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


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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Allene oxide synthase


Hydroperoxide lyase


Divinyl ether synthase


12-Oxo-phytodienoic acid


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


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


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


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


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


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


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




Cytochrome P450 family 74


Jasmonic acid


Jasmonic acid-isoleucine


Expressed sequence tag


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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).

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