title: Integration of promoters, inverted repeat sequences and proteomic data into a model for high silencing efficiency of coeliac disease related gliadins in bread wheat.
abstract: BACKGROUND
  Wheat gluten has unique nutritional and technological characteristics, but is also a major trigger of allergies and intolerances. One of the most severe diseases caused by gluten is coeliac disease. The peptides produced in the digestive tract by the incomplete digestion of gluten proteins trigger the disease. The majority of the epitopes responsible reside in the gliadin fraction of gluten. The location of the multiple gliadin genes in blocks has to date complicated their elimination by classical breeding techniques or by the use of biotechnological tools.As an approach to silence multiple gliadin genes we have produced 38 transgenic lines of bread wheat containing combinations of two endosperm-specific promoters and three different inverted repeat sequences to silence three fractions of gliadins by RNA interference.
  
  
  RESULTS
  The effects of the RNA interference constructs on the content of the gluten proteins, total protein and starch, thousand seed weights and SDSS quality tests of flour were analyzed in these transgenic lines in two consecutive years. The characteristics of the inverted repeat sequences were the main factor that determined the efficiency of silencing. The promoter used had less influence on silencing, although a synergy in silencing efficiency was observed when the two promoters were used simultaneously. Genotype and the environment also influenced silencing efficiency.
  
  
  CONCLUSIONS
  We conclude that to obtain wheat lines with an optimum reduction of toxic gluten epitopes one needs to take into account the factors of inverted repeat sequences design, promoter choice and also the wheat background used.
authors.affiliation: Instituto de Agricultura Sostenible, Consejo Superior de Investigaciones Científicas (IAS-CSIC), Córdoba E-14080, Spain. fpiston@ias.csic.es.
authors.forename: Fernando
authors.initials: F
authors.lastname: Pistón
authors.forename: Javier
authors.initials: J
authors.lastname: Gil-Humanes
authors.forename: Francisco
authors.initials: F
authors.lastname: Barro
mesh_terms: D002446:Celiac Disease / Q000378:metabolism*; D005903:Gliadin / Q000235:genetics*; D055029:Inverted Repeat Sequences / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D040901:Proteomics; D014908:Triticum / Q000235:genetics / Q000378:metabolism*
chemical_list: D005903:Gliadin
references.citation: J Exp Bot. 2011 Aug;62(12):4203-13
references.pmid: 21561951
references.citation: New Phytol. 2009 Dec;184(4):851-64
references.pmid: 19732349
references.citation: BMC Genomics. 2011 Jan 18;12:41
references.pmid: 21241522
references.citation: Aliment Pharmacol Ther. 2011 Feb;33(4):477-86
references.pmid: 21166832
references.citation: J Agric Food Chem. 2004 Apr 7;52(7):1958-64
references.pmid: 15053536
references.citation: PLoS One. 2012;7(2):e32850
references.pmid: 22393455
references.citation: Theor Appl Genet. 1991 Dec;83(2):209-16
references.pmid: 24202360
references.citation: Nature. 2004 Sep 16;431(7006):356-63
references.pmid: 15372043
references.citation: J Exp Med. 2000 Feb 21;191(4):603-12
references.pmid: 10684852
references.citation: Gastroenterology. 2003 Aug;125(2):337-44
references.pmid: 12891534
references.citation: J Agric Food Chem. 2007 Jul 25;55(15):6074-81
references.pmid: 17580876
references.citation: Theor Appl Genet. 1985 Sep;70(6):610-9
references.pmid: 24253118
references.citation: FEBS Lett. 2004 May 21;566(1-3):223-8
references.pmid: 15147899
references.citation: PLoS One. 2011;6(9):e24754
references.pmid: 21935456
references.citation: J Exp Bot. 2002 Apr;53(370):947-58
references.pmid: 11912237
references.citation: Prog Biophys Mol Biol. 1994;61(1):37-59
references.pmid: 8202599
references.citation: Proc Natl Acad Sci U S A. 2010 Sep 28;107(39):17023-8
references.pmid: 20829492
references.citation: Biom J. 2008 Jun;50(3):346-63
references.pmid: 18481363
references.citation: Theor Appl Genet. 2007 Jun;115(1):77-86
references.pmid: 17458535
references.citation: J Allergy Clin Immunol. 2008 May;121(5):1210-1218.e4
references.pmid: 18378288
references.citation: Gastroenterology. 2002 Sep;123(3):803-9
references.pmid: 12198706
references.citation: Theor Appl Genet. 1993 Nov;87(3):374-8
references.pmid: 24190265
references.citation: BMC Med. 2011 Mar 09;9:23
references.pmid: 21392369
references.citation: Nat Rev Immunol. 2002 Sep;2(9):647-55
references.pmid: 12209133
references.citation: Plant Physiol. 2010 Dec;154(4):1842-54
references.pmid: 20940349
references.citation: Plant Physiol. 2011 May;156(1):330-45
references.pmid: 21398260