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Gene expression profiling predicts processing properties in short-term imbibition of wheat

ABSTRACT: Objective: Germination of wheat maximizes phytochemical content and antioxidant activity while altering chemical composition, gluten content, and pasting properties. We previously reported marked changes in gene expression in soybean prior to germination before the radicle had grown from a seed. The present study investigated whether imibibition induces similar changes in wheat. Methods: Changes in gene expression profiles of wheat during short-term imbibition (0, 16, and 24 h) were evaluated by DNA microarray analysis. Gene Ontology (GO) analysis was carried out to categorize the function of genes with altered expression. The expression of genes encoding enzymes associated with starch breakdown was evaluated by quantitative real-time PCR, and changes in enzymatic activity were assessed with functional assays. Pasting properties of flour made from wheat seeds imbibed for different times were examined with a Rapid Visco Analyzer. The protein profile was determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis, and gluten content was quantified. Results: The GO analysis revealed that genes related to cellulose and cell wall synthesis were upregulated by imbibition for 16 h whereas those associated with polysaccharide catabolism and nucleosome assembly were upregulated in the subsequent 8 h. α-Amylase expression was highest after 24-h imbibition, with a corresponding increase in enzymatic activity. The pasting properties of wheat flour decreased when seeds were imbibed for over 16 h. Gluten was not degraded until 48 h imbibition. Conclusion: Short-term imbibition of wheat can produce a new type of starch with improved physical and functional properties that may be more appealing to consumers.

ORGANISM(S): Triticum aestivum

PROVIDER: GSE116933 | GEO | 2018/09/01

REPOSITORIES: GEO

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Project description:Within the complex wheat flour proteome, the gluten proteins have attracted most of the attention because of their importance in determining the functional properties of wheat flour doughs and their roles in human health conditions such as celiac disease and food allergies. However, certain non-gluten proteins also trigger immunological responses but may be present in flour in low amounts or obscured by the more abundant gluten proteins in two-dimensional gels of total protein preparations. Non-gluten proteins were preferentially extracted from the flour with a dilute salt solution and separated by two-dimensional gel electrophoresis. Proteins in 172 gel spots were identified by tandem mass spectrometry after cleavage with trypsin or chymotrypsin. Fifty-seven different types of non-gluten proteins were identified, including 14 types that are known or suspected immunogenic proteins. The predominant proteins in 18 gel spots were gluten proteins. Transgenic wheat lines in which specific groups of gluten proteins were suppressed by RNA interference were used to estimate the amount of carry-over of gluten proteins in the salt-soluble protein fraction. Analysis of salt-soluble proteins from a transgenic line missing omega-1,2 gliadins demonstrated that certain omega-1,2 gliadins were present in large amounts in the salt-soluble fraction and obscured relatively small amounts of beta-amylase and protein disulfide isomerase. In comparison, analysis of a transgenic line in which alpha gliadins were absent revealed that glyceraldehyde-3 phosphate dehydrogenase was a moderately abundant protein that co-migrated with several alpha gliadins. The proteomic map of the non-gluten protein fraction of wheat flour developed in this study complements a proteomic map of the total flour proteins developed previously for the same cultivar. Knowing the identities of low abundance proteins in the flour as well as proteins that are hidden by some of the major gluten proteins on two-dimensional gels is critical for studies aimed at assessing the immunogenic potential of wheat flour and determining how the growth conditions of the plants affect the levels of specific immunogenic proteins in the flour.

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Gene expression profiling predicts processing properties in short-term imbibition of wheat

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