Genome mapping of seed-borne allergens and immunoresponsive proteins in wheat

Juhász, Angéla; Belova, Tatiana; Florides, Chris; Maulis, Csaba; Fischer, Iris; Gell, Gyöngyvér; Birinyi, Zsófia; Ong, Jamie W.L.; Keeble‐Gagnère, Gabriel; Maharajan, Amudha Deepalakshmi; Ma, Wujun; Gibson, Peter R.; Jia, Jizeng; Lang, Daniel; Mayer, Klaus; Spannagl, Manuel; Tye–Din, Jason A.; Appels, R.; Olsen, Odd Arne

doi:10.1126/sciadv.aar8602

Cited by 121 publications

(150 citation statements)

References 45 publications

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“…Beside CD, a wide range of wheat, rye, and barley proteins are potential allergens or triggers of innate immunity in NCGS. The recently published reference sequence RefSeq v1.0 of the hexaploid common wheat genome (International Wheat Genome Sequencing Consortium (IWGSC), 2018) provides further insights as the first reference to which known immunoreactive gluten and non-gluten proteins can be annotated (Juhasz et al, 2018).…”

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confidence: 99%

Characterization and Relative Quantitation of Wheat, Rye, and Barley Gluten Protein Types by Liquid Chromatography–Tandem Mass Spectrometry

2019

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The consumption of wheat, rye, and barley may cause adverse reactions to wheat such as celiac disease, non-celiac gluten/wheat sensitivity, or wheat allergy. The storage proteins (gluten) are known as major triggers, but also other functional protein groups such as α-amylase/trypsin-inhibitors or enzymes are possibly harmful for people suffering of adverse reactions to wheat. Gluten is widely used as a collective term for the complex protein mixture of wheat, rye or barley and can be subdivided into the following gluten protein types (GPTs): α-gliadins, γ-gliadins, ω5-gliadins, ω1,2-gliadins, high-and lowmolecular-weight glutenin subunits of wheat, ω-secalins, high-molecular-weight secalins, γ-75k-secalins and γ-40k-secalins of rye, and C-hordeins, γ-hordeins, B-hordeins, and D-hordeins of barley. GPTs isolated from the flours are useful as reference materials for clinical studies, diagnostics or in food analyses and to elucidate disease mechanisms. A combined strategy of protein separation according to solubility followed by preparative reversed-phase high-performance liquid chromatography was employed to purify the GPTs according to hydrophobicity. Due to the heterogeneity of gluten proteins and their partly polymeric nature, it is a challenge to obtain highly purified GPTs with only one protein group. Therefore, it is essential to characterize and identify the proteins and their proportions in each GPT. In this study, the complexity of gluten from wheat, rye, and barley was demonstrated by identification of the individual proteins employing an undirected proteomics strategy involving liquid chromatography-tandem mass spectrometry of tryptic and chymotryptic hydrolysates of the GPTs. Different protein groups were obtained and the relative composition of the GPTs was revealed. Multiple reaction monitoring liquid chromatography-tandem mass spectrometry was used for the relative quantitation of the most abundant gluten proteins. These analyses also allowed the identification of known wheat allergens and celiac disease-active peptides. Combined with functional assays, Frontiers in Plant Science | www.frontiersin.org

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Characterization and Relative Quantitation of Wheat, Rye, and Barley Gluten Protein Types by Liquid Chromatography–Tandem Mass Spectrometry

2019

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“…Prolamin superfamily protein’s function may relate to protease inhibition or involvement in plant defence. As discussed in Juhasz, et al 2018 (77) , the hydrophobic-seed domain containing proteins include, cortical cell delineating (78) , hydropho-seed domain containing protein ( 79-83 ), glycine-rich protein ( 84-86 ) and proline-rich protein ( 87, 88 ), which are found to be included in the plant defence system and have antifungal properties. Lipid transfer protein ( 39, 40, 89-91 ) and non-specific lipid transfer protein (92) have a LTP-2 domain, and have antifungal properties.…”

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confidence: 99%

Wheat avenin-like protein and its significant Fusarium Head Blight resistant functions

Zhang

Cao

Juhász

et al. 2018

Preprint

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Wheat Avenin-like proteins (TaALP) are atypical storage proteins belonging to the Prolamin superfamily. Previous studies on ALPs have focused on the proteins' positive effects on dough strength, whilst no correlation has been made between TaALPs and the plant immune system. Here, we performed genome-wide characterization of ALP encoding genes in bread wheat. In silico analyses indicated the presence of critical peptides in TaALPs that are active in the plant immune system. Pathogenesis-related nucleotide motifs were also identified in the putative promoter regions of TaALP encoding genes. RT-PCR was performed on TaALP and previously characterised pathogenesis resistance genes in developing wheat caryopses under control and Fusarium graminearum infection conditions, respectively. The results showed 2 that TaALP and NMT genes were upregulated upon F. graminearum inoculation. mRNA insitu hybridization showed that TaALP genes were expressed in the embryo, aleurone and sub-aleurone layer cells. Seven TaALP genes were cloned for the expression of recombinant proteins in Escherichia coli, which displayed significant inhibitory function on F. graminearum under anti-fungal tests. In addition, FHB index association analyses showed that allelic variations of two ALP genes on chromosome 7A were significantly correlated with FHB symptoms. Over-expression of an ALP gene on chromosome 7A showed an enhanced resistance to FHB. Yeast two Hybridization results revealed that ALPs have potential proteases inhibiting effect on metacaspases and beta-glucosidases. A vital infection process related pathogen protein, F. graminearum Beta-glucosidase was found to interact with ALPs.Our study is the first to report a novel function for wheat storage protein in fungal resistance, which greatly advances our understanding of the biological roles of this protein class. The findings in this study is of great significance for future wheat breeding and production. Preparation of Albumin and Globulin Protein extractsAustralia spring bread wheat varieties Mace and Spitfire were used for albumin and globulin protein extraction. The albumin/globulin proteins were extracted from 100 mg of flour according to the procedure of Dupont et al. (48). Briefly, 100 mg of flour was extracted with 1 mL of 0.3 M NaI, 7.5% 1-propanol (NaI-propanol), and centrifuged at 4500 g for 10 min, After two extractions, the supernatant fractions were pooled in 15 ml tubes, precipitated with four volumes of ice-cold (-20°C) NH 4 Ac-MeOH (0.1 M ammonium acetate in 100% methanol), stored at -20 °C for at least 48 h, and centrifuged as above. The supernatant fluid was transferred into 50 ml tubes and precipitated with four volumes of ice-cold acetone and incubated at -20 °C overnight. Following incubation, the fluid was centrifuged as above to yield albumin/globulin fraction pellets. Determination of Individual ALPs of wheat cv. Mace and Spitfire via RP-HPLC Analysis coupled with SDS-PAGE, MALDI-TOF, and LC/MS RP-HPLC Freeze-dried protein pellets were dissolved in 500 µL 6 M guani...

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“…First, more experiments should be conducted under different temperature, CO 2 and precipitation conditions to systematically analyze the responses of water cycle and crop growth in GSPAC to drought and cold stress as well as elevated CO 2 . Second, scientists have cracked the DNA sequence code of wheat, which is a major breakthrough that could improve global food security and offer the opportunity to improve the adaptability of the world's most common corps such as maize to changes in the environment [44]. This will bring hope to farmers who suffer from drought, as the major crop can be genetically modified (bio-genetic-engineering) to be drought proof/resistant, fast-growing, high yield and less water demand.…”

Section: Discussionmentioning

confidence: 99%

Responses of Water Fluxes and Water-Use Efficiency of Maize to Warming Based on Water Transformation Dynamical Processes Experimental Device (WTDPED) Experiment

Song

et al. 2018

Water

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Evaluating the impacts of warming on water balance components in the groundwater-soilplant-atmosphere continuum (GSPAC) and crop growth are crucial for assessing the risk of water resources and food security under future global warming. A water transformation dynamical processes experimental device (WTDPED) was developed using a chamber coupled with a weighing lysimeter and groundwater supply system, which could simultaneously control both climatic and ground-water level conditions and accurately monitor water fluxes in the GSPAC. Two experiments with maize under increased temperature by 2 • C (T-warm) and ambient temperature (T-control) scenarios were conducted via the WTDPED. The duration of growing season decreased from 125 days under T-control to 117 days under 2 • C warming. There was little difference of total evapotranspiration (ET) (332.6 mm vs. 332.5 mm), soil water storage change (∆W) (−119.0 mm vs. −119.0 mm), drainage (D) (−13.6 mm vs. −13.5 mm) between T-control and T-warm experiments. The average daily ET for maize significantly increased by approximately 6.7% (p < 0.05) in the T-warm experiment, especially during the sixth leaf to tasseling-silking stage with an increase of 0.36 mm with respect to the T-control experiment. There were evident decreases in LAI (leaf area index), whereas non-significant decreases in mean stem diameter, crop height and leaf chlorophyll content under T-warm compared to T-control experiment. However, the chlorophyll content increased by 12% during the sixth leaf to tasseling-silking stage under 2 • C warming, which accelerated the photosynthesis and transpiration rate. The grain yield and water-use efficiency (WUE) for maize increased by 11.0% and 11.1% in the T-warm experiment, respectively, especially due to enhanced growth during the sixth leaf to tasseling-silking stage. This study provided important references for agricultural planting and water management to adapt to a warming environment.

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Genome mapping of seed-borne allergens and immunoresponsive proteins in wheat

Abstract: We present a comprehensive analysis of variation in genes and gene expression of allergens and immunoresponsive proteins in wheat.

Cited by 121 publications

References 45 publications

Characterization and Relative Quantitation of Wheat, Rye, and Barley Gluten Protein Types by Liquid Chromatography–Tandem Mass Spectrometry

Characterization and Relative Quantitation of Wheat, Rye, and Barley Gluten Protein Types by Liquid Chromatography–Tandem Mass Spectrometry

Wheat avenin-like protein and its significant Fusarium Head Blight resistant functions

Responses of Water Fluxes and Water-Use Efficiency of Maize to Warming Based on Water Transformation Dynamical Processes Experimental Device (WTDPED) Experiment

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