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Analyses of albumins, globulins and amphiphilic proteins by proteomic approach give new insights on waxy wheat starch metabolism
Authors:Cl  ment Debiton, Marielle Merlino, Christophe Chambon, Emmanuelle Bancel, M  lanie Decourteix, V  ronique Planchot,G  rard Branlard
Affiliation:a INRA UMR-1095, 234 avenue du Brézet, 63100 Clermont-Ferrand, France;b INRA UR-370 PFEM-Plateau Protéomique, 63122 Saint-Genès-Champanelle, France;c PIAF UMR-547, 24 avenue des Landais, 63177 Aubière, France;d INRA Unité Biopolymères Interactions Assemblages, 44316 Nantes, France
Abstract:Starch is composed of two types of glucose polymers: amylose and amylopectin. The Waxy (Wx) locus controls amylose synthesis in the wheat kernel. Hexaploid wheat has three Wx loci located on chromosomes 7A (Wx-A1), 4A (Wx-B1), and 7D (Wx-D1). Eight near isogenic lines (NILs) of Triticum aestivum cv. Tremie with one, two or three Wx null alleles were used. The albumin–globulin fraction, and amphiphilic proteins were separated using 2-dimensional electrophoresis (2DE) allowing the changes in the waxy kernel to be identified. Albumin–globulin fraction showed overexpression of sucrose synthases in the waxy NILs compared to the normal form of Tremie and a decrease in many proteins related to stress and defence metabolism such as serpins. A subunit of ADP-glucose pyrophosphorylase (AGPase), which is known to play a major role in starch synthesis, was also shown to be down regulated in the waxy NILs. Amphiphilic proteins confirmed the observations made on the albumin–globulin fraction with a decrease in a stress-related protein. These different regulations linked to observations made on wheat kernel (thousand kernel weight (TKW), protein amount per grain, size and distribution of the starch granules) led to formulation of the hypothesis that waxy endosperm does not reach maturity of the wild-type endosperm.
Keywords:Starch metabolism   Waxy   Bi-dimensional electrophoresis   Triticum aestivum
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