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      Proteomic Analysis of Proteins Responsive to Drought and Low Temperature Stress in a Hard Red Spring Wheat Cultivar

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          Abstract

          Drought stress is becoming more prevalent with global warming, and has been shown to have large effects on gluten proteins linked to wheat bread making quality. Likewise, low temperature stress can detrimentally affect proteins in wheat. This study was done to determine the differential abundance of high molecular weight (HMW) glutenin proteins in a drought and low temperature stressed high quality hard red spring wheat cultivar (PAN3478), against a control. The treatments were applied in the greenhouse at the soft dough stage. HMW glutenin proteins were extracted from the flour, and were separated by using two-dimensional gel electrophoresis. Protein spots that had p values lower than 0.05 and fold values equal to or greater than 1.2 were considered to be significantly differentially abundant. These proteins were further analyzed by using tandem mass spectrometry. There was a 1.3 to 1.8 fold change in 17 protein spots due to the cold treatment. The drought treatment caused a 1.3 to 3.8 fold change in 19 protein spots. These spots matched either HMW or low molecular weight (LMW) glutenin subunits. In the latter case, the C subunits of LMW glutenins were notably found to be up-regulated under both stress conditions. All the proteins that have been identified can directly influence dough characteristics. Data are available via ProteomeXchange with the identifier PXD017578.

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          Improved staining of proteins in polyacrylamide gels including isoelectric focusing gels with clear background at nanogram sensitivity using Coomassie Brilliant Blue G-250 and R-250.

          An improved procedure for staining of proteins following separation in polyacrylamide gels is described which utilizes the colloidal properties of Coomassie Brilliant Blue G-250 and R-250. The new method is based on addition of 20% v/v methanol and higher concentrations of ammonium sulfate to the staining solution previously described. The method combines the advantage of much shorter staining time with high sensitivity, a clear background not requiring destaining, stepwise staining, and stable fixation after staining. The method has been applied to staining of polyacrylamide gels after sodium dodecyl sulfate-electrophoresis and isoelectric focusing in carrier ampholyte-generated pH gradients.
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            The low-molecular-weight glutenin subunits of wheat gluten

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              A simplified SDS—PAGE procedure for separating LMW subunits of glutenin

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                Author and article information

                Journal
                Molecules
                Molecules
                molecules
                Molecules
                MDPI
                1420-3049
                17 March 2020
                March 2020
                : 25
                : 6
                : 1366
                Affiliations
                [1 ]Department of Plant Sciences, University of the Free State, Bloemfontein 9300, South Africa; avbiljon@ 123456ufs.ac.za
                [2 ]Department of Agricultural and Forestry Sciences, University of Tuscia, 01100 Viterbo, Italy; masci@ 123456unitus.it
                [3 ]Department of Land, Environment, Agriculture and Forestry (TESAF), Research Group in Plant Pathology, University of Padova, Viale dell’Università 16, 35020 Legnaro (PD), Italy; silvio.tundo@ 123456unipd.it
                [4 ]Department of Chemical Sciences, Organic Mass Spectrometry Laboratory, University of Catania, 95131 Catania, Italy; v.muccilli@ 123456unict.it (V.M.); rsaletti@ 123456unict.it (R.S.)
                Author notes
                [* ]Correspondence: labuscm@ 123456ufs.ac.za ; Tel.: +27-51-4012715
                Author information
                https://orcid.org/0000-0003-0593-2678
                https://orcid.org/0000-0003-2857-4498
                https://orcid.org/0000-0002-3638-7345
                https://orcid.org/0000-0002-6212-5154
                Article
                molecules-25-01366
                10.3390/molecules25061366
                7144396
                32192150
                37610071-4238-4ad3-8ae8-60be1a7cbaaa
                © 2020 by the authors.

                Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license ( http://creativecommons.org/licenses/by/4.0/).

                History
                : 03 February 2020
                : 08 March 2020
                Categories
                Article

                abiotic stress,bread wheat,glutenin,proteomics
                abiotic stress, bread wheat, glutenin, proteomics

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