Overexpression of sinapine esterase BnSCE3 in oilseed rape seeds triggers global changes in seed metabolism.
Sinapine (O-sinapoylcholine) is the predominant phenolic compound in a complex group of sinapate esters in seeds of oilseed rape (Brassica napus). Sinapine has antinutritive activity and prevents the use of seed protein for food and feed. A strategy was developed to lower its content in seeds by expressing an enzyme that hydrolyzes sinapine in developing rape seeds. During early stages of seedling development, a sinapine esterase (BnSCE3) hydrolyzes sinapine, releasing choline and sinapate. A portion of choline enters the phospholipid metabolism, and sinapate is routed via 1-O-sinapoyl-β-glucose into sinapoylmalate. Transgenic oilseed rape lines were generated expressing BnSCE3 under the control of a seed-specific promoter. Two distinct single-copy transgene insertion lines were isolated and propagated to generate homozygous lines, which were subjected to comprehensive phenotyping. Sinapine levels of transgenic seeds were less than 5% of wild-type levels, whereas choline levels were increased. Weight, size, and water content of transgenic seeds were significantly higher than those of wild-type seeds. Seed quality parameters, such as fiber and glucosinolate levels, and agronomically important traits, such as oil and protein contents, differed only slightly, except that amounts of hemicellulose and cellulose were about 30% higher in transgenic compared with wild-type seeds. Electron microscopic examination revealed that a fraction of the transgenic seeds had morphological alterations, characterized by large cavities near the embryonic tissue. Transgenic seedlings were larger than wild-type seedlings, and young seedlings exhibited longer hypocotyls. Examination of metabolic profiles of transgenic seeds indicated that besides suppression of sinapine accumulation, there were other dramatic differences in primary and secondary metabolism. Mapping of these changes onto metabolic pathways revealed global effects of the transgenic BnSCE3 expression on seed metabolism.
Department of Secondary Metabolism, Leibniz Institute of Plant Biochemistry, D-06120 Halle, Germany.
Kathleen
K
Clauss
Edda
E
von Roepenack-Lahaye
Christoph
C
Böttcher
Mary R
MR
Roth
Ruth
R
Welti
Alexander
A
Erban
Joachim
J
Kopka
Dierk
D
Scheel
Carsten
C
Milkowski
Dieter
D
Strack
D029688:Brassica napus / Q000201:enzymology* / Q000235:genetics / Q000648:ultrastructure; D002794:Choline / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism; D020090:Chromosome Segregation / Q000235:genetics; D004950:Esterases / Q000235:genetics / Q000378:metabolism*; D008401:Gas Chromatography-Mass Spectrometry; D018506:Gene Expression Regulation, Plant; D008055:Lipids / Q000032:analysis; D053858:Metabolic Networks and Pathways; D055442:Metabolome; D008969:Molecular Sequence Data; D010636:Phenols / Q000737:chemistry / Q000378:metabolism; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism*; D030821:Plants, Genetically Modified; D036226:Seedlings / Q000378:metabolism; D012639:Seeds / Q000378:metabolism* / Q000648:ultrastructure; D019265:Spectroscopy, Near-Infrared
D008055:Lipids; D010636:Phenols; D010940:Plant Proteins; C005685:sinapine; D004950:Esterases; D002794:Choline
Annu Rev Plant Biol. 2003;54:519-46
14503002
J Bacteriol. 1991 Jan;173(2):472-8
1987142
Metabolomics. 2010 Jun;6(2):322-333
20526350
BMC Bioinformatics. 2006 Mar 06;7:109
16519817
Proteomics. 2006 Aug;6(16):4586-98
16847873
Phytochemistry. 2006 Sep;67(17):1907-24
16843506
Plant Physiol. 2009 May;150(1):55-72
19329563
Curr Opin Plant Biol. 2002 Jun;5(3):224-9
11960740
Prog Lipid Res. 2004 Nov;43(6):534-52
15522763
Plant Physiol Biochem. 2008 Jul;46(7):647-54
18515127
Planta. 1985 Jul;164(4):507-11
24248224
Nutrition. 2000 Jul-Aug;16(7-8):669-71
10906592
Methods Mol Biol. 2007;358:19-38
17035678
Plant Cell. 2009 Jun;21(6):1830-45
19567706
J Biol Chem. 2000 Nov 24;275(47):36899-909
10934215
Transgenic Res. 1995 Jul;4(4):288-90
7655517
Anal Biochem. 2001 Aug 1;295(1):88-100
11476549
Bioinformatics. 2008 Mar 1;24(5):732-7
18204057
BMC Bioinformatics. 2008 Nov 28;9:504
19040729
Plant J. 2008 Mar;53(5):802-13
18036206
Physiol Plant. 2002 Jun;115(2):221-227
12060239
Phytochemistry. 2004 Mar;65(5):517-24
15003414
Plant J. 1998 Nov;16(4):487-96
9881168
Plant Physiol. 1993 Jan;101(1):267-276
12231682
J Biol Chem. 2002 Aug 30;277(35):31994-2002
12077151
Planta. 2008 Sep;228(4):663-74
18563436
Phytochemistry. 2010 Jul;71(10):1076-84
20451226
Planta. 2008 Jan;227(2):375-85
17882453
Plant Cell. 2010 Apr;22(4):1033-45
20371642
Metabolomics. 2009 Sep;5(3):277-291
19718268
Phytochemistry. 2003 Nov;64(5):1013-21
14561519
Planta. 2009 Oct;230(5):1057-69
19714359
Plant J. 2004 Apr;38(1):80-92
15053762
Plant Physiol. 1989 Oct;91(2):694-701
16667089
Nucleic Acids Res. 2004 Jan 1;32(Database issue):D277-80
14681412
Plant Physiol. 2000 Aug;123(4):1623-34
10938378
Phytochemistry. 2005 Jun;66(11):1334-45
15907956
Plant Physiol. 2000 Mar;122(3):747-56
10712538
Biochim Biophys Acta. 2004 Jun 11;1672(3):184-91
15182938
Theor Appl Genet. 2005 Nov;111(8):1553-62
16160820
Transgenic Res. 2009 Feb;18(1):31-44
18612839
Bioinformatics. 2004 Oct 12;20(15):2447-54
15087312