Isolation and Functional Characterization of a Methyl Jasmonate-responsive 3-carene Synthase from Lavandula X Intermedia

Overview

Journal Plant Mol Biol

Date 2017 Mar 5

PMID 28258552

Citations 8

Authors

Ayelign M Adal

Lukman S Sarker

Ashley D Lemke

Soheil S Mahmoud

Affiliations

Soon will be listed here.

Abstract

A methyl jasmonate responsive 3-carene synthase (Li3CARS) gene was isolated from Lavandula x intermedia and functionally characterized in vitro. Lavenders produce essential oils consisting mainly of monoterpenes, including the potent antimicrobial and insecticidal monoterpene 3-carene. In this study we isolated and functionally characterized a leaf-specific, methyl jasmonate (MeJA)-responsive monoterpene synthase (Li3CARS) from Lavandula x intermedia. The ORF excluding transit peptides encoded a 64.9 kDa protein that was expressed in E. coli, and purified with Ni-NTA agarose affinity chromatography. The recombinant Li3CARS converted GPP into 3-carene as the major product, with K and k of 3.69 ± 1.17 µM and 2.01 s respectively. Li3CARS also accepted NPP as a substrate to produce multiple products including a small amount of 3-carene. The catalytic efficiency of Li3CARS to produce 3-carene was over ten fold higher for GPP (k /K = 0.56 µMs) than NPP (k /K = 0.044 µMs). Production of distinct end product profiles from different substrates (GPP versus NPP) by Li3CARS indicates that monoterpene metabolism may be controlled in part through substrate availability. Li3CARS transcripts were found to be highly abundant in leaves (16-fold) as compared to flower tissues. The transcriptional activity of Li3CARS correlated with 3-carene production, and was up-regulated (1.18- to 3.8-fold) with MeJA 8-72 h post-treatment. The results suggest that Li3CARS may have a defensive role in Lavandula.

Citing Articles

Metabolic engineering of terpene metabolism in lavender.

Oseni O, Sajaditabar R, Mahmoud S Beni Suef Univ J Basic Appl Sci. 2024; 13(1):67.

PMID: 38988370 PMC: 11230991. DOI: 10.1186/s43088-024-00524-7.

Multi-omics analyses provide insights into the evolutionary history and the synthesis of medicinal components of the Chinese wingnut.

Zhang Z, Xia H, Yuan M, Gao F, Bao W, Jin L Plant Divers. 2024; 46(3):309-320.

PMID: 38798724 PMC: 11119516. DOI: 10.1016/j.pld.2024.03.010.

Lavandula Species, Their Bioactive Phytochemicals, and Their Biosynthetic Regulation.

Haban M, Korczyk-Szabo J, certekova S, Razna K Int J Mol Sci. 2023; 24(10).

PMID: 37240177 PMC: 10219037. DOI: 10.3390/ijms24108831.

Expression Profile and Ligand Screening of a Putative Odorant-Binding Protein, AcerOBP6, from the Asian Honeybee.

Zhao H, Peng Z, Huang L, Zhao S, Liu M Insects. 2021; 12(11).

PMID: 34821756 PMC: 8622152. DOI: 10.3390/insects12110955.

Metabolism and transcriptome profiling provides insight into the genes and transcription factors involved in monoterpene biosynthesis of borneol chemotype of induced by mechanical damage.

Yang Z, Xie C, Huang Y, An W, Liu S, Huang S PeerJ. 2021; 9:e11465.

PMID: 34249483 PMC: 8255067. DOI: 10.7717/peerj.11465.

References

van Schie C, Haring M, Schuurink R . Tomato linalool synthase is induced in trichomes by jasmonic acid. Plant Mol Biol. 2007; 64(3):251-63. PMC: 1876254. DOI: 10.1007/s11103-007-9149-8. View

Chen F, Tholl D, Bohlmann J, Pichersky E . 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. 2011; 66(1):212-29. DOI: 10.1111/j.1365-313X.2011.04520.x. View

Dudareva N, Klempien A, Muhlemann J, Kaplan I . Biosynthesis, function and metabolic engineering of plant volatile organic compounds. New Phytol. 2013; 198(1):16-32. DOI: 10.1111/nph.12145. View

Hoelscher D, Williams D, Wildung M, Croteau R . A cDNA clone for 3-carene synthase from Salvia stenophylla. Phytochemistry. 2003; 62(7):1081-6. DOI: 10.1016/s0031-9422(02)00674-x. View

Miller B, Madilao L, Ralph S, Bohlmann J . Insect-induced conifer defense. White pine weevil and methyl jasmonate induce traumatic resinosis, de novo formed volatile emissions, and accumulation of terpenoid synthase and putative octadecanoid pathway transcripts in Sitka spruce. Plant Physiol. 2004; 137(1):369-82. PMC: 548866. DOI: 10.1104/pp.104.050187. View

Heiling S, Schuman M, Schoettner M, Mukerjee P, Berger B, Schneider B . Jasmonate and ppHsystemin regulate key Malonylation steps in the biosynthesis of 17-Hydroxygeranyllinalool Diterpene Glycosides, an abundant and effective direct defense against herbivores in Nicotiana attenuata. Plant Cell. 2010; 22(1):273-92. PMC: 2828710. DOI: 10.1105/tpc.109.071449. View

Schuttelkopf A, van Aalten D . PRODRG: a tool for high-throughput crystallography of protein-ligand complexes. Acta Crystallogr D Biol Crystallogr. 2004; 60(Pt 8):1355-63. DOI: 10.1107/S0907444904011679. View

Wittstock U, Gershenzon J . Constitutive plant toxins and their role in defense against herbivores and pathogens. Curr Opin Plant Biol. 2002; 5(4):300-7. DOI: 10.1016/s1369-5266(02)00264-9. View

Bohlmann J, Croteau R . Plant terpenoid synthases: molecular biology and phylogenetic analysis. Proc Natl Acad Sci U S A. 1998; 95(8):4126-33. PMC: 22453. DOI: 10.1073/pnas.95.8.4126. View

10.

Vickers C, Gershenzon J, Lerdau M, Loreto F . A unified mechanism of action for volatile isoprenoids in plant abiotic stress. Nat Chem Biol. 2009; 5(5):283-91. DOI: 10.1038/nchembio.158. View

11.

Rivera S, Swedlund B, King G, Bell R, Hussey Jr C, Wrobel W . Chrysanthemyl diphosphate synthase: isolation of the gene and characterization of the recombinant non-head-to-tail monoterpene synthase from Chrysanthemum cinerariaefolium. Proc Natl Acad Sci U S A. 2001; 98(8):4373-8. PMC: 31842. DOI: 10.1073/pnas.071543598. View

12.

Frost C, Appel H, Carlson J, De Moraes C, Mescher M, Schultz J . Within-plant signalling via volatiles overcomes vascular constraints on systemic signalling and primes responses against herbivores. Ecol Lett. 2007; 10(6):490-8. DOI: 10.1111/j.1461-0248.2007.01043.x. View

13.

Livak K, Schmittgen T . Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods. 2002; 25(4):402-8. DOI: 10.1006/meth.2001.1262. View

14.

Demissie Z, Cella M, Sarker L, Thompson T, Rheault M, Mahmoud S . Cloning, functional characterization and genomic organization of 1,8-cineole synthases from Lavandula. Plant Mol Biol. 2012; 79(4-5):393-411. DOI: 10.1007/s11103-012-9920-3. View

15.

Demissie Z, Sarker L, Mahmoud S . Cloning and functional characterization of β-phellandrene synthase from Lavandula angustifolia. Planta. 2010; 233(4):685-96. DOI: 10.1007/s00425-010-1332-5. View

16.

Jullien F, Moja S, Bony A, Legrand S, Petit C, Benabdelkader T . Isolation and functional characterization of a τ-cadinol synthase, a new sesquiterpene synthase from Lavandula angustifolia. Plant Mol Biol. 2013; 84(1-2):227-41. DOI: 10.1007/s11103-013-0131-3. View

17.

Rynkiewicz M, Cane D, Christianson D . Structure of trichodiene synthase from Fusarium sporotrichioides provides mechanistic inferences on the terpene cyclization cascade. Proc Natl Acad Sci U S A. 2001; 98(24):13543-8. PMC: 61077. DOI: 10.1073/pnas.231313098. View

18.

Tholl D, Lee S . Terpene Specialized Metabolism in Arabidopsis thaliana. Arabidopsis Book. 2012; 9:e0143. PMC: 3268506. DOI: 10.1199/tab.0143. View

19.

Martin D, Tholl D, Gershenzon J, Bohlmann J . Methyl jasmonate induces traumatic resin ducts, terpenoid resin biosynthesis, and terpenoid accumulation in developing xylem of Norway spruce stems. Plant Physiol. 2002; 129(3):1003-18. PMC: 166496. DOI: 10.1104/pp.011001. View

20.

de Rapper S, Kamatou G, Viljoen A, van Vuuren S . The In Vitro Antimicrobial Activity of Lavandula angustifolia Essential Oil in Combination with Other Aroma-Therapeutic Oils. Evid Based Complement Alternat Med. 2013; 2013:852049. PMC: 3666441. DOI: 10.1155/2013/852049. View