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

Authors

    • Bioproducts and Bioprocesses, Research BranchAgriculture and Agri-Food Canada
    • Department of Biology, Ottawa–Carleton Institute of BiologyCarleton University
  • Ammar Saleem
    • Department of Biology and Center for Research in Biopharmaceuticals and BiotechnologyUniversity of Ottawa
  • Tara L. Rintoul
    • Bioproducts and Bioprocesses, Research BranchAgriculture and Agri-Food Canada
  • Daniel C. W. Brown
    • Southern Crop Protection and Food Research CentreAgriculture and Agri-Food Canada
  • John T. Arnason
    • Department of Biology and Center for Research in Biopharmaceuticals and BiotechnologyUniversity of Ottawa
  • Brian Miki
    • Bioproducts and Bioprocesses, Research BranchAgriculture and Agri-Food Canada
Original Paper

DOI: 10.1007/s11248-011-9566-y

Cite this article as:
Kovinich, N., Saleem, A., Rintoul, T.L. et al. Transgenic Res (2012) 21: 757. doi:10.1007/s11248-011-9566-y

Abstract

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, ANTHOCYANIDIN REDUCTASE1 (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.

Keywords

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

Supplementary material

11248_2011_9566_MOESM1_ESM.xlsx (12 kb)
Supplementary material 1 (XLSX 12 kb)
11248_2011_9566_MOESM2_ESM.xlsx (11 kb)
Supplementary material 2 (XLSX 10 kb)
11248_2011_9566_MOESM3_ESM.xls (26 kb)
Supplementary material 3 (XLS 25 kb)
11248_2011_9566_MOESM4_ESM.xls (24 kb)
Supplementary material 4 (XLS 23 kb)

Copyright information

© Her Majesty the Queen in Right of Canada 2011