Transgenic Research

, Volume 21, Issue 4, pp 757–771

Coloring genetically modified soybean grains with anthocyanins by suppression of the proanthocyanidin genes ANR1 and ANR2

  • Nik Kovinich
  • Ammar Saleem
  • Tara L. Rintoul
  • Daniel C. W. Brown
  • John T. Arnason
  • Brian Miki
Original Paper


Detection and quantification of the levels of adventitious presence of genetically modified (GM) soybeans in non-GM grain shipments currently requires sophisticated tests that can have issues with their reproducibility. We show here that pigment biosynthesis in the soybean seed coat can be manipulated to provide a distinct color that would enable the simple visible detection of the GM soybean grain. We observed that a distinct red-brown grain color could be engineered by the simultaneous suppression of two proanthocyanidin (PA) genes, ANTHOCYANIDINREDUCTASE1 (ANR1) and ANR2. Multiple reaction monitoring by liquid chromatography tandem mass spectrometry was used to quantify differentially accumulated seed coat metabolites, and revealed the redirection of metabolic flux into the anthocyanin pigment pathway and unexpectedly the flavonol-3-O-glucoside pathway. The upregulations of anthocyanin isogenes (DFR1 and GST26) and the anthocyanin/flavonol-3-O-glycosyltransferase (UGT78K2) were identified by quantitative RT-PCR to be endogenous feedback and feedforward responses to overaccumulation of upstream flavonoid intermediates resulting from ANR1 and ANR2 suppressions. These results suggested the transcription of flavonoid genes to be a key component of the mechanism responsible for the redirection of metabolite flux. This report identifies the suppression of PA genes to be a novel approach for engineering pigmentation in soybean grains.


Metabolic engineering Visual marker Soybean seed coat color Anthocyanin Proanthocyanidin ANTHOCYANIDIN REDUCTASE 

Supplementary material

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Supplementary material 1 (XLSX 12 kb)
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Supplementary material 4 (XLS 23 kb)


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Copyright information

© Her Majesty the Queen in Right of Canada 2011

Authors and Affiliations

  • Nik Kovinich
    • 1
    • 2
  • Ammar Saleem
    • 3
  • Tara L. Rintoul
    • 1
  • Daniel C. W. Brown
    • 4
  • John T. Arnason
    • 3
  • Brian Miki
    • 1
  1. 1.Bioproducts and Bioprocesses, Research BranchAgriculture and Agri-Food CanadaOttawaCanada
  2. 2.Department of Biology, Ottawa–Carleton Institute of BiologyCarleton UniversityOttawaCanada
  3. 3.Department of Biology and Center for Research in Biopharmaceuticals and BiotechnologyUniversity of OttawaOttawaCanada
  4. 4.Southern Crop Protection and Food Research CentreAgriculture and Agri-Food CanadaLondonCanada

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