An O-methyltransferase modifies accumulation of methylated anthocyanins in seedlings of tomato

M.V. Gomez Roldan, N.S. Outchkourov, A.M.M.L. van Houwelingen, M. Lammers, I. Romero Fuente, N. Ziklo, A. Aharoni, R.D. Hall, M.J. Beekwilder

Research output: Contribution to journalArticleAcademicpeer-review

7 Citations (Scopus)

Abstract

Anthocyanins contribute to the appearance of fruit by conferring to them a red, blue or purple colour. In a food context, they have also been suggested to promote consumer health. In purple tomato tissues, such as hypocotyls, stems and purple fruits, various anthocyanins accumulate. These molecules have characteristic patterns of modification, including hydroxylations, methylations, glycosylations and acylations. The genetic basis for many of these modifications has not been fully elucidated, and nor has their role in the functioning of anthocyanins. In this paper, AnthOMT, an O-methyltransferase (OMT) mediating the methylation of anthocyanins, has been identified and functionally characterized using a combined metabolomics and transcriptomics approach. Gene candidates were selected from the draft tomato genome, and their expression was subsequently monitored in a tomato seedling system comprising three tissues and involving several time points. In addition, we also followed gene expression in wild-type red and purple transgenic tomato fruits expressing Rosea1 and Delila transcription factors. Of the 57 candidates identified, only a single OMT gene showed patterns strongly correlating with both accumulation of anthocyanins and expression of anthocyanin biosynthesis genes. This candidate (AnthOMT) was compared to a closely related caffeoyl CoA OMT by recombinant expression in Escherichia coli, and then tested for substrate specificity. AnthOMT showed a strong affinity for glycosylated anthocyanins, while other flavonoid glycosides and aglycones were much less preferred. Gene silencing experiments with AnthOMT resulted in reduced levels of the predominant methylated anthocyanins. This confirms the role of this enzyme in the diversification of tomato anthocyanins.
LanguageEnglish
Pages695-708
JournalThe Plant Journal
Volume80
Issue number4
DOIs
Publication statusPublished - 2014

Fingerprint

Anthocyanins
methyltransferases
Methyltransferases
Lycopersicon esculentum
Seedlings
anthocyanins
tomatoes
seedlings
Fruit
caffeoyl-CoA O-methyltransferase
fruits
Genes
Hypocotyl
acylation
Metabolomics
genes
Acylation
metabolomics
hydroxylation
Gene Silencing

Keywords

  • plant transformation
  • transcription factor
  • petunia-hybrida
  • fruit
  • biosynthesis
  • expression
  • protein
  • system
  • metabolome
  • infection

Cite this

Gomez Roldan, M.V. ; Outchkourov, N.S. ; van Houwelingen, A.M.M.L. ; Lammers, M. ; Romero Fuente, I. ; Ziklo, N. ; Aharoni, A. ; Hall, R.D. ; Beekwilder, M.J. / An O-methyltransferase modifies accumulation of methylated anthocyanins in seedlings of tomato. In: The Plant Journal. 2014 ; Vol. 80, No. 4. pp. 695-708.
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title = "An O-methyltransferase modifies accumulation of methylated anthocyanins in seedlings of tomato",
abstract = "Anthocyanins contribute to the appearance of fruit by conferring to them a red, blue or purple colour. In a food context, they have also been suggested to promote consumer health. In purple tomato tissues, such as hypocotyls, stems and purple fruits, various anthocyanins accumulate. These molecules have characteristic patterns of modification, including hydroxylations, methylations, glycosylations and acylations. The genetic basis for many of these modifications has not been fully elucidated, and nor has their role in the functioning of anthocyanins. In this paper, AnthOMT, an O-methyltransferase (OMT) mediating the methylation of anthocyanins, has been identified and functionally characterized using a combined metabolomics and transcriptomics approach. Gene candidates were selected from the draft tomato genome, and their expression was subsequently monitored in a tomato seedling system comprising three tissues and involving several time points. In addition, we also followed gene expression in wild-type red and purple transgenic tomato fruits expressing Rosea1 and Delila transcription factors. Of the 57 candidates identified, only a single OMT gene showed patterns strongly correlating with both accumulation of anthocyanins and expression of anthocyanin biosynthesis genes. This candidate (AnthOMT) was compared to a closely related caffeoyl CoA OMT by recombinant expression in Escherichia coli, and then tested for substrate specificity. AnthOMT showed a strong affinity for glycosylated anthocyanins, while other flavonoid glycosides and aglycones were much less preferred. Gene silencing experiments with AnthOMT resulted in reduced levels of the predominant methylated anthocyanins. This confirms the role of this enzyme in the diversification of tomato anthocyanins.",
keywords = "plant transformation, transcription factor, petunia-hybrida, fruit, biosynthesis, expression, protein, system, metabolome, infection",
author = "{Gomez Roldan}, M.V. and N.S. Outchkourov and {van Houwelingen}, A.M.M.L. and M. Lammers and {Romero Fuente}, I. and N. Ziklo and A. Aharoni and R.D. Hall and M.J. Beekwilder",
year = "2014",
doi = "10.1111/tpj.12664",
language = "English",
volume = "80",
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An O-methyltransferase modifies accumulation of methylated anthocyanins in seedlings of tomato. / Gomez Roldan, M.V.; Outchkourov, N.S.; van Houwelingen, A.M.M.L.; Lammers, M.; Romero Fuente, I.; Ziklo, N.; Aharoni, A.; Hall, R.D.; Beekwilder, M.J.

In: The Plant Journal, Vol. 80, No. 4, 2014, p. 695-708.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - An O-methyltransferase modifies accumulation of methylated anthocyanins in seedlings of tomato

AU - Gomez Roldan, M.V.

AU - Outchkourov, N.S.

AU - van Houwelingen, A.M.M.L.

AU - Lammers, M.

AU - Romero Fuente, I.

AU - Ziklo, N.

AU - Aharoni, A.

AU - Hall, R.D.

AU - Beekwilder, M.J.

PY - 2014

Y1 - 2014

N2 - Anthocyanins contribute to the appearance of fruit by conferring to them a red, blue or purple colour. In a food context, they have also been suggested to promote consumer health. In purple tomato tissues, such as hypocotyls, stems and purple fruits, various anthocyanins accumulate. These molecules have characteristic patterns of modification, including hydroxylations, methylations, glycosylations and acylations. The genetic basis for many of these modifications has not been fully elucidated, and nor has their role in the functioning of anthocyanins. In this paper, AnthOMT, an O-methyltransferase (OMT) mediating the methylation of anthocyanins, has been identified and functionally characterized using a combined metabolomics and transcriptomics approach. Gene candidates were selected from the draft tomato genome, and their expression was subsequently monitored in a tomato seedling system comprising three tissues and involving several time points. In addition, we also followed gene expression in wild-type red and purple transgenic tomato fruits expressing Rosea1 and Delila transcription factors. Of the 57 candidates identified, only a single OMT gene showed patterns strongly correlating with both accumulation of anthocyanins and expression of anthocyanin biosynthesis genes. This candidate (AnthOMT) was compared to a closely related caffeoyl CoA OMT by recombinant expression in Escherichia coli, and then tested for substrate specificity. AnthOMT showed a strong affinity for glycosylated anthocyanins, while other flavonoid glycosides and aglycones were much less preferred. Gene silencing experiments with AnthOMT resulted in reduced levels of the predominant methylated anthocyanins. This confirms the role of this enzyme in the diversification of tomato anthocyanins.

AB - Anthocyanins contribute to the appearance of fruit by conferring to them a red, blue or purple colour. In a food context, they have also been suggested to promote consumer health. In purple tomato tissues, such as hypocotyls, stems and purple fruits, various anthocyanins accumulate. These molecules have characteristic patterns of modification, including hydroxylations, methylations, glycosylations and acylations. The genetic basis for many of these modifications has not been fully elucidated, and nor has their role in the functioning of anthocyanins. In this paper, AnthOMT, an O-methyltransferase (OMT) mediating the methylation of anthocyanins, has been identified and functionally characterized using a combined metabolomics and transcriptomics approach. Gene candidates were selected from the draft tomato genome, and their expression was subsequently monitored in a tomato seedling system comprising three tissues and involving several time points. In addition, we also followed gene expression in wild-type red and purple transgenic tomato fruits expressing Rosea1 and Delila transcription factors. Of the 57 candidates identified, only a single OMT gene showed patterns strongly correlating with both accumulation of anthocyanins and expression of anthocyanin biosynthesis genes. This candidate (AnthOMT) was compared to a closely related caffeoyl CoA OMT by recombinant expression in Escherichia coli, and then tested for substrate specificity. AnthOMT showed a strong affinity for glycosylated anthocyanins, while other flavonoid glycosides and aglycones were much less preferred. Gene silencing experiments with AnthOMT resulted in reduced levels of the predominant methylated anthocyanins. This confirms the role of this enzyme in the diversification of tomato anthocyanins.

KW - plant transformation

KW - transcription factor

KW - petunia-hybrida

KW - fruit

KW - biosynthesis

KW - expression

KW - protein

KW - system

KW - metabolome

KW - infection

U2 - 10.1111/tpj.12664

DO - 10.1111/tpj.12664

M3 - Article

VL - 80

SP - 695

EP - 708

JO - The Plant Journal

T2 - The Plant Journal

JF - The Plant Journal

SN - 0960-7412

IS - 4

ER -