Abstract
Artemisinin is currently the best therapeutic against both drug-resistant and cerebral-malaria-causing strains of Plasmodium falciparum. Allene oxide cyclase (AOC) is the key enzyme in the biosynthetic pathway of jasmonates. In this study, a full-length cDNA of AOC gene (named as AaAOC) was cloned from Artemisia annua L. AaAOC was 1,007 bp, containing an open reading frame (750 bp) encoding 249 amino acids. Comparative and bioinformatic analyses revealed that the deduced protein of AaAOC was highly homologous to AOC from other plant species. Phylogenetic analysis indicated that AaAOC was clustered in a closely related subgroup with AOC of Camptotheca acuminata. Southern blot analysis revealed that AaAOC was a multicopy gene. Reverse transcriptase-polymerase chain reaction (RT-PCR) and QPCR analysis showed that AaAOC mRNA accumulated most abundantly in alabastrums, in which the content of artemisinin was previously proven to be the highest. RT-PCR analysis revealed that MeJA, ABA, and ethylene treatments significantly enhanced AaAOC transcript expression. And the results of HPLC showed that the contents of artemisinin were greatly increased after the treatments of ABA and MeJA.
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References
Avanci NC, Luche DD, Goldman GH, Goldman MHS (2010) Jasmonates are phytohormones with multiple functions, including plant defense and reproduction. Genet Mol Res 9:484–505
Baldi A, Dixit VK (2008) Yield enhancement strategies for artemisinin production by suspension cultures of Artemisia annua. Bioresour Technol 99:4609–4614
Bendtsen JD, Nielsen H, von Heijne G, Brunak S (2004) Improved prediction of signal peptides: signalP 3.0. J Mol Biol 340:783–795
Browse J (2009) Jasmonate passes muster: a receptor and targets for the defense hormone. Annu Rev Plant Biol 60:183–205
Comber C, Blanchet C, Geourjon C, Deleage G (2000) NPS@: network protein sequence analysis. Trends Biochem Sci 25:147–150
Covello PS, Teoh KH, Polichuk DR, Reed DW, Nowak G (2007) Functional genomics and the biosynthesis of artemisinin. Phytochemistry 68:1864–1871
Dietrich JA, Yoshikuni Y, Fisher KJ, Woolard FX, Ockey D, McPhee DJ, Renninger NS, Chang MC, Baker D, Keasling JD (2009) A novel semi-biosynthetic route for artemisinin production using engineered substrate-promiscuous P450BM3. ACS Chem Biol 4:261–267
Firestone GL, Sundar SN (2009) Anticancer activities of artemisinin and its bioactive derivatives. Expert Rev Mol Med 11:e32
Graham IA, Besser K, Blumer S, Branigan CA, Czechowski T, Elias L, Guterman I, Harvey D, Isaac PG, Khan AM, Larson TR, Li Y, Pawson T, Penfield T, Rae AM, Rathbone DA, Reid S, Ross J, Smallwood MF, Segura V, Townsend T, Vyas D, Winzer T, Bowles D (2010) The genetic map of Artemisia annua L. Identifies loci affecting yield of the antimalarial drug artemisinin. Science 327:328–331
Guo XX, Yang XQ, Yang RY, Zeng QP (2010) Salicylic acid and methyl jasmonate but not Rose Bengal enhance artemisinin production through invoking burst of endogenous singlet oxygen. Plant Sci 178:390–397
Hause B, Stenzel I, Miersch O, Maucher H, Kramell R, Ziegler J, Wasternack C (2000) Tissue-specific oxylipin signature of tomato flowers: allene oxide cyclase is highly expressed in distinct flower organs and vascular bundles. Plant J 24:113–126
Hofmann E, Zerbe P, Schaller F (2006) The crystal structure of Arabidopsis thaliana allene oxide cyclase: insights into the oxylipin cyclization reaction. Plant Cell 18:3201–3217
Hong GJ, Hu WL, Li JX, Chen XY, Wang LJ (2009) Increased accumulation of artemisinin and anthocyanins in Artemisia annua expressing the Arabidopsis blue light receptor CRY1. Plant Mol Biol Rep 27:334–341
Jiang KJ, Liao ZH, Pi Y, Huang ZS, Hou R, Cao Y, Wang Q, Sun XF, Tang KX (2008) Molecular cloning and expression profile of a jasmonate biosynthetic pathway gene for allene oxide cyclase from Hyoscyamus niger. Mol Biol 42:381–390
Jiang KJ, Pi Y, Hou R, Zeng HN, Huang ZS, Zhang Z, Sun XF, Tang KX (2009) Molecular cloning and expression profiling of the first specific jasmonate, biosynthetic pathway gene allene oxide synthase from Lonicera japonica. Mol Biol Rep 36:487–493
Kong FJ, Li Y, Abe J, Liu B, Schaller F, Piotrowski M, Otagaki S, Takahashi K, Matsuura H, Yoshihara T, Nabeta K (2009) Expression of allene oxide cyclase from Pharbitis nil upon theobroxide treatment. Biosci Biotechnol Biochem 73(5):1007–1013
Korenromp E, Miller J, Nahlen B, Wardlaw T, Young M (2005) World malaria report 2005. World Health Organization (WHO), Roll Back Malaria Partnership, Geneva
Kumar S, Tamura K, Jakobsen IB, Nei M (2001) MEGA2: molecular evolutionary genetics analysis software. Bioinformatics 17:1244–1245
Levin EJ, Kondrashov DA, Wesenberg GE, Phillips GN (2007) Ensemble refinement of protein crystal structures: validation and application. Structure 15:1040–1052
Maucher H, Stenzel I, Miersch O, Stein N, Prasad M, Zierold U, Schweizer P, Dorer C, Hause B, Wasternack C (2004) The allene oxide cyclase of barley (Hordeum vulgare L.)—cloning and organ-specific expression. Phytochemistry 65:801–811
Olsson ME, Olofsson LM, Lindahl AL, Lundgren A, Brodelius M, Brodelius PE (2009) Localization of enzymes of artemisinin biosynthesis to the apical cells of glandular secretory trichomes of Artemisia annua L. Phytochemistry 70:1123–1128
Pi Y, Liao ZH, Jiang KJ, Huang BB, Deng ZX, Zhao DL, Zeng HN, Sun XF, Tang KX (2008) Molecular cloning, characterization and expression of a jasmonate biosynthetic pathway gene encoding allene oxide cyclase from Camptotheca acuminate. Biosci Rep 28:349–355
Putalun W, Luealon W, De-Eknamkul W, Tanaka H, Shoyama Y (2007) Improvement of artemisinin production by chitosan in hairy. Biotechnol Lett 29:1143–1146
Saitou N, Nei M (1987) The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 4:406–425
Wang HH, Ma CF, Li ZQ, Ma LQ, Wang H, Ye HC, Xu GW, Liu BY (2010) Effects of exogenous methyl jasmonate on artemisinin biosynthesis and secondary metabolites in Artemisia annua L. Ind Crops Prod 31:214–218
Weathers PJ, Elkholy S, Wobbe KK (2006) Artemisinin: the biosynthetic pathway and its regulation in Artemisia annua, a terpenoid-rich species. In Vitro Cell Dev Biol, Plant 42:309–317
Zhang L, Jing FY, Li FP, Li MY, Wang YL, Wang GF, Sun XF, Tang KX (2009) Development of transgenic Artemisia annua (Chinese wormwood) plants with an enhanced content of artemisinin, an effective anti-malarial drug, by hairpin-RNA-mediated gene silencing. Biotechnol Appl Biochem 52:199–207
Ziegler J, Stenzel I, Hause B, Maucher H, Miersch O, Hamberg M, Grimm M, Ganal M, Wasternack C (2000) Molecular cloning of allene oxide cyclase—the enzyme establishing the stereochemistry of octadecanoids and jasmonates. J Biol Chem 275:19132–19138
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This work was funded by China “973” Program, China “863” Program, Ministry of Education, Shanghai Science and Technology Committee and Shanghai Leading Academic Discipline Project (B209).
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Lu, X., Lin, X., Shen, Q. et al. Characterization of the Jasmonate Biosynthetic Gene Allene Oxide Cyclase in Artemisia annua L., Source of the Antimalarial Drug Artemisinin. Plant Mol Biol Rep 29, 489–497 (2011). https://doi.org/10.1007/s11105-010-0252-7
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DOI: https://doi.org/10.1007/s11105-010-0252-7