huang A, Osbourn A (2016) Plant metabolic clusters—from genetics to genomics. New Phytol 211(3):771–789

    Article  PubMed  PubMed Central  Google Scholar 

  • Raguso RA (2016) More lessons from linalool: insights gained from a ubiquitous floral volatile. Curr Opin Plant Biol 32:31–36

    Article  CAS  PubMed  Google Scholar 

  • Roy Choudhury S, Riesselman AJ, Pandey S (2014) Constitutive or seed-specific overexpression of Arabidopsis G-protein gamma subunit 3 (AGG3) results in increased seed and oil production and improved stress tolerance in Camelina sativa. Plant Biotechnol J 12(1):49–59. doi:10.1111/pbi.12115

    Article  CAS  PubMed  Google Scholar 

  • Small I, Peeters N, Legeai F, Lurin C (2004) Predotar: a tool for rapidly screening proteomes for N-terminal targeting sequences. Proteomics 4(6):1581–1590

    Article  CAS  PubMed  Google Scholar 

  • Sparkes IA, Runions J, Kearns A, Hawes C (2006) Rapid, transient expression of fluorescent fusion proteins in tobacco plants and generation of stably transformed plants. Nat Protoc 1(4):2019–2025

    Article  CAS  PubMed  Google Scholar 

  • Tholl D (2006) Terpene synthases and the regulation, diversity and biological roles of terpene metabolism. Curr Opin Plant Biol 9(3):297–304. doi:10.1016/j.pbi.2006.03.014

    Article  CAS  PubMed  Google Scholar 

  • Tholl D (2015) Biosynthesis and biological functions of terpenoids in plants. Biotechnology of isoprenoids. Springer International Publishing, Switzerland, pp 63–106

    Chapter  Google Scholar 

  • Tholl D, Lee S (2011) Terpene specialized metabolism in Arabidopsis thaliana. Arabidopsis Book Am Soc Plant Biol 9:e0143. doi:10.1199/tab.0143

    Article  Google Scholar 

  • Usher S, Han L, Haslam RP, Michaelson LV, Sturtevant D, Aziz M, Chapman KD, Sayanova O, Napier JA (2017) Tailoring seed oil composition in the real world: optimising omega-3 long chain polyunsaturated fatty acid accumulation in transgenic Camelina sativa. Sci Rep 7:6570

    Article  PubMed  PubMed Central  Google Scholar 

  • Wei G, Tian P, Zhang F, Qin H, Miao H, Chen Q, Hu Z, Cao L, Wang M, Gu X (2016) Integrative analyses of non-targeted volatile profiling and transcriptome data provide molecular insight into VOC diversity in cucumber plants (Cucumis sativus L.). Plant Physiol 172:603–618

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Winter D, Vinegar B, Nahal H, Ammar R, Wilson GV, Provart NJ (2007) An “Electronic Fluorescent Pictograph” browser for exploring and analyzing large-scale biological data sets. PloS one 2(8):e718

    Article  PubMed  PubMed Central  Google Scholar 

  • Xi J, Rossi L, Lin X, Xie DY (2016) Overexpression of a synthetic insect-plant geranyl pyrophosphate synthase gene in Camelina sativa alters plant growth and terpene biosynthesis. Planta 244(1):215–230. doi:10.1007/s00425-016-2504-8

    Article  CAS  PubMed  Google Scholar 

  • Yamada Y, Kuzuyama T, Komatsu M, Shin-Ya K, Omura S, Cane DE, Ikeda H (2015) Terpene synthases are widely distributed in bacteria. Proc Natl Acad Sci USA 112(3):857–862. doi:10.1073/pnas.1422108112

    Article  CAS  PubMed  Google Scholar 

  • Yuan L, Mao X, Zhao K, Ji X, Ji C, Xue J, Li R (2017) Characterisation of phospholipid: diacylglycerol acyltransferases (PDATs) from Camelina sativa and their roles in stress responses. Biol Open 6(7):1024–1034

    Article  PubMed  PubMed Central  Google Scholar 

  • Zhang Y, Yu L, Yung K-F, Leung DY, Sun F, Lim BL (2012) Over-expression of AtPAP2 in Camelina sativa leads to faster plant growth and higher seed yield. Biotechnol Biofuels 5(1):1

    Article  Google Scholar 

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Acknowledgements

We are grateful to Jing Xi for providing the plasmid containing the sequence of the small subunit of the RuBisco and to Eva Johannes for her support with the confocal analysis. This research was supported by the Advanced Research Projects Agency-Energy (ARPA-E Grant no. 554667-06858 to De-Yu Xie).

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Correspondence to De-Yu Xie.

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The original version of this article was revised: In the original publication, the order of figures and citations was incorrect and it has been corrected now.

A correction to this article is available online at https://doi.org/10.1007/s00425-017-2810-9.

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Borghi, M., Xie, DY. Cloning and characterization of a monoterpene synthase gene from flowers of Camelina sativa . Planta 247, 443–457 (2018). https://doi.org/10.1007/s00425-017-2801-x

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