Biochemical characterization of allene oxide synthases from the liverwort Marchantia polymorpha and green microalgae Klebsormidium flaccidum provides insight into the evolutionary divergence of the plant CYP74 family

Abstract

Main conclusion

Allene oxide synthases (AOSs) were isolated from liverworts and charophytes. These AOSs exhibited enzymatic properties similar to those of angiosperms but formed a distinct phylogenetic clade.

Allene oxide synthase (AOS) and hydroperoxide lyase (HPL) mediate the formation of precursors of jasmonates and carbon-six volatiles, respectively. AOS and HPL utilize fatty acid hydroperoxides and belong to the plant cytochrome P450 74 (CYP74) family that mediates plant defense against herbivores, pathogens, or abiotic stresses. Although members of the CYP74 family have been reported in mosses and other species, the evolution and function of multiple CYP74 genes in plants remain elusive. Here, we show that the liverwort Marchantia polymorpha belongs to a basal group in the evolution of land plants; has two closely related proteins (59 % identity), MpAOS1 and MpAOS2, that are similar to moss PpAOS1 (49 and 47 % identity, respectively); and exhibits AOS activity but not HPL activity. We also found that the green microalgae Klebsormidium flaccidum, consist of multicellular and non-branching filaments, contains an enzyme, KfAOS, that is similar to PpAOS1 (37 % identity), and converts 13-hydroperoxide of linolenic acid to 12-oxo-phytodienoic acid in a coupled reaction with allene oxide cyclase. Phylogenetic analysis showed two evolutionarily distinct clusters. One cluster comprised AOS and HPL from charophytic algae, liverworts, and mosses, including MpAOSs and KfAOS. The other cluster was formed by angiosperm CYP74. Our results suggest that plant CYP74 enzymes with AOS, HPL, and divinyl ether synthase activities have arisen multiple times and in the two different clades, which occurred prior to the divergence of the flowering plant lineage.

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Abbreviations

AOS:

Allene oxide synthase

HPL:

Hydroperoxide lyase

DES:

Divinyl ether synthase

OPDA:

12-Oxo-phytodienoic acid

9-HPOD:

9-Hydroperoxy-(E,Z)-10,12-octadecadienoic acid

9-HPOT:

9-Hydroperoxy-(E,Z,Z)-10,12,15-octadecatrienoic acid

13-HPOD:

13-Hydroperoxy-(E,Z)-9,11-octadecadienoic acid

13-HPOT:

13-Hydroperoxy-(E,Z,Z)-9,11,15-octadecatrienoic acid

12-HPETE:

12-Hydroperoxy-(Z,Z,E,Z)-5,8,10,14-eicosatetraenoic acid

15-HPETE:

15-Hydroperoxy-(Z,Z,Z,E)-5,8,11,13-eicosatetraenoic acid

15-HPEPE:

15-Hydroperoxy-(Z,Z,Z,E,Z)-5,8,11,13,17-eicosapentaenoic acid

HPO:

Hydroperoxide

CYP74:

Cytochrome P450 family 74

JA:

Jasmonic acid

JA-Ile:

Jasmonic acid-isoleucine

EST:

Expressed sequence tag

References

  1. Aleem AM, Jankun J, Dignam JD, Walther M, Kühn H, Svergun DI, Skrzypczak-Jankun E (2008) Human platelet 12-lipoxygenase, new findings about its activity, membrane binding and low-resolution structure. J Mol Biol 376:193–209

    CAS  Article  PubMed  Google Scholar 

  2. Arimura G, Matsui K, Takabayashi J (2009) Chemical and molecular ecology of herbivore-induced plant volatiles: proximate factors and their ultimate functions. Plant Cell Physiol 50:911–923

    CAS  Article  PubMed  Google Scholar 

  3. Buse T, Ruess L, Filser J (2013) New trophic biomarkers for Collembola reared on algal diets. Pedobiologia 56:153–159

    Article  Google Scholar 

  4. Fauconnier ML, Marlier M (1996) An efficient procedure for the production of fatty acid hydroperoxides from hydrolyzed flax seed oil and soybean lipoxygenase. Biotechnol Tech 10:839–844

    CAS  Article  Google Scholar 

  5. Galliard T, Phillips DR (1971) Lipoxygenase from potato tubers. Partial purification and properties of an enzyme that specifically oxygenates the 9-position of linoleic acid. Biochem J 124:431–438

    PubMed Central  CAS  Article  PubMed  Google Scholar 

  6. Gogolev YV, Gorina SS, Gogoleva NE, Toporkova YY, Chechetkin IR, Grechkin AN (2012) Green leaf divinyl ether synthase: gene detection, molecular cloning and identification of a unique CYP74B subfamily member. Biochim Biophys Acta 1821:287–294

    CAS  Article  PubMed  Google Scholar 

  7. Holopainen JK, Blande JD (2012) Molecular plant volatile communication. Adv Exp Med Biol 739:17–31

    CAS  Article  PubMed  Google Scholar 

  8. Hori K, Maruyama F, Fujisawa T, Togashi T, Yamamoto N, Seo M, Sato S, Yamada T, Mori H, Tajima N, Moriyama T, Ikeuchi M, Watanabe M, Wada H, Kobayashi K, Saito M, Masuda T, Sasaki-Sekimoto Y, Mashiguchi K, Awai K, Shimojima M, Masuda S, Iwai M, Nobusawa T, Narise T, Kondo S, Saito H, Sato R, Murakawa M, Ihara Y, Oshima-Yamada Y, Ohtaka K, Satoh M, Sonobe K, Ishii M, Ohtani R, Kanamori-Sato M, Honoki R, Miyazaki D, Mochizuki H, Umetsu J, Higashi K, Shibata D, Kamiya Y, Sato N, Nakamura Y, Tabata S, Ida S, Kurokawa K, Ohta H (2014) Klebsormidium flaccidum genome reveals primary factors for plant terrestrial adaptation. Nat Commun 5:3978

    PubMed Central  CAS  Article  PubMed  Google Scholar 

  9. Hughes RK, Belfield EJ, Ashton R, Fairhurst SA, Göbel C, Stumpe M, Feussner I, Casey R (2006) Allene oxide synthase from Arabidopsis thaliana (CYP74A1) exhibits dual specificity that is regulated by monomer-micelle association. FEBS Lett 580:4188–4194

    CAS  Article  PubMed  Google Scholar 

  10. Ishizaki K, Chiyoda S, Yamato KT, Kohchi T (2008) Agrobacterium-mediated transformation of the haploid liverwort Marchantia polymorpha L., an emerging model for plant biology. Plant Cell Physiol 49:1084–1091

    CAS  Article  PubMed  Google Scholar 

  11. Kajikawa M, Matsui K, Ochiai M, Tanaka Y, Kita Y, Ishimoto M, Kohzu Y, Shoji S, Yamato KT, Ohyama K, Fukuzawa H, Kohchi T (2008) Production of arachidonic and eicosapentaenoic acids in plants using bryophyte fatty acid Δ6-desaturase, Δ6-elongase, and Δ5-desaturase genes. Biosci Biotechnol Biochem 72:435–444

    CAS  Article  PubMed  Google Scholar 

  12. Kanamoto H, Takemura M, Ohyama K (2012) Cloning and expression of three lipoxygenase genes from liverwort, Marchantia polymorpha L., in Escherichia coli. Phytochemistry 77:70–78

    CAS  Article  PubMed  Google Scholar 

  13. Katoh K, Standley DM (2013) MAFFT multiple sequence alignment software version 7: improvements in performance and usability. Mol Biol Evol 30:772–780

    PubMed Central  CAS  Article  PubMed  Google Scholar 

  14. Kihara H, Tanaka M, Yamato KT, Horibata A, Yamada A, Kita S, Ishizaki K, Kajikawa M, Fukuzawa H, Kohchi T, Akakabe Y, Matsui K (2014) Arachidonic acid-dependent carbon-eight volatile synthesis from wounded liverwort (Marchantia polymorpha). Phytochemistry 107:42–49

    CAS  Article  PubMed  Google Scholar 

  15. Kishimoto K, Matsui K, Ozawa R, Takabayashi J (2007) Volatile 1-octen-3-ol induces a defensive response in Arabidopsis thaliana. J Gen Plant Pathol 73:35–37

    CAS  Article  Google Scholar 

  16. Koeduka T, Stumpe M, Matsui K, Kajiwara T, Feussner I (2003) Kinetics of barley FA hydroperoxide lyase are modulated by salts and detergents. Lipids 38:1167–1172

    CAS  Article  PubMed  Google Scholar 

  17. Koeduka T, Kajiwara T, Matsui K (2007) Cloning of lipoxygenase genes from a cyanobacterium, Nostoc punctiforme, and its expression in Eschelichia coli. Curr Microbiol 54:315–319

    CAS  Article  PubMed  Google Scholar 

  18. Koeduka T, Louie GV, Orlova I, Kish CM, Ibdah M, Wilkerson CG, Bowman ME, Baiga TJ, Noel JP, Dudareva N, Pichersky E (2008) The multiple phenylpropene synthases in both Clarkia breweri and Petunia hybrida represent two distinct protein lineages. Plant J 54:362–374

    PubMed Central  CAS  Article  PubMed  Google Scholar 

  19. Kongrit D, Jisaka M, Iwanaga C, Yokomichi H, Katsube T, Nishimura K, Nagaya T, Yokota K (2007) Molecular cloning and functional expression of soybean allene oxide synthases. Biosci Biotechnol Biochem 71:491–498

    CAS  Article  PubMed  Google Scholar 

  20. Lang I, Feussner I (2007) Oxylipin formation in Nostoc punctiforme (PCC73102). Phytochemistry 68:347–357

    Article  Google Scholar 

  21. Laudert D, Pfannschmidt U, Lottspeich F, Holländer-Czytko H, Weiler EW (1996) Cloning, molecular and functional characterization of Arabidopsis thaliana allene oxide synthase (CYP 74), the first enzyme of the octadecanoid pathway to jasmonates. Plant Mol Biol 31:323–335

    CAS  Article  PubMed  Google Scholar 

  22. Lee DS, Nioche P, Hamberg M, Raman CS (2008) Structural insights into the evolutionary paths of oxylipin biosynthetic enzymes. Nature 455:363–368

    CAS  Article  PubMed  Google Scholar 

  23. Matsui K, Ujita C, Fujimoto S, Wilkinson J, Hiatt B, Knauf V, Kajiwara T, Feussner I (2000a) Fatty acid 9- and 13-hydroperoxide lyases from cucumber. FEBS Lett 481:183–188

    CAS  Article  PubMed  Google Scholar 

  24. Matsui K, Miyahara C, Wilkinson J, Hiatt B, Knauf V, Kajiwara T (2000b) Fatty acid hydroperoxide lyase in tomato fruits: cloning and properties of a recombinant enzyme expressed in Escherichia coli. Biosci Biotechnol Biochem 64:1189–1196

    CAS  Article  PubMed  Google Scholar 

  25. Mwenda CM, Matsuki A, Nishimura K, Koeduka T, Matsui K (2015) Spatial expression of the Arabidopsis hydroperoxide lyase gene is controlled differently form that of the allene oxide synthase gene. J Plant Interact 10:1–10

    CAS  Article  Google Scholar 

  26. Ronquist F, Teslenko M, van der Mark P, Ayres DL, Darling A, Höhna S, Larget B, Liu L, Suchard MA, Huelsenbeck JP (2012) MrBayes 3.2: efficient Bayesian phylogenetic inference and model choice across a large model space. Syst Biol 61:539–542

    PubMed Central  Article  PubMed  Google Scholar 

  27. Scalschi L, Sanmartín M, Camañes G, Troncho P, Sánchez-Serrano JJ, García-Agustín P, Vicedo B (2015) Silencing of OPR3 in tomato reveals the role of OPDA in callose deposition during the activation of defense responses against Botrytis cinerea. Plant J 81:304–315

    CAS  Article  PubMed  Google Scholar 

  28. Scholz J, Brodhun F, Hornung E, Herrfurth C, Stumpe M, Beike AK, Faltin B, Frank W, Reski R, Feussner I (2012) Biosynthesis of allene oxides in Physcomitrella patens. BMC Plant Biol 12:228

    PubMed Central  CAS  Article  PubMed  Google Scholar 

  29. Shinmen Y, Katoh K, Shimizu S, Jareonkitmongkol S, Yamada H (1991) Production of arachidonic acid and eicosapentaenoic acids by Marchantia polymorpha in cell culture. Phytochemistry 30:3255–3260

    CAS  Article  Google Scholar 

  30. Splivallo R, Novero M, Bertea CM, Bossi S, Bonfante P (2007) Truffle volatiles inhibit growth and induce an oxidative burst in Arabidopsis thaliana. New Phytol 175:417–424

    CAS  Article  PubMed  Google Scholar 

  31. Stumpe M, Kandzia R, Göbel C, Rosahl S, Feussner I (2001) A pathogen-inducible divinyl ether synthase (CYP74D) from elicitor-treated potato suspension cells. FEBS Lett 507:371–376

    CAS  Article  PubMed  Google Scholar 

  32. Stumpe M, Bode J, Göbel C, Wichard T, Schaaf A, Frank W, Frank M, Reski R, Pohnert G, Feussner I (2006) Biosynthesis of C9-aldehydes in the moss Physcomitrella patens. Biochim Biophys Acta 1761:301–312

    CAS  Article  PubMed  Google Scholar 

  33. Stumpe M, Göbel C, Faltin B, Beike AK, Hause B, Himmelsbach K, Bode J, Kramell R, Wasternack C, Frank W, Reski R, Feussner I (2010) The moss Physcomitrella patens contains cyclopentenones but no jasmonates: mutations in allene oxide cyclase lead to reduced fertility and altered sporophyte morphology. New Phytol 188:740–749

    CAS  Article  PubMed  Google Scholar 

  34. Sugimoto K, Matsui K, Iijima Y, Akakabe Y, Muramoto S, Ozawa R, Uefune M, Sasaki R, Alamgir KM, Akitake S, Nobuke T, Galis I, Aoki K, Shibata D, Takabayashi J (2014) Intake and transformation to a glycoside of (Z)-3-hexenol from infested neighbors reveals a mode of plant odor reception and defense. Proc Natl Acad Sci USA 111:7144–7149

    PubMed Central  CAS  Article  PubMed  Google Scholar 

  35. Taki N, Sasaki-Sekimoto Y, Obayashi T, Kikuta A, Kobayashi K, Ainai T, Yagi K, Sakurai N, Suzuki H, Masuda T, Takamiya K, Shibata D, Kobayashi Y, Ohta H (2005) 12-oxo-phytodienoic acid triggers expression of a distinct set of genes and plays a role in wound-induced gene expression in Arabidopsis. Plant Physiol 139:1268–1283

    PubMed Central  CAS  Article  PubMed  Google Scholar 

  36. Talavera G, Castresana J (2007) Improvement of phylogenies after removing divergent and ambiguously aligned blocks from protein sequence alignments. Syst Biol 56:564–577

    CAS  Article  PubMed  Google Scholar 

  37. Tamura K, Stecher G, Peterson D, Filipski A, Kumar S (2013) MEGA6: molecular evolutionary genetics analysis version 6.0. Mol Biol Evol 30:2725–2729

    PubMed Central  CAS  Article  PubMed  Google Scholar 

  38. Toporkova YY, Gogolev YV, Mukhtarova LS, Grechkin AN (2008) Determinants governing the CYP74 catalysis: conversion of allene oxide synthase into hydroperoxide lyase by site-directed mutagenesis. FEBS Lett 582:3423–3428

    CAS  Article  PubMed  Google Scholar 

  39. Toporkova YY, Ermilova VS, Gorina SS, Mukhtarova LS, Osipova EV, Gogolev YV, Grechkin AN (2013) Structure-function relationship in the CYP74 family: conversion of divinyl ether synthases into allene oxide synthases by site-directed mutagenesis. FEBS Lett 587:2552–2558

    CAS  Article  PubMed  Google Scholar 

  40. Wu J, Hettenhausen C, Meldau S, Baldwin IT (2007) Herbivory rapidly activates MAPK signaling in attacked and unattacked leaf regions but not between leaves of Nicotiana attenuata. Plant Cell 19:1096–1122

    PubMed Central  CAS  Article  PubMed  Google Scholar 

  41. Yamamoto Y, Ohshika J, Takahashi T, Ishizaki K, Kohchi T, Matusuura H, Takahashi K (2015) Functional analysis of allene oxide cyclase, MpAOC, in the liverwort Marchantia polymorpha. Phytochemistry 116:48–56

  42. Yamato KT, Ishizaki K, Fujisawa M, Okada S, Nakayama S, Fujishita M, Bando H, Yodoya K, Hayashi K, Bando T, Hasumi A, Nishio T, Sakata R, Yamamoto M, Yamaki A, Kajikawa M, Yamano T, Nishide T, Choi SH, Shimizu-Ueda Y, Hanajiri T, Sakaida M, Kohno K, Takenaka M, Yamaoka S, Kuriyama C, Kohzu Y, Nishida H, Brennicke A, Shin-i T, Kohara Y, Kohchi T, Fukuzawa H, Ohyama K (2007) Gene organization of the liverwort Y chromosome reveals distinct sex chromosome evolution in a haploid system. Proc Natl Acad Sci USA 104:6472–6477

    PubMed Central  CAS  Article  PubMed  Google Scholar 

  43. Yanagi K, Sugimoto K, Matsui K (2011) Oxylipin-specific cytochrome P450s (CYP74s) in Lotus japonicus: their implications in response to mechanical wounding and nodule formation. J Plant Interact 6:255–264

    CAS  Article  Google Scholar 

  44. Zhu Z, Qian F, Yang R, Chen J, Luo Q, Chen H, Yan X (2015) A lipoxygenase from red alga Pyropia haitanensis, a unique enzyme catalyzing the free radical reactions of polyunsaturated fatty acids with triple ethylenic bonds. PLoS ONE 10:e0117351

    PubMed Central  Article  PubMed  Google Scholar 

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Acknowledgments

We wish to thank Dr. Mitsuo Jisaka and Dr. Kazushige Yokota (Shimane University) for providing the GmAOS construct and human platelet 12-lipoxygenase used in this study. We also thank Maya Tanaka (Yamaguchi University) for preparation of 12-HPETE. This work was supported by JSPS KAKENHI Grant Number 24580162.

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Correspondence to Takao Koeduka or Kenji Matsui.

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Koeduka, T., Ishizaki, K., Mwenda, C.M. et al. Biochemical characterization of allene oxide synthases from the liverwort Marchantia polymorpha and green microalgae Klebsormidium flaccidum provides insight into the evolutionary divergence of the plant CYP74 family. Planta 242, 1175–1186 (2015). https://doi.org/10.1007/s00425-015-2355-8

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Keywords

  • Allene oxide synthase
  • Plant evolution
  • CYP74
  • Hydroperoxide lyase
  • C8 volatiles