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Título : Lotus japonicusTriterpenoid Profile and Characterization oftheCYP716A51andLjCYP93E1Genes Involved in TheirBiosynthesis In Planta
Autor : Suzuki, Hayato
Fukushima, Ery Odette
Shimizu, Yuko
Seki, Hikaru
Fujisawa, Yukiko
Ishimoto, Masao
Osakabe, Keishi
Osakabe, Yuriko
Muranaka, Toshiya
Palabras clave : Betulinic acid
Ursolic acid.
Fecha de publicación : 2019
Editorial : Scopus
Citación : Suzuki, H., Fukushima, E. O., Shimizu, Y., Seki, H., Fujisawa, Y., Ishimoto, M., … Muranaka, T. (2019). Lotus japonicus Triterpenoid Profile and Characterization of the CYP716A51 and LjCYP93E1 Genes Involved in Their Biosynthesis in Planta. Plant and Cell Physiology, 60(11), 2496–2509. doi.org/10.1093/pcp/pcz145
Resumen : Lotus japonicusis an important model legume plant in sev-eral fields of research, such as secondary (specialized) me-tabolism and symbiotic nodulation. This plant accumulatestriterpenoids; however, less information regarding its com-position, content and biosynthesis is available comparedwithMedicago truncatulaandGlycine max. In this study,we analyzed the triterpenoid content and composition ofL. japonicus.Lotus japonicusaccumulated C-28-oxidized tri-terpenoids (ursolic, betulinic and oleanolic acids) and soya-sapogenols (soyasapogenol B, A and E) in a tissue-dependentmanner. We identified an oxidosqualene cyclase (OSC) andtwo cytochrome P450 enzymes (P450s) involved in triter-penoid biosynthesis using a yeast heterologous expressionsystem. OSC9 was the first enzyme derived fromL. japonicusthat showeda-amyrin (a precursor of ursolic acid)-produ-cing activity. CYP716A51 showed triterpenoid C-28oxidation activity. LjCYP93E1 convertedb-amyrin into24-hydroxy-b-amyrin, a metabolic intermediate of soyasapo-genols. The involvement of the identified genes in triterpen-oid biosynthesis inL. japonicusplants was evaluated byquantitative real-time PCR analysis. Furthermore, geneloss-of-function analysis ofCYP716A51andLjCYP93E1wasconducted. Thecyp716a51-mutantL. japonicushairy rootsgenerated by the genome-editing technique produced no C-28 oxidized triterpenoids. Likewise, the complete abolitionof soyasapogenols and soyasaponin I was observed in mutantplants harboringLotus retrotransposon 1(LORE1)inLjCYP93E1. These results indicate that the activities ofthese P450 enzymes are essential for triterpenoid biosynthe-sis inL. japonicus. This study increases our understanding oftriterpenoid biosynthesis in leguminous plants and providesinformation that will facilitate further studies of the physio-logical functions of triterpenoids usingL. japonicus.
URI : https://doi.org/10.1093/pcp/pcz145
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