Molecular characterization of a stable antisense chalcone synthase phenotype in strawberry (Fragaria ananassa)

S. Lunkenbein, H. Coiner, C.H. de Vos, J.G. Schaart, M.J. Boone, F.A. Krens, W. Schwab, E.M.J. Salentijn

Research output: Contribution to journalArticleAcademicpeer-review

54 Citations (Scopus)

Abstract

An octaploid (Fragaria × ananassa cv. Calypso) genotype of strawberry was transformed with an antisense chalcone synthase (CHS) gene construct using a ripening related CHS cDNA from Fragaria × ananassa cv. Elsanta under the control of the constitutive CaMV 35S promoter via Agrobacterium tumefaciens. Out of 25 transgenic lines, nine lines showed a reduction in CHS mRNA accumulation of more than 50% as compared to the untransformed cv. Calypso control. The antisense CHS construct was found to be integrated into the genome, with a copy number ranging from one to four. The pigmentation of the fruit was only affected when less than 5% of the control CHS expression level was detected. A stable antisense phenotype over a period of 4 years was obtained in the primary transgenic lines at a rate of 1:20. As a consequence of the reduced activity of CHS, the levels of anthocyanins, flavonols, and proanthocyanidins were downregulated and precursors of the flavonoid pathway were shunted to the phenylpropanoid pathway leading to highly increased levels of cinnamoyl glucose (520% of control), caffeoyl glucose (816% of control), and feruloyl glucose (1092% of control) as well as p-coumaryl alcohol (363% of control) and p-coumaryl-1-acetate (1079% of control), which occur only as trace components in untransformed control fruits. These results demonstrate that the introduction of an antisense CHS construct in strawberry results in an unpredictable biochemical phenotype, thereby confirming that CHS function is an important regulatory point of substrate flow between the flavonoid and the phenylpropanoid pathways
Original languageEnglish
Pages (from-to)2145-2153
JournalJournal of Agricultural and Food Chemistry
Volume54
Issue number6
DOIs
Publication statusPublished - 2006

Fingerprint

Fragaria
naringenin-chalcone synthase
Fragaria ananassa
strawberries
Phenotype
phenotype
Calypso
phenylpropanoids
Flavonoids
Glucose
glucose
Fruits
Fruit
flavonoids
octaploidy
genetically modified organisms
control components
Genes
Proanthocyanidins
Agrobacterium tumefaciens

Keywords

  • agrobacterium-mediated transformation
  • plant polyketide biosynthesis
  • shoot regeneration
  • phenolic-compounds
  • transgenic plants
  • dna microarrays
  • flower color
  • ms detection
  • glycine-max
  • gene

Cite this

@article{1f5d93a0f47b493997f3d48ce7b3b29c,
title = "Molecular characterization of a stable antisense chalcone synthase phenotype in strawberry (Fragaria ananassa)",
abstract = "An octaploid (Fragaria × ananassa cv. Calypso) genotype of strawberry was transformed with an antisense chalcone synthase (CHS) gene construct using a ripening related CHS cDNA from Fragaria × ananassa cv. Elsanta under the control of the constitutive CaMV 35S promoter via Agrobacterium tumefaciens. Out of 25 transgenic lines, nine lines showed a reduction in CHS mRNA accumulation of more than 50{\%} as compared to the untransformed cv. Calypso control. The antisense CHS construct was found to be integrated into the genome, with a copy number ranging from one to four. The pigmentation of the fruit was only affected when less than 5{\%} of the control CHS expression level was detected. A stable antisense phenotype over a period of 4 years was obtained in the primary transgenic lines at a rate of 1:20. As a consequence of the reduced activity of CHS, the levels of anthocyanins, flavonols, and proanthocyanidins were downregulated and precursors of the flavonoid pathway were shunted to the phenylpropanoid pathway leading to highly increased levels of cinnamoyl glucose (520{\%} of control), caffeoyl glucose (816{\%} of control), and feruloyl glucose (1092{\%} of control) as well as p-coumaryl alcohol (363{\%} of control) and p-coumaryl-1-acetate (1079{\%} of control), which occur only as trace components in untransformed control fruits. These results demonstrate that the introduction of an antisense CHS construct in strawberry results in an unpredictable biochemical phenotype, thereby confirming that CHS function is an important regulatory point of substrate flow between the flavonoid and the phenylpropanoid pathways",
keywords = "agrobacterium-mediated transformation, plant polyketide biosynthesis, shoot regeneration, phenolic-compounds, transgenic plants, dna microarrays, flower color, ms detection, glycine-max, gene",
author = "S. Lunkenbein and H. Coiner and {de Vos}, C.H. and J.G. Schaart and M.J. Boone and F.A. Krens and W. Schwab and E.M.J. Salentijn",
year = "2006",
doi = "10.1021/jf052574z",
language = "English",
volume = "54",
pages = "2145--2153",
journal = "Journal of Agricultural and Food Chemistry",
issn = "0021-8561",
publisher = "American Chemical Society",
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Molecular characterization of a stable antisense chalcone synthase phenotype in strawberry (Fragaria ananassa). / Lunkenbein, S.; Coiner, H.; de Vos, C.H.; Schaart, J.G.; Boone, M.J.; Krens, F.A.; Schwab, W.; Salentijn, E.M.J.

In: Journal of Agricultural and Food Chemistry, Vol. 54, No. 6, 2006, p. 2145-2153.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - Molecular characterization of a stable antisense chalcone synthase phenotype in strawberry (Fragaria ananassa)

AU - Lunkenbein, S.

AU - Coiner, H.

AU - de Vos, C.H.

AU - Schaart, J.G.

AU - Boone, M.J.

AU - Krens, F.A.

AU - Schwab, W.

AU - Salentijn, E.M.J.

PY - 2006

Y1 - 2006

N2 - An octaploid (Fragaria × ananassa cv. Calypso) genotype of strawberry was transformed with an antisense chalcone synthase (CHS) gene construct using a ripening related CHS cDNA from Fragaria × ananassa cv. Elsanta under the control of the constitutive CaMV 35S promoter via Agrobacterium tumefaciens. Out of 25 transgenic lines, nine lines showed a reduction in CHS mRNA accumulation of more than 50% as compared to the untransformed cv. Calypso control. The antisense CHS construct was found to be integrated into the genome, with a copy number ranging from one to four. The pigmentation of the fruit was only affected when less than 5% of the control CHS expression level was detected. A stable antisense phenotype over a period of 4 years was obtained in the primary transgenic lines at a rate of 1:20. As a consequence of the reduced activity of CHS, the levels of anthocyanins, flavonols, and proanthocyanidins were downregulated and precursors of the flavonoid pathway were shunted to the phenylpropanoid pathway leading to highly increased levels of cinnamoyl glucose (520% of control), caffeoyl glucose (816% of control), and feruloyl glucose (1092% of control) as well as p-coumaryl alcohol (363% of control) and p-coumaryl-1-acetate (1079% of control), which occur only as trace components in untransformed control fruits. These results demonstrate that the introduction of an antisense CHS construct in strawberry results in an unpredictable biochemical phenotype, thereby confirming that CHS function is an important regulatory point of substrate flow between the flavonoid and the phenylpropanoid pathways

AB - An octaploid (Fragaria × ananassa cv. Calypso) genotype of strawberry was transformed with an antisense chalcone synthase (CHS) gene construct using a ripening related CHS cDNA from Fragaria × ananassa cv. Elsanta under the control of the constitutive CaMV 35S promoter via Agrobacterium tumefaciens. Out of 25 transgenic lines, nine lines showed a reduction in CHS mRNA accumulation of more than 50% as compared to the untransformed cv. Calypso control. The antisense CHS construct was found to be integrated into the genome, with a copy number ranging from one to four. The pigmentation of the fruit was only affected when less than 5% of the control CHS expression level was detected. A stable antisense phenotype over a period of 4 years was obtained in the primary transgenic lines at a rate of 1:20. As a consequence of the reduced activity of CHS, the levels of anthocyanins, flavonols, and proanthocyanidins were downregulated and precursors of the flavonoid pathway were shunted to the phenylpropanoid pathway leading to highly increased levels of cinnamoyl glucose (520% of control), caffeoyl glucose (816% of control), and feruloyl glucose (1092% of control) as well as p-coumaryl alcohol (363% of control) and p-coumaryl-1-acetate (1079% of control), which occur only as trace components in untransformed control fruits. These results demonstrate that the introduction of an antisense CHS construct in strawberry results in an unpredictable biochemical phenotype, thereby confirming that CHS function is an important regulatory point of substrate flow between the flavonoid and the phenylpropanoid pathways

KW - agrobacterium-mediated transformation

KW - plant polyketide biosynthesis

KW - shoot regeneration

KW - phenolic-compounds

KW - transgenic plants

KW - dna microarrays

KW - flower color

KW - ms detection

KW - glycine-max

KW - gene

U2 - 10.1021/jf052574z

DO - 10.1021/jf052574z

M3 - Article

VL - 54

SP - 2145

EP - 2153

JO - Journal of Agricultural and Food Chemistry

JF - Journal of Agricultural and Food Chemistry

SN - 0021-8561

IS - 6

ER -