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A Digital PCR Assay to Quantify the Percentages of Hulled vs. Hulless Wheat in Flours and Flour-Based Products.

ABSTRACT: Several food products, made from hulled wheats, are now offered by the market, ranging from grains and pasta to flour and bakery products. The possibility of verifying the authenticity of wheat species used at any point in the production chain is relevant, in defense of both producers and consumers. A chip digital PCR assay has been developed to detect and quantify percentages of hulless (i.e., common and durum wheat) and hulled (i.e., einkorn, emmer and spelt) wheats in grains, flours and food products. The assay has been designed on a polymorphism in the miRNA172 target site of the AP2-5 transcription factor localized on chromosome 5A and involved in wheat spike morphogenesis and grain threshability. The assay has been evaluated even in a real-time PCR system to assess its applicability and to compare the analytical costs between dPCR and real-time PCR approaches.

SUBMITTER: Morcia C

PROVIDER: S-EPMC8614899 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:BACKGROUND:Wheat domestication is considered as one of the most important events in the development of human civilization. Wheat spikelets have undergone significant changes during evolution under domestication, resulting in soft glumes and larger kernels that are released easily upon threshing. Our main goal was to explore changes in transcriptome expression in glumes that accompanied wheat evolution under domestication. METHODS:A total of six tetraploid wheat accessions were selected for transcriptome profiling based on their rachis brittleness and glumes toughness. RNA pools from glumes of the central spikelet at heading time were used to construct cDNA libraries for sequencing. The trimmed reads from each library were separately aligned to the reference sub-genomes A and B, which were extracted from wheat survey sequence. Differentially expression analysis and functional annotation were performed between wild and domesticated wheat, to identity candidate genes associated with evolution under domestication. Selected candidate genes were validated using real time PCR. RESULTS:Transcriptome profiles of wild emmer wheat, wheat landraces, and wheat cultivars were compared using next generation sequencing (RNA-seq). We have found a total of 194,893 transcripts, of which 73,150 were shared between wild, landraces, and cultivars. From 781 differentially expressed genes (DEGs), 336 were down-regulated and 445 were up-regulated in the domesticated compared to wild wheat genotypes. Gene Ontology (GO) annotation assigned 293 DEGs (37.5 %) to GO term groups, of which 134 (17.1 %) were down-regulated and 159 (20.4 %) up-regulated in the domesticated wheat. Some of the down-regulated DEGs in domesticated wheat are related to the biosynthetic pathways that eventually define the mechanical strength of the glumes, such as cell wall, lignin, pectin and wax biosynthesis. The reduction in gene expression of such genes, may explain the softness of the glumes in the domesticated forms. In addition, we have identified genes involved in nutrient remobilization that may affect grain size and other agronomic traits evolved under domestication. CONCLUSIONS:The comparison of RNA-seq profiles between glumes of wheat groups differing in glumes toughness and rachis brittleness revealed a few DEGs that may be involved in glumes toughness and nutrient remobilization. These genes may be involved in processes of wheat improvement under domestication.

Project description:Amaranth is an underutilized pseudocereal that can be used to supplement wheat flour (WF) in order to improve the nutritional quality of bread. Bread digestibility is impacted by particle size which produces different nutritional properties. This research aims to evaluate the baking characteristics of optimal wheat-amaranth composite flour for each studied amaranth flour (AF) particle size at doses previously established based on an optimization process and to characterize from a physical, textural, nutritional, and sensorial point of view the obtained bread. The results revealed that the optimal wheat-amaranth composite flour with medium and small particle size, respectively showed a slightly lower α-amylase activity, while dough development time was significantly higher compared to the WF. A significant (p < 0.05) decrease was observed in the elasticity, deformation energy, and dynamic rheological parameters of the optimal composite dough for all the particle sizes, whereas fermentation parameters showed higher values compared to the control, indicating the ability of the gluten structure in large, medium, and small particle sizes of AF to hold the gas and to expand without collapsing. The physical, textural, and especially nutritional characteristics of the optimal WF-AF bread were enhanced. The sensory evaluation results revealed high scores (8.70) for the acceptability of optimal bread with a medium particle size as compared to wheat bread (8.25). The protein and ash content of the optimal breads with large, medium, and small AF particle sizes, respectively, increased significantly, from 8.92 to 10.58%, and 0.82 to 0.99%, respectively, relative to the wheat flour bread (8.35% and 0.72%, respectively). The mineral content was up to two times higher in the optimal breads compared to wheat flour bread. The findings generated from this study are helpful in bakery industry for designing products with enhanced nutritional properties and for introducing new products to the market.

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A Digital PCR Assay to Quantify the Percentages of Hulled vs. Hulless Wheat in Flours and Flour-Based Products.

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