Scopoletin 8-hydroxylase: a novel enzyme involved in coumarin biosynthesis and iron-deficiency responses in Arabidopsis

Joanna Siwińska , Kinga Siatkowska , Alexandre Olry , Jeremy Grosjean , Alain Hehn , Frederic Bourgaud , Andrew A. Meharg , Manus Carey , Ewa Łojkowska , Anna Ihnatowicz


Iron deficiency is a serious agricultural problem, particularly in alkaline soils. Secretion of coumarins by Arabidopsis thaliana roots is induced under iron deficiency. An essential enzyme for the biosynthesis of the major Arabidopsis coumarins, scopoletin and its derivatives, is Feruloyl-CoA 6′-Hydroxylase1 (F6′H1), which belongs to a large enzyme family of the 2-oxoglutarate and Fe2+-dependent dioxygenases. We have functionally characterized another enzyme of this family, which is a close homologue of F6′H1 and is encoded by a strongly iron-responsive gene, At3g12900. We purified At3g12900 protein heterologously expressed in Escherichia coli and demonstrated that it is involved in the conversion of scopoletin into fraxetin, via hydroxylation at the C8 position, and that it thus functions as a scopoletin 8-hydroxylase (S8H). Its function in plant cells was confirmed by the transient expression of S8H protein in Nicotiana benthamiana leaves, followed by metabolite profiling and biochemical and ionomic characterization of Arabidopsis s8h knockout lines grown under various iron regimes. Our results indicate that S8H is involved in coumarin biosynthesis, as part of mechanisms used by plants to assimilate iron.
Author Joanna Siwińska (IFB / IB / DBiotech)
Joanna Siwińska,,
- Department of Biotechnology
, Kinga Siatkowska (IFB)
Kinga Siatkowska,,
- Intercollegiate Faculty of Biotechnology UG
, Alexandre Olry - [Université de Lorraine]
Alexandre Olry,,
, Jeremy Grosjean - [Université de Lorraine]
Jeremy Grosjean,,
, Alain Hehn - [Université de Lorraine]
Alain Hehn,,
, Frederic Bourgaud - [Université de Lorraine]
Frederic Bourgaud,,
, Andrew A. Meharg - [Queen's University Belfast]
Andrew A. Meharg,,
, Manus Carey - [Queen's University Belfast]
Manus Carey,,
, Ewa Łojkowska (IFB / IB / DBiotech)
Ewa Łojkowska,,
- Department of Biotechnology
, Anna Ihnatowicz (IFB / IB / DBiotech)
Anna Ihnatowicz,,
- Department of Biotechnology
Journal seriesJournal of Experimental Botany, ISSN 0022-0957, (A 45 pkt)
Issue year2018
Publication size in sheets0.65
Keywords in Englishabiotic stress, Arabidopsis, enzyme activity, fraxetin, Fe- and 2OG-dependent dioxygenase, mineral nutrition, plant-environment interactions
ASJC Classification1110 Plant Science; 1314 Physiology
Languageen angielski
Score (nominal)45
Score sourcejournalList
ScoreMinisterial score = 45.0, 03-04-2020, ArticleFromJournal
Publication indicators WoS Citations = 9; Scopus Citations = 13; Scopus SNIP (Source Normalised Impact per Paper): 2018 = 1.589; WoS Impact Factor: 2018 = 5.36 (2) - 2018=6.305 (5)
Citation count*17 (2020-03-21)
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* presented citation count is obtained through Internet information analysis and it is close to the number calculated by the Publish or Perish system.
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