title: Coloring genetically modified soybean grains with anthocyanins by suppression of the proanthocyanidin genes ANR1 and ANR2.
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.
authors.affiliation: Bioproducts and Bioprocesses, Research Branch, Agriculture and Agri-Food Canada, Ottawa, ON, Canada. Nikola.Kovinich@AGR.GC.CA
authors.forename: Nik
authors.initials: N
authors.lastname: Kovinich
authors.forename: Ammar
authors.initials: A
authors.lastname: Saleem
authors.forename: Tara L
authors.initials: TL
authors.lastname: Rintoul
authors.forename: Daniel C W
authors.initials: DC
authors.lastname: Brown
authors.forename: John T
authors.initials: JT
authors.lastname: Arnason
authors.forename: Brian
authors.initials: B
authors.lastname: Miki
mesh_terms: D000872:Anthocyanins / Q000235:genetics*; D003116:Color; D018506:Gene Expression Regulation, Plant; D010641:Phenotype; D010858:Pigmentation; D010860:Pigments, Biological / Q000096:biosynthesis / Q000235:genetics; D030821:Plants, Genetically Modified; D044945:Proanthocyanidins / Q000235:genetics / Q000378:metabolism; D034622:RNA Interference; D012639:Seeds / Q000235:genetics / Q000378:metabolism; D030262:Soybean Proteins / Q000235:genetics; D013025:Glycine max / Q000235:genetics*
chemical_list: D000872:Anthocyanins; D010860:Pigments, Biological; D044945:Proanthocyanidins; D030262:Soybean Proteins; C013221:proanthocyanidin
references.citation: Plant Physiol. 1997 Apr;113(4):1437-45
references.pmid: 9112784
references.citation: Nat Biotechnol. 2008 Feb;26(2):161-2
references.pmid: 18259165
references.citation: Nat Biotechnol. 2009 Aug;27(8):700-1
references.pmid: 19668172
references.citation: Plant J. 1993 Dec;4(6):1003-10
references.pmid: 7904213
references.citation: Annu Rev Plant Biol. 2008;59:771-812
references.pmid: 18284373
references.citation: Plant Cell. 2011 Apr;23(4):1536-55
references.pmid: 21467581
references.citation: Plant Physiol. 2001 Jun;126(2):485-93
references.pmid: 11402179
references.citation: Plant Cell Rep. 2007 Nov;26(11):1951-9
references.pmid: 17639403
references.citation: Trends Plant Sci. 2010 Feb;15(2):72-80
references.pmid: 20006535
references.citation: Plant J. 2007 May;50(4):660-77
references.pmid: 17419845
references.citation: Plant Physiol. 2000 Jan;122(1):107-16
references.pmid: 10631254
references.citation: Plant Mol Biol. 2005 Jan;57(2):155-71
references.pmid: 15821875
references.citation: BMC Genomics. 2011 Jul 29;12:381
references.pmid: 21801362
references.citation: BMC Plant Biol. 2011 Apr 29;11:74
references.pmid: 21529361
references.citation: Nat Biotechnol. 2010 Feb;28(2):133-6
references.pmid: 20139947
references.citation: Planta. 1986 Mar;169(1):97-107
references.pmid: 24232434
references.citation: Biotechnol Lett. 2011 Mar;33(3):433-41
references.pmid: 21053046
references.citation: Eur J Biochem. 1976 Jan 2;61(1):199-206
references.pmid: 1245182
references.citation: Annu Rev Plant Biol. 2006;57:405-30
references.pmid: 16669768
references.citation: J Plant Physiol. 2009 Jan 1;166(1):32-9
references.pmid: 18423787
references.citation: Crit Rev Biotechnol. 2008;28(3):153-72
references.pmid: 18937106
references.citation: Plant J. 1999 Aug;19(4):387-98
references.pmid: 10504561
references.citation: Trends Biotechnol. 2006 Apr;24(4):155-62
references.pmid: 16488034
references.citation: Phytochemistry. 2010 Aug;71(11-12):1253-63
references.pmid: 20621794
references.citation: Nature. 1987 Dec 17-23;330(6149):677-8
references.pmid: 3683587
references.citation: Plant Mol Biol. 1992 Nov;20(4):743-7
references.pmid: 1450389
references.citation: Trends Plant Sci. 2005 Aug;10(8):390-6
references.pmid: 16023400
references.citation: Drug Metabol Drug Interact. 1995;12(3-4):359-68
references.pmid: 8820861
references.citation: Planta. 2006 Jun;224(1):96-107
references.pmid: 16395586
references.citation: Phytochemistry. 2003 Aug;63(7):753-63
references.pmid: 12877915
references.citation: Plant Cell. 2007 Jan;19(1):148-62
references.pmid: 17237352
references.citation: Science. 2003 Jan 17;299(5605):396-9
references.pmid: 12532018
references.citation: Plant Cell Physiol. 2004 Apr;45(4):490-5
references.pmid: 15111724
references.citation: Planta. 2009 Jan;229(2):427-45
references.pmid: 18998159
references.citation: Biotechnol Adv. 2009 Nov-Dec;27(6):879-894
references.pmid: 19647060
references.citation: Plant Biotechnol J. 2008 Apr;6(3):213-25
references.pmid: 18028290
references.citation: Plant Cell. 2004 Apr;16(4):819-35
references.pmid: 15064367
references.citation: J Agric Food Chem. 2012 Jan 18;60(2):574-84
references.pmid: 22107112
references.citation: Plant J. 1997 Feb;11(2):289-99
references.pmid: 9076994
references.citation: Plant Cell. 1991 Aug;3(8):829-40
references.pmid: 1820822