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Background. Due to its simplicity and good reproducibility, SDS-electrophoresis of seed proteins is widely used for investigating the gene pool of legumes and other plants, for species and varietal identification, analyzing the intraspecific variability, and registering collection material. The data obtained by this method agree well with the DNA analysis results complement them. Typically, legume seed proteins, including representatives of the genus Vicia L., show clear informative SDS electrophoretic profiles. When analyzing seed accessions of vetch species of the section Peregrinae Kupicha using standard approaches previously developed at VIR and approved by ISTA (the International Seed Testing Association), clear electrophoretic protein profiles could not be obtained for many accessions. This called into question the applicability of standard approaches to identifying vetch species in the section Peregrinae. The objective of the work was to clarify the nature of seed proteins degradation in representatives of the Peregrinae section and to find ways to prevent it to ensure the possibility of carrying out species identification and registration of all accessions in the vetch collection using a unified approach. Material and methods. Seed proteins of a number of vetch species Vicia L. from sections Bithynicae (B. Fedtsch.) Maxted, Hypechusa (Alef.) Aschers. et Graebner, Microcarinae Maxted and Peregrinae, members of the subgenus Vicia, were analyzed by SDS-electrophoresis using the standard method of protein extraction from flour with 0.025 M Tris-glycine buffer pH 8.3 at room temperature and its modifications, including heating the extract at 80°C or 100°C with or without the addition of 2-mercaptoethanol, as well as the addition of cysteine and serine protease inhibitors. Results and discussion. An analysis of seed proteins of representatives of most sections of the subgenus Vicia yielded informative species-specific protein profiles, whereas species of the section Peregrinae were characterized by the protein profiles, which indicated protein degradation, and species of this section differed in the frequency of such profile occurrence. While such profiles were obtained for all seeds of seven accessions of V. aintabensis Boiss & Hausskn. ex Boiss differing in geographical origin, year and place of regeneration, and 12 out of 13 of V. peregrina L. accessions demonstrated profiles of partially or completely degraded proteins, complete seed protein profiles were obtained for six out of nine V. michauxii Sprengel accessions. A change in conditions for protein isolation, namely replacement of their extraction from flour with Tris-glycine buffer pH 8.3 at room temperature with extraction in the same buffer by a short-term heating at 100°C in the presence of 2-mercaptoethanol, made it possible to obtain complete protein profiles for all accessions representing the section Peregrinae. The protein profiles of representatives of other vetch sections, as well as the profile of soybean proteins used as a standard for legume species identification, did not differ from the original ones under the modified conditions. Conclusions. The obtained results suggest that protein degradation in species of the Peregrinae section is associated with the abnormal activity of endogenous seed proteases under standard protein extraction conditions, and this trait is determined genotypically. A new modification of the method for isolating proteins from seeds makes it possible to apply the generally accepted approaches based on SDS-electrophoresis in the analysis of the gene pool of the Peregrinae section of the subgenus Vicia, as well as other vetch species.

Main Conclusion: A mutation was first found to cause the great generation of glutelin precursors (proglutelins) in rice (Oryza sativa L.) endosperm, and thus referred to as GPGG1. The GPGG1 was involved in synthesis and compartmentation of storage proteins. The PPR-like gene in GPGG1-mapped region was determined as its candidate gene. In the wild type rice, glutelins and prolamins are synthesized on respective subdomains of rough endoplasmic reticulum (ER) and intracellularly compartmentalized into different storage protein bodies. In this study, a storage protein mutant was obtained and characterized by the great generation of proglutelins combining with the lacking of 13 kD prolamins. A dominant genic-mutation, referred to as GPGG1, was clarified to result in the proteinous alteration. Novel saccular composite-ER was shown to act in the synthesis of proglutelins and 14 kD prolamins in the mutant. Additionally, a series of organelles including newly occurring several compartments were shown to function in the transfer, trans-plasmalemmal transport, delivery, deposition and degradation of storage proteins in the mutant. The GPGG1 gene was mapped to a 67.256 kb region of chromosome 12, the pentatricopeptide repeat (PPR)-like gene in this region was detected to contain mutational sites. [ABSTRACT FROM AUTHOR]

Among a vast number of polymorphisms in the wheat genome, of special interest are those that are markers for loci under selection. Such loci include storage protein loci, which directly determine the level of bread-making quality, and disease resistance genes. In this study we analyzed diversity of functional markers (seven storage protein loci and three disease resistance genes) in the group of East European winter common wheat genotypes (cultivars and lines of Poltava breeding). Using the UPGMA or NJ methods, the cultivars and lines were divided into two clusters differing in the Glu-1 quality score. The cultivars of cluster 1 with the lower mean Glu-1 quality score predominantly carried the combination of alleles Glu-B1c, Glu-D1a, and Gli-B1e, as well as the resistance allele of Lr34, whereas those of cluster 2 with the higher quality score mainly had the alleles Glu-B1u, Glu-D1d, and Gli-B1b. In the sample of Poltava cultivars, a high correlation between the Glu-1 quality score and the sedimentation value (0.84) was observed. On average for two years, the group of cultivars from cluster 1 showed higher grain protein content and some spike traits in comparison with the means of the sample from cluster 2. Significant associations of Lr34 alleles and alleles at some storage protein loci were revealed, in particular, the association of the resistance allele Lr34R and the allele Glu-D1a in both cultivar and line groups, indicating the adaptive value of such a combination for local soil and climate conditions. [ABSTRACT FROM AUTHOR]

Silybum marianum and Silybum eburneum are wild edible Mediterranean plants used in the human diet. This study presents the initial findings on the phytochemical characterization of Tunisian S. marianum and S. eburneum organs. It examined their mineral, sugar, organic acid, polyphenolic, and seed storage protein contents, as well as their antioxidant potential. In S. marianum, stems had high sodium and potassium contents, while the immature and mature seeds were rich in calcium and magnesium. However, S. eburneum had high potassium levels in stems and high sodium and calcium levels in the flowers. S. marianum showed substantial fructose variation among its organs. Conversely, S. eburneum exhibited significant heterogeneity in glucose, sucrose, and maltose levels across its organs, with maltose exclusively detected in the immature seeds. A notable organ-dependent distribution of organic acids was observed among the two species. Higher levels of phenolic contents were detected in both mature and immature seeds in both species compared to the other plant parts. The seeds possessed higher antioxidant activities than other plant organs. In both S. marianum and S. eburneum seeds, albumins and globulins were the predominant protein fractions. This study brings evidence supporting the important potential of Silybum organs as sources of nutrients with antioxidant properties for producing functional food. [ABSTRACT FROM AUTHOR]

Grain storage proteins (GSPs) play a pivotal role in determining wheat's food processing characteristics and commercial value. The composition of GSPs in wheat is exceptionally intricate, characterized by numerous alleles associated with each relevant gene locus. However, due to the absence of a straightforward and universally applicable evaluation method, the quantitative comparison of the effects of various GSPs on flour properties remains challenging. Extensive research has substantiated a close relationship between the content of free sulfhydryl groups and disulfide bonds in flour proteins and their impact on dough properties. In this investigation, we leveraged the highly significant positive correlation observed between the content of free sulfhydryl groups and a key dough strength parameter, known as stability time, to establish a simplified model for assessing the influence of GSPs on flour properties. This model is rooted in predicting free sulfhydryl groups and disulfide bonds. Subsequently, we applied this model to assess the contributions of distinct GSPs to flour properties. Our findings revealed that high molecular weight glutenin subunits (HMW-GSs) 1Dy10, DX5, and 1Dy3 garnered elevated scores. In contrast, low molecular weight glutenin subunits (LMW-GSs) Glu-B3 and Glu-D3 received a notably higher score (7.2) compared to 1Dy10 (6.3). Consequently, we postulate that the proteins Glu-B3 and Glu-D3 within the LMW-GS category are the primary contributors to dough strength, challenging previous assumptions that HMW-GSs hold primary sway. Furthermore, certain gliadins, avenin-like proteins, and gamma-secalin proteins were identified as vital storage proteins influencing dough strength. As a result, wheat breeding programs keen on manipulating dough quality should consider these proteins within the Glu-B3 and Glu-D3 categories, along with specific gliadins and avenin-like proteins. [ABSTRACT FROM AUTHOR]

IntroductionPeanut allergy (PA) in children is a major concern. There is a need for better biological material for both diagnosis and oral immunotherapy (OIT) treatments. The unique state of seeds at early reproductive stages may affect the allergenicity of storage proteins, and impact clinical diagnostic and OIT protocols. The objective of this study was to evaluate the major allergen content in sequential seed developmental stages and monitor allergenicity via specific IgE binding quantification and skin prick testing.MethodsSeeds were collected from peanut plants and sorted into five developmental stages: initial (S1), developing (S2), full-size without coloration (S3), full-size with coloration (S4), and fully mature (S5) seeds. Samples were characterized by RNA-Seq, ELISA, and immunohistochemistry. Lyophilized, ground preparations were used for evaluation of skin test responses in sixty challenge-proven PA children.ResultsGene expression, protein content, and specific IgE binding of allergenic proteins increased throughout seed maturation and development. An expression bias towards the less allergenic A-genome copy of the major allergen Ara h 2 was found in earlier stages, especially in stage S2. Immunohistochemical staining showed that Ara h 2 is more dispersed in the cell and less accumulated within organized bodies at stage S2 versus stage S4. Significant differences were found in mean wheal responses between the commercial peanut extract (equivalent to stage S5) and stages S1 and S2, but not with stage S4, upon skin prick testing in subjects with PA.DiscussionThe observed decrease in peanut-specific IgE binding of immature peanut seeds may be a result not only of decreased amounts of allergenic proteins, but also of profound changes in seed composition and conformation. This may be significant for developing a safer and more effective peanut OIT protocol.

Relevance. In Belarus, no full-fledged studies have previously been carried out to identify species diversity, to identify forms of vegetable beans that have similar genetic diversity. The purpose of the research is to determine the number and ratio of biotypes, to identify the level of genetic polymorphism in vegetable bean varieties by electrophoresis of storage proteins.Materials and Methods. The research was carried out at the BSAA in 2021–2022 on soddy-podzolic medium loamy soil. The experience was based on generally accepted methods and guidelines. The objects of research were 42 varieties of vegetable beans (34 - bush form) and (8 - climbing) of Belarusian and Russian selection. Analysis of seed storage proteins in vegetable bean varieties and identification of spectra were carried out according to the methods. To assess the differentiating positions (zones) of the spectrum, identify protein components, and estimate the molecular masses of proteins, standard marker solutions of proteins “Thermo Scientific” – Unstained Protein Ladder (range 5–112 kDa, number of identified proteins – 11) were used.Results. Differences between genotypes were revealed in internal genetic diversity and elements of the protein spectrum of globulins. It has been established that vegetable bean varieties are characterized by hidden genetic variability and a certain level of polymorphism with a different number of biotypes in the structure of the varietal population. In 60% of the analyzed forms, there is a direct relationship between the manifestation of rare, selection-significant components and the degree of complexity of the total component composition of the protein spectrum. This criterion has practical significance and can be used in breeding work with the vegetable bean crop.

Abscisic acid (ABA) and gibberellic acid (GA) antagonistically regulate many aspects of plant growth, including seed dormancy and germination. The effects of these hormones are mediated by a complex network of positive and negative regulators of transcription. The DELLA family of proteins repress GA response, and can promote an ABA response via interactions with numerous regulators, including the ABA-insensitive (ABI) transcription factors. The AFP family of ABI5 binding proteins are repressors of the ABA response. This study tested the hypothesis that the AFPs also interact antagonistically with DELLA proteins. Members of these protein families interacted weakly in yeast two-hybrid and bimolecular fluorescence complementation studies. Overexpression of AFPs in sleepy1, a mutant that over-accumulates DELLA proteins, suppressed DELLA-induced overaccumulation of storage proteins, hyperdormancy and hypersensitivity to ABA, but did not alter the dwarf phenotype of the mutant. The interaction appeared to reflect additive effects of the AFPs and DELLAs, consistent with action in convergent pathways.

The article is devoted to the study of the wheat samples of the 3rd classwith the same quality indicators according to DSTU 3768, but had different baking properties. In particular, the “strength” of flour (W) significantly differed. The quantitative and qualitative state of the gluten-protein complex is significant factor for obtaining grain and flour of high baking quality. But only the quality indicators of red winter wheat according to DSTU 3768 - protein content, amount and quality of gluten cannot objectively characterize the baking quality of a specific batch of grain. For the main indications of the Ukrainian standard for wheat, all batches of wheat are practically identical. Foreign buyers do not use our Ukrainian standards, they give precedence to international methods and one of the most integral of them is the “strength” (W). The method demonstrates the main characteristics of the physical properties of wheat dough, which maybe especially important during bread making: elasticity (P), extensibility (L) and alveogram configuration coefficient (P/L). It has been established that the reason is the difference in the genetically determined alleles of the storage proteins of the studied samples. In the last years "filler" and "weak" varieties of wheat of domestic and foreign selection have appeared in Ukraine. In the conditions of replacing traditionally grown varieties of "strong" wheat in Ukraine with new high-yielding varieties of domestic and foreign breeding, it is not enough to determine the parameters recorded in DSTU 3768 to determine the baking quality, it is necessary to introduce new, more objective research methods. It is concluded that for an objective assessment of the baking properties of wheat, international standards ISO 27971, ISO 21415, ISO 5529 should be used.

Silybum marianum and Silybum eburneum are wild edible Mediterranean plants used in the human diet. This study presents the initial findings on the phytochemical characterization of Tunisian S. marianum and S. eburneum organs. It examined their mineral, sugar, organic acid, polyphenolic, and seed storage protein contents, as well as their antioxidant potential. In S. marianum, stems had high sodium and potassium contents, while the immature and mature seeds were rich in calcium and magnesium. However, S. eburneum had high potassium levels in stems and high sodium and calcium levels in the flowers. S. marianum showed substantial fructose variation among its organs. Conversely, S. eburneum exhibited significant heterogeneity in glucose, sucrose, and maltose levels across its organs, with maltose exclusively detected in the immature seeds. A notable organ-dependent distribution of organic acids was observed among the two species. Higher levels of phenolic contents were detected in both mature and immature seeds in both species compared to the other plant parts. The seeds possessed higher antioxidant activities than other plant organs. In both S. marianum and S. eburneum seeds, albumins and globulins were the predominant protein fractions. This study brings evidence supporting the important potential of Silybum organs as sources of nutrients with antioxidant properties for producing functional food.

Improving and stabilizing the quality of seed proteins are of growing interest in the current food and agroecological transitions. Sulfur is a key determinant of this quality since it is essential for the synthesis of sulfur-rich proteins in seeds. A lack of sulfur provokes drastic changes in seed protein composition, negatively impacting the nutritional and functional properties of proteins, and leading in some cases to diseases or health problems in humans. Sulfur also plays a crucial role in stress tolerance through the synthesis of antioxidant or protective molecules. In the context of climate change, questions arise regarding the trade-off between seed yield and seed quality with respect to sulfur availability and use by crops that represent important sources of proteins for human nutrition. Here, we review recent work obtained in legumes, cereals, as well as in Arabidopsis, that present major advances on: (i) the interaction between sulfur nutrition and environmental or nutritional stresses with regard to seed yield and protein composition; (ii) metabolic pathways that merit to be targeted to mitigate negative impacts of environmental stresses on seed protein quality; and (iii) the importance of sulfur homeostasis for the regulation of seed protein composition and its interplay with seed redox homeostasis. [ABSTRACT FROM AUTHOR]

Rice protein is considered an important dietary protein source. Information regarding rice nutrient-related protein expression is still scarce, hence further study investigating this aspect is highly needed. Herein, we applied sequential window acquisition of all theoretical mass spectra (SWATH-MS) for a comparative proteomic analysis across six different Malaysian rice varieties. These consisted of black rice (BR: PH9 and BALI), red rice (RR: MRQ100 and MRM16), and white rice (WR: MRQ76 and MR297). This study aimed to unravel rice nutrient-related proteins and if their expressions were significantly different across varieties. A total of 4022 quantified proteins were found to be significantly expressed across all varieties with a false discovery rate (FDR) < 1% and p < 0.05. While among 1792 differentially expressed proteins (DEPs) that were identified, 74 DEPs had functions related to nutrient biosynthesis. There were significantly higher expressions of key enzymes for the carotenoid and amylopectin biosynthesis pathways and seed storage proteins, i.e., prolamins and glutelins in RR. Glycoproteins such as cupin and germin-like protein, as well as enzymes that are involved in the biosynthesis of thiamine and anthocyanin were abundantly found in BR. WR was particularly enriched with biosynthesis enzymes for essential amino acids (methionine and arginine), vitamin B, and unsaturated fatty acid. This study provides us insights into the differential expressions of storage and functional proteins with nutrient-related properties in shaping rice grain pigmentations and plant immunity, as well as in contributing diverse health benefits as daily functional food for human consumption.

A collection of 15 triticale varieties (Triticosecale) grown in Bulgaria, 17 varieties grown in Poland, 1 variety grown in France and 3 variety grown in Germany were analyzed using sodium dodecilsuphate polyacrylamide gel electrophoresis (SDS-PAGE) to describe allelic variability in the storage proteins encoded at the Glu-1 (Glu-A1, Glu-B1 and Glu-R1), Glu-3 (Glu-A3 and Glu-B3), Glu-B2 and Gli-R2 loci. Thirty-two alleles were identified. These alleles formed 40 allelic configurations including 37 specific for one cultivar each. Two new allelic forms for high-molecular weight (HMW) - secalins were found in Glu-R1 loci and one new allelic form of 75K γ-secalins (named "new") was found in Gli-R2 loci. 83% of the triticale cultivars were homogenous, only 17% showed two or more diagrams respectively in HMW glutenin subunits, low-molecular weight (LMW) glutenin subunits and 75K γ-secalins. Thus, the total number of genotypes in the study increased to fifty-four. Twenty-nine varieties possessed allele b and seven varieties commonly grown in Bulgaria possessed allele 'a' in Glu-A1 loci. These alleles are associated with good bread-making qualities. According to the data on the allelic composition, the cultivars were classified depending on their origin; it was found out that all analyzed accessions from Poland, Germany and France, as well as those developed in Bulgaria, fell within the same group, namely the group of triticale varieties of winter type developed through European germ plasma. [ABSTRACT FROM AUTHOR]

Rice protein is considered an important dietary protein source. Information regarding rice nutrient-related protein expression is still scarce, hence further study investigating this aspect is highly needed. Herein, we applied sequential window acquisition of all theoretical mass spectra (SWATH-MS) for a comparative proteomic analysis across six different Malaysian rice varieties. These consisted of black rice (BR: PH9 and BALI), red rice (RR: MRQ100 and MRM16), and white rice (WR: MRQ76 and MR297). This study aimed to unravel rice nutrient-related proteins and if their expressions were significantly different across varieties. A total of 4022 quantified proteins were found to be significantly expressed across all varieties with a false discovery rate (FDR) < 1% and p < 0.05. While among 1792 differentially expressed proteins (DEPs) that were identified, 74 DEPs had functions related to nutrient biosynthesis. There were significantly higher expressions of key enzymes for the carotenoid and amylopectin biosynthesis pathways and seed storage proteins, i.e., prolamins and glutelins in RR. Glycoproteins such as cupin and germin-like protein, as well as enzymes that are involved in the biosynthesis of thiamine and anthocyanin were abundantly found in BR. WR was particularly enriched with biosynthesis enzymes for essential amino acids (methionine and arginine), vitamin B, and unsaturated fatty acid. This study provides us insights into the differential expressions of storage and functional proteins with nutrient-related properties in shaping rice grain pigmentations and plant immunity, as well as in contributing diverse health benefits as daily functional food for human consumption. [ABSTRACT FROM AUTHOR]

Next to cow's milk and eggs, plant foods, i.e., legumes, tree nuts and cereal grains, most often sensitise atopic children. Storage proteins constitutes the most relevant protein fraction of plant foods, causing primary sensitisation. They exhibit strong allergenic properties and immunogenicity. Our goal was to analyse sensitisation to 26 plant storage proteins in a group of 76 children aged 0–5 years with chronic symptoms of atopic dermatitis using Allergy Explorer ALEX2 and to discover changes in serum protein–peptide patterns in allergic patients with the use of MALDI-TOF-MS. We reported that 25% of children were allergic to 2S albumins, 19.7% to 7S globulins, 13.2% to 11S globulins and 1.3% to cereal prolamins. The most common allergenic molecules were Ara h 1 (18.4%), Ara h 2 (17.1%), Ara h 6 (15.8%) and Ara h 3 (11.8%) from peanuts, and the mean serum sIgE concentrations in allergic patients were 10.93 kUA/L, 15.353 kUA/L, 15.359 kUA/L and 9.038 kUA/L, respectively. In children allergic to storage proteins compared to the other patients (both allergic and non-allergic), the cell cycle control protein 50A, testis-expressed sequence 13B, DENN domain-containing protein 5A and SKI family transcriptional corepressor 2 were altered. Our results indicate that the IgE-mediated allergy to storage proteins is a huge problem in a group of young, atopic children, and show the potential of proteomic analysis in the prediction of primary sensitisation to plant foods. It is the next crucial step for understanding the molecular consequences of allergy to storage proteins. [ABSTRACT FROM AUTHOR]

Introduction: Plant growth-promoting bacteria (PGPBs) could be developed as a sustainable strategy to promote plant growth and yield to feed the evergrowing global population with nutritious food. Foliar application of nano-zinc oxide (ZnO) is an environmentally safe strategy that alleviates zinc (Zn) malnutrition by improving biochemical attributes and storage proteins of grain. Methods: In this context, the current study aimed to investigate the combined effect of seed inoculation with PGPBs and foliar nano-ZnO application on the growth, biochemical attributes, nutrient metabolism, and yield of maize in the tropical savannah of Brazil. The treatments consisted of four PGPB inoculations [i.e., without inoculation, Azospirillum brasilense (A. brasilense), Bacillus subtilis (B. subtilis), Pseudomonas fluorescens (P. fluorescens), which was applied on the seeds] and two doses of Zn (i.e., 0 and 3 kg ha-1, applied from nano-ZnO in two splits on the leaf). Results: Inoculation of B. subtilis with foliar ZnO application increased shoot dry matter (7.3 and 9.8%) and grain yield (17.1 and 16.7%) in 2019-20 and 2020-2021 crop seasons respectively. Inoculation with A. brasilense increased 100-grains weight by 9.5% in both crop seasons. Shoot Zn accumulation was improved by 30 and 51% with inoculation of P. fluorescens in 2019-20 and 2020-2021 crop seasons. Whereas grain Zn accumulation was improved by 49 and 50.7% with inoculation of B. subtilis and P. fluorescens respectively. In addition, biochemical attributes (chlorophyll a, b and total, carotenoids, total soluble sugar and amino acids) were improved with inoculation of B. subtilis along with foliar nano ZnO application as compared to other treatments. Co-application of P. fluorescens with foliar ZnO improved concentration of grains albumin (20 and 13%) and globulin (39 and 30%). Also, co-application of B. subtilis and foliar ZnO improved concentration of grains glutelin (8.8 and 8.7%) and prolamin (15 and 21%) in first and second seasons. Discussion: Therefore, inoculation of B. subtilis and P. fluorescens with foliar nano-ZnO application is considered a sustainable and environmentally safe strategy for improving the biochemical, metabolic, nutritional, and productivity attributes of maize in tropical Savannah regions. [ABSTRACT FROM AUTHOR]

In this study were fractionated and characterized reserve endosperm proteins, hordeins of seven spring barley genotypes with different origin: Zernogradskii (Russia), Bodega, Fink, Scarlett, Barke (Germany), Josefin and Astoria (France) by SDSPAGE electrophoresis. On the basis of the obtained spectra are identified 19 bands (D + C + B) with different relative electrophoretic mobility and intensity. The electrophoresis profiles of the groups D-, C- and B-hordein are designated as separate types (models) using the index corresponding to hordein blocks. We have established one profile type for D-hordein (D1), two- for C-hordein (C1, C2), and five - for B hordein (B1, B2, B3, B4, B5). Based on these results are constructed hordein formulas (configurations) of accessions, which enable the expression of specific varietal characteristics and prove the existence of the inter-allelic variation (hordein polymorphism) due to the presence or absence of protein components and their different electrophoretic mobility in the profiles of D-, C- and B-hordein. [ABSTRACT FROM AUTHOR]

The article is devoted to the study of the wheat samples of the 3rd classwith the same quality indicators according to DSTU 3768, but had different baking properties. In particular, the “strength” of flour (W) significantly differed. The quantitative and qualitative state of the gluten-protein complex is significant factor for obtaining grain and flour of high baking quality. But only the quality indicators of red winter wheat according to DSTU 3768 - protein content, amount and quality of gluten cannot objectively characterize the baking quality of a specific batch of grain. For the main indications of the Ukrainian standard for wheat, all batches of wheat are practically identical. Foreign buyers do not use our Ukrainian standards, they give precedence to international methods and one of the most integral of them is the “strength” (W). The method demonstrates the main characteristics of the physical properties of wheat dough, which maybe especially important during bread making: elasticity (P), extensibility (L) and alveogram configuration coefficient (P/L). It has been established that the reason is the difference in the genetically determined alleles of the storage proteins of the studied samples. In the last years "filler" and "weak" varieties of wheat of domestic and foreign selection have appeared in Ukraine. In the conditions of replacing traditionally grown varieties of "strong" wheat in Ukraine with new high-yielding varieties of domestic and foreign breeding, it is not enough to determine the parameters recorded in DSTU 3768 to determine the baking quality, it is necessary to introduce new, more objective research methods. It is concluded that for an objective assessment of the baking properties of wheat. [ABSTRACT FROM AUTHOR]

The content and composition of wheat storage proteins are the major determinants of dough rheological properties and breadmaking quality and are influenced by cultivation conditions. This study aimed to investigate the effects of water deficit and high N-fertilizer application on wheat storage protein synthesis, gluten secondary structure, and breadmaking quality. Reverse-phase ultrahigh-performance liquid chromatography analysis showed that storage protein and gluten macropolymer accumulation was promoted under both independent applications and a combination of water-deficit and high N-fertilizer treatments. Fourier-transform infrared spectroscopy showed that water deficit and high N-fertilizer treatments generally improved protein secondary structure formation and lipid accumulation, and reduced flour moisture. In particular, high N-fertilizer application increased β-sheet content by 10.4% and the combination of water-deficit and high N-fertilizer treatments increased random coil content by 7.6%. These changes in gluten content and secondary structure led to improved dough rheological properties and breadmaking quality, including superior loaf internal structure, volume, and score. Our results demonstrate that moderately high N-fertilizer application under drought conditions can improve gluten accumulation, gluten secondary structure formation, and baking quality.

IntroductionPlant growth-promoting bacteria (PGPBs) could be developed as a sustainable strategy to promote plant growth and yield to feed the ever-growing global population with nutritious food. Foliar application of nano-zinc oxide (ZnO) is an environmentally safe strategy that alleviates zinc (Zn) malnutrition by improving biochemical attributes and storage proteins of grain.MethodsIn this context, the current study aimed to investigate the combined effect of seed inoculation with PGPBs and foliar nano-ZnO application on the growth, biochemical attributes, nutrient metabolism, and yield of maize in the tropical savannah of Brazil. The treatments consisted of four PGPB inoculations [i.e., without inoculation, Azospirillum brasilense (A. brasilense), Bacillus subtilis (B. subtilis), Pseudomonas fluorescens (P. fluorescens), which was applied on the seeds] and two doses of Zn (i.e., 0 and 3 kg ha−1, applied from nano-ZnO in two splits on the leaf). ResultsInoculation of B. subtilis with foliar ZnO application increased shoot dry matter (7.3 and 9.8%) and grain yield (17.1 and 16.7%) in 2019-20 and 2020-2021 crop seasons respectively. Inoculation with A. brasilense increased 100-grains weight by 9.5% in both crop seasons. Shoot Zn accumulation was improved by 30 and 51% with inoculation of P. fluorescens in 2019-20 and 2020-2021 crop seasons. Whereas grain Zn accumulation was improved by 49 and 50.7% with inoculation of B. subtilis and P. fluorescens respectively. In addition, biochemical attributes (chlorophyll a, b and total, carotenoids, total soluble sugar and amino acids) were improved with inoculation of B. subtilis along with foliar nano ZnO application as compared to other treatments. Co-application of P. fluorescens with foliar ZnO improved concentration of grains albumin (20 and 13%) and globulin (39 and 30%). Also, co-application of B. subtilis and foliar ZnO improved concentration of grains glutelin (8.8 and 8.7%) and prolamin (15 and 21%) in first and second seasons.DiscussionTherefore, inoculation of B. subtilis and P. fluorescens with foliar nano-ZnO application is considered a sustainable and environmentally safe strategy for improving the biochemical, metabolic, nutritional, and productivity attributes of maize in tropical Savannah regions.

Subcellular mRNA localization is an evolutionarily conserved mechanism to spatially and temporally drive local translation and, in turn, protein targeting. Hence, this mechanism achieves precise control of gene expression and establishes functional and structural networks during cell growth and development as well as during stimuli response. Since its discovery in ascidian eggs, mRNA localization has been extensively studied in animal and yeast cells. Although our knowledge of subcellular mRNA localization in plant cells lags considerably behind other biological systems, mRNA localization to the endoplasmic reticulum (ER) has also been well established since its discovery in cereal endosperm cells in the early 1990s. Storage protein mRNA targeting to distinct subdomains of the ER determines efficient accumulation of the corresponding proteins in different endosomal storage sites and, in turn, underlies storage organelle biogenesis in cereal grains. The targeting process requires the presence of RNA localization elements, also called zipcodes, and specific RNA-binding proteins that recognize and bind these zipcodes and recruit other factors to mediate active transport. Here, we review the current knowledge of the mechanisms and functions of mRNA localization to the ER in plant cells and address directions for future research. [ABSTRACT FROM AUTHOR]

Background and objectives: Rice is a good source of energy and has the highest protein efficiency among plant proteins. However, its protein content is relatively low compared to other cereals. Seed storage protein in rice classified into four groups: albumin and globulin in aleurone layer, and prolamine and glutelin in endosperm. Therefore, during the bleaching process of rice grains, albumin proteins and part of globulin are removed by its bran. Since the protein content of the grain and its amount will be directly related to its nutritional quality and due to the importance of rice grain in diet, seed storage proteins quality in a quantitative and qualitative breeding population of rice such as two new mutant cultivars, Roshan and Shahriar, with their parents (Nemat and Amol3 respectively) as well as qualitative varieties, Sang-tarom, along with two different varieties of brown and black rice was studied. Materials and methods: Qualitative properties of grain proteins were evaluated based on their solubility as well as the seed storage proteins For albumin, globulin, prolamine and glutelin proteins. Quantification of stored proteins was performed by Bradford reagent and quantification of their subunits by densitometric analysis. Results: The results showed that there is a significant difference for the amount of seed storage proteins of different rice genotypes. So that Sang)tarom and Roshan cultivars had the highest amount of albumin protein (19.53 and 22.40 mg/g respectively). This was while the brown grain variety with the lowest amount of albumin and globulin protein and the highest amount of glutelin and prolamine proteins has the highest amount of total protein (1640.43 mg/g). Evaluation of the banding pattern of stored storage proteins also showed that this banding pattern was the same in different genotypes except in Roshan (60 kDa) and Shahriar (13 kDa) mutant cultivars. Densitometric analysis of protein subunits also showed that the highest amount of glutelin subunits was present in Sang-tarom cultivar and the lowest in Shahriar cultivar and brown grain variety. Also, the highest amount of prolamine subunits was observed in Sang-tarom cultivar and brown grain variety and the lowest in Shahriar cultivar. As Shahriar cultivar was able to have the highest amount of glutelin to prolamine protein. Conclusion: Although the profiles of seed storage proteins in different varieties of rice are the same, but this pattern is different in the two cultivars of mutant Roshan and Shahriyar. So that, the absent of the 60kDa subunit of proglutelin and the 13kDa subunit of prolamine can be used as protein markers in both Roshan and Shahriar, respectively. Our study demonstrated that although there is a significant relationship between protein subunits and the amount of stored proteins, but there was no significant relationship between stored proteins with different quantitative and qualitative rice cultivars. Therefore it seems that improving the seed storage protein quality will probably not limit the improvement of new rice cultivars with desirable quantitative and qualitative characteristics. [ABSTRACT FROM AUTHOR]

Background. Bioactive peptides derived from food proteins have been the subject of considerable interest in recent years. Therefore, the present study aimed to evaluate the nutraceutical properties (antioxidant, antihypertensive, and antidiabetic activities) of bioactive peptides encrypted in seed storage proteins of Moringa oleifera using in vitro gastrointestinal digestion. Materials and methods. Seeds were ground and sieved. Next, the flour was defatted, washed, and dried. Seed storage proteins were obtained from moreinga seed flour using different solvents. Total proteins were extracted with sodium borato, centrifuged, and precipitated. Finally, hydrolysates were obtained with pepsin, trypsin, and chymotrypsin. The hydrolysates were fractionated by ultrafiltration with 10 and 3 kDa membranes. Results. Based on the obtained results, total globulin hydrolysate possessed a high percentage of angiotensinconverting enzyme inhibition (79.51 ±6.89%) and a-amylase inhibition activity (61.19 ±2.93%). On the other hand, the fraction F < 3 kDa had good antioxidant activity (290.66 ±26.9 µM). Conclusion. This study suggests that total globulin hydrolysates and F < 3 kDa had excellent in vitro nutraceutical potential. [ABSTRACT FROM AUTHOR]

Field experiments were conducted, aiming to evaluate whether the late-season foliar urea application (35 kg N ha−1) is effective in improving grain yield and protein-related quality in ten common wheat cultivars during two consecutive growing seasons. On average, late urea application significantly (p < 0.05) increased grain yields, thousand kernel, and hectolitre weight. The total grain protein significantly (p < 0.05) increased by 17.5% under urea application, while dough mixing behaviour was improved by increasing flour water absorption and reducing the degree of softening. Considering dough elastic properties, the changes in dough energy and maximum resistance under urea application were strongly cultivar dependent and their values compared to control varying from −12.7% to +42.4% and −25.1% to +7.7%, respectively. The distribution of grain storage proteins was significantly influenced by late urea application, but to a lesser extent than the total grain proteins. The proportion of total gliadins, α- and γ-gliadins increased by 5.3%, 5.8%, and 6.5%, respectively, while total glutenins and high-molecular weight glutenins decreased by 4.5% and 7.4%, respectively. In summary, the late-season application of urea has greatly improved the protein-related quality without considerable disruption of the storage proteins composition. [ABSTRACT FROM AUTHOR]

Summary: Protein abundance in cereal grains is determined by the relative rates of protein synthesis and protein degradation during grain development but quantitation of these rates is lacking.Through combining in vivo stable isotope labelling and in‐depth quantitative proteomics, we have measured the turnover of 1400 different types of proteins during wheat grain development.We demonstrate that there is a spatiotemporal pattern to protein turnover rates which explain part of the variation in protein abundances that is not attributable to differences in wheat gene expression. We show that c. 20% of total grain adenosine triphosphate (ATP) production is used for grain proteome biogenesis and maintenance, and nearly half of this budget is invested exclusively in storage protein synthesis. We calculate that 25% of newly synthesized storage proteins are turned over during grain development rather than stored.This approach to measure protein turnover rates at proteome scale reveals how different functional categories of grain proteins accumulate, calculates the costs of protein turnover during wheat grain development and identifies the most and the least stable proteins in the developing wheat grain. [ABSTRACT FROM AUTHOR]

Molecular and biochemical markers are used to analyze the intraspecific genetic diversity of crops. Prolamincoding loci are highly effective for assessing this indicator. On the basis of the Laboratory of Varietal Seed Identification of the State Agrarian University of the Northern Trans-Urals, 18 varieties of common oat included in the State Register of Selection Achievements in the Tyumen Region from the 1930s to 2019 were studied by electrophoresis in 2018–2019. The aim of the work was to study the dynamics of the genetic diversity of oat va rieties at avenin-coding loci. For the analysis, 100 grains of each variety were used. Electrophoresis was carried out in vertical plates of 13.2 % polyacrylamide gel at a constant vol tage of 500 V for 4.0–4.5 h. It was found that 44.4 % of the varieties are heterogeneous, each consisting of two biotypes. For three loci, 20 alleles were identified, 10 of which were detected for the first time. The allele frequency of avenin-coding loci varied with time. In the process of variety exchange, alleles that are characteristic of varieties of non-Russian origin were replaced by alleles present in domestic varieties and then in the varieties developed by local breeding institutions. The following alleles had the highest frequency in Tyumen varieties: Avn A4(50.0 %), A2(25.0 %), Avn B4(50.0 %), Bnew6(37.5 %), Avn C1(37.5 %), C2 and C5(25.0 %). These alleles are of great value as markers of agronomically and adaptively important characters for the region in question. The amount of genetic diversity of oats varied with time from 0.33 in 1929–1950 to up to 0.75 in 2019. The high value of genetic diversity in modern breeding varieties of the Scientific Research Institute of Agriculture of the Northern Trans-Urals and an increase in this indicator over the past 20 years are associated with the use of genetically heterogeneous source material in the breeding process. This allowed obtaining varieties with high adaptive potentials in the natural climatic conditions of the region.

Introduction: Peanut allergy (PA) in children is a major concern. There is a need for better biological material for both diagnosis and oral immunotherapy (OIT) treatments. The unique state of seeds at early reproductive stages may affect the allergenicity of storage proteins, and impact clinical diagnostic and OIT protocols. The objective of this study was to evaluate the major allergen content in sequential seed developmental stages and monitor allergenicity via specific IgE binding quantification and skin prick testing., Methods: Seeds were collected from peanut plants and sorted into five developmental stages: initial (S1), developing (S2), full-size without coloration (S3), full-size with coloration (S4), and fully mature (S5) seeds. Samples were characterized by RNA-Seq, ELISA, and immunohistochemistry. Lyophilized, ground preparations were used for evaluation of skin test responses in sixty challenge-proven PA children., Results: Gene expression, protein content, and specific IgE binding of allergenic proteins increased throughout seed maturation and development. An expression bias towards the less allergenic A-genome copy of the major allergen Ara h 2 was found in earlier stages, especially in stage S2. Immunohistochemical staining showed that Ara h 2 is more dispersed in the cell and less accumulated within organized bodies at stage S2 versus stage S4. Significant differences were found in mean wheal responses between the commercial peanut extract (equivalent to stage S5) and stages S1 and S2, but not with stage S4, upon skin prick testing in subjects with PA., Discussion: The observed decrease in peanut-specific IgE binding of immature peanut seeds may be a result not only of decreased amounts of allergenic proteins, but also of profound changes in seed composition and conformation. This may be significant for developing a safer and more effective peanut OIT protocol., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (© 2024 Cohen, Levy, Toscano-Rivero, Manasherova, Agmon-Levin, Kenett, Jean-Claude, Mazer, Hovav and Kidon.)

SUMMARY: As the frequency of extreme environmental events is expected to increase with climate change, identifying candidate genes for stabilizing the protein composition of legume seeds or optimizing this in a given environment is increasingly important. To elucidate the genetic determinants of seed protein plasticity, major seed proteins from 200 ecotypes of Medicago truncatula grown in four contrasting environments were quantified after one‐dimensional electrophoresis. The plasticity index of these proteins was recorded for each genotype as the slope of Finlay and Wilkinson's regression and then used for genome‐wide association studies (GWASs), enabling the identification of candidate genes for determining this plasticity. This list was enriched in genes related to transcription, DNA repair and signal transduction, with many of them being stress responsive. Other over‐represented genes were related to sulfur and aspartate family pathways leading to the synthesis of the nutritionally essential amino acids methionine and lysine. By placing these genes in metabolic pathways, and using a M. truncatula mutant impaired in regenerating methionine from S‐methylmethionine, we discovered that methionine recycling pathways are major contributors to globulin composition establishment and plasticity. These data provide a unique resource of genes that can be targeted to mitigate negative impacts of environmental stresses on seed protein composition. Significance Statement: In the context of climate change, legumes that produce seeds with high protein contents without nitrogen fertilizer are vital for human and livestock nutrition. Thus, optimizing and stabilizing their seed protein yield and composition is of crucial importance. GWASs using the natural genetic variation of Medicago truncatula showed that methionine recycling pathways are major determinants of the plastic response of seed protein composition to environmental changes. [ABSTRACT FROM AUTHOR]

Maize (Zea mays L.) is important cereal and its diversity research is essential from the point of breeding strategy. The aim of this work was to compare the protein profiles of 30 varieties (from the USA and Europe) and 34 lines (from Slovakia) using two electrophoretic methods (SDS-PAGE and A-PAGE) and to construct dendrograms of their genetic similarity. The highest amount of HMW-GS (high molecular weight glutelin subunits) was detected in the European varieties (7.35%), LMW-GS (low molecular weight glutelin subunits) was determined in the Slovak lines (45.42%) and most residual albumins and globulins were found in the American varieties (50.66%). According to the representation of the evaluable protein bands, 4 dendrograms were constructed. In the dendrograms, which were based on the polymorphism of glutelins, Line 15 was separated from other Slovak lines, while European and American varieties split into 2 main clusters. The highest genetic similarity were between varieties Wonderfull and Queen Anna; and Slovak lines 6 and 9. From the point of zein polymorphism, dendrograms separated the varieties and the lines into 2 main clusters. Fruhester Gelber and Maďarská cukrová were the most similar varieties, also the Slovak lines 21 and 22; 13 and 34. The findings of the study will be helpful for proper breeding strategy and further sustainable agricultural development.

This paper presents the results of research of Tripogon chinensis adaptation mechanisms to the environmental conditions. This relict cereal grows on petrophytic parts of the slopes, where it is affected by extreme environmental factors (temperature variations, lack of water, insolation, etc.). This cereal reproduces by seeds. Under such growth conditions, to be able to germinate and continue growing, the plant must effectively use the available moisture and the seed storage compounds. Experiments with seeds germinated under different moisture conditions showed the ability of T. chinensis seeds to germinate in substrates with widely varying moisture contents. Increased moisture adversely affected the process of germination and growth of the cereal. Seed storage proteins play a special role because they are used as source of amino acids for the synthesis of proteins necessary for the plant. Determination of the fraction composition of T. chinensis seed storage proteins showed the high content of albumins and globulins (45% of the total weight of storage proteins), which ensured germination under water deficit. The study showed that germinating T. chinensis seeds use prolamins (their concentration decreases fourfold during germination). The considerable prolamin content of seeds (37%) and prolamin dynamics during germination support the hypothesis on the adaptive role of prolamins, as they contain amino acids essential for the seedling. Thus, morphological, physiological and biochemical properties of this cereal characterize it as a xerophytic species, which developed due to a combination of environmental factors of the Eastern Transbaikal region

During barley germination, cysteine proteases are essential in the mobilization of storage compounds providing peptides and amino acids to sustain embryo growth until photosynthesis is completely established. Knockdown barley plants, generated by artificial miRNA, for the cathepsins B- and F-like HvPap-19 and HvPap-1 genes, respectively, showed less cysteine protease activities and consequently lower protein degradation. The functional redundancy between proteases triggered an enzymatic compensation associated with an increase in serine protease activities in both knockdown lines, which was not sufficient to maintain germination rates and behaviour. Concomitantly, these transgenic lines showed alterations in the accumulation of protein and carbohydrates in the grain. While the total amount of protein increased in both transgenic lines, the starch content decreased in HvPap-1 knockdown lines and the sucrose concentration was reduced in silenced HvPap-19 grains. Consequently, phenotypes of HvPap-1 and HvPap-19 artificial miRNA lines showed a delay in the grain germination process. These data demonstrate the potential of exploring the properties of barley proteases for selective modification and use in brewing or in the livestock feeding industry. [ABSTRACT FROM AUTHOR]

Field experiments were conducted, aiming to evaluate whether the late-season foliar urea application (35 kg N ha−1) is effective in improving grain yield and protein-related quality in ten common wheat cultivars during two consecutive growing seasons. On average, late urea application significantly (p < 0.05) increased grain yields, thousand kernel, and hectolitre weight. The total grain protein significantly (p < 0.05) increased by 17.5% under urea application, while dough mixing behaviour was improved by increasing flour water absorption and reducing the degree of softening. Considering dough elastic properties, the changes in dough energy and maximum resistance under urea application were strongly cultivar dependent and their values compared to control varying from −12.7% to +42.4% and −25.1% to +7.7%, respectively. The distribution of grain storage proteins was significantly influenced by late urea application, but to a lesser extent than the total grain proteins. The proportion of total gliadins, α- and γ-gliadins increased by 5.3%, 5.8%, and 6.5%, respectively, while total glutenins and high-molecular weight glutenins decreased by 4.5% and 7.4%, respectively. In summary, the late-season application of urea has greatly improved the protein-related quality without considerable disruption of the storage proteins composition.

The dynamics of interactions of viral proteins with their host are pivotal in establishing a successful infection and ensuring systemic spread. To uncover these, an in silico analysis of the interactions between the coat protein (CP) of Sesbania mosaic virus (SeMV), a group IV virus with single-stranded positive-sense RNA genome was carried out with the known crystal structures of proteins belonging to the Fabaceae family, which is its natural host. SeMV is an isometric plant virus which infects Sesbania grandiflora, a member of Fabaceae, and causes mosaic symptoms. Earlier results have indicated that the assembly and disassembly events of SeMV favor the formation of CP dimers. Hence, the ability and strength of interactions of CP dimer with the host proteins were assessed using in silico protein–protein docking approaches. A set of 61 unique crystal structures of native proteins belonging to Fabaceae were downloaded from the Protein Data Bank (PDB) and docked with the CP dimer of SeMV. From the docking scores and interaction analysis, the host proteins were ranked according to their strength and significance of interactions with the CP dimers. The leads that were identified present themselves as strong candidates for developing antivirals against not only SeMV but also other related viruses that infect Fabaceae. The study is a prototype to understand host protein interactions in viruses and hosts. [ABSTRACT FROM AUTHOR]

The effects of protein characteristics and the proportion of gluten on end-use quality in 13 Korean wheat cultivars for three years were verified in this study. Year, cultivar, and the interaction between the year and the cultivar influenced protein characteristics, the proportion of gluten except for γ- and ω-gliadin using RP-HPLC (reversed-phase high-performance liquid chromatography), and end-use quality. Protein characteristics and the proportion of gluten in Korean wheat cultivars were between those of Australian standard white (ASW) and hard wheat (AH). Korean wheat cultivars exhibited a higher average α+β gliadin proportion than imported wheat, a γ-gliadin proportion similar to that of dark northern spring wheat, and the same ω-gliadin proportion as AH. They showed a bread-loaf volume intermediate between those of ASW and AH and a texture of cooked noodles similar to that of soft white wheat, but less springiness than imported wheat. The cookie diameter of Korean wheat cultivars was similar to that of hard red winter wheat. There was a correlation between bread-loaf volume and protein characteristics, except for the protein content in Korean wheat cultivars. Springiness and cohesiveness of cooked noodles were not correlated with protein characteristics, while hardness was correlated with the protein content and water absorption of a mixograph. Cookie diameter was negatively correlated with the sodium dodecyl sulfate (SDS) sedimentation volume and water absorption of a mixograph. The end-use quality was not correlated with any proportion of gluten composition. Principal component analysis (PCA) showed that the proportion of gluten was not related to the quality of the bread (both PCs, 81.3%), noodle (77.7%), and cookie (82.4%). PCA explained that Keumkang is suitable for superior bread, while Uri is good for cooked noodles and cookies.

Wheat is one of the most important food crop in the world. Gluten proteins represent major protein fraction of the grain and are responsible for unique properties of the dough. The aim of this study was to analyze one genotype of the winter wheat (Triticum aestivum L.) and one genotype of hybrid wheat (Triticum aestivum L. x Triticum spelta L.) based on protein polymorphism and to compare their nutritional and technological quality. According to Kjeldahl and Golenkov, content of total nitrogen and protein fraction composition of analyzed samples were determined. Higher content of albumins and globulins (26.39%) was detected in hybrid wheat genotype PS Lubica with total nitrogen 2.02% and crude protein content 11.49%. Winter wheat genotype Elinor showed higher content of prolamins and glutelins (70.68%) and higher coefficient of nutritional quality (91.79%) than analyzed hybrid wheat. Higher total content of amino acids was determined in hybrid wheat genotype PS Lubica (122.02 mg*g-1 dry weight). For the separation of glutenin protein subunits, SDS-PAGE method by ISTA was used. On the basis of molecular weight, glutenins were separated in SDS-PAGE into HMW-GS (Elinor 10.7%; PS Lubica 3.78%) and LMW-GS (Elinor 58.4%; PS Lubica 67.28%). Electrophoretic profile of genotype Elinor was 0, 7+9, 5+10 with Glu Score 7 and composition of HMW-GS in genotype PS Lubica was 0, 20, 2+12 with Glu Score 4. According to the results, better nutritional and technological quality was proved in winter wheat genotype Elinor. According to identical results in 5 biological replicates, hybrid wheat genotype PS Lubica was not recommended for bread-making.

Abstract Background Cowpea (Vigna unguiculata) is an important source of protein supply for animal and human nutrition. The major storage globulins VICILIN and LEGUMIN (LEG) are synthesized from several genes including LEGA, LEGB, LEGJ and CVC (CONVICILIN). The current hypothesis is that the plant circadian core clock genes are conserved in a wide array of species and that primary metabolism is to a large extent controlled by the plant circadian clock. Our aim was to investigate a possible link between gene expression of storage proteins and the circadian clock. Results We identified cowpea orthologues of the core clock genes VunLHY, VunTOC1, VunGI and VunELF3, the protein storage genes VunLEG, VunLEGJ, and VunCVC as well as nine candidate reference genes used in RT-PCR. ELONGATION FACTOR 1-A (ELF1A) resulted the most suitable reference gene. The clock genes VunELF3, VunGI, VunTOC1 and VunLHY showed a rhythmic expression profile in leaves with a typical evening/night and morning/midday phased expression. The diel patterns were not completely robust and only VungGI and VungELF3 retained a rhythmic pattern under free running conditions of darkness. Under field conditions, rhythmicity and phasing apparently faded during early pod and seed development and was regained in ripening pods for VunTOC1 and VunLHY. Mature seeds showed a rhythmic expression of VunGI resembling leaf tissue under controlled growth chamber conditions. Comparing time windows during developmental stages we found that VunCVC and VunLEG were significantly down regulated during the night in mature pods as compared to intermediate ripe pods, while changes in seeds were non-significant due to high variance. The rhythmic expression under field conditions was lost under growth chamber conditions. Conclusions The core clock gene network is conserved in cowpea leaves showing a robust diel expression pattern except VunELF3 under growth chamber conditions. There appears to be a clock transcriptional reprogramming in pods and seeds compared to leaves. Storage protein deposition may be circadian regulated under field conditions but the strong environmental signals are not met under artificial growth conditions. Diel expression pattern in field conditions may result in better usage of energy for protein storage.

Featured Application: This work introduces a novel way of designing proteins for the treatment of diseases, especially metabolic syndrome. Metabolic syndrome comprises a cluster of diseases like hypertension, dyslipidemia, and insulin resistance, among others. Its treatment is based on lifestyle modification; however, this treatment often fails to improve metabolic syndrome indicators over the long term. In this work, sequences of some representative vegetable proteins were explored to find bioactive peptides with activity toward metabolic disorders of metabolic syndrome. Five proteins, i.e., legumin (chickpea), glutelin type A-2 (chickpea), glutelin type B-2 (rice), prolamin PPROL 17 (maize), and glutelin (rice) revealed a high potential to be effective against metabolic syndrome. We designed and evaluated in silico modifications to their amino acid sequence to release bioactive peptides after simulating gastrointestinal digestion (SGD). The approach presented here allows the design of proteins that could combat metabolic syndrome, for later production and study. In the future, these proteins can be used as functional foods. [ABSTRACT FROM AUTHOR]

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Abstract. In this study reserve endosperm proteins, hordeins of seven spring barley cultivars with different origin: Zernogradskii (Russia), Bodega, Fink, Scarlett and Barke (Germany), Josefin and Astoria (France) were fractionated and characterized by SDS-PAGE electrophoresis. On the basis of the obtained spectra 19 bands (D + C + B) with different relative electrophoretic mobility and intensity were identified. The electrophoresis profiles of the groups D-, C- and B- hordein are designated as separate types (models) using the index corresponding to hordein blocks. We have established one profile type for D-hordein (D1), two- for C-hordein (C1, C2), and five – for B-hordein (B1, B2, B3, B4, B5). Based on these results hordein formulas (configurations) of accessions are constructed, which enable the expression of specific varietal characteristics and prove the existence of the inter allelic variation (hordein polymorphism) due to the presence or absence of protein components and their different electrophoretic mobility in the profiles of D-, C- and B-hordein.

Abstract Background The extent of co-sensitization within and between food protein families in an adult population is largely unknown. This study aimed to identify the most frequently recognized components in the PR-10 and storage protein family, as well as patterns in (co-)sensitization, in a birch-endemic area. Methods Results of ImmunoCAP ISAC, performed during routine care in Dutch adult outpatients suspected of food allergy, were collected. Results A total of 305 patients were selected, aged 16–79 years (median 32 years). Sensitization to one or more PR-10 proteins was most frequent (74% of all subjects), followed by 35% to storage protein and 15% to nsLTPs. Within the PR-10 family, subjects were most often sensitized to Bet v 1 (73% of 305), Cor a 1.04 (72%) and Mal d 1 (68%). Sensitization to PR-10s from soy, celery and kiwi occurred distinctively less often ( 70%). Subjects sensitized to these ‘less common PR-10 proteins’ were sensitized to more food and inhalant components on the ISAC, compared to subjects sensitized to ‘common PR-10 proteins’ (median 22 vs 13 out of 112, p

Abstract Background IgE sensitization to storage proteins from nuts and seed is often related to severe allergic symptoms. There is a risk of immunological IgE cross-reactivity between storage proteins from different species. The potential clinical implication of such cross-reactivity is that allergens other than the known sensitizer can cause allergic symptoms. Previous studies have suggested that kiwi seed storage proteins may constitute hidden food allergens causing cross-reactive IgE-binding with peanut and other tree nut homologs, thereby mediating a potential risk of causing allergy symptoms among peanut ant tree nut allergic individuals. The objective of this study was to investigate the degree of sensitization towards kiwi fruit seed storage proteins in a cohort of peanut allergic individuals. Methods A cohort of 59 adolescents and adults with peanut allergy was studied, and self reported allergies to a number of additional foods were collected. Quantitative IgE measurements to seed storage proteins from kiwi and peanut were performed. Results In the cohort, 23 out of the 59 individuals were reporting kiwi fruit allergy (39%). The frequency of IgE sensitization to kiwi fruit and to any kiwi seed storage protein was higher among peanut allergic individuals also reporting kiwi fruit allergy (P = 0.0001 and P = 0.01). A positive relationship was found between IgE levels to 11S globulin (r = 0.65) and 7S globulin (r = 0.48) allergens from kiwi and peanut, but IgE levels to 2S albumin homologs did not correlate. Patients reporting kiwi fruit allergy also reported allergy to hazelnut (P = 0.015), soy (P

Cytochrome P450, CYP93A1, is involved in the synthesis of the phytoalexin glyceollin in soybean (Glycine max L. Merr). The gene encoding CYP93A1 has been used as defense marker in soybean cell cultures, however, little is known regarding how this gene is expressed in the intact plant. To further understand the tissue-specific role of CYP93A1 in soybean defense, we analyzed the expression of this gene in mechanically damaged leaves and stems. In leaves, CYP93A1 was constitutively expressed; its expression did not change in response to mechanical damage. In stems, however, expression of CYP93A1 was induced as quickly as 4 h after mechanical damage and remained upregulated for at least 48 h. The induction of CYP93A1 was associated with the synthesis of glyceollins. In comparison to several other defense-related genes encoding cysteine protease inhibitors L1 and R1 and storage proteins vspA and vspB, CYP93A1 was the most strongly induced by stem wounding. The induction of CYP93A1 was observed only locally, not systemically. Similar stem expression patterns were consistently observed among three different soybean genotypes. The strong induction of CYP93A1 in mechanically damaged stems suggests an important role in the soybean stem defense response; therefore, this study expands the use of CYP93A1 as a defense response marker to stems, not just soybean cell cultures.

ω-Secalin was an important factor influencing processing quality of wheat 1BL/1RS translocations. On the basis of four ω-secalin gene sequences cloned from Lankao 906 (a wheat cultivar with 1BL/1RS translocation) with putative transcription activity, a pair of primers with suitable restriction endonucleases added at their 5′ ends were designed to amplify the mature protein-coding regions of the four genes. After digestion with restriction endonucleases, the amplified products were ligated into the prokaryotic expression vector pET30a(+). The prokaryotically expressed recombinant proteins and gliadin isolated from the Lankao 906 seed were analyzed on the same acid polyacrylamide gel electrophoresis. All four prokaryotically expressed secalin bands had corresponding seed-expressed gliadin bands. The four corresponding gliadin bands were confirmed to be the expression products of the four ω-secalin genes by liquid chromatography tandem mass spectrometry (LC-MS/MS). This conclusion was further confirmed because the four ω-secalin bands could be detected in all 14 wheat 1BL/1RS translocation cultivars used in the study, although there was some interference for the detection of one ω-secalin band from nearby wheat gliadin bands. The sequence information of ω-secalin genes with expression activity will be helpful for improving the processing quality of wheat with 1BL/1RS translocations by using RNA interference method to silence the expression of the ω-secalin genes.

Winter wheat is especially used for bread-making. The specific composition of the grain storage proteins and the representation of individual subunits determines the baking quality of wheat. The aim of this study was to analyze 15 slovak varieties of the winter wheat (Triticum aestivum L.) based on protein polymorphism and to predict their technological quality. SDS-PAGE method by ISTA was used to separate glutenin protein subunits. Glutenins were separated into HMW-GS (15.13%) and LMW-GS (65.89%) on the basis of molecular weight in SDS-PAGE. At the locus Glu-A1 was found allele Null (53% of genotypes) and allele 1 (47% of genotypes). The locus Glu-B1 was represented by the HMW-GS subunits 6+8 (33% of genotypes), 7+8 (27% of genotypes), 7+9 (40% of genotypes). At the locus Glu-D1 were detected two subunits, 2+12 (33% of genotypes) and 5+10 (67% of genotypes) which is correlated with good bread-making properties. The Glu – score was ranged from 4 (genotype Viglanka) to 10 (genotypes Viola, Vladarka). According to the representation of individual glutenin subunits in samples, the dendrogram of genetic similarity was constructed. By the prediction of quality the results showed that the best technological quality was significant in the varieties Viola and Vladarka which are suitable for use in food processing.

Phytochromes comprise a small family of photoreceptors with which plants gather environmental information that they use to make developmental decisions, from germination to photomorphogenesis to fruit development. Most phytochromes are activated by red light and de-activated by far-red light, but phytochrome A (phyA) is responsive to both and plays an important role during the well-studied transition of seedlings from dark to light growth. The role of phytochromes during skotomorphogenesis (dark development) prior to reaching light, however, has received considerably less attention although previous studies have suggested that phytochrome must play a role even in the dark. We profiled proteomic and transcriptomic seedling responses in tomato during the transition from dark to light growth and found that phyA participates in the regulation of carbon flux through major primary metabolic pathways, such as glycolysis, beta-oxidation, and the tricarboxylic acid (TCA) cycle. Additionally, phyA is involved in the attenuation of root growth soon after reaching light, possibly via control of sucrose allocation throughout the seedling by fine-tuning the expression levels of several sucrose transporters of the SWEET gene family even before the seedling reaches the light. Presumably, by participating in the control of major metabolic pathways, phyA sets the stage for photomorphogenesis for the dark grown seedling in anticipation of light.