Overexpression of the MYB29 transcription factor affects aliphatic glucosinolate synthesis in Brassica oleracea

Diana L. Zuluaga*, Neil S. Graham, Annett Klinder, Elaine A. van Ommen Kloeke, Angelo R. Marcotrigiano, Carol Wagstaff, Ruud Verkerk, Gabriella Sonnante, Mark G.M. Aarts

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

Key message: Overexpression of BoMYB29 gene up-regulates the aliphatic glucosinolate pathway in Brassica oleracea plants increasing the production of the anti-cancer metabolite glucoraphanin, and the toxic and pungent sinigrin. Abstract: Isothiocyanates, the bio-active hydrolysis products of glucosinolates, naturally produced by several Brassicaceae species, play an important role in human health and agriculture. This study aims at correlating the content of aliphatic glucosinolates to the expression of genes involved in their synthesis in Brassica oleracea, and perform functional analysis of BoMYB29 gene. To this purpose, three genotypes were used: a sprouting broccoli, a cabbage, and a wild genotype (Winspit), a high glucosinolate containing accession. Winspit showed the highest transcript level of BoMYB28, BoMYB29 and BoAOP2 genes, and BoAOP2 expression was positively correlated with that of the two MYB genes. Further analyses of the aliphatic glucosinolates also showed a positive correlation between the expression of BoAOP2 and the production of sinigrin and gluconapin in Winspit. The Winspit BoMYB29 CDS was cloned and overexpressed in Winspit and in the DH AG1012 line. Overexpressing Winspit plants produced higher quantities of alkenyl glucosinolates, such as sinigrin. Conversely, the DH AG1012 transformants showed a higher production of methylsulphinylalkyl glucosinolates, including glucoraphanin, and, despite an up-regulation of the aliphatic glucosinolate genes, no increase in alkenyl glucosinolates. The latter may be explained by the absence of a functional AOP2 gene in DH AG1012. Nevertheless, an extract of DH AG1012 lines overexpressing BoMYB29 provided a chemoprotective effect on human colon cells. This work exemplifies how the genetic diversity of B. oleracea may be used by breeders to select for higher expression of transcription factors for glucosinolate biosynthesis to improve its natural, health-promoting properties.

Original languageEnglish
Pages (from-to)65-79
JournalPlant Molecular Biology
Volume101
Issue number1-2
Early online date12 Jun 2019
DOIs
Publication statusPublished - Sep 2019

Fingerprint

Glucosinolates
Brassica
glucosinolates
Brassica oleracea
Transcription Factors
transcription factors
synthesis
sinigrin
glucoraphanin
Genes
genes
Up-Regulation
Genotype
gluconapin
Isothiocyanates
Brassicaceae
Gene Expression
health promotion
gene overexpression
genotype

Keywords

  • Antigenotoxic effect
  • Brassicaceae plants
  • Glucosinolates
  • Isothiocyanates
  • MYB29 transcription factor
  • Sinigrin

Cite this

Zuluaga, D. L., Graham, N. S., Klinder, A., van Ommen Kloeke, E. A., Marcotrigiano, A. R., Wagstaff, C., ... Aarts, M. G. M. (2019). Overexpression of the MYB29 transcription factor affects aliphatic glucosinolate synthesis in Brassica oleracea. Plant Molecular Biology, 101(1-2), 65-79. https://doi.org/10.1007/s11103-019-00890-2
Zuluaga, Diana L. ; Graham, Neil S. ; Klinder, Annett ; van Ommen Kloeke, Elaine A. ; Marcotrigiano, Angelo R. ; Wagstaff, Carol ; Verkerk, Ruud ; Sonnante, Gabriella ; Aarts, Mark G.M. / Overexpression of the MYB29 transcription factor affects aliphatic glucosinolate synthesis in Brassica oleracea. In: Plant Molecular Biology. 2019 ; Vol. 101, No. 1-2. pp. 65-79.
@article{3194b4d0f4504440b5d7ee303ebe6f3e,
title = "Overexpression of the MYB29 transcription factor affects aliphatic glucosinolate synthesis in Brassica oleracea",
abstract = "Key message: Overexpression of BoMYB29 gene up-regulates the aliphatic glucosinolate pathway in Brassica oleracea plants increasing the production of the anti-cancer metabolite glucoraphanin, and the toxic and pungent sinigrin. Abstract: Isothiocyanates, the bio-active hydrolysis products of glucosinolates, naturally produced by several Brassicaceae species, play an important role in human health and agriculture. This study aims at correlating the content of aliphatic glucosinolates to the expression of genes involved in their synthesis in Brassica oleracea, and perform functional analysis of BoMYB29 gene. To this purpose, three genotypes were used: a sprouting broccoli, a cabbage, and a wild genotype (Winspit), a high glucosinolate containing accession. Winspit showed the highest transcript level of BoMYB28, BoMYB29 and BoAOP2 genes, and BoAOP2 expression was positively correlated with that of the two MYB genes. Further analyses of the aliphatic glucosinolates also showed a positive correlation between the expression of BoAOP2 and the production of sinigrin and gluconapin in Winspit. The Winspit BoMYB29 CDS was cloned and overexpressed in Winspit and in the DH AG1012 line. Overexpressing Winspit plants produced higher quantities of alkenyl glucosinolates, such as sinigrin. Conversely, the DH AG1012 transformants showed a higher production of methylsulphinylalkyl glucosinolates, including glucoraphanin, and, despite an up-regulation of the aliphatic glucosinolate genes, no increase in alkenyl glucosinolates. The latter may be explained by the absence of a functional AOP2 gene in DH AG1012. Nevertheless, an extract of DH AG1012 lines overexpressing BoMYB29 provided a chemoprotective effect on human colon cells. This work exemplifies how the genetic diversity of B. oleracea may be used by breeders to select for higher expression of transcription factors for glucosinolate biosynthesis to improve its natural, health-promoting properties.",
keywords = "Antigenotoxic effect, Brassicaceae plants, Glucosinolates, Isothiocyanates, MYB29 transcription factor, Sinigrin",
author = "Zuluaga, {Diana L.} and Graham, {Neil S.} and Annett Klinder and {van Ommen Kloeke}, {Elaine A.} and Marcotrigiano, {Angelo R.} and Carol Wagstaff and Ruud Verkerk and Gabriella Sonnante and Aarts, {Mark G.M.}",
year = "2019",
month = "9",
doi = "10.1007/s11103-019-00890-2",
language = "English",
volume = "101",
pages = "65--79",
journal = "Plant Molecular Biology",
issn = "0167-4412",
publisher = "Springer Verlag",
number = "1-2",

}

Zuluaga, DL, Graham, NS, Klinder, A, van Ommen Kloeke, EA, Marcotrigiano, AR, Wagstaff, C, Verkerk, R, Sonnante, G & Aarts, MGM 2019, 'Overexpression of the MYB29 transcription factor affects aliphatic glucosinolate synthesis in Brassica oleracea', Plant Molecular Biology, vol. 101, no. 1-2, pp. 65-79. https://doi.org/10.1007/s11103-019-00890-2

Overexpression of the MYB29 transcription factor affects aliphatic glucosinolate synthesis in Brassica oleracea. / Zuluaga, Diana L.; Graham, Neil S.; Klinder, Annett; van Ommen Kloeke, Elaine A.; Marcotrigiano, Angelo R.; Wagstaff, Carol; Verkerk, Ruud; Sonnante, Gabriella; Aarts, Mark G.M.

In: Plant Molecular Biology, Vol. 101, No. 1-2, 09.2019, p. 65-79.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - Overexpression of the MYB29 transcription factor affects aliphatic glucosinolate synthesis in Brassica oleracea

AU - Zuluaga, Diana L.

AU - Graham, Neil S.

AU - Klinder, Annett

AU - van Ommen Kloeke, Elaine A.

AU - Marcotrigiano, Angelo R.

AU - Wagstaff, Carol

AU - Verkerk, Ruud

AU - Sonnante, Gabriella

AU - Aarts, Mark G.M.

PY - 2019/9

Y1 - 2019/9

N2 - Key message: Overexpression of BoMYB29 gene up-regulates the aliphatic glucosinolate pathway in Brassica oleracea plants increasing the production of the anti-cancer metabolite glucoraphanin, and the toxic and pungent sinigrin. Abstract: Isothiocyanates, the bio-active hydrolysis products of glucosinolates, naturally produced by several Brassicaceae species, play an important role in human health and agriculture. This study aims at correlating the content of aliphatic glucosinolates to the expression of genes involved in their synthesis in Brassica oleracea, and perform functional analysis of BoMYB29 gene. To this purpose, three genotypes were used: a sprouting broccoli, a cabbage, and a wild genotype (Winspit), a high glucosinolate containing accession. Winspit showed the highest transcript level of BoMYB28, BoMYB29 and BoAOP2 genes, and BoAOP2 expression was positively correlated with that of the two MYB genes. Further analyses of the aliphatic glucosinolates also showed a positive correlation between the expression of BoAOP2 and the production of sinigrin and gluconapin in Winspit. The Winspit BoMYB29 CDS was cloned and overexpressed in Winspit and in the DH AG1012 line. Overexpressing Winspit plants produced higher quantities of alkenyl glucosinolates, such as sinigrin. Conversely, the DH AG1012 transformants showed a higher production of methylsulphinylalkyl glucosinolates, including glucoraphanin, and, despite an up-regulation of the aliphatic glucosinolate genes, no increase in alkenyl glucosinolates. The latter may be explained by the absence of a functional AOP2 gene in DH AG1012. Nevertheless, an extract of DH AG1012 lines overexpressing BoMYB29 provided a chemoprotective effect on human colon cells. This work exemplifies how the genetic diversity of B. oleracea may be used by breeders to select for higher expression of transcription factors for glucosinolate biosynthesis to improve its natural, health-promoting properties.

AB - Key message: Overexpression of BoMYB29 gene up-regulates the aliphatic glucosinolate pathway in Brassica oleracea plants increasing the production of the anti-cancer metabolite glucoraphanin, and the toxic and pungent sinigrin. Abstract: Isothiocyanates, the bio-active hydrolysis products of glucosinolates, naturally produced by several Brassicaceae species, play an important role in human health and agriculture. This study aims at correlating the content of aliphatic glucosinolates to the expression of genes involved in their synthesis in Brassica oleracea, and perform functional analysis of BoMYB29 gene. To this purpose, three genotypes were used: a sprouting broccoli, a cabbage, and a wild genotype (Winspit), a high glucosinolate containing accession. Winspit showed the highest transcript level of BoMYB28, BoMYB29 and BoAOP2 genes, and BoAOP2 expression was positively correlated with that of the two MYB genes. Further analyses of the aliphatic glucosinolates also showed a positive correlation between the expression of BoAOP2 and the production of sinigrin and gluconapin in Winspit. The Winspit BoMYB29 CDS was cloned and overexpressed in Winspit and in the DH AG1012 line. Overexpressing Winspit plants produced higher quantities of alkenyl glucosinolates, such as sinigrin. Conversely, the DH AG1012 transformants showed a higher production of methylsulphinylalkyl glucosinolates, including glucoraphanin, and, despite an up-regulation of the aliphatic glucosinolate genes, no increase in alkenyl glucosinolates. The latter may be explained by the absence of a functional AOP2 gene in DH AG1012. Nevertheless, an extract of DH AG1012 lines overexpressing BoMYB29 provided a chemoprotective effect on human colon cells. This work exemplifies how the genetic diversity of B. oleracea may be used by breeders to select for higher expression of transcription factors for glucosinolate biosynthesis to improve its natural, health-promoting properties.

KW - Antigenotoxic effect

KW - Brassicaceae plants

KW - Glucosinolates

KW - Isothiocyanates

KW - MYB29 transcription factor

KW - Sinigrin

U2 - 10.1007/s11103-019-00890-2

DO - 10.1007/s11103-019-00890-2

M3 - Article

VL - 101

SP - 65

EP - 79

JO - Plant Molecular Biology

JF - Plant Molecular Biology

SN - 0167-4412

IS - 1-2

ER -

Zuluaga DL, Graham NS, Klinder A, van Ommen Kloeke EA, Marcotrigiano AR, Wagstaff C et al. Overexpression of the MYB29 transcription factor affects aliphatic glucosinolate synthesis in Brassica oleracea. Plant Molecular Biology. 2019 Sep;101(1-2):65-79. https://doi.org/10.1007/s11103-019-00890-2